Enterobacters

Saudi Med J, 2004 May, 25(5), 570 - 4
Prevalence of extended spectrum beta-lactamase among multidrug resistant gram-negative isolates from a general hospital in Saudi Arabia; Kader AA et al.; OBJECTIVE: To determine the prevalence of extended spectrum beta-lactamase (ESBL) among multidrug resistant isolates of enterobacteriaceae and non-fermenting gram-negative bacilli . METHODS: This study was carried out at the Almana General Hospital, Eastern Province, Kingdom of Saudi Arabia, during the period March 2002 through to June 2003 . Multidrug resistant gram-negative isolates from patients admitted to the surgical, medical, pediatric, long-term care and intensive care units were studied for the presence of the ESBL enzyme . RESULTS: A total of 3231 gram-negative organisms were studied for the presence of multidrug resistance and ESBLs . Of these, 197 (6%) isolates were multidrug resistant (MDR), and 156 (4.8%) were positive for ESBL . Seventy nine percent of the MDR strains were positive for ESBL . The most frequent isolates were Escherichia coli (1116) and Klebsiella pneumoniae (687) and ESBL was detected in 72 (6.5%) and 37 (5.4%) of these isolates . The MDR strains that produced ESBL were most commonly isolated from surgical care patients with diabetic fascitis (83%) and patients with indwelling Foley's catheter (79%) . Extended spectrum beta-lactamase producing strains showed the highest susceptibility to imipenem and meropenem (86%) . The non-beta-lactam antibiotics with greatest activity against these ESBL strains in vitro were ciprofloxacin (72%), amikacin (70%), tobramycin (67%) and gentamicin (56%) . CONCLUSION: The majority (79%) of the MDR enterobacteriaceae and non-fermenting gram-negative bacilli tested over 15-months were positive for ESBL . Imipenem, meropenem, ciprofloxacin and amikacin showed the highest activity against these ESBL-producing organisms . Due to the growing problem of infection with ESBL-producing bacteria, which are frequently resistant to many classes of antibiotics resulting in difficult-to-treat infections, clinicians need to be familiar with the clinical significance of these enzymes and potential strategies for dealing with them.

FEMS Microbiol Lett, 2004 May 15, 234(2), 209 - 13
Novel GES/IBC extended-spectrum beta-lactamase variants with carbapenemase activity in clinical enterobacteria; Vourli S et al.; Two clinical isolates, an Escherichia coli and a Klebsiella pneumoniae, with decreased susceptibility to carbapenems were studied . This phenotype was associated with production of novel GES/IBC variant beta-lactamases, designated GES-3 (from E . coli) and GES-4 (from K . pneumoniae), exhibiting carbapenemase activity . Both enzymes possessed Ser at Ambler's position 170 instead of Gly found in the beta-lactamases GES-1 and IBC-1 that lack carbapenemase activity . Additionally, position 104 in GES-4 was occupied by a Lys as in IBC-1 . bla(GES-3) and bla(GES-4) occurred as gene cassettes in the variable regions of class 1 integrons carried by plasmids . The structure of the GES-4-encoding integron was similar to that of previously described IBC-1 integrons . The GES-3-encoding integron was, most likely, truncated at the 3' conserved segment .

FEMS Microbiol Lett, 2004 May 15, 234(2), 201 - 7
Diversity of genetic environment of bla(CTX-M) genes; Lartigue MF et al.; Twenty-four non-clonally related enterobacterial isolates producing the emerging CTX-M-type extended-spectrum beta-lactamases were recovered from several countries including France, India, Poland, and Turkey . They had been isolated from 2000 to 2003 . beta-Lactamases CTX-M-2, CTX-M-3, CTX-M-10, CTX-M-14 and CTX-M-15 were identified . Most of the isolates produced beta-lactamase CTX-M-15 . Insertion sequence ISEcp1 was found upstream of bla(CTX-M-3), bla(CTX-M-10), bla(CTX-M-14) and bla(CTX-M-15) genes . A sequence similar to the inverted right repeat of ISEcp1 was identified downstream of bla(CTX-M-3), bla(CTX-M-10) and bla(CTX-M-15) genes suggesting the mobilization of these beta-lactamase genes by transposition events . In addition, Orf513 was identified upstream of the bla(CTX-M-2) gene . This work further underlined widespread of bla(CTX-M-15) gene associated with ISEcp1 .

Spectrochim Acta A Mol Biomol Spectrosc, 2004 May, 60(6), 1279 - 89
Chromophoric spin-labeled beta-lactam antibiotics for ENDOR structural characterization of reaction intermediates of class A and class C beta-lactamases; Mustafi D et al.; The chromophoric spin-label substrate 6-N-{3-(2,2,5,5-tetramethyl-1-oxypyrrolin-3-yl)-propen-2-oyl}penicillanic acid (SLPPEN) was synthesized by acylation of 6-aminopenicillanic acid with the acid chloride of 3-(2,2,5,5-tetramethyl-1-oxypyrrolinyl)-2-propenoic acid and characterized by physical methods . By application of angle-selected electron nuclear double resonance (ENDOR), we have determined the molecular structure of SLPPEN in solution . SLPPEN exhibited UV absorption properties that allowed accurate monitoring of the kinetics of its enzyme-catalyzed hydrolysis . The maximum value of the (substrate-product) difference extinction coefficient was 2824 M(-1) cm(-1) at 275 nm compared to 670 M(-1) cm(-1) at 232 nm for SLPEN {J . Am . Chem . Soc . 117 (1995) 6739} . For SLPPEN, the steady-state kinetic parameters kcat and kcat/KM, determined under initial velocity conditions, were 637 +/- 36 s(-1) and 13.8 +/- 1.4 x 10(6) M(-1) s(-1), respectively, for hydrolysis catalyzed by TEM-1 beta-lactamase of E . coli, and 0.5 +/- 0.04 s(-1) and 3.9 +/- 0.4 x 10(4) M(-1) s(-1) for hydrolysis catalyzed by the beta-lactamase of Enterobacter cloacae P99 . We have also observed "burst kinetics" for the hydrolysis of SLPPEN with P99 beta-lactamase, indicative of formation of an acylenzyme reaction intermediate . In DMSO:H2O (30:70, v:v) cryosolvent mixtures buffered to pH* 7.0, the half-life of the acylenzyme intermediate formed with the P99 enzyme at -5 degrees C was > or = 3 min, suitable for optical characterization . The observation of burst kinetics in the hydrolysis of SLPPEN catalyzed by P99 beta-lactamase suggests that this chromophoric spin-labeled substrate is differentially sensitive to active site interactions underlying the cephalosporinase and penicillinase reactivity of this class C enzyme.

Microbiology, 2004 May, 150(Pt 5), 1439 - 46
Unique organization and regulation of the mrx fimbrial operon in Xenorhabdus nematophila; He H et al.; Xenorhabdus nematophila, a Gram-negative bacterium belonging to the Proteus clade of the family Enterobacteriaceae, forms a mutualistic association with the soil nematode Steinernema carpocapsae . The nematode invades insects and releases Xenorhabdus into the haemolymph, where it participates in insect killing . To begin to understand the role of fimbriae in the unique life cycle of Xenorhabdus, the organization and expression of the mrx fimbrial operon was analysed . The mrx operon contained only five structural genes (mrxACDGH), making it one of the smallest chaperone-usher fimbrial operons studied to date . Unlike the mrp operon of Proteus mirabilis, a site-specific recombinase was not linked to the mrx operon . The intergenic region between the major fimbrial gene (mrxA) and the usher gene (mrxC) lacked a mrpB-like gene, but contained three tandem inverted repeat sequences located downstream of mrxA . A 940 nt mrxA-containing mRNA was the major transcript produced in cells growing on agar, while an mrx polycistronic mRNA was produced at low levels . A canonical sigma(70) promoter, identified upstream of mrxA, was not subject to promoter inversion . Fimbriae were not produced in an lrp-mutant strain, suggesting that the leucine-responsive regulatory protein, Lrp, plays a role in the regulation of the mrx operon . These findings show that the genetic organization and regulation of the mrx operon is in several respects distinct from other chaperone-usher fimbrial operons.

J Biol Chem, 2004 Jul 16, 279(29), 30563 - 72 Epub 2004 May 05.
Atomic resolution structures and solution behavior of enzyme-substrate complexes of Enterobacter cloacae PB2 pentaerythritol tetranitrate reductase . Multiple conformational states and implications for the mechanism of nitroaromatic explosive degradation; Khan H et al.; The structure of pentaerythritol tetranitrate (PETN) reductase in complex with the nitroaromatic substrate picric acid determined previously at 1.55 A resolution indicated additional electron density between the indole ring of residue Trp-102 and the nitro group at C-6 of picrate . The data suggested the presence of an unusual bond between substrate and the tryptophan side chain . Herein, we have extended the resolution of the PETN reductase-picric acid complex to 0.9 A . This high-resolution analysis indicates that the active site is partially occupied with picric acid and that the anomalous density seen in the original study is attributed to the population of multiple conformational states of Trp-102 and not a formal covalent bond between the indole ring of Trp-102 and picric acid . The significance of any interaction between Trp-102 and nitroaromatic substrates was probed further in solution and crystal complexes with wild-type and mutant (W102Y and W102F) enzymes . Unlike with wild-type enzyme, in the crystalline form picric acid was bound at full occupancy in the mutant enzymes, and there was no evidence for multiple conformations of active site residues . Solution studies indicate tighter binding of picric acid in the active sites of the W102Y and W102F enzymes . Mutation of Trp-102 does not impair significantly enzyme reduction by NADPH, but the kinetics of decay of the hydride-Meisenheimer complex are accelerated in the mutant enzymes . The data reveal that decay of the hydride-Meisenheimer complex is enzyme catalyzed and that the final distribution of reaction products for the mutant enzymes is substantially different from wild-type enzyme . Implications for the mechanism of high explosive degradation by PETN reductase are discussed.

Appl Environ Microbiol, 2004 May, 70(5), 3013 - 23
Genomic diversity of Erwinia carotovora subsp . carotovora and its correlation with virulence; Yap MN et al.; We used genetic and biochemical methods to examine the genomic diversity of the enterobacterial plant pathogen Erwinia carotovora subsp . carotovora . The results obtained with each method showed that E . carotovora subsp . carotovora strains isolated from one ecological niche, potato plants, are surprisingly diverse compared to related pathogens . A comparison of 23 partial mdh sequences revealed a maximum pairwise difference of 10.49% and an average pairwise difference of 2.13%, values which are much greater than the maximum variation (1.81%) and average variation (0.75%) previously reported for Escherichia coli . Pulsed-field gel electrophoresis analysis of I-CeuI-digested genomic DNA revealed seven rrn operons in all E . carotovora subsp . carotovora strains examined except strain WPP17, which had only six copies . We identified 26 I-CeuI restriction fragment length polymorphism patterns and observed significant polymorphism in fragment sizes ranging from 100 to 450 kb for all strains . We detected large plasmids in two strains, including the model strain E . carotovora subsp . carotovora 71 . The two least virulent strains had an unusual chromosomal structure, suggesting that a particular pulsotype is correlated with virulence . To compare chromosomal organization of multiple enterobacterial genomes, several genes were mapped onto I-CeuI fragments . We identified portions of the genome that appear to be conserved across enterobacteria and portions that have undergone genome rearrangements . We found that the least virulent strain, WPP17, failed to oxidize cellobiose and was missing several hrp and hrc genes . The unexpected variability among isolates obtained from clonal hosts in one region and in one season suggests that factors other than the host plant, potato, drive the evolution of this common environmental bacterium and key plant pathogen.

Appl Environ Microbiol, 2004 May, 70(5), 2959 - 65
Isolation, characterization, and U(VI)-reducing potential of a facultatively anaerobic, acid-resistant Bacterium from Low-pH, nitrate- and U(VI)-contaminated subsurface sediment and description of Salmonella subterranea sp . nov; Shelobolina ES et al.; A facultatively anaerobic, acid-resistant bacterium, designated strain FRCl, was isolated from a low-pH, nitrate- and U(VI)-contaminated subsurface sediment at site FW-024 at the Natural and Accelerated Bioremediation Research Field Research Center in Oak Ridge, Tenn . Strain FRCl was enriched at pH 4.5 in minimal medium with nitrate as the electron acceptor, hydrogen as the electron donor, and acetate as the carbon source . Clones with 16S ribosomal DNA (rDNA) sequences identical to the sequence of strain FRCl were also detected in a U(VI)-reducing enrichment culture derived from the same sediment . Cells of strain FRCl were gram-negative motile regular rods 2.0 to 3.4 micro m long and 0.7 to 0.9 microm in diameter . Strain FRCl was positive for indole production, by the methyl red test, and for ornithine decarboxylase; it was negative by the Voges-Proskauer test (for acetylmethylcarbinol production), for urea hydrolysis, for arginine dihydrolase, for lysine decarboxylase, for phenylalanine deaminase, for H(2)S production, and for gelatin hydrolysis . Strain FRCl was capable of using O(2), NO(3)(-), S(2)O(3)(2-), fumarate, and malate as terminal electron acceptors and of reducing U(VI) in the cell suspension . Analysis of the 16S rDNA sequence of the isolate indicated that this strain was 96.4% similar to Salmonella bongori and 96.3% similar to Enterobacter cloacae . Physiological and phylogenetic analyses suggested that strain FRCl belongs to the genus Salmonella and represents a new species, Salmonella subterranea sp . nov.

Rev Saude Publica, 2004 Apr, 38(2), 326 - 8 Epub 2004 Apr 26.
{Antibacterial activity of essential oils on microorganisms isolated from urinary tract infection}; Pereira RS et al.; The antibacterial activity of essential oils extracted from medicinal plants (Ocimum gratissimum, L., Cybopogum citratus (DC) Stapf., and Salvia officinalis, L.) was assessed on bacterial strains derived from 100 urine samples . Samples were taken from subjects diagnosed with urinary tract infection living in the community . Microorganisms were plated on Muller Hinton agar . Plant extracts were applied using a Steers replicator and petri dishes were incubated at 37 degrees C for 24 hours . Salvia officinalis, L . showed enhanced inhibitory activity compared to the other two herbs, with 100% efficiency against Klebsiella and Enterobacter species, 96% against Escherichia coli, 83% against Proteus mirabilis, and 75% against Morganella morganii.

Int J Antimicrob Agents, 2004 May, 23(5), 506 - 9
Resistance of uropathogens in symptomatic urinary tract infections in León, Nicaragua; Matute AJ et al.; Management of urinary tract infections (UTI) in Central America and especially Nicaragua, is complicated by the lack of knowledge about the antibiotic resistance of uropathogens . We conducted a prevalence study to gain more insight into the aetiology, bacterial resistance and risk factors for symptomatic UTI in the region of Leon, Nicaragua . In 2002, all consecutive patients with UTI symptoms and pyuria >/=10 WBC/hpf were admitted to the study . Positive cultures from midstream urine specimens were defined as >/=10(5) cfu/ml of a single uropathogen . Susceptibility tests were performed with disc diffusion tests using the Kirby-Bauer method and broth microdilution using National Committee for Clinical Laboratory Standards criteria both in Leon and a reference laboratory in Utrecht . A positive culture was present in 62 of 208 study subjects (30%) . Escherichia coli (56%), Klebsiella spp . (18%) and Enterobacter spp . (11%) were the most frequent pathogens isolated . Presence of cystocele, incontinence and increasing age were risk factors for bacterial UTI . E . coli was least resistant to ceftriaxone, amikacin and nitrofurantoin (>90% susceptible) . We observed high resistance rates in E . coli to amoxicillin (82%, MIC(90) 128 mg/l), trimethoprim-sulphamethoxazole (TMP-SMX) (64%, MIC(90) 32 mg/l), cephalothin (58%, MIC(90), 32 mg/l), ciprofloxacin (30%; MIC(90), 32 mg/l), amoxicillin/clavulanate (21%, MIC(90) 8 mg/l) and gentamicin (12%, MIC(90) 2 mg/l) . Our results suggests that community acquired uropathogens in Nicaragua are highly resistant to many antimicrobial agents . The use of amoxicillin, trimethoprim-sulphamethoxazole and cephalothin against uropathogens needs to be reconsidered . High quinolone resistance rates among E . coli in Nicaragua gives cause for great concern.

Int J Antimicrob Agents, 2004 May, 23(5), 480 - 6
Antimicrobial susceptibility of clinical isolates of Enterobacteriaceae producing complex beta-lactamase patterns including extended-spectrum enzymes; Segatore B et al.; The antimicrobial susceptibility of 103 clinical isolates of Enterobacteriaceae to 11 antibiotics, was investigated, using a conventional inoculum size (5 x 10(5) CFU) and a higher inoculum size (5 x 10(8) CFU) . All the isolates produced complex beta-lactamase patterns, including an extended-spectrum beta-lactamase (ESBL) of the TEM- or SHV-type plus other enzymes (a TEM-type or an SHV-type non-ESBL and/or a class C enzyme) . The following repertoire of ESBLs was produced by the isolates: TEM-15, TEM-19, TEM-26, TEM-52, TEM-72, TEM-87, TEM-92, SHV-2a, SHV-5 and SHV-12, as assessed by sequencing . Production of the other enzymes was showed by analytical isoelectric focusing . Overall, meropenem was the most active agent and less influenced by inoculum size, while other beta-lactams showed a lower activity and a significant inoculum size effect . In conclusion, from its in vitro performance, meropenem could be considered as the last resource drug against strains producing complex beta-lactamase patterns including an ESBL.

J Endotoxin Res, 2004, 10(2), 107 - 12
Enzymology of lipid A palmitoylation in bacterial outer membranes; Bishop RE et al.; The enzymology of palmitate addition to lipid A can be traced to the early discovery of monosaccharide lipid A precursors, but the functional importance of lipid A palmitoylation in bacterial resistance to the host immune response has emerged only recently . Lipid A palmitoylation in enterobacteria is determined by a PhoP/PhoQ-activated gene pagP, which encodes an unusual outer membrane enzyme of lipid A biosynthesis . PagP structure and dynamics have now been elucidated by both NMR spectroscopy and X-ray crystallography . PagP is an 8-stranded antiparallel beta-barrel preceded by an N-terminal amphipathic alpha-helix . The PagP barrel axis is uniquely tilted by 30 degrees with respect to the membrane normal . An interior hydrophobic pocket in the upper half of the molecule functions as a hydrocarbon ruler, which allows the enzyme to distinguish palmitate from other acyl chains found in phospholipids . Internalization of a phospholipid palmitoyl group within the barrel appears to occur by lateral diffusion from the outer leaflet through non-hydrogen bonded regions between beta-strands . The MsbA-dependent trafficking of lipids from the inner membrane to the outer membrane outer leaflet is necessary for lipid A palmitoylation in vivo . Efforts to determine the PagP catalytic mechanism may lead to the development of inhibitors for the treatment of infections.

Scand J Infect Dis, 2004, 36(3), 165 - 73
Gram-negative bacillary meningitis after cranial surgery or trauma in adults; Briggs S et al.; In order to assess the clinical features, aetiology, treatment and outcome of post-neurosurgical and post-traumatic Gram-negative bacillary meningitis (GNBM) we performed a retrospective review of all adult patients admitted to the Department of Neurosurgery who had Gram-negative bacilli cultured from cerebrospinal fluid (CSF) following a neurosurgical procedure or traumatic head/spinal injury . During the 12 y of the review 33 patients had CSF isolates of Gram-negative bacilli that were thought to be significant . The median patient age was 47 y (range 22-77 y) and 21 (64%) were male . Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli were the most common isolates . Minimal inhibitory concentrations (MIC) measured for half the patients' isolates resulted in 5 regimen changes, including 2 patients with E . cloacae meningitis in whom cephalosporin susceptibility decreased during cephalosporin treatment . Our recommended initial treatment was intravenous ceftriaxone and amikacin, subsequently tailored by susceptibility results; approximately half the patients remained on the antibiotics they started and half were changed to an alternate regimen, most often a carbapenem . Five patients (15%) died, 1 dying after cure of his GNBM . There were no failures in those who received more than 12 d of appropriate treatment: treatment for at least 14 d after the last positive CSF culture guaranteed cure . Initial ceftriaxone and amikacin subsequently changing to susceptibility driven alternatives, often a carbapenem, resulted in cure of 85% of our patients with GNBM.

Jpn J Infect Dis, 2004 Apr, 57(2), 37 - 40
An epidemiological analysis of Stenotrophomonas maltophilia strains in a university hospital; Caylan R et al.; The aim of this investigation was to evaluate the epidemiology of Stenotrophomonas maltophilia in a university hospital of Turkey . From June 2000 to December 2001, S . maltophilia strains were collected, clinical presentations were noted, and MIC determinations were performed by means of E-test . Enterobacterial repetitive intergenic consensus sequences-PCR (ERIC-PCR) was used for molecular typing of the strains . Forty-four strains of S . maltophilia were isolated from 41 hospitalized patients in a teaching hospital . The majority of specimens were from the blood and respiratory tract . Antimicrobial sensitivities of these strains were as follows: 97.7 % trimethoprim-sulfamethoxazole, 15.9% ticarcillin, and 95.4% ticarcillin- clavulanate . The strains were evaluated using the ERIC-PCR method . It was of interest to note that epidemiological typing revealed three small outbreaks that were caused by a total of 12 strains . The remaining isolates generated singular DNA patterns . DNA amplification was possible in 38 isolates and yielded 26 different patterns in a period of 20 months, leading to the suggestion that commensal bacteria becomes selected in the presence of a suitable host.

J Antimicrob Chemother, 2004 Jun, 53(6), 1090 - 4 Epub 2004 Apr 29.
In vitro susceptibility of recent antibiotic-resistant urinary pathogens to ertapenem and 12 other antibiotics; Alhambra A et al.; Background: The treatment of complicated urinary tract infections may require the use of a parenteral antibiotic with potent activity against the most common urinary pathogens . Ertapenem is a broad-spectrum 1beta-methyl carbapenem with a long plasma half-life that allows administration of a single daily dose . METHODS: The purpose of this work was to test the in vitro susceptibility to ertapenem, ampicillin, cefazolin, cefuroxime, cefotaxime, co-amoxiclav, piperacillin/tazobactam, imipenem, gentamicin, amikacin, fosfomycin, ciprofloxacin and co-trimoxazole of 482 strains of urinary pathogens of the family Enterobacteriaceae isolated from patients in the community of Madrid (40% from males) . The distribution was as follows: Escherichia coli (n = 315), Proteus mirabilis (n = 42), Klebsiella spp . (n = 14) and AmpC-producing Enterobacteriaceae (n = 111) . The strains studied were selected based on their resistance to quinolones and aminoglycosides, and their production of extended-spectrum beta-lactamases (ESBLs) or AmpC-type beta-lactamases . RESULTS: All the strains were susceptible to ertapenem, imipenem and amikacin . The MIC(90) of ertapenem ranged from a minimum of 0.03 mg/L for Proteus vulgaris and a maximum of 1 mg/L for Enterobacter spp . Ertapenem was the most active of all drugs tested in all cases . On comparing antibiotic resistance among ESBL-producing strains of E . coli (n = 35) and E . coli strains not producing ESBLs (n = 280), statistically significant differences were obtained for ciprofloxacin (P = 0.002) and gentamicin (P = 0.011) . Regarding ertapenem, only a slight increase in MIC(50) was seen, the value being 0.015 mg/L for strains not producing ESBLs versus 0.03 mg/L for ESBL-producing strains . CONCLUSIONS: In view of its significant antibiotic potency against antibiotic-resistant Enterobacteriaceae, ertapenem may constitute a good therapeutic alternative in urinary infections caused by these pathogens.

Am J Physiol Gastrointest Liver Physiol, 2004 Sep, 287(3), G638 - 46 Epub 2004 Apr 29.
Muscularis inflammation and the loss of interstitial cells of Cajal in the endothelin ETB receptor null rat; Suzuki T et al.; Endothelin receptor null rats {ETB(-/-)} are a model for long-segment Hirschsprung's disease . These animals have significant intestinal distension (megaileum) proximal to a constricted region of the gastrointestinal tract lacking enteric ganglia . Experiments were performed to determine the pathophysiological changes that occur in these animals and to examine the tunica muscularis as a unique, immunologically active compartment . We observed abnormal intestinal flora in ETB(-/-) rats, which included a marked increase in gram-negative aerobes (Enterobacteriaceae) and anaerobes (Bacteroidaceae) in the distended region of the small intestine . Histochemical observations showed that neutrophilic infiltration was rarely or not observed, but the number of ED2 positive macrophages was increased in the tunica muscularis . Expression of IL-1beta and IL-6 mRNA was also significantly increased, and the level of CD14 (LPS receptors) were increased significantly in the tunica muscularis . Spontaneous phasic contractions were irregular in the distended intestinal regions of ETB(-/-) rats, and this was associated with an increased number of macrophages and damage to interstitial cells of Cajal (ICC) as revealed by using Kit-like immunoreactivity and electron microscopy . These results suggest that ED2-positive resident macrophages may play an important role in the inflammation of tunica muscularis in ETB(-/-) rats . Increased numbers and activation of macrophages may result in damage to ICC networks leading to disordered intestinal rhythmicity in regions of the gut in which myenteric ganglia are intact.

Crit Rev Microbiol, 2004, 30(1), 25 - 32
Extended-spectrum beta-lactamases (ESbLs): characterization, epidemiology and detection; Shah AA et al.; Beta-lactamases of Enterobacteriaceae are the most important mechanism of resistance against beta-lactam drugs . Two types of beta-lactamases can confer resistance against 3rd generation cephalosporins . Chromosomally mediated beta-lactamases are inducible and are not inhibited by clavulanic acid . Resistance due to these enzymes is non-transferable . The 2nd type of enzyme is plasmid-mediated beta-lactamases, which are inhibited by clavulanic acid . These enzymes are more important clinically as these can be transferred between various species of Enterobacteriaceae . These enzymes are called extended-spectrum beta-lactamases (ESBLs) . ESBL-producing Enterobacteriaceae have been responsible for numerous outbreaks of infection throughout the world and pose challenging infection control issues . Antibacterial choice is often complicated by multi-resistance . ESBLs can confer resistance against all beta-lactam drugs except carbapenems and cephamycins . Nursing home patients may be an important reservoir of ESBL-containing multiple antibiotic-resistant organisms . Use of broad-spectrum oral antibiotics and probably poor infection control practices may facilitate spread of this plasmid-mediated resistance . In addition to known populations at risk, ambulatory patients with chronic conditions represent another patient population that may harbor ESBL-producing organisms . Various methods can be used for detection of ESBLs in the laboratory . These tests include double disc diffusion test, Vitek ESBL test, E Tests, MIC Determination, genetic method, and isoelectric focusing (IEF).

Jpn J Antibiot, 2004 Feb, 57(1), 70 - 104
{Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2002}; Yamaguchi K et al.; The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 899 strains of Gram-positive bacteria, 1500 strains of Gram-negative bacteria, and 158 strains of anaerobic bacteria obtained from 28 medical institutions during 2002 was measured . The results were as follows; 1 . MEPM was more active than other carbapenem antibiotics against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae . MIC90 of MEPM against Pseudomonas aeruginosa was the lowest of the drugs tested . MEPM showed low cross-resistant rate against both imipenem-resistant P . aeruginosa and ciprofloxacin-resistant P . aeruginosa . MEPM was active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE) . 2 . The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (4 strains) in Escherichia coli and 1.9% (2 strains) in Klebsiella pneumoniae . Carbapenems including MEPM were active against these ESBL strains . In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem; at present, 7 years after available for commercial use.

Eur J Immunol, 2004 May, 34(5), 1461 - 71
Necrosis-like cell death induced by bacteria in mouse macrophages; Kirschnek S et al.; The death of individual cells is a frequent and physiological event in the mammalian immune system and most often occurs by apoptosis . It is becoming increasingly clear that cell death is also induced during bacterial infections . Here we report that, in addition to the apoptotic form already established, a necrosis-like form of cell death is induced by pyogenic bacteria (Enterobacteriaceae, Pseudomonas, enterococci) in mouse macrophages . Necrosis could be separated from apoptosis as it did not require phagocytosis of bacteria and occurred when apoptosis was inhibited by caspase blockade or by Bcl-2 . Furthermore, ligands that stimulate Toll-like receptors were also found to have the capacity to induce necrosis . Strikingly, this form of cell death was sufficient for the uptake of dead cells by either mouse bone marrow-derived DC or a cell line derived from DC, possibly by virtue of the externalization of phosphatidylserine . Since the loading with bacteria-carrying cells is likely to impact on DC function, this form of necrosis may have a previously unsuspected role in the development of an immune response.

Clin Microbiol Infect, 2004 May, 10(5), 436 - 40
Prevalence of colonisation with third-generation cephalosporin-resistant Enterobacteriaceae in ICU patients of Heidelberg University Hospitals; von Baum H et al.; The aim of this study was to assess colonisation and transmission of third-generation cephalosporin-resistant Enterobacteriaceae (CRE) from patients in 16 intensive care units . A prospective, repetitive point prevalence survey was performed over 6 months, involving samples from 1851 patients . CRE were isolated from 186 (10%) patients, with Enterobacter spp . being the most common . Mean point prevalence rates were significantly higher for paediatric wards (22.5%) compared to surgical (8.1%) and medical (5.5%) units . All CRE isolates were typed by pulsed-field gel electrophoresis . Non-outbreak nosocomial transmission rates of these pathogens were calculated as 12.8% for paediatric patients, compared to 6.8% for adult patients, which may reflect differences in sensitivity to overgrowth with resistant bacteria and contact with health care workers.

Clin Microbiol Infect, 2004 May, 10(5), 416 - 20
Evaluation of the OSIRIS video reader as an automated measurement system for the agar disk diffusion technique; Kolbert M et al.; Measurement of inhibition zones by the automated OSIRIS system was compared with manual measurement . In total, 14 176 measurements were made with 352 staphylococcal and 80 Enterobacteriaceae isolates, involving four panels of antibiotics on round and square Mueller-Hinton agar plates, according to the German DIN 58940 recommendations . Variations of +/- 3 mm in zone size measurements were defined as tolerable . Very major errors (i.e., classification of a resistant isolate as susceptible by the OSIRIS system) occurred in < 1% of tests . With staphylococci, the best concordance was recorded for rifampicin (91.3%), moxifloxacin (88.1%), and gentamicin (86.3%), while the concordance on square plates for vancomycin, pristinamycin and kanamycin was 97.2%, 96.1% and 96.0%, respectively . The poorest concordance was for cefuroxime (43.7%) and novobiocin (47.0%) on round plates, and fosfomycin (36.5%) and chloramphenicol (84.0%) on square plates . With Enterobacteriaceae, 100% concordance was recorded for ampicillin, gentamicin and ciprofloxacin on round agar plates, and for gentamicin, cefoxitin and nalidixic acid on square plates . The poorest results were recorded for nalidixic acid (32.5%) and piperacillin (82.5%) on round plates, and for nitrofurantoin (72.5%) and amoxycillin (82.5%) on square plates . It was concluded that the OSIRIS system was a rapid and reliable system for measuring disk susceptibility test results on round and square agar plates.

Arch Biochem Biophys, 2004 May 15, 425(2), 184 - 92
Flavoenzyme-catalyzed redox cycling of hydroxylamino- and amino metabolites of 2,4,6-trinitrotoluene: implications for their cytotoxicity; Sarlauskas J et al.; The toxicity of 2,4,6-trinitrotoluene (TNT), a widespread environmental contaminant, is exerted through its enzymatic redox cycling and/or covalent binding of its reduction products to proteins and DNA . In this study, we examined the possibility of another cytotoxicity mechanism of the amino- and hydroxylamino metabolites of TNT, their flavoenzyme-catalyzed redox cycling . The above compounds acted as redox-cycling substrates for single-electron transferring NADPH:cytochrome P-450 reductase (P-450R) and ferredoxin:NADP(+) reductase (FNR), as well as substrates for the two-electron transferring flavoenzymes rat liver NAD(P)H:quinone oxidoreductase (NQO1) and Enterobacter cloacae NAD(P)H:nitroreductase (NR) . Their reactivity in P-450R-, FNR-, and NR-catalyzed reactions increased with an increase in their single-electron reduction potential (E(1)(7)) or the decrease in the enthalpy of free radical formation . The cytotoxicity of the amino- and hydroxylamino metabolites of TNT towards bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) was partly prevented by the antioxidant N,N'-diphenyl-p-phenylene diamine and desferrioxamine, and potentiated by 1,3-bis-(2-chloroethyl)-1-nitrosourea, thus pointing to the involvement of oxidative stress . In general, their cytotoxicity increased with an increase in their electron accepting properties, or their reactivity towards the single-electron transferring FNR and P-450R . Thus, our data imply that the flavoenzyme-catalyzed redox cycling of amino and hydroxylamino metabolites of TNT may be an important factor in their cytotoxicity.

Int J Food Microbiol, 2004 Apr 15, 92(2), 217 - 25
Microbial contamination on beef in relation to hygiene assessment based on criteria used in EU Decision 2001/471/EC; McEvoy JM et al.; Thirty-six carcasses were sampled over a 12-month period at an Irish beef abattoir . Between one and five carcass sites (including the hock, brisket, cranial back, bung, inside round and outside round) were sampled after hind leg skinning, hide removal, bung tying, evisceration, splitting, washing, chilling for 24 h and boning, using a wet and dry, cotton wool swab technique . For each sample, total viable counts (TVC), Escherichia coli, total coliforms and Enterobacteriaceae were enumerated . The results are considered in relation to European Union Decision 2001/471/EC which sets performance criteria for TVCs and Enterobacteriaceae in samples taken by excision . Though not explicitly stated in the Decision, it has been proposed that microbiological performance criteria for samples taken by swabbing be set at 20% of the values set for excision samples . Therefore, log mean TVCs in carcass swab samples taken before chilling are acceptable, marginal and unacceptable when they are <2.8, 2.8-4.30 and >4.30 cm(-2), respectively . By these criteria, TVCs on carcasses in the present study were in the marginal range . The marginal result for TVCs was due in the most part to hide removal operations, particularly at the hock and brisket sites . Bacterial contamination on post-chill carcasses was similar or lower to that on pre-chill carcasses, while boning resulted in general increases in TVCs and in E . coli, total coliform and Enterobacteriaceae numbers . In Decision 2001/471/EC, the effects of chilling and boning are not included in the assessment of process control . Data from this study indicate that performance criteria based on log mean Enterobacteriaceae values are unsuitable because of the infrequent occurrence of these organisms on the carcass.

Int J Food Microbiol, 2004 Apr 15, 92(2), 181 - 7
Antimicrobial and antioxidative enrichment of oak (Quercus robur) bark by rotation planar extraction using ExtraChrom; Andrensek S et al.; The multifunctional ExtraChrom instrument was used in the extraction of antimicrobial and radical scavenging components from oak (Quercus robur L.) bark . Milled and sieved oak bark was extracted with 80% (v/v) methanol solution in water on the ExtraChrom instrument using step-gradient in the preparative separation . Extracts were tested using agar diffusion method on Staphylococcus aureus, Enterobacter aerogenes and Candida albicans . Some extracts showed moderate bactericidal, fungicidal, bacteriostatic and fungistatic activity . The composition related to activity of the fractions and extracts was screened simultaneously by thin-layer chromatography (TLC) detected by UV and by spraying the plate with radical scavenging reagent 1,1-diphenyl-2-picrylhydrazyl (DPPH) to detect antioxidant activity . Thus, we could demonstrate the antiradical and antimicrobial activity of oak beneficial in the storage of wine against the oxidation and human microbial exposure.

Proc Natl Acad Sci U S A, 2004 May 4, 101(18), 7118 - 22 Epub 2004 Apr 23.
Gram-positive bacteria are a major reservoir of Class 1 antibiotic resistance integrons in poultry litter; Nandi S et al.; Reversing the spread of antibiotic multiresistant bacteria is hampered by ignorance of the natural history of resistance genes, the mobile elements carrying them, and the bacterial hosts harboring them . Using traditional cultivation and cultivation-independent molecular techniques, we quantified antibiotic resistance genes and mobile elements called integrons in poultry house litter from commercial poultry farms . Unexpectedly, the major reservoir for Class 1 integrons in poultry litter is not their previously identified hosts, Gram-negative Enterobacteriaceae such as Escherichia coli . Rather, integrons and associated resistance genes abound in several genera of Gram-positive bacteria that constitute >85% of the litter community compared with Enterobacteriaceae that comprise <2% of this ecosystem . This finding warrants reexamination of our assumptions about the persistence and spread of antibiotic resistance genes.

Presse Med, 2004 Apr 10, 33(7), 460 - 6
{Etiology of bacterial infections in febrile neutropenic patients: the role of the laboratory in the diagnosis}; Poyart C et al.; EPIDEMIOLOGICAL EVOLUTION: Until the mid-eighties, infectious complications (pneumonia, septicemia) observed in neutropenic patients were, in 70% of cases, of bacterial origin with Gram negative bacillae (Escherichia coli, Klebsiella sp, Pseudomonas aeruginosa) isolated 8 times out of 10 . Among the Gram positive bacteria, Staphylococcus aureus predominated . The etiological profile of bacterial infections has since evolved with a predominance (60 to 70%) of Gram positive bacteria (coagulase-negative staphylococci, viridans streptococci) and a change in the epidemiology of the Gram positive bacteria notably with a lesser frequency of P . aeruginosa infections . THE GRAM POSITIVE BACTERIA: Coagulase-negative staphylococci are among the first germs responsible for nosocomial bacteremia (central venous catheters) and they are usually multiresistant . Viridans streptococci are a frequent cause of bacteremia; they are generally sensitive to antibiotics active on Gram positive bacteria, but the incidence of resistant strains is increasing . Enterococci are in majority responsible for colonisation in neutropenic patients and less frequently for infections; they raise the problem of resistance to antibiotics, notably to glycopeptides . Other Gram positive bacteria can be responsible for infections in neutropenic patients; it is crucial that they be identified because they require treatment with an appropriate antibiotic . GRAM NEGATIVE BACTERIA: Among the enterobacteria, Escherichia coli is predominantly isolated and raises the problem of the increasing incidence of resistance to fluoroquinolone . Pseudomonas aeruginosa, less frequently responsible today, remains associated with a far greater rate of mortality than that observed with the other microorganisms . Other Gram negative bacteria can be identified; they require an adapted antibiotherapy because they are often naturally multiresistant to antibiotics . THE ROLE OF THE LABORATORY: For the diagnosis of infections in neutropenic patients, the microbiology laboratory has a determinating role . The laboratory ensures the analysis of various biological examinations: blood cultures, methods permitting the diagnosis of an infection on a permanent catheter, copro-cultures (research for common enteropathogens, quantification in the case of digestive decontamination, screening for multiresistant bacteria), cytobacteriological examination of urine, samples of respiratory origin, cytobacteriological examination of cerebro-spinal fluid...).

Antimicrob Agents Chemother, 2004 May, 48(5), 1719 - 26
Peritoneal fluid titer test for peritoneal dialysis-related peritonitis; Strijack C et al.; Standard microbiological tests (i.e., MIC) do not account for the unique factors of peritoneal dialysis (PD)-related peritonitis which can significantly influence treatment response . Our goals were to develop a peritoneal fluid titer (PFT) test and to conduct a pilot study of its association with clinical outcome . The methodology was developed by using spent dialysate collected from patients with bacterial PD-related peritonitis prior to the initiation of antibiotics . Dialysate was processed and spiked with antibiotic to simulate two standard intraperitoneal regimens: cefazolin plus tobramycin and cefazolin alone . Thirty-six clinical isolates, including Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa, were tested . In the pilot study, dialysate was collected from 14 patients with bacterial PD-related peritonitis . Titers were determined by using each patient's dialysate and infecting pathogen . Titers were highly reproducible, with discrepancies in only 1% of cases . Overall, PFTs were notably higher against gram-positive bacteria (P < 0.0001) . The addition of tobramycin increased titers significantly from zero to values of 1/16 to 1/64 against E . cloacae and P . aeruginosa (P < 0.0001) . In the pilot study, peritoneal fluid inhibitory titers were significantly associated with clinical outcome, with a median value of 1/96 for patients who were cured compared to 1/32 for those who failed treatment (P = 0.036) . In conclusion, this study provides preliminary support for the PFT as a pharmacodynamic index specific to the treatment of PD-related peritonitis . With further characterization and validation in patients, the PFT test may advance the study of antibiotic therapies for PD-related peritonitis.

Antimicrob Agents Chemother, 2004 May, 48(5), 1688 - 98
Comparative study of the effects of ceftizoxime, piperacillin, and piperacillin-tazobactam concentrations on antibacterial activity and selection of antibiotic-resistant mutants of Enterobacter cloacae and Bacteroides fragilis in vitro and in vivo in mixed-infection abscesses; Stearne LE et al.; The effects of ceftizoxime (CZX), piperacillin (PIP), and PIP-tazobactam (PT) concentrations on the antibacterial activity and selection of resistant mutants of Bacteroides fragilis and Enterobacter cloacae were investigated in vitro in a mixed-culture anaerobic time-kill study and in vivo in a mixed-infection abscess model . Mixed cultures were incubated for 24 h with 0.125 to 512 micro g of CZX per ml or 0.125 to 2,048 micro g of PIP or PT per ml . Mice were treated every 2 h for 24 h with CZX at 6 to 1,536 mg/kg/day or with PIP or PT at 24 to 6,144 mg/kg/day starting 30 min before inoculation with different B . fragilis-E . cloacae combinations . There was a good correlation between the in vitro and in vivo activities of the antibiotics and their MICs obtained with high inocula (10(8) CFU/ml) . The respective 50% effective doses (milligrams per kilogram per day) with B . fragilis and E . cloacae 22491 were 771 and 521 for CZX, 416 and 643 for PIP, and 85 and 554 for PT, and with the B . fragilis-E . cloacae 032349 combination, they were 81 and 21 for CZX and 77 and 766 for PT . Resistant mutants of E . cloacae 22491 were preferentially selected in vitro with 2 to 64 micro g of CZX per ml and in vivo with CZX at 12 to 384 mg/kg/day . There was no preferential selection of CZX-resistant B . fragilis or E . cloacae 032349 . For CZX-resistant E . cloacae 22491, we found a 16- to 512-fold increase in the MIC of CZX and increased MICs of other expanded-spectrum cephalosporins, owing in part to the production of a stably derepressed cephalosporinase . In vitro and in vivo, PT did not select resistant mutants of E . cloacae and B . fragilis . Results demonstrate the adverse microbiological outcome of choosing an expanded-spectrum cephalosporin like CZX for empirical treatment of mixed infections involving a susceptible Enterobacter strain.

Med Dosw Mikrobiol, 2003, 55(4), 351 - 6
{Antibiotic sensitivity of Enterobacteriaceae isolated from women vagina and uterine cervix}; Nolewajka-Lasak I et al.; Based on performed investigation in the group of 200 women treated for recurrent and chronic vaginitis and cervicitis, the characteristic of isolated microorganism was done . There were found series of drug-resistant bacteria in the vagina and uterine cervical canal in women with recurrent vaginitis and chronic cervicitis . In 42.6% of patients with diagnosed chronic cervicitis (Cervicitis chronica n = 50) Escherichia coli strains were isolated, 5% of which produced extended spectrum beta-lactamases (ES beta L) . In women with recurrent vaginitis (Colpitis recidivans n = 150) Escherichia coli strains where isolated in 45.6% . Among them 4.5% produced ES beta L . Expression of beta-lactamases with broadened substrate spectrum was done in double-disc-test . CONCLUSIONS: 1 . Study of antibiotic sensitivity of isolated bacteria should be a diagnostic standard in bacterial infections of uterine cervix and vagina . 2 . The choice of appropriate antibiotics should take into consideration the drug-resistance mechanisms of isolated bacteria . 3 . When drug-resistant bacteria are isolated, combined therapy should be applied.

Med Dosw Mikrobiol, 2003, 55(4), 343 - 9
{Evaluation of relations between plasmids and phage host range among clinical isolates of Enterobacter cloacae}; Nieradko J et al.; The aim of this study was evaluation the plasmid influence on phage host range of clinical strains of Enterobacter cloacae . We found that strains included in restrictive pattern A, displayed reduced host range . Such reduced sensitivity make these strains excellent candidates for search restrictive-modification systems . High discriminative efficacy of isolated phages (specific for strains Enterobacter cloacae) make them useful tool for phage typing in epidemiological investigations.

Phytother Res, 2004 Mar, 18(3), 208 - 11
Antimicrobial effects of Quercus ilex L . extract; Gulluce M et al.; The antimicrobial activities of the methanol extract of Quercus ilex L . (Pirnal oak) leaves were tested in vitro against a wide range of human and plant-associated microorganisms . A total of 132 microbial organisms belonging to 55 bacteria and five fungi and yeast species were studied using a disc-diffusion method and microdilution assays . The results were evaluated as inhibition zones around the disc impregnated with Q . ilex extract at a concentration of 300 micro L/mL . The results showed that Q . ilex did not have any antifungal activities against Alterneria alternata, Aspergillus flavus, Fusarium oxysporum, Penicillum spp., whereas there were inhibition effects on the growth of all Candida albicans isolates . In total 97 bacterial strains (74%) were found to be resistant to Q . ilex extract . The remaining 35 (27%) strains of seven different bacteria genera including Brucella, Bacillus, Enterobacter, Neisseria, Pseudomonas and Escherichia were susceptible to the extract tested . The minimum inhibitory concentrations (MIC) of the extract ranged from 125 to 500 micro L/mL . These results suggest that Q . ilex possesses compounds with antibacterial and anticandidal properties .

Ann Acad Med Singapore, 2004 Mar, 33(2), 228 - 34
Septic arthritis after arthroscopic anterior cruciate ligament reconstruction; Fong SY et al.; INTRODUCTION: A retrospective review of postoperative infected anterior cruciate ligament (ACL) reconstruction was done on 472 consecutive cases in one institution . The purpose was to assess the incidence, diagnosis, treatment and outcome factors . MATERIALS AND METHODS: Out of 472 arthroscopic-assisted ACL reconstructions performed between 1999 and 2002, 7 (1%) postoperative deep intra-articular infections were detected . Seven males with a mean age of 23 years (range, 19 to 30 years) formed the study group; 3 had undergone prior knee surgery . RESULTS: Four patients had acute infection (<2 weeks), 3 had subacute infection (2 weeks to 2 months) and none had late infection (>2 months) . All were admitted within 24 hours of onset of symptoms and underwent immediate arthroscopic lavage, incision and drainage of abscess, debridement with graft retention and intravenous (8 to 31 days) followed by oral (4 to 6 weeks) antibiotics . Staphylococcus aureus was present in 4 patients, Peptostreptococcus in 3, Klebsiella in 1, and Enterobacter in 1 . The patients underwent an average of 1.4 arthroscopic procedures (range, 1 to 3 procedures), with an average hospital stay of 17.3 days per patient . All were evaluated at an average of 11.7 months (range, 5 to 26 months) . In all cases, the infection resolved with stable knees and with all grafts and implants retained . Although rare, early diagnosis and prompt treatment of infection can result in successful eradication without sacrificing the graft.

Environ Pollut, 1987, 48(4), 311 - 9
Bacterial bioabsorption of nickel from industrial cooling water; Kasan HC et al.; Three bacterial strains tolerant to the presence of 100 mg litre(-1) nickel ions were isolated from a water reclamation system . Each organism was tested for ability to accumulate nickel at the above-mentioned concentration . The organism capable of maximum nickel accumulation was identified as an Enterobacter sp . and intracellular nickel deposition by this microorganism was determined using energy dispersive X-ray analysis (EDAX) and transmission electron microscopy . A metal-staining technique for light microscopy was developed . Further studies revealed that growth and glucose utilisation by this isolate was inhibited in culture by nickel.

Environ Pollut, 1990, 65(1), 1 - 17
Crude oil and hydrocarbon-degrading strains of Rhodococcus rhodochrous isolated from soil and marine environments in Kuwait; Sorkhoh NA et al.; Soil and marine samples collected from different localities in Kuwait were screened for microorganisms capable of oil degradation . Both fungi and bacteria were isolated . The fungal flora consisted of Aspergillus terreus, A . sulphureus, Mucor globosus, Fusarium sp . and Penicillum citrinum . Mucor globosus was the most active oil degrading fungus isolated . Bacterial isolates included Bacillus spp . Enterobacteriaceae, Pseudomonas spp., Nocardia spp., Streptomyces spp.,and Rhodococcus spp . Among these Rhodococcus strains were the most efficient in oil degradation and, relatively speaking, the most abundant . Bacterial and fungal isolates differed in their ability to degrade crude oil, with Rhodococcus isolates being more active that fungin in n-alkane biodegradation, particularly in the case of R . rhodochrous . In addition to medium chain n-alkanes, fungi utilized one or more of the aromatic hydrocarbons studied, while bacteria failed to do so . R . rhodochorous KUCC 8801 was shown by GLC and post-growth studies to be more efficient in oil degradation than isolates known to be active oil degraders.

Am J Med Sci, 2004 Mar, 327(3), 118 - 22
Simultaneous presence of blaTEM and blaSHV genes on a large conjugative plasmid carried by extended-spectrum beta-lactamase-producing Klebsiella pneumoniae; Araque M et al.; BACKGROUND: Among members of the family Enterobacteriaceae, the production of plasmid-mediated extended-spectrum beta-lactamases (ESbetaLs) has emerged as an important mechanism of the resistance to beta-lactams . METHODS: The molecular basis of extended-spectrum beta-lactamase in 48 strains of Klebsiella pneumoniae, recovered from neonatal patients with nosocomial septicemia during an outbreak that occurred in November 2000 at a Neonatal Intensive Care Unit of The Andes University Hospital in Venezuela, were investigated . RESULTS: The isolates were resistant to expanded-spectrum cephalosporins, aztreonam, gentamicin, kanamycin, tetracycline, and chloramphenicol, but remained susceptible to cefoxitin, imipenem, amikacin, and tobramycin . Production of ESbetaL activity was confirmed by restoring susceptibility to ceftazidime in the presence of clavulanic acid . All isolates harbored an 87-kilobase plasmid . Analysis of the outer membrane protein patterns did not reveal relevant changes of the porins profile . All resistance markers were transferable to Escherichia coli by conjugation and were lost en bloc after treatment with acridine orange . Isoelectric focusing for beta-lactamases was performed on transconjugants, obtaining 2 bands with isoelectric points of 5.4 and 8.2 . Genes encoding both enzymes are located on the single, self-transferable 87-kilobase plasmid pKAM542 . Analysis of the plasmid by hybridization revealed the presence of both blaTEM and blaSHV determinants . Cloning and sequencing identified them as blaTEM-1 and blaSHV-5, respectively; the latter was responsible for the ESbetaL activity among nosocomial isolates of K pneumoniae . CONCLUSIONS: Microbiologists, epidemiologists, and clinicians must be aware of potential ESbetaL-encoding organisms to assess proper antimicrobial managements and effective infection controls.

J Biol Chem, 2004 Jul 2, 279(27), 27928 - 40 Epub 2004 Apr 15.
Biosynthesis of a novel 3-deoxy-D-manno-oct-2-ulosonic acid-containing outer core oligosaccharide in the lipopolysaccharide of Klebsiella pneumoniae; Frirdich E et al.; The core oligosaccharide region of Klebsiella pneumoniae lipopolysaccharide contains some novel features that distinguish it from the corresponding lipopolysaccharide region in other members of the Enterobacteriaceae family, such as Escherichia coli and Salmonella . The conserved Klebsiella outer core contains the unusual trisaccharide 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo)-(2,6)-GlcN-(1,4)-GalUA . In general, Kdo residues are normally found in the inner core, but in K . pneumoniae, this Kdo residue provides the ligation site for O polysaccharide . The outer core Kdo residue can also be non-stoichiometrically substituted with an l-glycero-d-manno-heptopyranose (Hep) residue, another component more frequently found in the inner core . To understand the genetics and biosynthesis of core oligosaccharide synthesis in Klebsiella, the gene products involved in the addition of the outer core GlcN (WabH), Kdo (WabI), and Hep (WabJ) residues as well as the inner core HepIII residue (WaaQ) were identified . Non-polar mutations were created in each of the genes, and the resulting mutant lipopolysaccharide was analyzed by mass spectrometry . The in vitro glycosyltransferase activity of WabI and WabH was verified . WabI transferred a Kdo residue from CMP-Kdo onto the acceptor lipopolysaccharide . The activated precursor required for GlcN addition has not been identified . However, lysates overexpressing WabH were able to transfer a GlcNAc residue from UDP-GlcNAc onto the acceptor GalUA residue in the outer core.

J Bacteriol, 2004 May, 186(9), 2540 - 7
Characterization of the alternative sigma factor sigma54 and the transcriptional regulator FleQ of Legionella pneumophila, which are both involved in the regulation cascade of flagellar gene expression; Jacobi S et al.; We cloned and analyzed Legionella pneumophila Corby homologs of rpoN (encoding sigma(54)) and fleQ (encoding sigma(54) activator protein) . Two other genes (fleR and pilR) whose products have a sigma(54) interaction domain were identified in the genome sequence of L . pneumophila . An rpoN mutant strain was nonflagellated and expressed very small amounts of the FlaA (flagellin) protein . Like the rpoN mutant, the fleQ mutant strain of L . pneumophila was also nonflagellated and expressed only small amounts of FlaA protein compared to the amounts expressed by the wild type . In this paper we show that the sigma(54) factor and the FleQ protein are involved in regulation of flagellar gene operons in L . pneumophila . RpoN and FleQ positively regulate the transcription of FliM and FleN, both of which have a sigma(54)-dependent promoter consensus sequence . However, they seemed to be dispensable for transcription of flaA, fliA, or icmR . Our results confirmed a recently described model of the flagellar gene regulation cascade in L . pneumophila (K . Heuner and M . Steinert, Int . J . Med . Microbiol . 293:133-145, 2003) . Flagellar gene regulation was found to be different from that of Enterobacteriaceae but seems to be comparable to that described for Pseudomonas or Vibrio spp.

Di Yi Jun Yi Da Xue Xue Bao, 2004 Apr, 24(4), 439 - 40, 444
{Drug-resistance analysis of Gram-negative bacilli producing extended-spectrum beta-lactamases in lower respiratory tract infection}; Long J et al.; OBJECTIVE: To analyze occurrence of Gram-negative bacteria producing extended-spectrum beta-lactamase (ESBL) in patients with lower respiratory tract infection and assess their drug sensitivity to provide evidence for clinical management of lower respiratory tract infection with drug-resistant ESBL-producing bacteria . METHODS: A total of 312 Gram-negative bacterial strains were isolated from patients with lower respiratory tract infection and identified for the presence of ESBL by Vitek ESBL detection test and double-disk synergy tests as well as standard disk diffusion test . The effects of 10 antibacterial drugs on ESBL-producing strains were compared . RESULTS: A rate of 20.1% of the isolated strains were identified to produce ESBLs, including Klebsiella Pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginasa and Klebsiella oxytocla (accounting for 30.1%, 29.1%, 27.0%, 11.3%, and 2.5%, respectively, of the ESBL-producing strains) . Imipenem possessed the most powerful antibacterial activity against ESBL-producing strains . CONCLUSIONS: Attention should be given to lower respiratory tract infections with ESBL-producing Gram-negative bacilli, and imipenem can be the primary choice for fighting such infections.

Cancer Res, 2004 Apr 15, 64(8), 2853 - 7
Efficient cancer therapy with a nanobody-based conjugate; Cortez-Retamozo V et al.; Nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain . Nanobodies are strictly monomeric, very stable, and highly soluble entities . We identified a nanobody with subnanomolar affinity for the human tumor-associated carcinoembryonic antigen . This nanobody was conjugated to Enterobacter cloacae beta-lactamase, and its site-selective anticancer prodrug activation capacity was evaluated . The conjugate was readily purified in high yields without aggregation or loss of functionality of the constituents . In vitro experiments showed that the nanobody-enzyme conjugate effectively activated the release of phenylenediamine mustard from the cephalosporin nitrogen mustard prodrug 7-(4-carboxybutanamido) cephalosporin mustard at the surface of carcinoembryonic antigen-expressing LS174T cancer cells . In vivo studies demonstrated that the conjugate had an excellent biodistribution profile and induced regressions and cures of established tumor xenografts . The easy generation and manufacturing yield of nanobody-based conjugates together with their potent antitumor activity make nanobodies promising vehicles for new generation cancer therapeutics.

Cir Cir, 2004 Jan-Feb, 72(1), 41 - 6
{Nosocomial infection surveillance: experience at a cardiology hospital in Mexico}; Valenzuela-Flores AA et al.; OBJECTIVE: To describe the results of the Nosocomial Infection surveillance program at the Cardiology Hospital in Centro Medico Nacional Siglo XXI of the Mexican Social Security Institute . METHODOLOGY: To inform of the epidemiologic follow-up results from January 2000 to July 2003 . Global frequency, infection rates by infection site, hospital services, and frequency of most common microorganisms were estimated . RESULTS: During this period, global incidence showed that in every 100 discharges, there were 4.3 infections (4.3/100) . Frequency of infection was slightly higher in the surgical intensive care unit . Average infection rate in lower respiratory tract infections was 27/1,000; in surgical-site infections: 8/1,000 (mediastinitis 0.8/1,000); it was found that in urinary tract infection, rate was 6.6/1,000 . Most commonly isolated microorganisms were: coagulase-negative Staphylococcus (25%), Enterobacter sp . (17%), Candida albicans (13%), S . aureus (9%), P . aeruginosa (9%) and K . pneumoniae (6%) . CONCLUSION: This study describes the epidemiology of nosocomial infections in a Cardiology Hospital . The information is obtained through epidemiologic follow-up programs . This information is very important to develop specific strategies for control of infections.

Bacteriol Virusol Parazitol Epidemiol, 2002 Jul-Dec, 47(3-4), 185 - 91
{Beta-lactam resistance in aquatic Enterobacter cloacae strains using phenotypic and genotypic criteria}; Lazar V et al.; Bacterial resistance by producing of beta-lactamases represents an increasing problem of infections chemotherapy . beta-lactam hydrolyzing activities are detected in virtually all bacteria, from witch Enterobacter cloacae produce chromosomal beta-lactamases included in inducible class AmpC beta-lactamases . The purpose of this preliminary study was to investigate the antibiotic susceptibility of 7 inducible beta-lactamase-producing Enterobacter cloacae strains isolated from aquatic sources (river and polluted waters) . The identification to the species level was performed with the API 32E system and susceptibility to antimicrobial agents was tested by the disk diffusion method according to NCCLS recommendations . The following antibiotics were tested: ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cefamandole, cephaclor, imipenem, amikacin, gentamycin, kanamycin, tobramycin, ciprofloxacin, norfloxacin, ofloxacin, nalidixic acid, tetracycline and chloramphenicol . Minimum inhibitory concentrations (MICs) were performed using dilution method in Mueller-Hinton broth with a 0.06-64 micrograms/ml concentration range for all antimicrobials and bacterial inoculum of about 1-2 x 10(8) cfu/ml . For the data analysis NCCLS breakpoints for resistance and sensitivity were used . Interaction of beta-lactamase inhibitor clavulanate with cefotaxime was performed by double-disk synergy test . Detection of inducible beta-lactamase expression was performed by the inductibility disk diffusion test using cefotaxime, ceftazidime and imipenem . Genomic DNA was isolated using CTAB technique and bacterial plasmid isolation was performed by an alkaline lysis method . Genetic characterization was performed by agarose gel electrophoresis and spectrophotometric analysis . The majority of examined E . cloacae strains were sensitive to imipenem, cefamandole, amikacin and quinolones (norfloxacin and ofloxacin), a higher moderate resistance being observed only to nalidixic acid (higher than 50%) and ciprofloxacin (15%) . The percentage of resistant strains ranged from 72% (to kanamycin) to 87% (to gentamicin) . The best results (resistance about 99%) were obtained with ampicillin, amoxycillin/clavulanic acid, ceftazidime, cefotaxime, cephalotin, cephaclor, tobramycin, tetracycline and chloramphenicol . The disk diffusion tests showed the absence of extended-spectrum beta-lactamases production and the expression of inducible beta-lactamases . Electrophoretic patterns point out the presence of plasmid DNA . Plasmid profile revealed the presence of several different plasmids ranging from 2.5 kpb to more than 30 kpb . The presence of inducible beta-lactamase E . cloacae strains in aquatic media (river and polluted waters) and the closely related pattern of susceptibility among these strains reflect the possible contamination of these sources and the common origin of them.

Bacteriol Virusol Parazitol Epidemiol, 2002 Jul-Dec, 47(3-4), 137 - 42
{Enterobacteria isolated from different pathological products and their sensitivity to antibiotics}; Idomir M et al.; The objectives of the study we have enhanced have focused on identifying the etiological implication of the Enterobacteria isolated from different pathological products and determining their sensitivity to the main antibiotics . We observed that from pharyngeal exudate and sputum the Enterobacter spp . strains were isolated the most frequently while from purulent secretion, urine, puncture liquids and blood, E . coli strains were the most frequently implicated . For all the categories etiologically implicated, there were selected strains resistant to one or more of the antibiotics tested, showing once again the importance of antibiogram in the initiation and control of infections disease treatment.

J Food Prot, 2004 Apr, 67(4), 658 - 65
Escherichia coli O157 prevalence and enumeration of aerobic bacteria, Enterobacteriaceae, and Escherichia coli O157 at various steps in commercial beef processing plants; Arthur TM et al.; The effectiveness of current antimicrobial interventions used in reducing the prevalence or load of Escherichia coli O157 and indicator organisms on cattle hides and carcasses at two commercial beef processing plants was evaluated . Sponge sampling of beef cattle was performed at five locations from the initial entry of the animals to the slaughter floor to the exit of carcasses from the "hotbox" cooler . For each sample, E . coli O157 prevalence was determined and total aerobic bacteria, Enterobacteriaceae, and E . coli O157 were enumerated . E . coli O157 was found on 76% of animal hides coming into the plants, but no carcasses leaving the cooler were identified as contaminated with E . coli O157 . A positive relationship was seen between the incidence of E . coli O157 in hide samples and that in preevisceration samples . Aerobic plate counts and Enterobacteriaceae counts averaged 7.8 and 6.2 log CFU/100 cm2, respectively, on hides, and 1.4 and 0.4 log CFU/100 cm2, respectively, on chilled carcasses . Aerobic plate counts and Enterobacteriaceae counts on preevisceration carcasses were significantly related to the respective levels on the corresponding hides; the carcasses of animals whose hides carried higher numbers of bacteria were more likely to carry higher numbers of bacteria . Implementation of the sampling protocol described here would allow processors to evaluate the efficacy of on-line antimicrobial interventions and allow industrywide benchmarking of hygienic practices.

J Food Prot, 2004 Apr, 67(4), 646 - 50
Prevalence of Escherichia coli O157 and levels of aerobic bacteria and Enterobacteriaceae are reduced when hides are washed and treated with cetylpyridinium chloride at a commercial beef processing plant; Bosilevac JM et al.; The objective of this experiment was to test the potential of a combined water wash and cetylpyridinium chloride (CPC) treatment as a hide intervention applied to cattle in the holding pens of a processing plant immediately before stunning . Over 2 processing days, 149 control and 139 treated cattle were tested . Control cattle were processed in the normal manner . The treatment group was prewashed with water the day before harvest . Immediately before stunning, these cattle were sprayed twice with 1% CPC, first for 3 min, then for 1 min . Hides and preevisceration carcasses were sampled to determine aerobic plate counts, Enterobacteriaceae counts (EBC), and Escherichia coli O157 prevalence . The treatment reduced the prevalence of E . coli O157 on hides from 56% to 34% and the prevalence on preevisceration carcasses from 23% to 3% . The treatment decreased aerobic plate counts from 4.9 log CFU/100 cm2 to 3.4 log CFU/100 cm2 and EBC from 3.1 log CFU/100 cm2 to 2.0 log CFU/100 cm2 on preevisceration carcasses . The treatment of hides did not result in any detectable CPC contamination of the chilled carcasses . These data indicated that a 1% CPC treatment preceded by a water wash was capable of reducing hide prevalence of E . coli O157 from as high as 80% to less than 50%, resulting in preevisceration carcass prevalence of 5% or less . We conclude that water washing followed by an antimicrobial treatment, such as CPC, has great potential as an effective hide intervention step and should be further evaluated for implementation as a processing step after stunning and before hide removal.

Biochem Biophys Res Commun, 2004 May 7, 317(3), 851 - 6
Resistance to imipenem, cefepime, and cefpirome associated with mutation in Omp36 osmoporin of Enterobacter aerogenes; Thiolas A et al.; Enterobacter aerogenes develops increased multidrug resistance via a functional alteration of outer-membrane permeability associated with a decrease in porin function . We have sequenced the gene coding the major porin of Enterobacter aerogenes, omp36 . The sequence shows a high similarity with the Klebsiella pneumoniae ompK36 gene and is closely related to the enterobacterial OmpC family . Sequence analysis of several Omp36 issued from clinical strains indicated variability in putative cell-surface exposed domains . Interestingly, substitution Gly112Asp was observed in the conserved eyelet L3 region of the porin produced by two strains, C and 3 . This substitution is associated with a high general beta-lactam resistance observed in these isolates and with alteration of pore properties previously described in strain 3 porin {Mol . Microbiol . 41 (2001) 189} . This is the first genetic identification of impermeability-mediated resistance to beta-lactams in various clinical E . aerogenes strains.

Mil Med, 2004 Mar, 169(3), 194 - 7
Management of postirradiation infection: lessons learned from animal models; Brook I et al.; Ionizing radiation depresses host defenses and enhances susceptibility to local and systemic infection due to endogenous or exogenous microorganisms . Exposure of mice to a lethal dose of ionizing 60Co-gamma radiation induces a dose-related reduction in the number of both aerobic and anaerobic bacteria from 10(10-12) to 10(4-6) per gram of stool within 4 days . The number of anaerobic bacteria stays low, but the number of Enterobacteriaceae per gram of stool increases significantly up to 10(9) by the 12th day after irradiation . This increase is associated with bacterial translocation of these organisms and fatal bacteremia . The use of quinolones in the irradiated animals was effective in controlling systemic endogenous Gram-negative infection after irradiation . Supplementation with penicillin prevented treatment failures due to Streptococcus spp . and increased survival . Quinolones given for 21 days also were effective in management of systemic exogenous infections due to orally ingested Klebsiella pneumoniae and Pseudomonas aeruginosa . Effectiveness of quinolones may be attributed to inhibition of exogenous organism growth within the gut lumen while preserving the anaerobic gut flora as well as their systemic antibacterial activity . Based on these findings, antimicrobial agents recommended for therapy of infection after exposure to irradiation are: ciprofloxacin, levofloxacin, ceftriaxone, cefepime, gentamicin +/- amoxicillin, or vancomycin.

Emerg Infect Dis, 2004 Jan, 10(1), 94 - 9
Fluoroquinolones protective against cephalosporin resistance in gram-negative nosocomial pathogens; Schwaber MJ et al.; In a matched case-control study, we studied the effect of prior receipt of fluoroquinolones on isolation of three third-generation cephalosporin-resistant gram-negative nosocomial pathogens . Two hundred eighty-two cases with a third-generation cephalosporin-resistant pathogen (203 with Enterobacter spp., 50 with Pseudomonas aeruginosa, and 29 with Klebsiella pneumoniae) were matched on length of stay to controls in a 1:2 ratio . Case-patients and controls were similar in age (mean 62 years) and sex (54% male) . Variables predicting third-generation cephalosporin resistance were surgery (p = 0.005); intensive care unit stay (p < 0.001); and receipt of a b-lactam/b-lactamase inhibitor (p < 0.001), a ureidopenicillin (p = 0.002), or a third-generation cephalosporin (p < 0.001) . Receipt of a fluoroquinolone was protective against isolation of a third-generation cephalosporin-resistant pathogen (p = 0.005) . Interventional studies are required to determine whether replacing third-generation cephalosporins with fluoroquinolones will be effective in reducing cephalosporin resistance and the effect of such interventions on fluoroquinolone resistance.

Pediatr Dermatol, 2004 Mar-Apr, 21(2), 113 - 6
Microbiology of infected hemangiomas in children; Brook I; Bacterial infections are a common complication of hemangiomas in children . The objective of this study was to establish the aerobic and anaerobic microbiology of infected hemangiomas . A retrospective 8-year review of clinical and microbiology laboratory records from patients with secondarily infected hemangiomas was carried out . Specimens from infected sites were processed for the presence of aerobic and anaerobic bacteria . Bacterial growth was present in 32 of 38 specimens . Aerobic bacteria alone were recovered in 12 infected hemangiomas (37.5%), anaerobic bacteria alone in 8 (33%), and mixed aerobic and anaerobic flora in 12 (37.5%) . A total of 80 isolates (47 aerobes and 33 anaerobes) were recovered, giving an average of 2.5 isolates per specimen (1.5 aerobes and 1.0 anaerobes) . The highest number of isolates were recovered in infections of the perineum (3.7 per site) and the legs (2.8 per site) . The predominant aerobic isolates were Staphylococcus aureus, group A beta-hemolytic streptococci, and Enterobacteriaceae . The predominant anaerobes were Peptostreptococcus sp., gram-negative bacilli, and Fusobacterium sp . Organisms that belong to the mucous membranes close to the lesions predominated in infections next to those membranes . The polymicrobial etiology of secondarily infected hemangiomas and the association of bacterial flora with the anatomic site of the lesions is thereby demonstrated.

J Chin Med Assoc, 2004 Jan, 67(1), 15 - 20
Neonatal bacteremia in a neonatal intensive care unit: analysis of causative organisms and antimicrobial susceptibility; Lee NC et al.; BACKGROUND: Infections cause significant mortality and morbidity in neonates, especially the premature ones . Although there are various antibiotics can be used to combat neonatal infection, resistant strains have subsequently emerged . In an epidemiological survey, we analyzed bacterial isolates and their antibiotic susceptibilities for cases of bacteremia in a neonatal intensive care unit (NICU) of a teaching hospital . METHODS: From November 1999 to October 2001, 623 neonates admitted to the NICU were enrolled . The incidence and incidence density of bacteremia, morbidity and mortality of sepsis, as well as antibiotic susceptibility, were investigated . RESULTS: Totally, 754 blood cultures were done on 623 patients . Fifty-eight patients experienced 85 episodes of bacteremia, with 87 isolates cultured . The incidence of bacteremia in our NICU was 9.31% (58/623) with an incidence density of 10.98/1000 patient-days . The overall mortality rate was 7.22% . The case fatality rate of bacteremia was 20.7% (12/58) . The bacterial pathogens encountered, in order of frequency, were coagulase-negative Staphylococcus (29%), Staphylococcus aureus (22%), and Enterobacter cloacae (17%) . All of the gram-positive bacteria were susceptible to vancomycin, while the gram-negative bacteria were susceptible to imipenem, amikacin and ciprofloxacin . Oxacillin-resistant S . epidermidis, oxacillin-resistant S . aureus, and multi-drug resistant enterobacteriae were the leading microorganisms causing bacteremia in our NICU . CONCLUSIONS: It is an endless struggle to combat neonatal infection . Periodic evaluation of bacterial antibiotic susceptibility is necessary . More judicious selection of antibiotics and rotating antibiotic regimens should be kept in mind to reduce the resurgence of multidrug resistant strains.

J Physiol Pharmacol, 2003 Dec, 54 Suppl 4, 165 - 82
Role of COX inhibition in pathogenesis of NSAID-induced small intestinal damage; Takeuchi K et al.; Nonsteroidal antiinflammatory drugs (NSAIDs) such as indomethacin decrease mucosal PGE(2) production by inhibiting cyclooxygenase (COX) activity and produce damage in the small intestine . The development of intestinal lesions as induced by indomethacin was accompanied by increases in intestinal motility, enterobacterial invasion, and myeloperoxidase (MPO) as well as inducible nitric oxide synthase (iNOS) activity, together with the up-regulation of COX-2 and iNOS mRNA expression . Neither the selective COX-1 inhibitor, SC-560, nor the selective COX-2 inhibitor, rofecoxib, alone caused intestinal damage, but their combined administration produced lesions . SC-560, but not rofecoxib, caused intestinal hypermotility, bacterial invasion and the expression of COX-2 as well as iNOS mRNAs, yet the iNOS and MPO activity was increased only when rofecoxib was administered together with SC-560 . Although SC-560 inhibited the PG production, the level of PGE(2) was recovered, in a rofecoxib-dependent manner . The intestinal hypermotility response to indomethacin was prevented by both 16,16-dimethyl PGE(2) and atropine but not ampicillin, yet all these agents inhibited not only the bacterial invasion but also the expression of COX-2 as well as the iNOS activity in the intestinal mucosa following indomethacin treatment, resulting in preventing the intestinal lesions . These results suggest that inhibition of COX-1, despite causing intestinal hypermotility, bacterial invasion and iNOS expression, up-regulates the expression of COX-2, and the PGE(2) derived from COX-2 counteracts deleterious events caused by COX-1 inhibition and maintains the mucosal integrity . These sequences of events explain why intestinal damage occurs when both COX-1 and COX-2 are inhibited.

Arq Bras Cardiol, 2004 Mar, 82(3), 287 - 90 Epub 2004 Apr 05.
{Vascular prosthesis infection in thoracic aorta surgery: review of the experience and a case report illustrating treatment with an unconventional technique}; Fontes RD et al.; We report the case of a 37-year-old-female patient who had undergone a Bentall procedure at our service and returned with intense chest pain and acute aortic dissection type III, which was diagnosed and clinically treated . One year after this episode, this dissection expanded, and the patient underwent surgery with interposition of a Dacron graft in the descending aorta . In the immediate postoperative period, the patient experienced left bronchopneumonia and was discharged afebrile and in good condition . One month after discharge, she returned with fever and toxemia . Pleural empyema was diagnosed, and she underwent an exploratory thoracotomy that did not confirm this diagnosis, but revealed intense effusion thickening . Four months after the exploratory thoracotomy, Klebsiella pneumoniae and Enterobacter sp were isolated in a blood culture . Magnetic resonance imaging revealed shapes compatible with perigraft infection . With this clinical and laboratory picture, graft removal was indicated as was axillo-bifemoral grafting . Surgery was successfully performed, the patient was discharged in good condition, and remains well after a 57-month follow-up without complications . The methods used for diagnosis and treatment of prosthesis infection in thoracic aorta surgery are discussed.

J Clin Microbiol, 2004 Apr, 42(4), 1542 - 6
Evaluation of the automated phoenix system for potential routine use in the clinical microbiology laboratory; Donay JL et al.; A comparative study was designed to evaluate the identification (ID) and antimicrobial susceptibility testing (AST) performances of the BD Phoenix Automated Microbiology System (Becton Dickinson Diagnostic Systems {BD}, Pont de Claix, France) . A total of 305 single clinical isolates were collected, and comparisons were made with routine manual methods in use in our microbiology laboratories . The percentages of correct IDs were 93.3, 89.4, 91.8, and 85.7% for enterobacteria, nonfermenting gram-negative bacilli, staphylococci, and streptococci-enterococci, respectively . The median ID time was 3 h, and the median time for AST was 10 h 30 min . AST results showed variable percentages of errors for the different antibiotics . None of the enterobacteria and 0.3% of Pseudomonas aeruginosa isolates showed a very major error (VME) . Only one strain of Staphylococcus aureus showed a VME with oxacillin . We demonstrate here the efficiency of the Phoenix system, which can be used for the majority of strains encountered in a university-based laboratory, for ID and AST.

Infect Control Hosp Epidemiol, 2004 Mar, 25(3), 226 - 30
Nosocomial pediatric bacteremia: the role of intravenous set contamination in developing countries; Macias AE et al.; OBJECTIVE: To assess the rate of bacterial contamination of intravenous administration sets at their rubber injection ports and matching infusates . DESIGN: Cultures of injection ports and infusate during 26 visits to 4 hospitals . SETTING: Four public general pediatric hospitals in Mexico City with substandard care practices . PATIENTS: Hospitalized pediatric patients receiving intravenous solutions . RESULTS: Overall, 176 of 251 injection ports were contaminated (70.1%; 95% confidence interval {CI95}, 64.5% to 75.8%), 35 (13.9%; CI95, 9.7% to 18.2%) with gram-negative rods, primarily of the tribe Klebsielleae . Cultures of infusates were positive in 17 cases (6.8%, CI95, 3.7% to 9.9%), 5 of which grew gram-negative rods (2%; CI95, 0.6% to 4.6%) . In 3 cases (1.2%), the same species with gram-negative rods was found in the infusates and on the injection ports . During one visit, 8 clustered cases of injection port contamination with a clonal Enterobacter cloacae were found; this agent was also found in the blood culture, intravenous fluid, and parenteral nutrition of one patient . Inadequate chlorination of tap water, a potential risk factor, was recorded during 22 visits (84.6%) . CONCLUSION: These data suggest that external contamination of the intravenous administration set could play a role in infusate contamination.

Infect Control Hosp Epidemiol, 2004 Mar, 25(3), 192 - 7
Dynamics of bacterial hand contamination during routine neonatal care; Pessoa-Silva CL et al.; OBJECTIVE: To evaluate the dynamics of bacterial contamination of healthcare workers' (HCWs) hands during neonatal care . SETTING: The 20-bed neonatal unit of a large acute care teaching hospital in Geneva, Switzerland . METHODS: Structured observation sessions were conducted . A sequence of care began when the HCW performed hand hygiene and ended when the activity changed or hand hygiene was performed again . Alcohol-based handrub was the standard procedure for hand hygiene . An imprint of the five fingertips of the dominant hand was obtained before and after hand hygiene and at the end of a sequence of care . Regression methods were used to model the final bacterial count according to the type and duration of care and the use of gloves . RESULTS: One hundred forty-nine sequences of care were observed . Commensal skin flora comprised 72.4% of all culture-positive specimens (n = 360) . Other microorganisms identified were Enterobacteriaceae (n = 55, 13.8%); Staphylococcus aureus (n = 10, 2.5%); and fungi (n = 7, 1.8%) . Skin contact, respiratory care, and diaper change were independently associated with an increased bacterial count; the use of gloves did not fully protect HCWs' hands from bacterial contamination . CONCLUSIONS: These data confirm that hands become progressively contaminated with commensal flora and potential pathogens during neonatal care, and identify activities at higher risk for hand contamination . They also reinforce the need for hand hygiene after a sequence of care, before starting a different task, and after glove removal.

J Microbiol Immunol Infect, 2004 Feb, 37(1), 35 - 8
Characteristics of nosocomial bacterial meningitis in children; Lin PC et al.; Nosocomial meningitis is uncommon in children . We reviewed the medical records of all children who developed bacterial meningitis at least 72 hours after admission to Mackay Memorial Hospital for the period July 1992 through June 2000 . Clinical manifestations, predisposing factors, pathogens, and outcomes were analyzed . Twenty-two cases of nosocomial meningitis were identified, comprising 9.2% (22/239) of all pediatric cases of bacterial meningitis during the study period . The male-to-female ratio was 14:8 . All patients were younger than 6 months of age except for one, who was 7 years old . The mean duration between admission and onset of meningitis was 15.3 days (range, 3 to 58 days) . Twenty-two organisms were isolated, including 13 Gram-negative bacteria (59%) and 9 Gram-positive bacteria (41%) . The most common pathogen was Escherichia coli (5 cases), followed by Enterobacter cloacae (3), Staphylococcus aureus (3), and Chryseobacterium meningosepticum (3) . Seventeen patients (77%) had concomitant bacteremia . Predisposing factors for acquisition of nosocomial meningitis included previous treatment with broad-spectrum antibiotics (68%), prematurity with very low birth weight (41%), and total parenteral nutrition (32%) . Two patients (9%) had previous neurosurgical intervention . Four patients (18%) died, 3 of whom were low birth weight premature infants . Nine patients (41%) had sequelae, including developmental delay, hydrocephalus, hearing impairment, and epilepsy . Neurosurgery was not a significant risk factor for the development of nosocomial meningitis, while very low birth weight played an important role . Previous intraventricular hemorrhage or hydrocephalus, prematurity with very low birth weight, infection with Gram-negative bacteria, and prior broad-spectrum antibiotic administration were associated with poor outcome.

J Microbiol Immunol Infect, 2004 Feb, 37(1), 1 - 7
Effects of PNPG on cell growth cycle, motility machinery and quorum sensing in Serratia marcescens; Wei JR et al.; p-Nitrophenylglycerol (PNPG) effectively inhibits swarming of the enterobacterium Proteus mirabilis . The underlying mechanism of inhibition is unclear . We have now found that both PNPG also inhibits motility and swarming in another enterobacterium, Serratia marcescens . While the peak promoter activities of the flagellar master operon (flhDCSm), the flagellin structural gene (hagSm) and the nuclease gene (nucASm) in S . marcescens increased with increasing PNPG concentration, the expression of these genes was delayed in accordance with the reduced growth rate . As the quorum-sensing system is involved in the regulation of swarming in S . marcescens, we also examined the effect of PNPG on the production of quorum-sensing signal molecules and found that their expression was delayed with a reduced level . PNPG, therefore, had a pleiotropic effect on all aspects of S . marcescens physiology relating to swarming . The underlying molecular mechanism remains to be elucidated.

Spinal Cord, 2004 Apr, 42(4), 230 - 4
Bacteriological investigation of infected pressure ulcers in spinal cord-injured patients and impact on antibiotic therapy; Heym B et al.; STUDY DESIGN: Retrospective . OBJECTIVES: To improve the use of bacteriological results for treating spinal cord-injured patients with infected pressure ulcers . SETTING: Microbiology and Orthopaedics Department, Ambroise Pare University Hospital, Boulogne-Billancourt, France . METHODS: Tissue specimens, sampled at the end of the surgical intervention from unbridled and cleaned ulcers were analysed . Drainage liquids were cultured at day 1 (D1) and day 5 (D5) postsurgery . For part of the patients, a presurgery superficial sample was analysed and compared with the surgical and postsurgical samples . RESULTS: In all, 168 surgical samples from 101 patients, 183 D1 and 104 D5 wound drainage liquids were included in this study . Out of the 168 surgical samples 17 (10%) had a negative culture, whereas 151 (90%) had a positive culture . For drainage liquids, the culture was negative in 48% and 56% of the samples at D1 and D5, respectively . The most frequently isolated species were enterobacteria, followed by staphylococci and streptococci . CONCLUSION: Culturing deep tissue specimens sampled from the surgically cleaned and unbridled ulcers allows for the isolation of the bacterial species that are really involved in the ulcer infection . As the identification of these bacteria and their antibiotic susceptibility are available, when the culture results of the D1 postsurgical drainage liquid is also available, it is easier to choose targeted antibiotic treatment.

Zhonghua Nei Ke Za Zhi, 2004 Feb, 43(2), 112 - 6
{The impact of gastric colonization on the pathogenesis of ventilator-associated pneumonia}; Li HY et al.; OBJECTIVE: To investigate the risk factors for gastric bacterial colonization and its role in the endogenous pathogenesis of ventilator-associated pneumonia (VAP) . METHODS: The type and concentration of gastric colonized bacteria and its relationship with the time when samples were collected, and with the type and occurrence order of the pathogens detected in samples from lower respiratory tract after the onset of VAP were analyzed dynamically in the patients with tracheal-intubation or tracheotomy in intensive care unit (ICU) . RESULTS: (1) The isolation rate of colonized bacteria in gastric cavity was associated with the pH of gastric juice . When the pH of gastric juice was > 4, the isolation rate of Gram-negative bacilli (GNB) in gastric cavity markedly increased, achieving 52.5% in VAP group, and the incidence of VAP was higher (P = 0.017) . The pH value of gastric juice was positively correlated with the logarithmic concentration of GNB in gastric cavity (P = 0.001) . (2) As the duration of intubation prolonged, the isolation rate of enterobacteriaceae in VAP group increased, which was 45.2% on the fifth day of intubation . In contrast, the isolation rate in non-VAP group was 11.1% (P < 0.01) . (3) The colonization of enterobacteriaceae in gastric cavity was 1 - 2 days earlier than that in oropharynx . The order was statistically significant (P = 0.015) . (4) The reverse order of stomach-pharynx-lower respiratory tract colonization was found in 12 cases of the total 52 VAP patients and the order of stomach to lower respiratory tract colonization was found in 3 cases . CONCLUSIONS: (1) The pH value of gastric juice proved to be the major factor which influenced the colonization of bacteria especially GNB in gastric cavity . (2) The gastric cavity was probably a colonization place of GNB especially enterobacteriaceae . (3) The enterobateriaceae in gastric cavity tended to colonize reversely to oropharynx . (4) The phenotypic analysis of the pathogens showed that the reverse stomach-pharynx-lower respiratory tract infection route existed in VAP patients.

Zhonghua Gan Zang Bing Za Zhi, 2004 Mar, 12(3), 167 - 9
{Dynamic variability of intestinal flora and endotoxin in rat with fulminate hepatic failure}; Li LJ et al.; OBJECTIVE: To investigate the dynamic variability of intestinal flora and endotoxins in rats with fulminate hepatic failure . METHODS: Establishing the fulminate hepatic failure models by intraperitoneal injection of Galactosamine . Forty Sprague-Dawley rats were divided into three groups: group A (n=10) were killed at the beginning of the experiment as control; while Group B (n=12) and C (n=18), the fulminate hepatic failure models, were killed 24 and 48 hours respectively after successful induction . Then, the contents of the jejunum, ileum and colon descendents were collected and a quantitative analysis was made about intestinal flora . Meanwhile, the concentrations of endotoxin in portal vein and right ventricle were determined and so were those in contents of ileums and colons . RESULTS: Our experiments showed that the livers of rats in group B were injured most seriously among three groups, and a minor recovery of hepatic function was observed in group C with the decrease of total bile acids (P< 0.05) . Analysis on intestinal flora show: the intestinal enterobacteriacea increase and the lactobacillus decrease in group B (P< 0.01 in jejunum and ileum and P<0.05 in colon) . The comparisons between group C and B showed that the enterobacteriacea in the former decreased in both jejunum and colon (P< 0.05) while the number of lactobacillus recovered in the jejunum of group C (P<0.05) . Quantitative analysis on endotoxins showed that the ileum endotoxin increased in group B (P< 0.05) and in group C, endotoxins in ileum and colons also increased (vs . control, P<0.01); portal endotoxin in group B showed higher level than that in group A and C (P< 0.01) . CONCLUSION: The alteration of intestinal flora was observed in fulminate hepatic failure rats . Abnormal intestinal flora might lead to incline of endotoxin in ileum, colon and portal vein, while the recovery of normal intestinal flora would decrease the level of portal endotoxin.

J Pak Med Assoc, 2004 Jan, 54(1), 20 - 4
Intraprostatic tissue infection in catheterised patients in comparison to controls; Talpur AN et al.; OBJECTIVE: To determine the effect of indwelling urinary catheter on frequency of intraprostatic tissue infection and posto-perative morbidity in patients with benign prostatic hyperplasia undergoing transurethral resection of prostate (TURP) . METHODS: Frequency of intraprostatic tissue and urinary infection, prevalent organisms, histopathology of prostatic tissue, post-operative morbidity were analyzed for 25 consecutive patients' of clinically diagnosed benign prostatic hyperplasia who underwent TURP in catheterized and non-catheterized groups . RESULTS: Patients mean age was 62.2+7.9 years . In non-catheterized group nocturia and frequency were the most common symptoms . Ninety two percent and 28% patients acquired intraprostatic tissue infection in catheterized and non-catheterized group respectively, while 80% of catheterized patients and 24% of the non-catheterized patients had bacteriuria . Catheterized patients had E . coli as prevalent organism both in intaprostatic tissue and urine (34.8% and 40% respectively) . E . coli Serratia and enterobacter were equally prevalent in intraprostatic tissue of non-catheterized patients . Enterobacter was the prevalent organism in urine (50%) of these patients . There was no significant difference in the presence of non-specific inflammatory cells in the two groups . Catheterized group showed significantly high frequency of fever >38.5 degrees C and hematuria for more than 24 hours . CONCLUSION: Catheterization significantly increases the frequency of intraprostatic tissue infection as well as morbidity of TURP.

Curr Microbiol, 2004 Feb, 48(2), 124 - 9
Decolorization of azo dyes with Enterobacter agglomerans immobilized in different supports by using fluidized bed bioreactor; Moutaouakkil A et al.; Immobilized cells of Enterobacter agglomerans, able to reduce azo dyes enzymatically, were used as a biocatalyst for the decolorization of synthetic medium containing the toxic azo dye methyl red (MR) . This bacterial strain exhibits high ability to completely decolorize 100 mg/L of MR after only 6 h of incubation under aerobic conditions . Cells of E . agglomerans were immobilized in calcium alginate, polyacylamide, cooper beech, and vermiculite, and were used for the decolorization of MR from synthetic water by using a fluidized bed bioreactor . The highest specific decolorization rate was obtained when E . agglomerans was entrapped in calcium alginate beads and was of about 3.04 mg MR/g cell/h with a 50% conversion time ( t(1/2)) of about 1.6 h . Moreover, immobilized cells in calcium alginate continuously decolorized MR even after seven repeated experiments without significant loss of activity, while polyacrylamide-, cooper beech-, and vermiculite-immobilized cells retained only 62, 15, and 13% of their original activity, respectively.

Curr Microbiol, 2004 Mar, 48(3), 167 - 74
Cloning and characterization of the gene encoding for OMP-PD porin: the major Photobacterium damsela outer membrane protein; Gribun A et al.; The outer membrane protein of Photobacterium damsela (OMP-PD) and the gene encoding for this porin protein were isolated and characterized . The deduced amino acid sequence of the OMP-PD monomer has 338 amino acids and a calculated molecular weight of 36,951 Da . This sequence includes a 22-amino acid signal peptide at the N-terminal, which is not found when the monomer is located in the outer membrane . Native OMP-PD protein forms a trimeric structure of approximately 110 kDa . It exhibits resistance to proteases, and it can be cleaved only following denaturation by SDS . The degree of identity of the OMP-PD amino acid sequence to porins from the Enterobacteriaceae was only 24% . Identity to Vibrio or Photobacterium porins was 38% and 48%, respectively . Nevertheless, the multiple alignment of this sequence with other structurally defined Enterobacteria porins demonstrated that the location of the 16 beta-strands and eight external loops, including a larger external L3 loop, are conserved in OMP-PD . These results, together with the previously known ability of OMP-PD to form an ion channel in artificial liposomes, strongly support its role as a porin in P . damsela and will help further investigations into the role of OMP-PD in P . damsela pathogenicity.

Roum Arch Microbiol Immunol, 2002 Oct-Dec, 61(4), 285 - 91
Resistance pattern of extended-spectrum beta-lactamase producing Enterobacteriaceae isolates; Tuchilus C et al.; Gram-negative pathogens harboring extended-spectrum beta-lactamases (ESBL) are becoming an increasing therapeutic problem in many wards . The aim of our work was to study ESBL production by Enterobacteriaceae strains from Eastern Romania and their antimicrobial resistance . We selected 54 clinical isolates among 1068 enterobacteria according to their susceptibility spectrum (National Committee for Clinical Laboratory Standards, 1999) . Antimicrobial susceptibility tests were performed using the Rapid ATB E gallery of mini API system (BioMerieux) and by a macrodilution method in Mueller-Hinton agar following standard procedure of the National Committee for Clinical Laboratory Standards (NCCLS) . ESBL production was established by using both double disk synergy test (DDT) and Expert computer program of mini API . The isoelectric point (pI) was determined by isoelectric focusing in polyacrylamide gel and revealed by nitrocefin . As references we used beta-lactamases with known pI . The Expert computer program of mini API confirms the positive DDT test for all selected strains . Almost all strains displayed resistance to ampicillin, ampicillin/sulbactam or third generation cephalosporins and aztreonam . By IEF we identified 51 strains which have a unique enzyme . IEF pattern showed presence of two enzymes in three Escherichia coli strains . According to our results, the ESBL TEM-type are the most common for the studied isolates . The production of extended-spectrum beta-lactamases and the presence of the multiresistant of antimicrobial agents reflect, probably, the over use of third generation cephalosporins in Eastern Romania.

Arch Immunol Ther Exp (Warsz), 2004 Jan-Feb, 52(1), 43 - 9
Serological characterization of the O-specific polysaccharide of Providencia alcalifaciens O23; Torzewska A et al.; INTRODUCTION: The genus Providencia belongs to the Enterobacteriaceae family and currently consists of five species: P . alcalifaciens, P . heimbachae, P . rettgerii, P . rustigianii and P . stuartii . The serological classification scheme of P . alcalifaciens, P . rustigianii and P . stuartii includes 63 O-serogroups and 30 H-serogroups . The O-antigenic specificity is defined by the structure of the O-antigen (O-specific polysaccharide--OPS), a part of the lipopolysaccharide (LPS, endotoxin), one of the major components of the outer membrane of gram-negative bacteria and an important virulence factor of these bacteria . Among the bacteria of the Enterobacteriaceae family, the genus Providencia is one of the least studied in respect to its LPS structure and antigenic specificity . Studies of the chemical structures and the serological specificity of the O-antigens aim at the elucidation of the molecular basis of the serological classification of Providencia sp . MATERIALS AND METHODS: LPS and alkali-treated LPS of P . alcalifaciens O23 and serologically related P . rustigianii O14, P . mirabilis O13 and P . myxofaciens as well as O-antiserum against P . alcalifaciens O23 were used . Serological characterization of P . alcalifaciens O23 O-specific polysaccharide was done by use enzyme immunosorbent assay (EIA), passive hemolysis test (PHT) as well as by inhibition and sodium deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE) of LPS and Western blot . RESULTS AND CONCLUSIONS: The OPS of P . alcalifaciens, O23, contains an N-(D-glucuronoyl)-N-{(R)-1-carboxyethyl}-L-lysine residue (GlcAAlaLys) . The LPS of P . alcalifaciens, O23, and other LPSs containing AlaLys from Providencia and Proteus strains were tested with rabbit anti-P . alcalifiaciens O23 serum . The serological data showed that a GlcAAlaLys-associated epitope plays a role as an antigenic determinant in the P . alcalifaciens O23 OPS and revealed the particular importance of glucuronic acid and the carboxyethyl group for the binding of O23-specific antibodies.

Poult Sci, 2004 Mar, 83(3), 384 - 91
Salmonella in commercially manufactured feeds; Jones FT et al.; We collected 886 samples (68 feed ingredient samples, 189 dust samples, and 629 feed samples) from 3 feed mills each of which produced between 100,000 and 400,000 tons of feed a year . Samples were collected on 3 d (Monday, Wednesday, and Friday), during 2 seasons (early spring and summer), and between 0700 and 1700 h approximately once per hour . Samples were collected from 5 locations within each mill: ingredient receiving, at the mixer, at the pellet mill, from pellet coolers, and at load-out . Temperatures were taken of the samples obtained at the pellet mill immediately following collection . All samples were analyzed for Enterobacteriaceae counts (EC) and Salmonella . The data confirm that feed ingredients and dust can be a major source of Salmonella contamination in feed mills . There were no differences (P < 0.05) in the Salmonella contamination rates of samples collected in spring as compared with samples collected in summer . Salmonella contamination rates were observed to be higher in samples collected on Friday compared with samples collected on Monday or Wednesday, an effect that may be management related . Data collected at the pellet mill clearly illustrate the uneven distribution of Salmonella contamination in feed as well as the need for control of dust around the pellet mill . Feed samples (both mash and pellets) contaminated with Salmonella contained significantly higher EC than samples not contaminated with Salmonella . Thus, EC may provide some indication of the likelihood of Salmonella contamination in feed samples.

Otolaryngol Pol, 2003, 57(6), 813 - 7
{Long term administration of itraconazole with surgical treatment in fungal and bacterial infections of the paranasal sinuses}; Jordan J et al.; A case of chronic paranasal sinuses with recurrent polyposis caused by miscellaneous infection--fungal (Aspergillus, Candida) and bacterial (Staphylococcus aureus, Enterobacter, Streptococcus) is described . The patient underwent 5 times surgical treatment (polypectomies, sinus operations) . Good result was achieved after 2-years application of itraconazole and local Amphotericin B.

Rev Soc Bras Med Trop, 2003 Nov-Dec, 36(6), 711 - 7
{Spontaneous bacterial peritonitis}; Strauss E et al.; Spontaneous bacterial peritonitis occurs in 30% of patients with ascites due to cirrhosis leading to high morbidity and mortality rates . The pathogenesis of spontaneous bacterial peritonitis is related to altered host defenses observed in end-stage liver disease, overgrowth of microorganisms, and bacterial translocation from the intestinal lumen to mesenteric lymph nodes . Clinical manifestations vary from severe to slight or absent, demanding analysis of the ascitic fluid . The diagnosis is confirmed by a number of neutrophils over 250/mm3 associated or not to bacterial growth in culture of an ascites sample . Enterobacteriae prevail and Escherichia coli has been the most frequent bacterium reported . Mortality rates decreased markedly in the last two decades due to early diagnosis and prompt antibiotic treatment . Third generation intravenous cephalosporins are effective in 70% to 95% of the cases . Recurrence of spontaneous bacterial peritonitis is common and can be prevented by the continuous use of oral norfloxacin . The development of bacterial resistance demands the search for new options in the prophylaxis of spontaneous bacterial peritonitis; probiotics are a promising new approach, but deserve further evaluation . Short-term antibiotic prophylaxis is recommended for patients with cirrhosis and ascites shortly after an acute episode of gastrointestinal bleeding.

Mem Inst Oswaldo Cruz, 2003 Dec, 98(8), 1093 - 5 Epub 2004 Mar 09.
Detection of pathogenic bacteria in skin lesions of patients with chiclero's ulcer . Reluctant response to antimonial treatment; Isaac-Marquez AP et al.; We investigated the bacterial flora present in skin lesions of patients with chiclero's ulcer from the Yucatan peninsula of Mexico using conventional culture methods (11 patients), and an immunocolorimetric detection of pathogenic Streptococcus pyogenes (15 patients) . Prevalence of bacteria isolated by culture methods was 90.9% (10/11) . We cultured, from chiclero's ulcers (60%), pathogenic bacterial such as Staphylococcus aureus (20%), S . pyogenes (1.6%), Pseudomonas aeruginosa (1.6%), Morganella morganii (1.6%), and opportunist pathogenic bacteria such as Klebsiella spp . (20.0%), Enterobacter spp . (20%), and Enterococcus spp . (20%) . We also cultured coagulase-negative staphylococci in 40% (4/10) of the remaining patients . Micrococcus spp . and coagulase-negative staphylococci constituted the bacterial genuses more frequently isolated in the normal skin of patients with chiclero's ulcer and healthy individuals used as controls . We also undertook another study to find out the presence of S . pyogenes by an immunocolorimetric assay . This study indicated that 60% (9/15) of the ulcerated lesions, but not normal controls, were contaminated with S . pyogenes . Importantly, individuals with purulent secretion and holding concomitant infections with S . pyogenes, S . aureus, P . aeruginosa, M . morganii, and E . durans took longer to heal Leishmania (L.) mexicana infections treated with antimonial drugs . Our results suggest the need to eliminate bacterial purulent infections, by antibiotic treatment, before starting antimonial administration to patients with chiclero's ulcer.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1384 - 96
In vitro antimicrobial activity of doripenem, a new carbapenem; Ge Y et al.; The doripenem MICs at which 90% of the tested strains were inhibited ranged from 0.03 to 1 microg/ml for 10 species of Enterobacteriaceae (n = 351), from 0.03 to 0.12 microg/ml for oxacillin-susceptible staphylococci (n = 119), from 4 to 32 microg/ml for oxacillin-resistant staphylococci (n = 64), from < or =0.008 to 0.06 microg/ml for penicillin-susceptible streptococci (n = 132), and from 1 to 4 microg/ml for penicillin-resistant streptococci (n = 51) . Overall, doripenem demonstrated in vitro activity similar to that of meropenem against gram-negative pathogens and to that of imipenem against gram-positive pathogens.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1249 - 55
Dissemination of CTX-M-type beta-lactamases among clinical isolates of Enterobacteriaceae in Paris, France; Eckert C et al.; We analyzed 19 clinical isolates of the family Enterobacteriaceae (16 Escherichia coli isolates and 3 Klebsiella pneumoniae isolates) collected from four different hospitals in Paris, France, from 2000 to 2002 . These strains had a particular extended-spectrum cephalosporin resistance profile characterized by a higher level of resistance to cefotaxime and aztreonam than to ceftazidime . The bla(CTX-M) genes encoding these beta-lactamases were involved in this resistance, with a predominance of bla(CTX-M-15) . Ten of the 19 isolates produced both TEM-1- and CTX-M-type enzymes . One strain (E . coli TN13) expressed CMY-2, TEM-1, and CTX-M-14 . bla(CTX-M) genes were found on large plasmids . In 15 cases the same insertion sequence, ISEcp1, was located upstream of the 5' end of the bla(CTX-M) gene . In one case we identified an insertion sequence designated IS26 . Examination of the other three bla(CTX-M) genes by cloning, sequencing, and PCR analysis revealed the presence of a complex sul1-type integron that includes open reading frame ORF513, which carries the bla gene and the surrounding DNA . Five isolates had the same plasmid DNA fingerprint, suggesting clonal dissemination of CTX-M-15-producing strains in the Paris area.

Ann Clin Microbiol Antimicrob . 2004 Mar 25;3(1):3.
Prevalence of antimicrobial resistance in bacteria isolated from central nervous system specimens as reported by U.S . hospital laboratories from 2000 to 2002; Jones ME et al.; BACKGROUND: Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy . The widespread emergence of resistance among bacterial species is a cause for concern . Current antibacterial susceptibility data among central nervous system (CNS) pathogens is important to define current prevalence of resistance . METHODS: Antimicrobial susceptibility of pathogens isolated from CNS specimens was analyzed using The Surveillance Database (TSN(R)) USA Database which gathers routine antibiotic susceptibility data from >300 US hospital laboratories . A total of 6029 organisms derived from CNS specimen sources during 2000-2002, were isolated and susceptibility tested . RESULTS: Staphylococcus aureus (23.7%) and Streptococcus pneumoniae (11.0%) were the most common gram-positive pathogens . Gram-negative species comprised approximately 25% of isolates . The modal patient age was 1 or <1 year for most organisms . Prevalence of MRSA among S . aureus from cerebrospinal fluid (CSF) and brain abscesses were 29.9-32.9% . Penicillin resistance rates were 16.6% for S . pneumoniae, 5.3% for viridans group streptococci, and 0% for S . agalactiae . For CSF isolates, ceftriaxone resistance was S . pneumoniae (3.5%), E . coli (0.6%), Klebsiella pneumoniae (2.8%), Serratia marcescens (5.6%), Enterobacter cloacae (25.0%), Haemophilus influenzae (0%) . Listeria monocytogenes and N . meningitidis are not routinely susceptibility tested . CONCLUSIONS: Resistance is commonly detected, albeit still at relatively low levels for key drugs classes such as third-generation cephalosporins . This data demonstrates the need to consider predominant resistance phenotypes when choosing empiric therapies to treat CNS infections.

Biochem J, 2004 Jul 15, 381(Pt 2), 527 - 36
Combinational clustering of receptors following stimulation by bacterial products determines lipopolysaccharide responses; Triantafilou M et al.; The innate immune system has the capacity to recognize a wide range of pathogens based on conserved PAMPs (pathogen-associated molecular patterns) . In the case of bacterial LPS (lipopolysaccharide) recognition, the best studied PAMP, it has been shown that the innate immune system employs at least three cell-surface receptors: CD14, TLR4 (Toll-like receptor 4) and MD-2 protein . CD14 binds LPS from Enterobacteriaceae and then transfers it to MD-2, leading to TLR4 aggregation and signal transduction . LPS analogues such as lipid IVa seem to act as LPS antagonists in human cells, but exhibit LPS mimetic activity in mouse cells . Although TLR4 has been shown to be involved in this species-specific discrimination, the mechanism by which this is achieved has not been elucidated . The questions that remain are how the innate immune system can discriminate between LPS from different bacteria as well as different LPS analogues, and whether or not the structure of LPS affects its interaction with the CD14-TLR4-MD-2 cluster . Is it possible that the 'shape' of LPS induces the formation of different receptor clusters, and thus a different immune response? In the present study, we demonstrate using biochemical as well as fluorescence-imaging techniques that different LPS analogues trigger the recruitment of different receptors within microdomains . The composition of each receptor cluster as well as the number of TLR4 molecules that are recruited within the cluster seem to determine whether an immune response will be induced or inhibited.

Infect Immun, 2004 Apr, 72(4), 2254 - 62
Growth control of small-colony variants by genetic regulation of the hemin uptake system; Roggenkamp A et al.; Small-colony variants (SCVs) are slow-growing variants of human bacterial pathogens . They are associated with chronic persistent infections, and their biochemical identification and antimicrobial treatment are impaired by altered metabolic properties . To contribute to the understanding of SCV-mediated infections, we analyzed a clinical SCV isolate derived from a chronic prosthetic hip infection . A sequence analysis of housekeeping genes identified an Enterobacter hormaechei-like organism . The SCV phenotype, with growth as microcolonies, was caused by a block within the heme biosynthesis pathway through deletion of the hemB locus, as shown by hybridization and complementation experiments . At a low frequency, large-colony variants (LCVs) arose that were dependent on exogenous hemin . To investigate this phenomenon, we cloned and sequenced the 5.8-kb hemin uptake system, denoted ehu . Gene expression analysis indicated regulation of this locus in wild-type bacteria by the global iron regulator Fur . Inactivation of Fur in LCVs caused the derepression of ehu expression and facilitated bacterial growth . Genetic alterations of the fur locus in LCVs were identified as insertions of IS1A elements and point mutations . In contrast, SCVs could utilize exogenous hemin only in the absence of iron . Thus, we provide the first molecular characterization of the growth properties of a clinical SCV isolate, which may help to improve the diagnostic and therapeutic management of patients with chronic persistent infections.

Infect Immun, 2004 Apr, 72(4), 1983 - 90
An abundance of Escherichia coli is harbored by the mucosa-associated bacterial flora of interleukin-2-deficient mice; Schuppler M et al.; Mice deficient in interleukin-2 are well suited for use as an animal model for inflammatory bowel disease . Raised under specific-pathogen-free conditions, interleukin-2-deficient mice develop an inflammatory bowel disease resembling ulcerative colitis in humans . The finding that colitis was attenuated when the mice were kept under germfree conditions implies that the resident intestinal flora is involved in the pathogenesis of colitis . The present study addresses the composition of the mucosa-associated bacterial flora in colon samples from interleukin-2-deficient mice that developed colitis . This was investigated by comparative 16S ribosomal DNA (rDNA) sequence analysis and fluorescence in situ hybridization using rRNA-targeted fluorescent probes to quantify the bacterial populations of the mucosa-associated flora . The investigations revealed distinct differences in the bacterial composition of the mucosa-associated flora between interleukin-2-deficient mice and healthy controls . Fluorescence in situ hybridization identified up to 10% of the mucosa-associated flora in interleukin-2-deficient mice as Escherichia coli, whereas no E . coli was detected in the mucosa from healthy wild-type mice . This finding was consistent with the results from comparative 16S rDNA analysis . About one-third of the clones analyzed from 16S rDNA libraries of interleukin-2-deficient mice represented Enterobacteriaceae, whereas none of the clones analyzed from the healthy controls harbored 16S rDNA from Enterobacteriaceae . The abundance of E . coli in the colonic mucosa of interleukin-2-deficient mice strongly suggests a participation in the pathogenesis of colitis in the interleukin-2-deficient mouse model for inflammatory bowel disease.

Curr Opin Microbiol, 2004 Feb, 7(1), 62 - 6
How do neutrophils and pathogens interact?
Mayer-Scholl A, Averhoff P, Zychlinsky A.
Many pathogens can manipulate macrophages after phagocytosis yet are efficiently killed by neutrophils . This poses the question of whether neutrophils have mechanisms that enable them to specifically recognise pathogens and have pathogens evolved mechanisms to modulate neutrophil function? Here, we review recent work on neutrophils and their interaction with four different bacteria: Staphylococcus aureus, Helicobacter pylori, Anaplasma phagocytophilum and members of the Enterobacteriae family.

J Food Prot, 2004 Mar, 67(3), 607 - 9
Biogenic amines in restructured beef steaks as affected by added walnuts and cold storage; Ruiz-Capillas C et al.; This article evaluates changes in biogenic amines and how these relate to microbiological growth in chilled, fresh restructured beef steaks containing transglutaminase as a cold binding agent and different amounts of walnut . Added walnut and chilling favored higher total and lactic acid bacteria counts during storage, whereas Enterobacteriaceae were not affected . The highest initial biogenic amine concentrations were identified as spermidine, spermine, and tyramine . Both added walnut and cold storage generally favored the formation of amines (tyramine, histamine, putrescine, and cadaverine), which was more obviously apparent by the end of the storage period . Agmatine, on the other hand, was not generally affected by the walnut.

J Food Prot, 2004 Mar, 67(3), 567 - 73
Volatile compounds produced in cheese by Enterobacteriaceae strains of dairy origin; Morales P et al.; The formation of volatile compounds in fresh cheese by 10 Enterobacteriaceae strains of dairy origin (4 Hafnia alvei, 2 Serratia liquefaciens, 1 Enterobacter cloacae, 1 Enterobacter sakazakii, and 2 Escherichia coli strains) was investigated . Small cheeses were made from pasteurized cow's milk separately inoculated with 1-3 x 10(3) CFU/ml of each of the Enterobacteriaceae strains, with glucono-8-lactone added to achieve a pH value of 5.2 in the curds . All strains reached counts close to 10(8) CFU/g in 1-day-old cheeses and survived well from day 1 to day 8 . Cheeses were analyzed for volatile compounds by gas chromatography-mass spectroscopy, after extraction by dynamic headspace using a purge and trap apparatus . Sixty-one volatile compounds were determined in cheeses, 31 of which were further investigated . Significant increases of aldehydes, sulfur compounds, and aromatic compounds were recorded from 2-h curd to 1-day-old cheese, and of ketones, alcohols, and acids from 2-h curd to 8-day-old cheese . Acetaldehyde, 2-methyl propanal, and 3-methyl butanal predominated among aldehydes; 2,3-butanedione, 2,3-pentanedione, and 3-hydroxy 2-butanone among ketones; ethanol, 2-methyl propanol, and 3-methyl butanol among alcohols; and ethyl acetate among esters . Hierarchical cluster analysis of strains using the data of 31 volatile compounds separated clearly the strain of E . sakazakii, which produced high amounts of volatile compounds, from the other Enterobacteriaceae strains.

Infez Med, 1995 Mar, 3(1), 45 - 7
{Pyrogen reaction to bacterial endotoxins in dialysis patients}; Camporese A; The author describes three cases of pirogenic reaction in patients in treatment with hemodialysis: Pseudomonas aeruginosa and Enterobacter cloacae were isolated from water of dyalisis machines . He, moreover, analyses different antibacterial decontamination systems

J Biol Chem, 2004 May 28, 279(22), 23661 - 7 Epub 2004 Mar 18.
A pitfall in diagnosis of human prion diseases using detection of protease-resistant prion protein in urine . Contamination with bacterial outer membrane proteins; Furukawa H et al.; Because a definite diagnosis of prion diseases relies on the detection of the abnormal isoform of prion protein (PrPSc), it has been urgently necessary to establish a non-invasive diagnostic test to detect PrPSc in human prion diseases . To evaluate diagnostic usefulness and reliability of the detection of protease-resistant prion protein in urine, we extensively analyzed proteinase K (PK)-resistant proteins in patients affected with prion diseases and control subjects by Western blot, a coupled liquid chromatography and mass spectrometry analysis, and N-terminal sequence analysis . The PK-resistant signal migrating around 32 kDa previously reported by Shaked et al . (Shaked, G . M., Shaked, Y., Kariv-Inbal, Z., Halimi, M., Avraham, I., and Gabizon, R . (2001) J . Biol . Chem . 276, 31479-31482) was not observed in this study . Instead, discrete protein bands with an apparent molecular mass of approximately 37 kDa were detected in the urine of many patients affected with prion diseases and two diseased controls . Although these proteins also gave strong signals in the Western blot using a variety of anti-PrP antibodies as a primary antibody, we found that the signals were still detectable by incubation of secondary antibodies alone, i.e . in the absence of the primary anti-PrP antibodies . Mass spectrometry and N-terminal protein sequencing analysis revealed that the majority of the PK-resistant 37-kDa proteins in the urine of patients were outer membrane proteins (OMPs) of the Enterobacterial species . OMPs isolated from these bacteria were resistant to PK and the PK-resistant OMPs from the Enterobacterial species migrated around 37 kDa on SDS-PAGE . Furthermore, nonspecific binding of OMPs to antibodies could be mistaken for PrPSc . These findings caution that bacterial contamination can affect the immunological detection of prion protein . Therefore, the presence of Enterobacterial species should be excluded in the immunological tests for PrPSc in clinical samples, in particular, urine.

Hautarzt, 2004 Mar, 55(3), 280 - 8
{Bacterial colonization of chronic wounds . Studies on outpatients in a university dermatology clinic with special consideration of ORSA}; Dissemond J et al.; In this retrospective investigation, we documented the bacterial colonization of 79 patients with chronic wounds, who had been treated between January 2002 and May 2003 in an outpatient wound healing clinic of a university dermatology program . We isolated 106 facultative pathogenic bacterial strains of which 56 were Staphylococcus aureus, 19 Pseudomonas aeruginosa, 11 Escherichia coli, 4 Proteus mirabilis, 4 Enterobacter cloacae, 2 Serratia marcescens, 2 Streptococcus group G und 8 further species . 68 of these bacterial strains were gram-positive and 46 gram-negative . Moreover we identified one patient with Candida parapsilosis . Therefore, 70.8% of all patients showed Staphylococcus aureus in their chronic wounds . Determination of the specific resistances showed 17 patients to be colonized with oxacillin- resistant Staphylococcus aureus (ORSA) strain; this corresponds to 21.5% of all patients . Consequently, 30.4% of all Staphylococcus aureus isolates were ORSA strains . All of the ORSA isolates were sensitive to vancomycin . Sensitivity to tetracycline was documented in 15, to amikacin in 13, to clindamycin in 7, to gentamicin and erythromycin in 6 of the ORSA-positive patients . In the case of trimethoprim/sulfamethoxazole, 10 were sensitive and 3 were intermediate in sensitivity . Beside the obligate resistance to oxacillin, penicillin G, ampicillin, cefuroxime and imipenem, none of the ORSA was sensitive to ofloxacin . The results of our investigations demonstrate the actual spectrum of bacterial colonization in chronic wounds of patients in an university dermatologic wound clinic and underline the growing problem of ORSA.

J Bacteriol, 2004 Apr, 186(7), 1933 - 44
Complete genomic sequence of the virulent Salmonella bacteriophage SP6; Dobbins AT et al.; We report the complete genome sequence of enterobacteriophage SP6, which infects Salmonella enterica serovar Typhimurium . The genome contains 43,769 bp, including a 174-bp direct terminal repeat . The gene content and organization clearly place SP6 in the coliphage T7 group of phages, but there is approximately 5 kb at the right end of the genome that is not present in other members of the group, and the homologues of T7 genes 1.3 through 3 appear to have undergone an unusual reorganization . Sequence analysis identified 10 putative promoters for the SP6-encoded RNA polymerase and seven putative rho-independent terminators . The terminator following the gene encoding the major capsid subunit has a termination efficiency of about 50% with the SP6-encoded RNA polymerase . Phylogenetic analysis of phages related to SP6 provided clear evidence for horizontal exchange of sequences in the ancestry of these phages and clearly demarcated exchange boundaries; one of the recombination joints lies within the coding region for a phage exonuclease . Bioinformatic analysis of the SP6 sequence strongly suggested that DNA replication occurs in large part through a bidirectional mechanism, possibly with circular intermediates.

Int J Syst Evol Microbiol, 2004 Mar, 54(Pt 2), 429 - 37
The leucine-responsive regulatory protein (lrp) gene for characterization of the relationship among Xanthomonas species; Cubero J et al.; Characterization of strains of Xanthomonas axonopodis pv . citri by using DNA fingerprints that were generated from primers for enterobacterial repetitive intergenic consensus (ERIC) elements led to the discovery of differential sequences for a leucine-responsive regulatory protein (lrp) gene in two subgroups of strains with different host ranges on Citrus spp . DNA hybridization and PCR-based assays that used different sets of primers were designed to detect the core sequence, as well as to obtain the entire sequence of the lrp gene for several Xanthomonas species and pathovars . Higher variability was observed at the nucleotide level than at the amino acid level among the different species and pathovars, revealing selection pressure on the lrp gene, which is presumably due to an essential role of the gene in bacterial metabolism . Moderate variability in the 3' and 5' domains was used to study relationships among different species within the genus XANTHOMONAS: Species of this genus that were isolated from citrus, as well as other pathovars of X . axonopodis, showed highly similar lrp gene sequences, whereas other Xanthomonas species, especially Xanthomonas campestris, had sequences that were more dissimilar to that of X . axonopodis . Thus, the lrp gene sequence is useful to distinguish X . axonopodis pv . citri groups and promising for polyphasic taxonomic analysis of the genus XANTHOMONAS: Data from analysis of lrp gene sequences support the current concepts for classification of xanthomonads, which are based on other approaches.

Diagn Microbiol Infect Dis, 2004 Mar, 48(3), 167 - 72
Spread of bla(VIM-1)-producing E . coli in a university hospital in Greece . Genetic analysis of the integron carrying the bla(VIM-1) metallo-beta-lactamase gene; Scoulica EV et al.; Bla(VIM-1) gene was detected in four Escherichia coli clinical isolates with both reduced susceptibility to carbapenems and an ESBL phenotype . The VIM-1 determinant was located within the variable region of a Class I integron along with a 6'-N-aminoglycoside acetyltransferase gene (aac(6')-Ib) and it could be transferred by conjugation . In all four clinical isolates the VIM-1 gene cassette presented a characteristic duplication of the 3' end coding 153 nucleotides followed by the first 14 nucleotides of the 59 base element (59be) that however did not seem to affect either the integrity of the coding sequence or the 59be of the gene cassette . These clinical isolates not only harbored the same Class I integron, but they also shared the same discrete ribotype-pattern, indicative for their clonal origin . Spread of carbapenem resistance genes among Enterobacteriaceae in hospital is a matter of great concern.

Klin Med (Mosk), 2004, 82(1), 42 - 4
{Clinical characteristics of respiratory infection in patients with hematologic malignancy and neutropenia}; Domnikova NP et al.; The aim of the study was a comparative assessment of clinical and laboratory findings characterizing the course of pneumonia or bronchitis in patients with hematological malignancy (HM) and neutropenia . The course of the respiratory infections (RI) was studied in 47 HM patients . RI in HM were found to develop in patients with a large tumor, in bone marrow involvement, in patients without a complete remission, more frequently pneumonia arises in acute leukemia, bronchitis--in patients with lymphoproliferative diseases . Among causative agents Gram-negative and Gram-positive microorganisms were detected at the same rate (44.2 and 45.9%, respectively), fungi were diagnosed in 9.8% cases . Enterobacteriaceae microorganisms were found more often in patients with pneumonia . Gram-positive agents occurred more frequently in bronchitis . Such clinical markers in neutropenic patients as dyspnea, dullness in the lung, weakened respiration, moist rales in the lungs and fever 39 degrees C and higher point to pneumonia . If the above markers are absent but there is cough, fever 38-39 degrees C, tough respiration, dry rales both pneumonia and bronchitis are possible.

Immunol Lett, 2004 Feb 15, 91(2-3), 103 - 11
Contribution of mast cells to bacterial clearance and their proliferation during experimental cystitis induced by type 1 fimbriated E . coli; Malaviya R et al.; Bacterial infections of the urinary bladder are very common, and the role of mast cells in these infections is invariably thought of as a detrimental one . However, recent studies have shown that mast cells play a key role in host defense against various enterobacterial infections . In this manuscript, using mast cell-deficient (WBB6F1 - W/Wv) and mast cell-sufficient (WBB6F1 - +/+) mice we have investigated the protective role of mast cells in urinary bladder infections in vivo . Our findings show that (i) the mast cells are activated by FimH-expressing E . coli, and release large amount of histamine in the urinary bladder; (ii) the number of surviving bacteria in the urine is dependent on the presence of mast cells, and (iii) mast cell number in the bladder increases following uropathogenic infection in mice which is likely due to an increase in the mast cell growth-promoting cytokine IL-3 in bacteria-activated mast cells . Taken together, these observations suggest a beneficial role of mast cells in urinary bladder infections in mice.

Bioorg Med Chem, 2004 Mar 15, 12(6), 1537 - 42
Kinetic and structural consequences of the leaving group in substrates of a class C beta-lactamase; Ahn YM et al.; The class C beta-lactamase of Enterobacter cloacae P99 is known to catalyze the hydrolysis of certain acyclic (thio)esters . Previous experiments have employed thioglycolate, m-hydroxybenzoate, and phenylphosphate leaving groups . The relative effectiveness of these leaving groups has now been quantitatively assessed by employment of a series of compounds with common acyl groups, and found to rank in the order phenylphosphate >m-hydroxybenzoate >thioglycolate . Structural models suggest that these leaving groups interact during acylation principally with Tyr 150, Lys 315, and Thr 316 of the beta-lactamase active site . The positions of the leaving group carboxylates in these models is compared with those in published crystal structures of complexes of class C beta-lactamases with beta-lactams . The particular effectiveness of the acyl phosphate indicates the positions of two oxyanions that strongly interact with the active site . This information should be useful in the design of inhibitors of class C beta-lactamases.

J Biomol NMR, 2004 Jun, 29(2), 199 - 204
Cyclic enterobacterial common antigen: potential contaminant of bacterially expressed protein preparations; Erbel PJ et al.; We have previously reported the identification of the cyclic enterobacterial common antigen (ECA(CYC)) polysaccharide in E . coli strains commonly used for heterologous protein expression (PJA Erbel et al., J . Bacteriol . 185 (2003): 1995) . Following this initial report, interactions among several NMR groups established that characteristic N -acetyl signals of ECA(CYC) have been observed in (15)N-(1)H HSQC spectra of samples of various bacterially-expressed proteins suggesting that this water-soluble carbohydrate is a common contaminant . We provide NMR spectroscopic tools to recognize ECA(CYC) in protein samples, as well as several methods to remove this contaminant . Early recognition of ECA-based NMR signals will prevent time-consuming analyses of this copurifying carbohydrate.

Int J Antimicrob Agents, 2004 Feb, 23(2), 175 - 80
Dissemination of CTX-M type beta-lactamases in Enterobacteriaceae isolates in the People's Republic of China; Munday CJ et al.; Previously there have been a number of reports of extended spectrum beta-lactamase (ESBL) producing isolates of the family Enterobacteriaceae in Asia . We first reported the occurrence of bla(CTX-M) in Guangzhou, China, subsequently there have been reports of bla(CTX-M) from a number of other south Asian countries . Initial surveillance study data suggested that bla(CTX-M) might be widely distributed in China . This study examines the type of bla(CTX-M) occurring in other major population centres in China . Initial disk diffusion method susceptibility testing (NCCLS) selected ESBL producing Escherichia coli and Klebsiella pneumoniae isolates from Beijing and near Wuhan, PRC . After screening in both China and the UK, 13 isolates producing CTX-M ESBLs were identified and studied, 11 also produced TEM-1, and 4 also produced SHV-1 . Sequence analysis of the bla(CTX-M) containing isolates revealed these isolates contained two different bla(CTX-M), three with bla(CTX-M-3) and 10 with bla(CTX-M-14) . After comparison with other previously published studies in the English language, we conclude that the most prevalent bla(CTX-M) so far reported in Asia are bla(CTX-M-14) and bla(CTX-M-3).

J Appl Microbiol, 2004, 96(4), 853 - 60
Application of molecular methods for the differentiation of acetic acid bacteria in a red wine fermentation; Gonzalez A et al.; AIMS: To apply rapid and reliable molecular techniques for typing acetic acid bacteria and studying their population dynamics during wine-making processes . METHODS AND RESULTS: We tested the usefulness of the Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) and Repetitive Extragenic Palindromic-PCR (REP-PCR) techniques with reference strains of most of the species of acetic acid bacteria . We obtained exclusive patterns for each strain with the ERIC-PCR technique, proving the utility for characterizing below species level . REP-PCR technique was not as adequate for this purpose because some strains yielded identical fingerprint . One hundred twenty isolates from a commercial red wine fermentation were fingerprinted using both techniques . We detected a high degree of strain diversity in the first stage of fermentation that decreased throughout the process . However, several strains and species were dominant in the alcoholic fermentation phases . The identification of different strains or genotypes at the species level was carried out by restriction analysis of the 16S ribosomal DNA gene . Gluconobacter oxydans dominated the fresh must, while Acetobacter aceti was the only isolated species at the end of the process . Gluconacetobacter hansenii and G . liquefaciens were also isolated in significant numbers at the beginning of fermentation . CONCLUSIONS: ERIC-PCR and REP-PCR techniques proved useful for characterizing strains of acetic acid bacteria . SIGNIFICANCE AND IMPACT OF THE STUDY: The availability of molecular techniques for a fast and reliable genotypic characterization should increase our knowledge of the ecology of acetic acid bacteria and determine more accurately their growth behaviour during various stages of vinification.

Clin Lab Sci, 2004 Winter, 17(1), 25 - 9
Yersinia pestis: still a plague in the 21st century; Josko D; Yersinia pestis, the causative agent of plague, is an aerobic, non-motile, gram-negative bacillus belonging to the family Enterobacteriacea . It is a zoonotic infection transmitted to humans via the bite of a flea . Three clinical forms of human plague exist: bubonic, pneumonic, and septicemic . Many important virulence factors associated with this organism are responsible for its extreme pathogenicity and high mortality rates . The bubonic form of plague is usually not transmitted human to human but the pneumonic form is--through inhalation of contaminated aerosol droplets . The pneumonic plague would be the form most likely implicated in the event of an intentional attack . Inhalation of aerosols can cause devastating consequences resulting in many casualties . Unless antibiotics are administered within 24 hours of the initial symptoms, death is inevitable . Its potential for use as a biological weapon is of major concern to public health officials.

Rev Invest Clin, 2003 Nov-Dec, 55(6), 600 - 5
{Increasing trend of antimicrobial drug-resistance in organisms causing bacteremia at a tertiary-care hospital: 1995 to 2000}; Kato-Maeda M et al.; We described the trends of drug-resistant organisms isolated in blood cultures from patients detected in a teaching hospital from 1995 to 2000 . We found an increase in the number of clinical isolates of Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter spp, Serratia spp, Staphylococcus aureus, S . epidermidis and Enterococcus spp, resistant to antibiotics commonly used to treat infections caused by these organisms . The frequency of gram-negative bacilli resistant to third-generation cephalosporins and quinolones increased during the period of study, and in 2000 more than 20% of the isolates were resistant . In contrast, the frequency of resistance to aminoglycosides and carbapenems was less than 20% . The frequency of resistant staphylococci increased exuberantly fifty fold to quinolones and five fold to oxacillin during the study period, therefore in 2000, 26.1% of S . aureus isolates and 61% of S . epidermidis were resistant to oxacillin . The frequency of resistant enterococci also increased, and in 2000, 50% were resistant to ampicillin, and 37.5% to gentamicin . The increase of drug resistant organisms isolated in blood had a direct impact in the empirical treatment of severely infected patients in our hospital . It is important to continuously supervise antibiotic use, and to adopt more strict control measures to decrease the frequency of infections caused by drug resistant organisms.

Jpn J Antibiot, 2003 Dec, 56(6), 697 - 704
{Application of the MIC breakpoints based on pharmacokinetics and pharmacodynamics parameter in the clinical laboratory}; Komatsu M et al.; The effectiveness of time-dependent antibiotics such as beta-lactams is related to the time above the MIC (TAM, %) . We constructed a program to calculate the TAMs of beta-lactams using the pharmacokinetic parameters of the Japanese dosing regimen of a phase I study of the Japanese Society for Antimicrobial Chemotherapy (JSAC), and compared them with the MIC breakpoints published by the National Committee for Clinical Laboratory Standards (NCCLS) and JSAC . If the effective TAM was assumed to be more than 40% of the dosing interval, the pharmacokinetic/pharmacodynamic (PK/PD) breakpoints calculated by our program were in agreement with the JSAC breakpoints for pneumonia within 1 dilution MIC . When comparing with the NCCLS breakpoints for Enterobacteriaceae or Staphylococcus, the PK/PD breakpoints dosing three times per day of ampicillin (1 g, intravenous dose; i.v.), piperacillin (2 g, i.v.), cefotaxime (1 g, i.v.) and cefmetazole (1 g, i.v.) were calculated to be less than 2-fold dilution MIC, and those of amoxicillin (0.25 g, oral dose; p.o.) and cefaclor (0.5 g, p.o.) were calculated to be less than 3- to 4-fold dilution of MIC . Our program could calculate TAMs and PK/PD breakpoints by inputting the two factors of MIC and dosing interval . If this information is routinely reported to physicians from clinical laboratories, an appropriate dosing schedule could be proposed for various infectious cases.

Ann Trop Paediatr, 2004 Mar, 24(1), 41 - 4
Neonatal bacterial septicaemia at the Mount Hope Women's Hospital, Trinidad; Ali Z; This retrospective study to determine the incidence of bacterial septicaemia in neonates at the Mount Hope Women's Hospital, Trinidad during a 2-year period, 1996 to 1997, included all neonates whose blood or cerebrospinal fluid cultured positive for bacteria . There were 9866 live births (LB), 102 of whom were diagnosed with bacterial sepsis, an incidence of 10/1000 LB . Thirty-one neonates had a positive culture for group B Streptococcus, an incidence of 3/1000 LB . Gram-negative organisms accounted for 63% of positive cultures . There were three outbreaks of nosocomial infection, two caused by Enterobacter spp with mortality rates of 37% and 50% and one outbreak caused by Pseudomonas aeruginosa with a mortality rate of 25% . The overall mortality rate was 27% (27/102), 63% were boys and 58% were preterm . The incidence of neonatal bacterial sepsis of 10/1000 LB is the highest recorded for the Caribbean and indicates that infection might be an important cause of the high perinatal mortality rate.

Regul Pept, 2004 May 15, 118(3), 135 - 41
A family of brevinin-2 peptides with potent activity against Pseudomonas aeruginosa from the skin of the Hokkaido frog, Rana pirica; Conlon JM et al.; Nine peptides displaying varying degrees of antimicrobial activity were extracted from the skin of the Hokkaido frog, Rana pirica . Five structurally related peptides were identified as members of the brevinin-2 family . These peptides were active against reference strains of Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Klebsiella pneumoniae) and Gram-positive (Staphlococcus aureus) bacteria but displayed relatively low hemolytic activity . The most abundant peptide, brevinin-2PRa (680 nmol/g weight of dry skin) showed high potency {minimal inhibitory concentration (MIC) values between 6 and 12 microM} against a range of clinical isolates of P . aeruginosa . In addition, activity was unaffected by NaCl concentrations up to 200 mM . Cladistic analysis based on the primary structures of brevinin-2 peptides supports a close phylogenetic relationship between R . pirica and Japanese mountain brown frog Rana ornativentris . One peptide of the ranatuerin-2 family and one strongly hemolytic peptide of the brevinin-1 family were also isolated from the extract along with two members of the temporin family, temporin-1PRa (ILPILGNLLNGLL.NH(2)) and temporin-1PRb (ILPILGNLLNSLL.NH(2)) that atypically lacked basic amino acid residues and showed only very weak antimicrobial and hemolytic activity.

Plasmid, 2004 Mar, 51(2), 116 - 26
Analysis of pVU3695, a plasmid encoding glutathione-dependent formaldehyde dehydrogenase activity and formaldehyde resistance in the Escherichia coli VU3695 clinical strain; Dorsey CW et al.; The formaldehyde resistance of Escherichia coli VU3695 is due to the expression of glutathione-dependent formaldehyde dehydrogenase (GSH-FDH) activity, which is encoded by the adhC gene located on the plasmid pVU3695 . Conjugation of this plasmid to an unrelated PolA deficient strain of E . coli indicated that it encodes its own replication initiation protein and does not confer resistance to several other antimicrobial agents tested in this work . In addition, pVU3695 has homology with replicons that belong to the IncL/M plasmid incompatibility group, which are widely distributed among the Enterobacteriaceae . Curing of pVU3695 abolished the expression of formaldehyde resistance and the presence of a 46-kDa periplasmic protein immunologically related to GSH-FDH . However, the curing of pVU3695 reduced drastically but did not abolish the expression of a protein with similar electrophoretic motility, which was associated with the expression of GSH-FDH activity still present in the cytoplasm of the plasmidless derivative . The data demonstrate that E . coli VU3695 contains a chromosomal and a plasmid copy of adhC actively expressed, with the latter being involved in resistance to exogenous formaldehyde.

Plasmid, 2004 Mar, 51(2), 75 - 86
Escherichia coli R-factors unstable in Salmonella typhi are deleted before being segregated in this host; Mendoza-Medellin A et al.; Although resistant Salmonella typhi strains are found in the environment, many epidemiological data indicate that most isolates are multisensitive . Plasmids are not common in S . typhi, contrasting with other enterobacteria . Since S . typhi is able to receive R plasmids from other enterobacteria, such plasmidless condition may be due to destabilization of the plasmids acquired . The segregation process of three plasmids known to behave unstably in S . typhi strains was analyzed, and evidence was obtained that the three of them lost an antibiotic-resistance determinant due to small deletions before plasmid loss occurred . Once deleted, the plasmids entered a segregation phase with particular kinetic features in each case . Selective pressure prevented plasmid deletion and segregation . Like their parental plasmids, the deleted plasmids were unstable in S . typhi but stable in Escherichia coli, suggesting that deletions imply loss of genetic information relevant for a plasmid stability mechanism operative in S . typhi, but not in E . coli.

Pathol Biol (Paris), 2004 Mar, 52(2), 82 - 8
{Antimicrobial susceptibility and frequency of occurrence of clinical blood isolates in Sfax-Tunisia (1993-1998)}; Ben Jemaa Z et al.; Antimicrobial susceptibility and frequency of occurrence of clinical blood isolates in Sfax-Tunisia (1993-1998) . The choice of antimicrobial therapy for the treatment of bacteremia is often empirical and based on the knowledge of susceptibility profiles of the most common bacteria causing such infections . This study determines the bacterial etiology of bacteremic episodes and antimicrobial susceptibility patterns recorded at a teaching hospital, from January 1993 to December 1998 . We collected 2979 strains responsible for bacteremia . Gram negative bacteria were predominant (60%) . The organisms recovered most frequently were Staphylococcus aureus (18.9%), Escherichia coli (14.7%), Klebsiella pneumoniae (14%) and Pseudomonas aeruginosa (7.6%) . The incidence of resistance to methicillin were 17.4% for Staphylococcus aureus and 26.8% for coagulase negative Staphylococcus . No resistance to glycopeptides was observed among the enterococci and staphylococci studied . 27.7% of enterobacteriaceae were resistant to third generation cephalosporins . Imipenem was the most active agent against gram negative bacteria . To carry out a surveillance of bacteremic episodes occurring at every hospital, it is necessary to provide valuable information which should be the basis for effective empiric therapy.

Microb Drug Resist, 2003 Winter, 9(4), 317 - 22
New dfr2 gene as a single-gene cassette in a class 1 integron from a trimethoprim-resistant Escherichia coli isolate; Grape M et al.; Resistance to trimethoprim among Enterobacteriaceae is increasing in spite of a stable or decreasing use of the drug . Integrons are common among these bacteria and many of them contain dfr gene cassettes . A clinical isolate of Escherichia coli from a urine specimen obtained at the Karolinska Hospital in Stockholm was resistant to trimethoprim, ampicillin, sulfonamides, chloramphenicol, streptomycin, and norfloxacin . PCR analysis with primers specific for the 5' and 3' sequences flanking the cassettes in class 1 integrons, detected a very short cassette region of no more than approximately 400 bp . Sequence analysis of the entire integron was performed, revealing a single-gene cassette of 411 bp encoding a new dihydrofolate reductase . The gene cassette comprised all sequences between the flanking conserved sequences and encoded only 78 amino acids . By homology it belongs to the unique group of dfr2 gene cassettes and being the fourth described gene in this group the new gene is called dfr2d . The enzymes encoded by the dfr2 genes are 67% identical and are not related to any other known dfr gene products.

J Chemother, 2003 Dec, 15(6), 568 - 73
Modification of patients' endogenous bacterial flora during hospitalization in a large teaching hospital in Naples; Esposito S et al.; The increasing attention addressed to methicillin-resistant staphylococci, vancomycin-resistant enterococci (VRE) and Extended Spectrum beta-Lactamases (ESbetaL)-producing enterobacteria is due to their etiologic role especially in nosocomial infections . In March 2001 we started an 8-month microbiological prospective surveillance of patients in the General Surgery, Orthopedic and Obstetric & Gynecology wards of the Azienda Universitaria Policlinico, 2nd University of Naples, Italy, to monitor the possible changes in endogenous flora during patients' hospital stay and the possible emergence of bacterial resistance . Data concerning antibiotic surgical prophylaxis (antimicrobial agent and duration) and length of hospitalization (pre- and post-surgery) were also collected . All patients underwent a microbiological screening by culturing nasal, pharyngeal and rectal swabs performed at admission and during hospitalization . Overall, 526 nasal swabs, 506 pharyngeal swabs and 482 rectal swabs were performed . Methicillin-resistant staphylococci were isolated from nasal swabs at admission in 2.1% of patients and in 7.5% of patients during hospitalization (day-14) . VRE and ESbetaL-producing strains were isolated from rectal swabs in 1.9 and 4.7% of patients, respectively, with no change during hospital stay . Nasal and pharyngeal flora significantly changed after 7-14 days of hospitalization, Gram-negative microorganisms being isolated more frequently following hospitalization . The authors conclude that excessive hospital stay duration, along with the inappropriate duration of surgical antibiotic prophylaxis could be important causes of bacterial flora modification.

Acupunct Electrother Res, 2003, 28(3-4), 201 - 6
The importance of Bi-Digital O-Ring Test in the treatment of multiple hepatic abscesses: a case history; Iwasa S et al.; The Bi-Digital O-Ring Test has been very useful in the identification of bacterial and viral infections, as well as other etiological agents, in difficult clinical cases . Case report of a patient with multiple hepatic abscesses (pylephlebitis induced hepatic abscess is the most difficult abscess to treat), in which the etiological agent was suggested through Bi-Digital O-Ring Test with excellent clinical evolution after modification of previously ineffective multi anti-microbial treatment is presented . 45 years old, female with a history of pain at right hypochondria for 15 days, with jaundice, oscillating fever and shivering . Computerized tomography showed liver with multiples nodules in the parenchyma with additional appendicitis . An appendectomy was performed with drainage of intra abdominal abscesses . Treatment with metronidazol, ceftazidim and amicacine was performed with no improvement while the general condition of the patient was deteriorating progressively in the following 3 weeks . Bi-Digital O-Ring Test was then performed to determine the etiological agent and the drug compatibility test among effective antimicrobial agents . Based on the Bi-Digital O-Ring Test, the main etiological agent was found to be Enterobacter aerogenes . Amongst the three antibiotics that were being used, only metronidazol was effective and the other 2 was cancelled its effect . Based on Bi-Digital O-Ring Test findings two new antibiotics (cefadroxil and imipenen), were added to metronidazol and additional cilantro tablets by Hayashibara, Japan was given, and Selective Drug Uptake Enhancement Method performed, with excellent clinical, laboratory testing and tomography improvement within 10 days . Bi-Digital O-Ring Test suggested the etiological agent, and effective and mutually compatible antibiotics for treating the abscesses which resulted in a good clinical evolution, characterized by relief of fever and reduction of the hepatic abscesses in a short period and followed complete disappearance of hepatic abscess.

Expert Opin Pharmacother, 2004 Feb, 5(2), 237 - 46
The use of fluoroquinolones in the treatment of skin infections; Martin SJ et al.; Fluoroquinolones have been studied for both uncomplicated and complicated skin and skin structure infections . Their broad spectrum, rapid bactericidal activity, extensive tissue penetration, excellent bioavailability and ease of administration have made these drugs a common choice for many infectious diseases, including skin infections . Extensive research has shown the fluoroquinolones to be as effective as beta-lactam antibiotics in managing a spectrum of diseases including erysipelas, cellulitis, impetigo, surgical wounds and diabetic foot infections . However, resistance to the fluoroquinolones has increased among the staphylococci, streptococci, Enterobacteriaceae and other important Gram-negative bacilli . Resistance has been linked directly to the widespread use of these compounds . Despite their appeal in the treatment of both uncomplicated and complicated skin infections, the fluoroquinolones should be reserved as alternatives to beta-lactams and other antibiotics or as empirical therapy in complicated infections until pathogens have been identified and drug regimens can be focused.

Int J Food Microbiol, 2004 Mar 1, 91(2), 205 - 8
Decontamination of broilers with hydrogen peroxide stabilised with glycerol during processing; Wagenaar CL et al.; Experiments were conducted to assess the effects of different concentrations of hydrogen peroxide stabilised by glycerol solution in potable water on the bacteriological and organoleptic quality of freshly slaughtered broiler carcasses . Skin-pH and colour were measured 3.5 and 24 h after treatment and compared to untreated carcasses . Bacterial colonisation was determined 3.5 and 24 h and, 7 days after treatment, carcasses being stored at 1 degrees C . None of the concentrations used affected the appearance and "bloom" of the carcasses as could be measured by colorimeter and changing of the acidity . Mean microbial counts were significantly reduced (P<0.01) when treated and control broilers were compared . Average reductions of 0.3 up to1.4 log N for the mesophilic aerobic counts were achieved and from 0.4 up to 1.2 log N for Enterobacteriaceae . A 3% w/w solution made from a commercially available stock solution (Glyroxyl), which consists of 44% hydrogen peroxide, 44% demineralised water and 12% glycerol proved to lower colonisation more effectively than a 2% solution.

Transfusion, 2004 Mar, 44(3), 359 - 63
Evaluation of a new generation of plastic culture bottles with an automated microbial detection system for nine common contaminating organisms found in PLT components; Brecher ME et al.; BACKGROUND: A microbial detection system (BacT/ALERT 3D, bioMerieux {formerly Organon Teknika}) has previously been validated with a variety of bacterial contaminants in PLTs . The recovery of nine organisms seeded into PLTs with new plastic culture bottles was studied in comparison to the current glass bottles . The use of plastic instead of glass would be expected to reduce the risk of injury . STUDY DESIGN AND METHODS: Isolates of Bacillus cereus, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Serratia marcescens, Streptococcus viridans, and Propionibacterium acnes were inoculated into Day 2 (>24 hr <48 hr) apheresis PLT units to 10 and 100 CFUs per mL . Replicate samples (4 mL) were inoculated into both current- and new-generation standard aerobic and anaerobic bottles . RESULTS: All organisms (with the exception of P . acnes) were detected in a mean time of 9.3 to 18.9 hours (10 CFUs/mL) or 8.7 to 18.2 hours (100 CFUs/mL) . In aggregate (with the exception of P . acnes), the plastic and glass aerobic bottles had a mean difference in detection of 1.2 hours (p < 0.0001), and the plastic and glass anaerobic bottles had a mean difference of 3.3 hours (p < 0.0001) . In all cases, the mean detection time was superior or clinically comparable (within 0.1 hr) with the new plastic bottles . P . acnes (an anaerobic organism) was detected with the new and current anaerobic bottles in a mean of 72.8 and 90.4 hours (10 CFUs/mL) or 64.0 and 80.8 hours (100 CFUs/mL), respectively . The narrower bottle neck and smaller inoculation septum present with the new-generation plastic bottles were inoculated with comparable ease to that of the glass bottles . CONCLUSIONS: These data demonstrate that the new plastic bottles are clinically comparable or superior to the current glass standard aerobic and anaerobic culture bottles.

Biochemistry, 2004 Mar 9, 43(9), 2664 - 72
Kinetics of turnover of cefotaxime by the Enterobacter cloacae P99 and GCl beta-lactamases: two free enzyme forms of the P99 beta-lactamase detected by a combination of pre- and post-steady state kinetics; Kumar S et al.; Third-generation cephalosporins bearing oximino side chains are resistant to hydrolysis by class C beta-lactamases such as that from Enterobacter cloacae P99 . For example, steady state parameters for hydrolysis of cefotaxime by this enzyme are as follows: k(cat) = 0.41 s(-1), K(m) = 17.2 microM, and k(cat)/K(m) = 2.3 x 10(4) s(-1) M(-1) . On the other hand, however, the K(i) value for cefotaxime as an inhibitor of cephalothin hydrolysis is 27 nM . The discrepancy between K(m) and K(i) indicated that a real steady state had not been achieved in at least one of these experiments . Analysis indicated that only two to three cefotaxime turnovers occurred during the K(i) determination . This suggested that the first few turnovers of cefotaxime by the P99 beta-lactamase may be different from those in the subsequent steady state . A direct pre-steady state experiment confirmed this hypothesis . The simplest reaction scheme that fitted the data involved replacement of the initial enzyme form, E, which bound cefotaxime tightly, with a second more weakly binding form, E', by partitioning of the acyl-enzyme intermediate during the first few turnovers . Steady state turnover of cefotaxime then largely involved E' as the free enzyme form . E' slowly reverted to E in the post-steady state regime . Further evidence for this scheme included quantitative analysis of the post-steady state and observation of a difference in the catalytic activity of E and E' in 2 M ammonium sulfate . The kinetics of P99 beta-lactamase-catalyzed hydrolysis of an acyclic depsipeptide substrate bearing a third-generation cephalosporin side chain showed that the side chain is necessary but not sufficient for production of resistance to beta-lactamase; a combination of the side chain and the dihydrothiazine ring of a cephalosporin is required . The beta-lactamase of E . cloacae GC1, an extended spectrum mutant of the P99 enzyme, rapidly hydrolyzes third-generation cephalosporins, without the structural transition described above . The flexibility of the extended Omega loop of the GC1 enzyme probably leads to this situation . Conformational restriction of the loop in the P99 enzyme is probably responsible for the long-lived acyl-enzyme intermediate and the transition to E' induced by cefotaxime.

Eur J Immunol, 2004 Mar, 34(3), 695 - 704
Demonstration of strong enterobacterial reactivity of CD4+CD25- T cells from conventional and germ-free mice which is counter-regulated by CD4+CD25+ T cells; Gad M et al.; Unfractionated CD4+ T cells from the gut-associated lymphoid tissue (GALT) and peripheral lymph nodes are unresponsive when exposed to enterobacterial antigens in vitro . Under similar conditions, CD4+ T cells depleted in vivo or in vitro of CD4+CD25+ T cells proliferate extensively . The CD4+CD25- T cell reactivity depends on MHC class II presentation, specific TCR stimulation, CD4 ligation, and antigen processing by antigen-presenting cells . The CD4+CD25- T cells respond to autologous and heterologous enterobacterial antigens, but not to antigens from the feces of germ-free mice . Surprisingly, CD4+CD25- T cells obtained from the GALT of germ-free mice also proliferate when exposed to enterobacterial antigens, and adding back the conventional or germ-free CD4+CD25+ T cells to the enteroantigen-stimulated CD4+CD25- T cells abolishes proliferation . As judged from carboxyfluorescein diacetate succinimidyl ester-labeling experiments, 4-5% of the CD4+CD25- T cells respond to enteroantigen . The data show for the first time that CD4+CD25- T cells with reactivity towards the enterobacterial flora and regulatory CD4+CD25- T cells are present in both conventional and germ-free mice . The data suggest that a significant proportion of the peripheral pool of CD4+CD25- T cells express anti-enterobacterial reactivity, which, due to the presence of regulatory CD4+CD25+ T cells, is kept in a quiescent state.

Appl Microbiol Biotechnol, 2004 Sep, 65(4), 465 - 72 Epub 2004 Feb 26.
Stability and activity of an Enterobacter aerogenes-specific bacteriophage under simulated gastro-intestinal conditions; Verthe K et al.; A bacteriophage, designated UZ1 and showing lytic activity against a clinically important strain (BE1) of Enterobacter aerogenes was isolated from hospital sewage . The stability and lytic activity against this strain under simulated gastro-intestinal conditions was evaluated . After addition of bacteriophage UZ1 to a liquid feed at gastric pH 2, the phage was immediately inactivated and could not be recovered . However, by use of an antacid to neutralize stomach acidity, no significant changes in phage titer were observed after 2 h incubation at 37 degrees C . After supplementing pancreatic juice and further incubation for 4 h, the phage titer remained stable . The persistence of UZ1 in a mixed microbial ecosystem that was representative for the large intestine was monitored using an in vitro simulation of the human intestinal microbial ecosystem . A pulse administration of bacteriophage UZ1 at a concentration of 10(5) plaque-forming units (PFU)/ml to reactor 3 (which simulates the ascending colon) showed that, in the absence of the host, bacteriophage UZ1 persisted for 13 days in the simulated colon, while the theoretical washout was calculated at 16 days . To assess its lytic activity in an intestinal microbial ecosystem, a green fluorescent protein (gfp)-labeled E . aerogenes BE1 strain was constructed and gfp-specific primers were designed in order to quantify the host strain using real-time PCR . It was observed that bacteriophage UZ1 was able to replicate and showed lytic activity against E . aerogenes BE1/ gfp in an intestinal microbial ecosystem . Indeed, after 17 h a 2 log unit reduction of E . aerogenes BE1/ gfp was measured as compared with the assay without bacteriophage UZ1, while the phage titer increased by 2 log units at an initial multiplicity of infection of 0.07 PFU/colony-forming unit . This is the first report of an in vitro model to study bacteriophage activity in the complex intestinal microbial community.

An Pediatr (Barc), 2004 Mar, 60(3), 271 - 3
{Nosocomial Hafnia alvei sepsis in a neonatal intensive care unit}; Amil Perez B et al.; Hafnia alvei is a member of the Enterobacteriaceae family, which is rarely associated with infection in pediatric patients and exceptionally in the neonatal period . Infections caused by this organism are usually opportunistic . H . alvei bacteremias are mostly associated with abdominal disease or immunosuppression . We report four cases of nosocomial sepsis in preterm infants with positive blood cultures to H . alvei that were treated in our institution within a short period of time.

Clin Infect Dis, 2004 Mar 1, 38(5), 655 - 62 Epub 2004 Feb 17.
Longitudinal trends in fluoroquinolone resistance among Enterobacteriaceae isolates from inpatients and outpatients, 1989-2000: differences in the emergence and epidemiology of resistance across organisms; Lautenbach E et al.; We conducted a 12-year study to identify and compare trends in annual prevalence of fluoroquinolone (FQ) resistance among Enterobacteriaceae isolates obtained from inpatients and outpatients in our health care system . A total of 46,070 clinical Enterobacteriaceae isolates underwent susceptibility testing . Although there were significant increases in inpatient FQ resistance for all Enterobacteriaceae, FQ resistance trends differed significantly across Enterobacteriaceae (P<.001) . For isolates obtained from outpatients, only Escherichia coli and Proteus mirabilis demonstrated significant increases in FQ resistance (P<.001 for each) . Trends in outpatient FQ resistance also differed significantly across Enterobacteriaceae (P<.001) . There were significant differences between inpatient and outpatient FQ resistance trends for all Enterobacteriaceae except P . mirabilis and Enterobacter cloacae . Although hospital-wide use of certain antibiotics correlated significantly with inpatient FQ resistance, these correlations differed substantially across organisms . Efforts to elucidate the epidemiology of FQ resistance and identify targets for intervention must recognize and account for the variability of FQ resistance across organisms and clinical settings.

Eur J Clin Microbiol Infect Dis, 2004 Mar, 23(3), 185 - 93 Epub 2004 Feb 19.
Antibiotic resistance rates and phenotypes among isolates of Enterobacteriaceae in French extra-hospital practice; Quentin C et al.; Antibiotic resistance among members of the family Enterobacteriaceae was prospectively surveyed by eight French private laboratories over a 5-month period in 1999 . A total of 2,599 consecutive and nonduplicate strains were collected, mainly (60.9%) from patients in the community . Most strains (82.9%) derived from urine . Escherichia coli was the predominant (73.9%) organism isolated . The overall rates of antibiotic resistance were as follows: amoxicillin, 53.4%; amoxicillin-clavulanic acid, 27.3%; ticarcillin, 44.2%; piperacillin-tazobactam, 3.2%; cephalothin, 29.2%; cefuroxime, 14.7%; cefoxitin, 11.5%; ceftazidime, 3.6%; cefotaxime, 2.8%; cefepime, 0.3%; imipenem, 0.1%; gentamicin (G), 3.8%; tobramycin (T), 5.0%; netilmicin (Nt), 3.7%; amikacin (A), 0.7%; nalidixic acid, 14.3%; ofloxacin, 10.4%; cotrimoxazole, 21.1%; nitrofurantoin, 12.7%; fosfomycin, 5.2%; tetracycline, 50.1%; and colistin, 12.5% . Beta-lactam resistance phenotypes essentially comprised penicillinase production (33.9%), overexpression of chromosomal cephalosporinase (4.6%), and synthesis of inhibitor-resistant TEM/OXA enzymes (1.5%) or extended-spectrum beta-lactamases (1.5%) . Aminoglycoside resistance phenotypes consisted of GTNt (93 strains), TNtA (68 strains), GTNtA (14 strains), T (4 strains), GT (3 strains), G (1 strain), and reduced uptake/permeability (3 strains) . Most of the nalidixic acid-resistant strains were resistant to ofloxacin (72.8%) . Antibiotic resistance rates and phenotypes varied widely according to the bacterial group and the source of the strains . Significantly higher rates were observed in private healthcare centers than in the community, due to a higher proportion of both resistant species and resistant strains . However, multidrug-resistant isolates, including five extended-spectrum beta-lactamase-producing strains, were also recovered from the community.

J Perinatol, 2004 Mar, 24(3), 169 - 74
Postconception age and other risk factors associated with mortality following Gram-negative rod bacteremia; Benjamin DK Jr et al.; BACKGROUND: Neonatal nosocomial Gram-negative rod bacteremia (GNR-b) is considered ominous . DESIGN: Multi-center cohort study of premature infants (N=6172) who had a blood culture after day of life 3 and whose birthweight was < or =1250 g . RESULTS: A total of 437 neonates developed GNR-b; most commonly with Klebsiella (122/437; 28%), Enterobacter (97/437; 22%), Escherichia coli (90/437; 21%), Pseudomonas (63/437; 14%), and Serratia (49/437; 11%) . Neonates infected with Pseudomonas were more likely to die (21/63; 33%) than infants infected with other GNR (50/374; 13%) . In multivariable logistic regression, infection with Pseudomonas, mechanical ventilation, and race were associated with subsequent mortality . Postconception age (PCA) was most strongly associated with mortality . Using neonates with >34 weeks PCA at the time of the first blood culture as the reference category, mortality was higher in neonates <26 weeks PCA (odds ratio (OR)=9.21; 95% confidence interval (CI)=2.79, 30.44), and in neonates 26 to 28 weeks PCA (OR=3.94; 95% CI=1.29, 12.03) . CONCLUSIONS: Among premature infants, much of the mortality experienced in GNR-b is due to infection with Pseudomonas rather than enteric GNR . Race, the need for mechanical ventilation, and younger PCA when the blood culture was obtained were also strongly associated with mortality.

J Trop Pediatr, 2004 Feb, 50(1), 48 - 50
Neonatal sepsis and meningitis in Haiti; Desinor OY et al.; The aim of the study was to determine the etiology of meningitis and sepsis in the newborn at the State University Hospital of Haiti and evaluate the susceptibility 'in vitro' of the pathogens to the antibiotics commonly used . This was a prospective case series study over a 10-month period (May 1997-February 1998) of 42 newborns with sepsis and/or meningitis . Besides the clinical signs, a positive blood culture and/or a positive culture of cerebrospinal fluid was present in each case . Gram-negative bacteria were most commonly found as a cause of early onset sepsis, with Enterobacter aerogenes as the most common agent . There were no such difference between gram-negative and gram-positive in late onset sepsis . Group B Streptococcus was associated with neonatal meningitis (44 per cent of cases) which was more related to gram-positive bacteria (66 per cent) . Risk factors were vaginal discharge and dysuria in mothers, and low apgar score in newborns . Thirty-three per cent of the pathogens found, among them Klebsiella pneumoniae, were resistant 'in vitro' to ampicillin and gentamycin . All were susceptible to amikacin . Enterobacter aerogenes is an important pathogen in the etiology of early onset sepsis in the newborn at the State University Hospital of Haiti, while Group B Streptococcus is the leading cause of meningitis in that age group . Resistance to gentamycin should be taken into consideration for the treatment of sepsis and meningitis in the newborn.

Antimicrob Agents Chemother, 2004 Mar, 48(3), 1043 - 6
Inhibitors of antibiotic efflux in resistant Enterobacter aerogenes and Klebsiella pneumoniae strains; Chevalier J et al.; In Enterobacter aerogenes and Klebsiella pneumoniae, efflux provides efficient extrusion of antibiotics and contributes to the multidrug resistance phenotype . One of the alkoxyquinoline derivatives studied here, 2,8-dimethyl-4-(2'-pyrrolidinoethyl)-oxyquinoline, restores noticeable drug susceptibility to resistant clinical strains . Analyses of energy-dependent chloramphenicol efflux indicate that this compound inhibits the efflux pump mechanism and improves the activity of structurally unrelated antibiotics on multidrug-resistant E . aerogenes and K . pneumoniae isolates.

Antimicrob Agents Chemother, 2004 Mar, 48(3), 1040 - 2
Selection during cefepime treatment of a new cephalosporinase variant with extended-spectrum resistance to cefepime in an Enterobacter aerogenes clinical isolate; Barnaud G et al.; Enterobacter aerogenes resistant to cefepime (MIC, 32 microg/ml) was isolated from a patient treated with cefepime for an infection caused by a strain of E . aerogenes overproducing its AmpC beta-lactamase (MIC of cefepime, 0.5 microg/ml) . The AmpC beta-lactamase of the resistant strain had an L-293-P amino acid substitution and a high k(cat)/K(m) ratio for cefepime . Both of these modifications were necessary for resistance to cefepime.

Infect Immun, 2004 Mar, 72(3), 1786 - 94
Identification and molecular characterization of EatA, an autotransporter protein of enterotoxigenic Escherichia coli; Patel SK et al.; Enterotoxigenic Escherichia coli (ETEC) strains remain a formidable cause of diarrheal disease . To identify novel surface proteins of ETEC, we performed TnphoA mutagenesis of prototype ETEC strain H10407 and discovered a secreted protein not previously recognized in ETEC . DNA sequencing of the interrupted locus in mutant TnphoA.977 revealed a candidate 4,095-bp open reading frame without significant homology to commensal E . coli K-12 genomic DNA . Translation of this sequence revealed that it encoded a predicted peptide of 147.7 kDa that bears significant homology to members of the autotransporter family of bacterial virulence factors, particularly the serine protease autotransporters of the Enterobacteriaceae proteins . The gene identified in H10407, eatA (ETEC autotransporter A), encodes a potential serine protease motif (GDSGSP) in the secreted amino-terminal domain, and the predicted peptide shows more than 80% homology with SepA, a virulence protein secreted by Shigella flexneri . DNA hybridization and PCR demonstrated that eatA resides on the 92-kDa pCS1 virulence plasmid of H10407 and that it is present in multiple clinical ETEC strains . Immunoblots with antisera directed against a recombinant EatA passenger protein fragment identified a 110-kDa protein in supernatants purified from H10407 but not from the TnphoA.977 mutant or H10407-P, which lacks pCS1 . EatA possesses serine protease activity that is abolished by mutations within a serine protease catalytic triad formed by residues H(134), D(162), and S(267) . Finally, interruption of the eatA gene retarded fluid accumulation in the rabbit ileal loop model, suggesting that this autotransporter contributes to the virulence of ETEC.

Folia Microbiol (Praha), 2003, 48(5), 563 - 71
Alternative sources of biologically active substances; Behal V; The majority of antibiotics and substances with diverse biological activity used in medicine are produced by actinomycetes, nonfilamentous bacteria and fungi . Other microorganisms, such as myxobacteria, pseudomonads, nocardias, basidiomycetes, marine microorganisms, enterobacteria, halobacteria, hyperthermophiles etc . are investigated for new biologically active metabolites.

Virus Genes, 2004 Mar, 28(2), 207 - 14
Isolation, molecular characterisation and genome sequence of a bacteriophage (Chp3) from Chlamydophila pecorum; Garner SA et al.; Chlamydiae are obligate intracellular pathogens that have a unique developmental cycle . Thirty nine viable isolates representing all nine currently recognised chlamydial species were screened by immunofluorescence with a cross-reacting chlamydiaphage monoclonal antibody . A novel chlamydiaphage (Chp3) was detected in C . pecorum, a chlamydial species not previously known to carry bacteriophages . Chp3 belongs to the Microviridae, members of this virus family are characterised by circular, single-stranded DNA genomes and small T = 1 icosahedral capsids . Double-stranded replicative form Chp3 DNA was purified from elementary bodies and used as a template to determine the complete genome sequence . The genome of Chp3 is 4,554 base pairs and encodes eight open reading frames organised in the same genome structure as other chlamydiaphages . An unrooted phylogenetic tree was constructed based on the major coat proteins of 11 members of the Microviridae and Chp3 . This showed that the Microviridae are clearly divided into two discrete sub-families; those that infect the Enterobacteriaceae e.g . OX174 and the bacteriophages that infect obligate intracellular bacteria or mollicutes including SpV4 (Spiroplasma melliferum), OMH2K (Bdellovibrio bacteriovorus) and the chlamydiaphages . Comparative analyses demonstrate that the chlamydiaphages can be further subdivided into two groupings, one represented by Chp2/Chp3 and the other by OCPG1/OCPAR39.

Virology, 2004 Jan 5, 318(1), 245 - 66
The genome and proteome of coliphage T1; Roberts MD et al.; The genome of enterobacterial phage T1 has been sequenced, revealing that its 50.7-kb terminally redundant, circularly permuted sequence contains 48,836 bp of nonredundant nucleotides . Seventy-seven open reading frames (ORFs) were identified, with a high percentage of small genes located at the termini of the genomes displaying no homology to existing phage or prophage proteins . Of the genes showing homologs (47%), we identified those involved in host DNA degradation (three endonucleases) and T1 replication (DNA helicase, primase, and single-stranded DNA-binding proteins) and recombination (RecE and Erf homologs) . While the tail genes showed homology to those from temperate coliphage N15, the capsid biosynthetic genes were unique . Phage proteins were resolved by 2D gel electrophoresis, and mass spectrometry was used to identify several of the spots including the major head, portal, and tail proteins, thus verifying the annotation.

Biochemistry (Mosc), 2004 Jan, 69(1), 103 - 7
D- and L-aspartic acids: new non-sugar components of bacterial polysaccharides; Kocharova NA et al.; For the first time in bacterial polysaccharides, residues of D- and L-aspartic acids were identified as N-acyl substituents of 4-amino-4,6-dideoxy-D-glucose in the O-antigens of enterobacteria of the genera Providencia and Proteus.

J Food Prot, 2004 Feb, 67(2), 303 - 9
Protocol for evaluating the efficacy of cetylpyridinium chloride as a beef hide intervention; Bosilevac JM et al.; The objective of this study was to establish the necessary protocols and assess the efficacy of cetylpyridinium chloride (CPC) as an antimicrobial intervention on beef cattle hides . Experiments using CPC were conducted to determine (i) the methods of neutralization needed to obtain valid efficacy measurements, (ii) the effect of concentration and dwell time after treatment, (iii) the effect of CPC on hide and carcass microbial populations when cattle were treated at a feedlot and then transported to a processing facility for harvest, and (iv) the effectiveness of spray pressure and two-spray combinations of CPC and water to reduce hide microbial populations . Residual CPC in hide sponge samples prevented bacterial growth . Dey-Engley neutralization media at 7.8% and a centrifugation step were necessary to overcome this problem . All dwell times, ranging from 30 s to 4 h, after 1% CPC application to cattle hides resulted in aerobic plate counts and Enterobacteriaceae counts 1.5 log CFU/100 cm2 lower than controls . The most effective dose of CPC was 1%, which reduced aerobic plate counts and Enterobacteriaceae counts 2 and 1 log CFU/100 cm2, respectively . Low-pressure application of 1% CPC at the feedlot, transport to the processing facility, and harvest within 5 h of application resulted in no effect on Escherichia coli O157 prevalence on hides or preevisceration carcasses . Two high-pressure CPC washes lowered aerobic plate counts and Enterobacteriaceae counts by 4 log CFU/100 cm2, and two medium-pressure CPC washes were only slightly less effective . These results indicate that under the proper conditions, CPC may still be effective for reducing microbial populations on cattle hides . Further study is warranted to determine if this effect will result in reduction of hide-to-carcass contamination during processing.

Rev Esp Salud Publica, 2003 Nov-Dec, 77(6), 749 - 60
{Microbiological study of the meals served in school lunch rooms on the island of Tenerife, Spain}; Campos Diaz J et al.; BACKGROUND: School lunchrooms and catered meals are of major importance from the Public Health standpoint . This study is aimed at evaluating the microbiological quality of the meals served in school lunchrooms for the purpose of ascertaining whether it is suitable or, to the contrary, the intake thereof may involve a serious health problem for this high-risk group . METHODS: A transversal descriptive epidemiological study . An analysis was conducted of a total of 898 food samples collected from the lunchrooms at 101 schools in Tenerife, selected by a stratified random probabillistic sampling procedure, fifty-eight of which were prepared at the school proper (direct management) and 43 involving meals served by a catering firm (prepared under contract) . RESULTS: No disease-causing Salmonella spp . or Listeria monocytogenes bacteria were isolated from any of the samples . A total 79% of the foods studies showed counts for this parameter, (91%) in salads and (85%) in main courses . A total 15% of the samples analyzed tested positive for total Enterobacteriaceae . Escherichia coli was isolated in 24% of the salads, in 4% of the side dishes and in 1% of the main dishes . Staphylococcus aureus having in isolated in three foods . The highest counts were found for the total aerobic mesophyllic microorganisms . A total 8.24% of the samples analyzed exceeded one or more of the limits stipulated for the parameters studies . CONCLUSIONS: The microbiological quality of the meals served in these school lunchrooms is acceptable, although due to a certain percentage of the foods having exceeded the stipulated limits for microorganisms indicative of and revealing a lack of hygiene, and school-children being a high-risk group, a revision of the surveillance related to critical checkpoints will be necessary.

Acta Paediatr Taiwan, 2003 Sep-Oct, 44(5), 300 - 2
Congenital nephrotic syndrome: report of an infant with diffuse mesangial sclerosis; Chang JW et al.; A case of congenital nephrotic syndrome (CNS) caused by diffuse mesangial sclerosis (DMS) is presented . A female baby weighting 2,680 gm was delivered at 35 weeks' gestation . She had early onset of generalized edema, heavy proteinuria, oligouria, uncorrectable hypoalbuminemia, and rapid progression to renal failure . Even after being treated with strong antibiotics (teicoplanin and ceftazidine), the infant died of septic shock with Enterobacter cloacea, only sensitive to imipenem, at the age of 7 days . The necropsy showed diffuse mesangial sclerosis . This case demonstrates that prematurity, low birth body weight and early onset of symptoms are not pathognomonic of the congenital nephrotic syndrome of the Finnish type (CNF) . It can also occur in DMS . Besides, empirical antibiotics should be started promptly and should cover the major hospital strains of bacteria if the patient is not well.

Genetika, 2003 Dec, 39(12), 1606 - 11
{Application of pM3, a broad host range IncP-9 plasmid, for genetic analysis in Enterobacteriaceae}; Titok MA; The possibility of using a transposon-carrying variant of broad host range plasmid pM3 (IncP-9) as a universal vector for transposon mutagenesis and as a chromosome-mobilizing factor was demonstrated in bacteria of the family Enterobacteriaceae.

Eur J Intern Med, 2003 Dec, 14(8), 484 - 487
The incidence of oral Gram-negative bacteria in patients with Parkinson's disease; Gosney M et al.; Background: Parkinson's disease is a common neurodegenerative disorder that affects an increasing number of older people every year . Dysphagia is not only a common feature, but one that results in poor nutrition and an increased risk of bronchopneumonia . Previous work has suggested that the oral flora is altered in patients with oral pathology . Methods: Fifty patients were assessed to quantify the incidence of oral Gram-negative bacteria . Results: Sixteen of the patients with Parkinson's disease were found to have six different Gram-negative bacilli in their oral cavities . The 20 different Gram-negative bacteria present were Escherichia coli (n=7), Klebsiella spp . (n=3), Kluyvera spp . (n=3), Serratia spp . (n=3), Proteus spp . (n=2) and Enterobacter spp . (n=2) . We found that the oral cavity of 16 (32%) of the patients with Parkinson's disease was abnormally colonised with Gram-negative bacteria and that Gram-negative bacteria were more likely to occur in those patients in whom oromuscular dysfunction was present (88% vs . 21%; p<0.05) . Conclusion: Further work is required to determine the association between oral flora and the pathogenic organisms found in aspiration pneumonia as well as work on innovative treatments to reduce oral Gram-negative bacteria in those patients at particular risk of aspiration pneumonia.

Rev Esp Quimioter, 2003 Dec, 16(4), 394 - 7
{Class 1 integrons in Enterobacteriaceae and its association with multidrug resistance and conjugative plasmids}; Alvarez-Fernandez M et al.; The prevalence of antimicrobial resistance genes is a cause of concern . The combination of antimicrobial resistance genes and mobile genetic elements leads to their widespread presence in different bacterial species, in which integrons are a new and important element . We studied the presence of integrons in 123 unrelated enterobacteria and identified them in 20.3% of the strains . The combination of integrons and multidrug resistance was shown to be statistically significant (p <0.001) . Integron-positive isolates were statistically (p <0.05) more likely to be resistant to amoxicillin-clavulanic acid, quinolones and trimethoprim-sulfamethoxazole . All the integrons were identified in conjugative plasmids . The prevalence of integrons increased from 21.2% in 1992-1994 to 72% in 1995-1997 (p <0.001) . The aacC1 and aacC2 genes were identified in 80% of the integrons . The relationship between integrons and conjugative plasmids is a matter of concern because it could contribute to the dissemination of antimicrobial resistance genes among different bacterial populations.

Pediatr Surg Int, 2004 Apr, 20(4), 267 - 70 Epub 2004 Feb 10.
Critical assessment of the methods used for detection of bacterial translocation; Hernandez Oliveros F et al.; AIM: Bacterial translocation (BT) can be demonstrated by blood and lymph node cultures and also by polymerase chain reaction (PCR) detection of DNA of enteric bacteria . Aiming at investigating BT after gastrointestinal operations we assessed it on two endpoints after ischemia-reperfusion (IR) or sham operation (SO) . METHODS: 2 groups of 200-g Brown Norway male rats were treated as follows: SO animals ( n=12) had laparotomy alone and IR animals ( n=12) had successively 15 min clamping of the portal vein and the mesenteric artery . Half the animals in each group were killed on postoperative (p.o.) day 2 the other half on p.o . day 7 . Under sterile conditions regional lymph nodes and vena cava and portal vein blood samples were recovered and cultured for aerobes and anaerobes . Escherichia coli beta-galactosidase DNA was assessed in blood samples by PCR . The findings in the two groups were compared by means of chi(2) tests . RESULTS: Post-hepatic (peripheral blood) BT was detected by cultures of gram-negative bacteria in 16% and 0% of SO and IR animals, respectively, on p.o . day 2 and in 16% and 50% on p.o . day 7 . These differences were not significant (ns) . E . coli DNA was found in one SO rat . Pre-hepatic BT (portal blood and/or lymph nodes) of gram-negative bacteria was found in 16% and 33%, respectively, on day 2 and in 16% and 16% on day 7 (ns) . However, if gram-positive cultures were taken into account, the figures were 66% and 66% on day 2 and 66% and 83% on day 7 (ns) . No anaerobes could be cultured . CONCLUSIONS: (1) BT is frequent in surgically manipulated animals . (2) To limit the assessment of BT to Enterobacteriaceae is probably misleading, since consistent amounts of gram-positive bacteria are found in the pre-hepatic territory . (3) PCR tests limited to E . coli DNA alone are likely incomplete . (4) Short periods of vascular clamping do not increase BT on the two endpoints selected in comparison with SO animals.

Mol Microbiol, 2004 Feb, 51(4), 1117 - 28
Metabolic block at early stages of the glycolytic pathway activates the Rcs phosphorelay system via increased synthesis of dTDP-glucose in Escherichia coli; El-Kazzaz W et al.; A mutational block in the early stages of the glycolytic pathway facilitates the degradation of the ptsG mRNA encoding the major glucose transporter IICBGlc in Escherichia coli . The degradation is RNase E dependent and is correlated with the accumulation of either glucose-6-P or fructose-6-P (Kimata et al., 2001, EMBO J 20: 3587-3595; Morita et al., 2003, J Biol Chem 278: 15608-15614) . In this paper, we investigate additional physiological effects resulting from the accumulation of glucose-6-P caused by a mutation in pgi encoding phosphoglucose isomerase, focusing on changes in gene expression . The addition of glucose to the pgi strain caused significant growth inhibition, in particular in the mlc background . Cell growth then gradually resumed as the level of IICBGlc decreased . We found that the transcription of the cps operon, encoding a series of proteins responsible for the synthesis of colanic acid, was markedly but transiently induced under this metabolic stress . Both genetic and biochemical studies revealed that the metabolic stress induces cps transcription by activating the RcsC/YojN/RcsB signal transduction system . Overexpression of glucose-6-P dehydrogenase eliminated both growth inhibition and cps induction by reducing the glucose-6-P level . Mutations in genes responsible for the synthesis of glucose-1-P and/or dTDP-glucose eliminated the activation of the Rcs system by the metabolic stress . Taken together, we conclude that an increased synthesis of dTDP-glucose activates the Rcs phosphorelay system, presumably by affecting the synthesis of oligosaccharides for enterobacterial common antigen and O-antigen.

Pesqui Odontol Bras, 2003 Jul-Sep, 17(3), 228 - 33 Epub 2003 Dec 16.
Effect of a 0.5% chlorhexidine gel on dental plaque superinfecting microorganisms in mentally handicapped patients; Pannuti CM et al.; A randomized clinical trial was conducted to investigate the effect of a 0.5% chlorhexidine (CHX) gel on dental plaque superinfecting microorganisms in mentally handicapped patients . Thirty inmates from the institution "Casas Andre Luiz" were assigned to either test group (CHX gel, n = 15) or control group (placebo gel, n = 15) . The gel was administered over a period of 8 weeks . Supragingival plaque samples were collected at baseline, after gel use (8 weeks) and 16 weeks after baseline . The presence of Gram-negative Enterobacteriaceae, Staphylococcus and yeasts was evaluated . No significant growth of any superinfecting microorganism was observed in the CHX group, when compared to the placebo group . The results indicated that the 0.5% chlorhexidine gel did not produce an undesirable shift in these bacterial populations.

Diagn Microbiol Infect Dis, 2004 Jan, 48(1), 73 - 5
In vitro activity of BAL9141 against clinical isolates of gram-negative bacteria; Issa NC et al.; The minimum inhibitory concentration and minimum bactericidal concentration of BAL9141 against 150 clinical isolates of gram-negative bacteria were determined and compared to those of cefepime and ceftriaxone . BAL9141 exhibited comparable activity to cefepime and ceftriaxone against Haemophilus influenzae, Klebsiella pneumoniae, and extended-spectrum beta-lactamase nonproducing Enterobacter cloacae, and comparable activity to cefepime against Pseudomonas aeruginosa.

J Eur Acad Dermatol Venereol, 2003 Nov, 17(6), 702 - 5
Langerhans' cell histiocytosis and haemophagocytic lymphohistiocytosis in an elderly patient; Ioannidou D et al.; We present a case of a 78-year-old man suffering from a chronic psoriasiform eruption, with rapid deterioration over the previous 8 weeks . Langerhans' cell histiocytosis with skin and bone involvement was diagnosed, and there was evidence of liver and lung dysfunction . The patient was treated with prednisolone and etoposide, and initially experienced a partial improvement . Three weeks later, haemophagocytic lymphohistiocytosis and subsequently a large pulmonary abscess with sepsis attributed to opportunistic gram-negative enterobacteriaceae Serratia marcescens developed, and the patient died . The present case of Langerhans' cell histiocytosis is of particular interest because of the previously unreported development of haemophagocytic lymphohistiocytosis in the elderly population.

FEMS Microbiol Lett, 2004 Jan 30, 230(2), 197 - 202
Diversity of chromosomal AmpC beta-lactamases from Enterobacter cloacae isolates in a Portuguese hospital; Conceicao T et al.; Six clinical isolates of Enterobacter cloacae isolated in a Portuguese hospital, between April 1999 and November 2000, demonstrated resistance to almost all broad-spectrum cephalosporins, except to cefepime . These isolates were susceptible to quinolones and to aminoglycosides . Isoelectric focusing demonstrated production of beta-lactamases with pIs > 8.0 and by all six isolates, exhibiting a cephalosporinase phenotype . The results of pulsed field gel electrophoresis revealed that these isolates were genetically unrelated . The amino acid sequence of six AmpC beta-lactamases (Eclo1FF, Eclo6FF, Eclo9FF, Eclo10FF, Eclo11FF and Eclo15FF) shared 97-99% homology with the chromosomal AmpC beta-lactamase from E . cloacae P99 and 86-87% homology with those of two plasmid-mediated AmpC beta-lactamases, MIR-1 and ACT-1 . This is the first report of chromosomal AmpC beta-lactamase production by E . cloacae isolates in a Portuguese hospital.

FEMS Microbiol Lett, 2004 Jan 30, 230(2), 167 - 70
Permeabilizing effects of sub-inhibitory concentrations of microcystin on the growth of Escherichia coli; Dixon RA et al.; Microcystin, a cyanotoxin produced by Microcystis aeruginosa, lacks potent antibacterial activity . When tested in combination, in vitro, inhibitory values for a range of hydrophobic antibiotics were significantly reduced in the presence of at least 1/20 x the minimum inhibitory concentration of microcystin . The degree of inhibition was equivalent to that of a well-characterised permeabilizing agent, polymyxin B nonapeptide . The permeabilizing ability of sub-inhibitory concentrations of microcystin to affect the envelope of Escherichia coli was demonstrated by a rapid and sustained reduction in absorbance values of lysozyme-treated cells and by enhanced uptake of crystal violet in microcystin-treated cultures . Direct effects of appropriate concentrations of microcystin on the integrity of bacterial outer and inner membranes were measured by release of specific enzyme markers . Although the exact mechanism for permeabilizing E . coli with microcystin has not been elucidated, the effects were consistent with permeability changes to the enterobacterial outer membrane caused by polymyxin B nonapeptide.

Curr Microbiol, 2003 Dec, 47(6), 457 - 61
Cloning and expression of pyrroloquinoline quinone (PQQ) genes from a phosphate-solubilizing bacterium Enterobacter intermedium; Kim CH et al.; A grass rhizosphere bacterium, Enterobacter intermedium (60-2G), has a strong ability to solubilize insoluble phosphate . Certain phosphate-solubilizing bacteria secrete gluconic acid for this process . The gluconic acid is produced by direct extracellular oxidation of glucose by a glucose dehydrogenase equipped with pyrroloquinoline quinone (PQQ) as a cofactor . A pqq gene cluster producing PQQ was detected in E . intermedium and this sequence conferred phosphate-solubilizing activity to Escherichia coli DH5alpha . The 6,783-bp pqq sequence had six open reading frames (pqqA, B, C, D, E, and F) and showed 50-95% homology to pqq genes of other bacteria . E . coli DH5alpha expressing the E . intermedium pqq genes solubilized phosphate from hydroxyapatite after a pH drop to pH 4.0, which paralleled in time the secretion of gluconic acid . We speculate that production of PQQ in E . coli DH5alpha expressing the pqq cluster activates an endogenous glucose dehydrogenase to permit gluconic acid secretion that solubilizes the insoluble phosphate.

Infect Control Hosp Epidemiol, 2004 Jan, 25(1), 65 - 71
Genetic analysis of Pseudomonas aeruginosa by enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) and arbitrarily primed PCR: gel analysis compared with microchip gel electrophoresis; Jamasbi RJ et al.; OBJECTIVES: To assess the applicability of a newly emerging microchip gel electrophoresis for rapid strain differentiation among clinical isolates of Pseudomonas aeruginosa, and to compare this technique with the traditional gel method for DNA separation . METHODS: One hundred clinical strains of P . aeruginosa obtained from a hospital in northwestern Ohio were tested for reactivity to 3 serotype-specific monoclonal antibodies by enzyme-linked immunosorbent assay . Twelve strains (4 from each serogroup) were selected for DNA analysis by polymerase chain reaction (PCR)-based, single primer DNA fingerprinting methods with 3 different primers: 1 enterobacterial repetitive intergenic consensus PCR and 2 arbitrarily primed PCRs . The PCR products were analyzed by agarose slab gel and microchip gel electrophoresis . RESULTS: Of the 100 clinical isolates tested, 39% (4%, 14%, and 21%) were found to be serotypes 0:3, 0:6, and 0:11, respectively . Twelve strains were chosen for DNA analysis by PCR . The PCR products were analyzed by agarose slab gel electrophoresis and on microchips to determine interspecies diversity . Both methods demonstrated that different serotypes exhibited different electrophoretic patterns . Two strains (clinical strains 6 and 7, serotype 0:6) showed identical patterns, indicating a high degree of relatedness . CONCLUSION: In all cases, there was concordance between the electrophoretic patterns detected by the two methods . The capability of conducting both PCR and microchip gel electrophoresis offers an opportunity for an automated and rapid method for genetic analysis and differentiation among strains of P . aeruginosa and other microorganisms.

Rev Med Chir Soc Med Nat Iasi, 2003 Jul-Sep, 107(3), 595 - 8
{Activity of fourth generation cephalosporins against clinical isolates of Enterobacteriaceae}; Tuchilus C et al.; Cefpirome and cefepime are a novel group of cephalosporins which contain a positively charged quaternary ammonium at carbon 3 of the dihidrothiazone ring . The antimicrobial agents cefpirome, cefepime, cefotaxime, ceftriaxone, ceftazidime, cefoperazone and imipenem were tested against clinical isolates of Escherichia coli (n = 302) and Klebsiella spp . (n = 62) obtained during september-december 2002 from patients of Galati Emergency Hospital . The fourth generation cephalosporins cefpirome and cefepime have similar in vitro activities to the third generation cephalosporins . E . coli showed the comparable resistance rates for all cephalosporins . Against Klebsiella spp . strains cefpirome was less active (35.5% resistance) than cefepime (25.8% resistance) . As expected, imipenem had excellent activity (100% susceptibility).

Surg Endosc, 2004 May, 18(5), 755 - 6 Epub 2004 Feb 02.
Experience with routine intraabdominal cultures during laparoscopic gastric bypass with implications for antibiotic prophylaxis; Williams MD et al.; BACKGROUND: Techniques for laparoscopic Roux-en-Y gastric bypass that do not use bowel cross-clamping raise a question of increased risk for infectious complications . However, to the authors' knowledge, no studies have recorded routine intraoperative peritoneal cultures . This article reports the authors' experience with routine peritoneal cultures during laparoscopic Roux-en-Y gastric bypass and the role of antibiotic prophylaxis . METHODS: From January 2000 to March 2000, 66 consecutive patients undergoing a laparoscopically divided proximal Roux-en-Y gastric bypass had peritoneal fluid aspirated for routine culture . No mechanical or oral antibiotic bowel preparation was used . All the patients received preoperative intravenous antibiotic prophylaxis with Levoquin 500 mg and Flagyl 500 mg . Peritoneal fluid was aspirated from the left gutter near the site of the enteroenterostomy before irrigation with 1,000 ml of normal saline containing 50,000 U of bacitracin and 1 of kanamycin . RESULTS: The follow-up period averaged 9 months for 100% of patients . For 15 patients (22.7%), the culture results were positive . The 22 organisms cultured involved 15 streptococcus species, 4 anaerobes, 2 staphylococcus species, and 1 enterobacter . None of the patients experienced a clinical infection or required an extension of antibiotics beyond the first 24 h . CONCLUSIONS: This study demonstrated frequent peritoneal contamination during laparoscopic gastric bypass . Prophylactic intravenous antibiotics and antibiotic irrigation may have reduced the risk of clinically significant infections in this small uncontrolled series.

J Microbiol Methods, 2004 Feb, 56(2), 277 - 86
Detection of clinically relevant antibiotic-resistance genes in municipal wastewater using real-time PCR (TaqMan); Volkmann H et al.; Real-time PCR assays were developed for the quantifiable detection of the antibiotic-resistance genes vanA of enterococci, ampC of Enterobacteriaceae, and mecA of staphylococci in different municipal wastewater samples . Primer and probe designs for these resistance genes were constructed and optimised for application in standardised TaqMan PCR assays . Using reference strains, the linear measurement ranges of the assays were defined and covered concentration ranges of five to seven exponential values . Wastewater isolates of vancomycin-resistant enterococci (VRE) and beta-lactam-resistant Enterobacteriaceae were cultivated from municipal wastewaters in order to verify the specificity and sensitivity of the primer-probe systems . Additionally, clinical strains of staphylococci resistant to methicillin (MRSA) confirmed the applicability of the mecA-specific detection system . Total DNAs were extracted from five different wastewater treatment plants and used for direct TaqMan PCR detection of the resistance genes without prior cultivation . In municipal wastewater, the resistance gene vanA was detected in 21% of the samples, and ampC in 78% . The gene mecA was not found in municipal wastewater, but in two clinical wastewater samples.

Pediatr Infect Dis J, 2004 Jan, 23(1), 41 - 6
Diversity and sharing of Haemophilus influenzae strains colonizing healthy children attending day-care centers; Farjo RS et al.; BACKGROUND: Children attending day-care centers (DCCs) are at risk for Haemophilus influenzae nasopharyngeal colonization and acute otitis media . The degree to which a given strain circulates within a day-care center and the heterogeneity of strains among DCCs in a geographic area are not well-characterized . This study describes the prevalence rates of H . influenzae colonization in a large number of children attending day-care centers and examines the genetic diversity of colonizing strains and the degree of sharing among children . METHODS: Throat cultures were collected from 198 healthy children <3 years old attending 16 day-care centers in Michigan . All H . influenzae isolates were genetically typed by enterobacterial repetitive intergenic consensus PCR as the initial screening technique to identify unique strains within each child . Pulsed field gel electrophoresis was used subsequently to examine the genetic diversity of strains between children . RESULTS: There were 127 (64%) children colonized with H . influenzae . Wide variation in rates of colonization was identified among day-care centers (0 to 95%) . A total of 179 genetically unique H . influenzae strains were isolated, and 47 children (37%) were colonized with 2 or more genetically distinct H . influenzae organisms . Evidence of sharing of the same strain in different children was found in 13 of 15 colonized DCCs and 23% of genotypically unique strains were shared . CONCLUSION: The degree of sharing of H . influenzae among children in this study suggests transmission of these potentially pathogenic microorganisms in day-care centers.

Int J Syst Evol Microbiol, 2004 Jan, 54(Pt 1), 107 - 13
Erwinia papayae sp . nov., a pathogen of papaya (Carica papaya); Gardan L et al.; Bacterial canker of papaya (Carica papaya) emerged during the 1980s in different islands of the Caribbean . Nineteen strains of Gram-negative, rod-shaped, non-spore-forming bacteria isolated from papaya were compared to 38 reference and type strains of phytopathogenic Enterobacteriaceae and related bacteria . Phylogenetic analysis of 16S rRNA gene sequences showed that the papaya strains belonged to the genus Erwinia . The DNA G+C content of strain CFBP 5189T, 52.5 mol%, is in the range of the genus Erwinia . The 19 papaya strains were all pathogenic to papaya and were differentiated clearly from type or reference strains of phytopathogenic enterobacteria and related bacteria by phenotypic tests . The papaya strains constituted a discrete DNA hybridization group, indicating that they belonged to a unique genomic species . Thus, strains pathogenic to papaya belong to a novel species for which the name Erwinia papayae sp . nov . is proposed, with the type strain CFBP 5189T (=NCPPB 4294T).

Antimicrob Agents Chemother, 2004 Feb, 48(2), 648 - 50
Emergence in Klebsiella pneumoniae and Enterobacter cloacae clinical isolates of the VIM-4 metallo-beta-lactamase encoded by a conjugative plasmid; Luzzaro F et al.; Resistance to carbapenems is an emerging problem among gram-negative hospital pathogens . A transferable plasmid encoding the VIM-4 metallo-beta-lactamase was detected in isolates of Klebsiella pneumoniae and Enterobacter cloacae obtained from a single patient under carbapenem therapy . Thus, enterobacteria appear to increasingly contribute to the spread of VIM-type enzymes.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 505 - 13
Genetic analysis and complete primary structure of microcin L; Pons AM et al.; Escherichia coli LR05, in addition to producing MccB17, J25, and D93, secretes microcin L, a newly discovered microcin that exhibits strong antibacterial activity against related Enterobacteriaceae, including Salmonella enterica serovars Typhimurium and Enteritidis . Microcin L was purified using a two-step procedure including solid-phase extraction and reverse-phase C(18) high-performance liquid chromatography . A 4,901-bp region of the DNA plasmid of E . coli LR05 was sequenced revealing that the microcin L cluster consists of four genes, mclC, mclI, mclA, and mclB . The structural gene mclC encoded a 105-amino-acid precursor with a 15-amino-acid N-terminal extension ending with a Gly-Ala motif upstream of the cleavage site . This motif is typical of the class II microcins and other gram-positive bacteriocins exported by ABC transporters . The mclI immunity gene was identified upstream of the mclC gene and encodes a 51-amino-acid protein with two potential transmembrane domains . Located on the reverse strand, two genes, mclA and mclB, encoded the proteins MclA and MclB, respectively . They bear strong relatedness with the ABC transporter proteins and accessory factors involved in the secretion of microcins H47, V, E492, and 24 . The microcin L genetic system resembles the genetic organization of MccV . Furthermore the MccL primary structure has been determined . It is a 90-amino-acid peptide of 8,884 Da with two disulfide bridges . The N-terminal region has significant homologies with several gram-positive bacteriocins . The C-terminal 32-amino-acid sequence is 87.5% identical to that of MccV . Together, these results strongly indicate that microcin L is a gram-negative class II microcin.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 491 - 6
Plasmid-mediated 16S rRNA methylase in Serratia marcescens conferring high-level resistance to aminoglycosides; Doi Y et al.; Serratia marcescens S-95, which displayed an unusually high degree of resistance to aminoglycosides, including kanamycins and gentamicins, was isolated in 2002 from a patient in Japan . The resistance was mediated by a large plasmid which was nonconjugative but transferable to an Escherichia coli recipient by transformation . The gene responsible for the aminoglycoside resistance was cloned and sequenced . The deduced amino acid sequence of the resistance gene shared 82% identity with RmtA, which was recently identified as 16S rRNA methylase conferring high-level aminoglycoside resistance in Pseudomonas aeruginosa . Histidine-tagged recombinant protein showed methylation activity against E . coli 16S rRNA . The novel aminoglycoside resistance gene was therefore designated rmtB . The genetic environment of rmtB was further investigated . The sequence immediately upstream of rmtB contained the right end of transposon Tn3, including bla(TEM), while an open reading frame possibly encoding a transposase was identified downstream of the gene . This is the first report describing 16S rRNA methylase production in S . marcescens . The aminoglycoside resistance mechanism mediated by production of 16S rRNA methylase and subsequent ribosomal protection used to be confined to aminoglycoside-producing actinomycetes . However, it is now identified among pathogenic bacteria, including Enterobacteriaceae and P . aeruginosa in Japan . This is a cause for concern since other treatment options are often limited in patients requiring highly potent aminoglycosides such as amikacin and tobramycin.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 477 - 83
Mechanism of action of NB2001 and NB2030, novel antibacterial agents activated by beta-lactamases; Stone GW et al.; Two potent antibacterial agents designed to undergo enzyme-catalyzed therapeutic activation were evaluated for their mechanisms of action . The compounds, NB2001 and NB2030, contain a cephalosporin with a thienyl (NB2001) or a tetrazole (NB2030) ring at the C-7 position and are linked to the antibacterial triclosan at the C-3 position . The compounds exploit beta-lactamases to release triclosan through hydrolysis of the beta-lactam ring . Like cephalothin, NB2001 and NB2030 were hydrolyzed by class A beta-lactamases (Escherichia coli TEM-1 and, to a lesser degree, Staphylococcus aureus PC1) and class C beta-lactamases (Enterobacter cloacae P99 and E . coli AmpC) with comparable catalytic efficiencies (k(cat)/K(m)) . They also bound to the penicillin-binding proteins of S . aureus and E . coli, but with reduced affinities relative to that of cephalothin . Accordingly, they produced a cell morphology in E . coli consistent with the toxophore rather than the beta-lactam being responsible for antibacterial activity . In biochemical assays, they inhibited the triclosan target enoyl reductase (FabI), with 50% inhibitory concentrations being markedly reduced relative to that of free triclosan . The transport of NB2001, NB2030, and triclosan was rapid, with significant accumulation of triclosan in both S . aureus and E . coli . Taken together, the results suggest that NB2001 and NB2030 act primarily as triclosan prodrugs in S . aureus and E . coli.

Bioorg Med Chem Lett, 2004 Feb 9, 14(3), 649 - 51
Biosynthesis of agglomerin A: stereospecific incorporation of pro-R- and pro-S-hydrogens at sn-C-3 of glycerol into the branched C(3) moiety; Mashimo Y et al.; The biosynthetic origin of the C(3) branched unit of agglomerin A has been investigated . Feeding of sn-(3R)- and sn-(3S)-{3-(2)H}glycerols to Enterobacter agglomerans PB-6042 followed by (2)H NMR analysis of the resulting agglomerin A revealed that pro-R and pro-S hydrogens at sn-C-3 of glycerol were incorporated stereospecifically into 5E and 5Z hydrogens of agglomerin A, respectively . These results imply that the immediate precursor of the C(3) branched unit is not pyruvate, but 1,3-bisphosphoglyceric acid or its biological equivalent.

Med Pregl, 2003 Sep-Oct, 56(9-10), 460 - 4
{Catheter-related urinary infections at the Clinical Center in Banja Luka}; Verhaz A et al.; INTRODUCTION: Catheter-associated urinary tract infections are the most common nosocomial infections of the urinary tract, and among the most common nosocomial infections in general . The major problems of these infections include antibiotic resistance and enormous direct and indirect cost of treatment . MATERIAL AND METHODS: A retrospective study on major causes of infections and antibiotic resistance was conducted at four clinics of the Clinical Center of Banja Luka . An anonymous questionnaire was distributed to nursing staff dealing with urinary catheters in order to get an overview of their clinical performance . RESULTS: The results showed that in 89% of cases (out of 198 patients with developed catheter-associated urinary tract infection) infections were caused by gram-negative bacteria, in 7% by gram-positive bacteria and in 4% by candida . The most common bacteria were: Escherichia coli (33.6%), Pseudomonas aeruginosa (14.1%), Proteus mirabilis (13.3%), and Enterobacter (10.5%) . Majority of bacteria presented with extremely high resistance (72-100%) to ampicillin, gentamycin and cotrimoxazole, and in some cases a significant resistance to ciproflaxacine, nalidixic acid, ceftriaxone and ceftazidime . The questionnaire showed that nursing staff did not follow guidelines for medical care of patients with urinary catheters . CONCLUSION: It can be concluded that poor hygienic and epidemiological conditions, as well as irrational use of antibiotics contribute to uncontrolled development of urinary tract infections in catheterized patients.

J Pak Med Assoc, 2003 Nov, 53(11), 534 - 6
Comparison of double disc and combined disc method for the detection of extended spectrum beta lactamases in enterobacteriaceae; Jabeen K et al.; OBJECTIVE: To compare double disc approximation and combined disc method for their ability to detect extended spectrum b lactamase (ESBL) production in enterobacteriaceae and determine the percentage of isolates which are falsely reported as sensitive in absence of ESBL detection, in a clinical microbiology laboratory of a tertiary care hospital between September-October 2002 . METHODS: Selected isolates were identified according to standard biochemical tests . Disc susceptibility tests were performed according to NCCLS . ESBL detection by combined disc {cefotaxime (30 ug) versus cefotaxime plus clavulanate (30+10 ug)} was compared with detection using double discs {amoxy-clavulanic acid (20+10 ug) and aztreonam (30 ug) applied 10 mm apart} . Results were interpreted according to NCCLS and analysed on SPSS version 10 . RESULTS: ESBL production was detected in 140 (30%) isolates by combined disc method and 139 (29.5%) by double disc method . There was no significant difference between two methods . Of the ESBL positive isolates 41 (29%) gave zone diameters that were within the sensitivity range cutoff and would have been falsely reported as being beta lactam sensitive in absence of ESBL detection . CONCLUSION: ESBL detection should be routinely performed in clinical laboratories as false reporting would result in treatment failure despite in vitro sensitivity . No difference was found between the combined disc and double disc methods hence either of two could be used.

FEMS Microbiol Lett, 2004 Jan 15, 230(1), 73 - 83
PicU, a second serine protease autotransporter of uropathogenic Escherichia coli; Parham NJ et al.; Escherichia coli is the major aetiological agent of urinary tract infections (UTI) . Like diarrhoeagenic strains of E . coli, uropathogenic isolates possess virulence determinants that distinguish them from commensal strains and allow them to produce the clinical manifestations associated with UTI . Several autotransporter proteins have been associated with the ability of E . coli, and other Gram-negative bacteria, to cause disease . Recently, we described the existence within uropathogenic E . coli (UPEC) strains of Sat, a toxin of the serine protease autotransporter of Enterobacteriaceae (SPATE) subfamily . Using features common to proteins secreted via the autotransporter pathway we have identified nine additional autotransporter proteins from the genomic sequence data of UPEC CFT073 . Surprisingly, two additional members of the SPATE subfamily were identified . One protein, designated PicU, was homologous to the Pic protein identified in Shigella flexneri and enteroaggregative E . coli . The PicU protein was expressed and investigated for functional activity.

Mol Microbiol, 2004 Feb, 51(3), 837 - 48
A novel integrative and conjugative element (ICE) of Escherichia coli: the putative progenitor of the Yersinia high-pathogenicity island; Schubert S et al.; Diversification of bacterial species and pathotypes is largely caused by horizontal transfer of diverse DNA elements such as plasmids, phages and genomic islands (e.g . pathogenicity islands, PAIs) . A PAI called high-pathogenicity island (HPI) carrying genes involved in siderophore-mediated iron acquisition (yersiniabactin system) has previously been identified in Yersinia pestis, Y . pseudotuberculosis and Y . enterocolitica IB strains, and has been characterized as an essential virulence factor in these species . Strikingly, an orthologous HPI is a widely distributed virulence determinant among Escherichia coli and other Enterobacteriaceae which cause extraintestinal infections . Here we report on the HPI of E . coli strain ECOR31 which is distinct from all other HPIs described to date because the ECOR31 HPI comprises an additional 35 kb fragment at the right border compared to the HPI of other E . coli and Yersinia species . This part encodes for both a functional mating pair formation system and a DNA-processing region related to plasmid CloDF13 of Enterobacter cloacae . Upon induction of the P4-like integrase, the entire HPI of ECOR31 is precisely excised and circularised . The HPI of ECOR31 presented here resembles integrative and conjugative elements termed ICE . It may represent the progenitor of the HPI found in Y . pestis and E . coli, revealing a missing link in the horizontal transfer of an element that contributes to microbial pathogenicity upon acquisition.

Zhonghua Nan Ke Xue, 2003 Dec, 9(9), 690 - 2, 696
{Distribution and resistance trends of pathogens from urinary tract infections and impact on management}; Shao HF et al.; OBJECTIVE: To assess the bacterial profile and pattern of antibiotic resistance of urinary tract infections (UTIs) pathogens and to determine its clinical impact on management . METHODS: Midstream urine samples were submitted for culture from 1998 to 2002, and 798 isolates were obtained for antimicrobial susceptibility testing including amikacin (AMK), ampicillin (AMP), cefzolin (CFZ), cefuroxime (CXM), ceftriaxone (CRO), ceftaxime (CTX), ceftazidime (CAZ), nalidixoc acid (NAL), ciprofloxacin (CIP), trimethoprim/sulfamethoxazole (SXT), nitrofurantoin (NIT) for Gram-negative bacteria and oxcillin (OXA), ampicillin (AMP), cefzolin (CFZ), ciprofloxacin (CIP), gentamicin (Gen), vancomycin (VAN), trimethoprim/sulfamethoxazole (SXT), nitrofurantoin (NIT) for Gram-positive cocci . beta-lactamases and ESBLs were tested when needed . RESULTS: Enterobacteriaceae was the most frequently isolated pathogen . Among all the isolates, Escherichia coli accounted for 66.0%, followed by Enterococcus (6.5%), Klebsiella spp . (6.0%), Staphylococcus (5.4%) . High resistance rates to CIP (56.0%), SXT (67.0%) and AMP (78.9%) were observed among the E . coli . CIP-resistant E . coli strains are being isolated with increasing frequency . From 1998 to 2002 the incidence of CIP-resistant increased steadily from 46.6% to 59.4% . A higher resistance rate to NAL was apparent . In contrast, NIT displayed a resistance rate of 8.9%, and AMK 4.9% . The ESBLs positive rate was 12.9% among the E . coli and 33.3% among the Klebsiella spp . respectively . A high resistance rate to CIP was also observed among the Staphylococcus (38.1%), Enterococcus (61.5%) and Streptococcus (85.0%), and the beta-lactamases positive rate was 95.2% among the Staphylococcus, but a lower resistance rate to NIT among Staphylococcus (2.4%) and Enterococcus (11.5%) . CONCLUSIONS: Resistance rates among common uropathogens continue to evolve and appear to be increasing to many commonly used agents especially to quinolones . Continued surveillance of resistance rates among uropathogens is needed to ensure appropriate recommendations for the treatment of the infections . Currently, the most appropriate agent for the empirical management of UTIs seems to be nitrofurantoin.

Clin Infect Dis, 2004 Feb 1, 38(3), 329 - 34 Epub 2004 Jan 06.
Multidrug-resistant Escherichia coli clonal groups causing community-acquired pyelonephritis; Manges AR et al.; From October 1999 through January 2000, an Escherichia coli clonal group (designated "CgA") was isolated from the urine of nearly one-half of all women with urinary tract infections (UTIs) caused by trimethoprim-sulfamethoxazole (TMP-SMZ)-resistant E . coli in a California community . This study describes the prevalence of pyelonephritis caused by CgA in the same community . E . coli isolates were characterized by enterobacterial repetitive intergenic consensus (ERIC2) polymerase chain reaction (PCR), serogrouping, and pulsed-field gel electrophoresis . Fourteen (11%) of 130 women with UTIs received a diagnosis of pyelonephritis . CgA was associated with 4 (57%) of the 7 pyelonephritis cases caused by TMP-SMZ-resistant E . coli and was associated with none of the cases caused by TMP-SMZ-susceptible E . coli (P<.02) . Six (86%) of these TMP-SMZ-resistant E . coli isolates belonged to 2 distinct ERIC2 PCR-defined clonal groups, whereas all of the TMP-SMZ-susceptible E . coli strains had unique fingerprints (P<.001) . The prevalence of antimicrobial-resistant pyelonephritis in a community may be affected by a limited number of E . coli clonal groups.

J Am Acad Dermatol, 2004 Feb, 50(2), 315 - 8
Infectious eccrine hidradenitis caused by Nocardia; Antonovich DD et al.; Neutrophilic eccrine hidradenitis is a nonspecific clinical reaction pattern classified as a neutrophilic dermatosis that typically occurs in the setting of chemotherapy for hematologic malignant disease . Neutrophilic eccrine hidradenitis more rarely has been reported in association with infectious agents, including Serratia, Enterobacter, Staphylococcus, and HIV . We describe the first case of infectious eccrine hidradenitis occurring in a patient with cutaneous Nocardia infection.

Lancet, 2004 Jan 3, 363(9402), 39 - 40
Occurrence of Enterobacter sakazakii in food production environments and households; Kandhai MC et al.; Enterobacter sakazakii occasionally causes illness in premature babies and neonates . Contamination of infant formulae during factory production or bottle preparation is implicated . Advice to health-care professionals focuses on bottle preparation, but the effectiveness of prevention depends on the degree of contamination and contamination sites, which are generally unknown . To keep contamination to a minimum in the finished product depends on knowledge of the occurrence of E sakazakii . We used a refined isolation and detection method to investigate the presence of this micro-organism in various food factories and households . Environmental samples from eight of nine food factories and from five of 16 households contained E sakazakii . The widespread nature of this micro-organism needs to be taken into account when designing preventive control measures.

Am J Respir Med, 2003, 2(1), 75 - 107
Cefepime: a review of its use in the management of hospitalized patients with pneumonia; Chapman TM et al.; Cefepime (Maxipime), Maxcef, Cepimax, Cepimex, Axepim, a parenteral fourth-generation cephalosporin, is active against many organisms causative in pneumonia . Cefepime has in vitro activity against Gram-positive organisms including Staphylococcus aureus and penicillin-sensitive, -intermediate and -resistant Streptococcus pneumoniae similar to that of cefotaxime and ceftriaxone . Cefepime also has good activity against Gram-negative organisms, including Pseudomonas aeruginosa, similar to that of ceftazidime . Importantly, cefepime is stable against many of the common plasmid- and chromosome-mediated beta-lactamases and is a poor inducer of AmpC beta-lactamases . As a result, it retains activity against Enterobacteriaceae that are resistant to third-generation cephalosporins, such as derepressed mutants of Enterobacter spp . Cefepime may be hydrolyzed by the extended-spectrum beta-lactamases produced by some members of the Enterobacteriaceae, but to a lesser extent than the third-generation cephalosporins . Monotherapy with cefepime 1 or 2g, usually administered intravenously twice daily, was as effective for clinical and bacteriological response as ceftazidime, ceftriaxone or cefotaxime monotherapy (1 or 2g two or three times daily) in a number of randomized, clinical trials in hospitalized adult, or less commonly, pediatric, patients with generally moderate to severe community-acquired or nosocomial pneumonia . More limited data indicated that monotherapy with cefepime 2g three times daily was also as effective in treating patients with nosocomial pneumonia as imipenem/cilostatin 0.5g four times daily, and when combined with amikacin, cefepime was as effective as ceftazidime plus amikacin . Patients with pneumonia who failed to respond to previous antibacterial therapy with penicillins or other cephalosporins responded to treatment with cefepime . Cefepime is generally well tolerated, with a tolerability profile similar to those of other parenteral cephalosporins . In clinical trials, the majority of adverse events experienced by cefepime recipients were mild to moderate and reversible . The most common adverse events with a causal relationship to cefepime reported in clinical trials included rash and diarrhea . Other, less common, adverse events included pruritus, urticaria, nausea, vomiting oral candidiasis, colitis, headache, fever, erythema and vaginitis . CONCLUSION: Cefepime is an established and generally well tolerated parenteral drug with a broad spectrum of antibacterial activity which, when administered twice daily, provides coverage of most of the pathogens that may be causative in pneumonia . In randomized clinical trials in hospitalized patients with generally moderate to severe community-acquired or nosocomial pneumonia, cefepime monotherapy exhibited good clinical and bacteriological efficacy . Cefepime may become a preferred antibacterial agent for infections caused by Enterobacter spp . With prudent use in order to prevent the emergence of resistant organisms, cefepime will continue to be a suitable option for the empiric treatment of pneumonia.

Biotechnol Lett, 2003 Nov, 25(22), 1893 - 8
Treatment of winery effluent with upflow anaerobic sludge blanket (UASB)--granular sludges enriched with Enterobacter sakazakii; Keyser M et al.; Three upflow anaerobic sludge blankets (UASBs) were evaluated for the treatment of winery wastewater: the first was seeded with granular sludge enriched with Enterobacter sakazakii and reached a 90% COD removal within 17 d at hydraulic retention time of 24 h; the second was seeded with brewery granules and achieved 85% COD removal within 50 d, the third was seeded with just sludge and showed the typical problems encountered with conventional sludge seeding and had continuously to be re-seeded . A PCR-based technique was developed for the rapid detection of E . sakazakii in the granular sludge.

J Food Prot, 2004 Jan, 67(1), 185 - 8
Bacterial contamination of recirculating brine used in the commercial production of moisture-enhanced pork; Greer GG et al.; In a commercial process for the production of moisture-enhanced pork, boneless pork loins were conveyed through a recirculating injection apparatus, and brine (sodium phosphate, sodium chloride, and lemon juice solids) was pumped into the meat through banks of needles inserted automatically into the upper surfaces of cuts . Brine samples were collected at intervals during the production process and analyzed to determine the total plate count and the numbers of lactic acid bacteria, pseudomonads, Brochothrix thermosphacta, and Enterobacteriaceae . Listeria monocytogenes numbers in the brine were determined using a PCR with primers for the hemolysin gene in combination with a most probable numbers determination . Maximum numbers of bacteria (log CFU/ml) recovered from the brine after 2.5 h of recirculation were as follows: total plate count, 4.50; lactic acid bacteria, 2.99; pseudomonads, 3.95; B . thermosphacta, 2.79; and enterics, 3.01 . There was an increase in the number of L . monocytogenes in the recirculating brine with time, reaching a maximum of 2.34 log CFU/100 ml after 2.5 h of moisture-enhanced pork production . Thus, recirculating brines can harbor large populations of spoilage bacteria and L . monocytogenes and are an important source of contamination for moisture-enhanced pork.

J Food Prot, 2004 Jan, 67(1), 60 - 3
Thermal inactivation of Enterobacter sakazakii in rehydrated infant formula; Edelson-Mammel SG et al.; The presence of low levels of Enterobacter sakazakii in dried infant formula have been linked to outbreaks of meningitis, septicemia, and necrotizing enterocolitis in neonates, particularly those who are premature or immunocompromised . In the current study, the ability of 12 strains of E . sakazakii to survive heating in rehydrated infant formula was determined at 58 degrees C with a submerged coil apparatus . The observed D58-values ranged from 30.5 to 591.9 s, with the strains appearing to fall into two distinct heat resistance phenotypes . The z-value of the most heat-resistant strain was 5.6 degrees C . When dried infant formula containing this strain was rehydrated with water preequilibrated to various temperatures, a more than 4-log reduction in E . sakazakii levels was achieved by preparing the formula with water at 70 degrees C or greater.

Ann Urol (Paris), 2003 Dec, 37(6), 322 - 38
{Orchi-epididymitis}; Delavierre D; The term orchiepididymitis encompasses inflammation of the epididymis and/or testis, i.e . epididymitis, orchitis, and true orchiepididymitis . Epididymitis is defined as inflammation of the epididymis . Young adults are predominantly affected, with a frequency peak between 20 and 40 years of age . The cause is usually an infectious agent, and the main route of access to the epididymis is retrograde propagation through the vas deferens . From puberty to 35 years of age, many cases are sexually transmitted . The main causative agents are Chlamydia trachomatis and Neisseria gonorrhoeae . In prepubertal children and in adults older than 35 years of age, epididymitis is among the commonplace genitourinary infections usually caused by enterobacteria . A urinary tract abnormality, most notably an obstruction of the distal urinary tract, is often the cause of the infection . Orchitis, a less common condition, is defined as inflammation of the testis . Again, most cases are related to an infection . Dissemination of the organism occurs either via the bloodstream, particularly with viruses (the most classic example being orchitis due to mumps) or by direct spread from a focus in the epididymis (producing true orchiepididymitis) . In patients younger than 35 years of age who have urethritis and suspected sexually transmitted disease, tetracyclines are the best agents and can be given intravenously at first if needed . Tetracyclines are effective not only on C . trachomatis but also on N . gonorrhoeae . This last agent also responds to other antimicrobials, such as ceftriaxone . Macrolides and second-generation quinolones are also effective on C . trachomatis . Typically, treatment is given for 3 weeks . Sexual partners should be evaluated and treated . In patients older than 35 years who have positive urine cultures for bacteria, urinary tract symptoms, a prior diagnosis of a urinary tract abnormality, or a history of a recent endourethral procedure, treatment can be given orally provided the symptoms are of moderate intensity . Either extra-strength cotrimoxazole or second-generation quinolones should be used . Patients with severe disease should be admitted for parenteral therapy with an aminoglycoside and a cephalosporin in combination, followed by oral cotrimoxazole or a second-generation quinolone . If needed, the antibiotics should be changed according to antibiotic susceptibility test results.

J Clin Microbiol, 2004 Jan, 42(1), 294 - 8
Utility of NCCLS guidelines for identifying extended-spectrum beta-lactamases in non-Escherichia coli and Non-Klebsiella spp . of Enterobacteriaceae; Schwaber MJ et al.; NCCLS screening and confirmation methods for detecting extended-spectrum beta-lactamases (ESBLs) apply only to Escherichia coli and Klebsiella spp., yet ESBLs have been found in other members of the family Enterobacteriaceae . We evaluated the effectiveness of NCCLS methods for detecting ESBLs in 690 gram-negative isolates of Enterobacteriaceae that excluded E . coli, Klebsiella pneumoniae, and Klebsiella oxytoca . Isolates were collected between January 1996 and June 1999 from 53 U.S . hospitals participating in Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology) . The antimicrobial susceptibility patterns of the isolates were determined by using the NCCLS broth microdilution method (BMD), and those isolates for which the MIC of ceftazidime, cefotaxime, ceftriaxone, or aztreonam was >or=2 microg/ml or the MIC of cefpodoxime was >or=8 microg/ml (positive ESBL screen test) were further tested for a clavulanic acid (CA) effect by BMD and the disk diffusion method (confirmation tests) . Although 355 (51.4%) of the isolates were ESBL screen test positive, only 15 (2.2%) showed a CA effect . Since 3 of the 15 isolates were already highly resistant to the five NCCLS indicator drugs, ESBL detection would have an impact on the reporting of only 1.7% of the isolates in the study . Only 6 of the 15 isolates that showed a CA effect contained a bla(TEM), bla(SHV), bla(CTX-M), or bla(OXA) beta-lactamase gene as determined by PCR (with a corresponding isoelectric focusing pattern) . Extension of the NCCLS guidelines for ESBL detection to Enterobacteriaceae other than E . coli and Klebsiella spp . does not appear to be warranted in the United States at present, since the test has poor specificity for this population and would result in changes in categorical interpretations for only 1.7% of Enterobacteriaceae tested.

J Clin Microbiol, 2004 Jan, 42(1), 220 - 8
Typing of clinical and environmental Aeromonas sp . strains by random amplified polymorphic DNA PCR, repetitive extragenic palindromic PCR, and enterobacterial repetitive intergenic consensus sequence PCR; Szczuka E et al.; A collection of 120 strains isolated from stool specimens collected from humans suffering from gastroenteritis and from environmental samples were analyzed by random amplified polymorphic DNA PCR (RAPD), repetitive extragenic palindromic PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) . Species of Aeromonoas hydrophila, A . bestiarum, A . salmonicida, A . caviae, A . media, and A . veronii revealed clonal structure . There was no dominant clone causing gastroenteritis in humans . Moreover, there was no genetic similarity between clinical and environmental strains of Aeromonas sp . isolated from different geographical areas as well as from the same geographical area . Some clones colonized specific ecosystems, e.g., drinking water distribution systems . RAPD and ERIC-PCR methods had the same discriminatory power and proved to be useful for epidemiological investigation and population genetic analysis of Aeromonas spp., whereas REP-PCR was less effective for differentiating the isolates of Aeromonas spp.

J Clin Microbiol, 2004 Jan, 42(1), 60 - 4
Practical bench comparison of BBL CHROMagar Orientation and standard two-plate media for urine cultures; D'Souza HA et al.; A total of 1023 urine samples sent for routine culture were plated onto sheep blood and MacConkey agars and a BBL CHROMagar Orientation (CO; Becton Dickinson, Cockeysville, Md.) plate, and the results were compared . Of these, 250 urine samples (24%) grew >10000 CFU of one or two putative pathogens/ml and 773 showed no growth (NG), mixed growth of <10000 CFU/ml, or three or more strains (mixed) . The CO and conventional medium results agreed completely for 595 cultures with NG or <10000 CFU/ml . An additional 178 urine samples yielded clinically insignificant differences . Both medium sets essentially agreed on quantities and identification for 400 single-pathogen cultures and 9 mixed cultures . With the caveat that CO cannot differentiate Klebsiella, Enterobacter, and Serratia spp., enteric pathogens were identified only by morphology and color on CO . Direct visual differentiation of group B streptococci from lactobacilli is not possible, but lactobacillus cells always exhibited easily recognizable morphology on Gram stain . Of 108 paired organism susceptibility results encompassing 2268 drug-pathogen combinations, there were 3% errors and only 1% very major errors . Use of CO allowed a >50% reduction in inoculation time and a >20% reduction in work-up time . For our laboratory, with 50% "no growth" and ca . 25% significant results (50% Escherichia coli), CO allowed time and workup cost savings for a majority of cultures . A cost analysis (time and supplies for our laboratory) showed that if CO is used alone, the break-even level for CO pricing is US dollars 1.78; if CO and blood agar are both used, the break-even pricing for CO is US dollars 1.53.

J Clin Microbiol, 2004 Jan, 42(1), 7 - 11
Identification and susceptibility testing of Enterobacteriaceae and Pseudomonas aeruginosa by direct inoculation from positive BACTEC blood culture bottles into Vitek 2; Bruins MJ et al.; Inoculation of an automated system for rapid identification (ID) and antimicrobial susceptibility testing (AST) directly from positive blood culture bottles will reduce the turnaround time of laboratory diagnosis of septicemic patients, which benefits clinical outcome and decreases patient costs . Direct test results, however, must always be confirmed by testing a pure overnight culture, which is the "gold standard." We studied the accuracy of direct testing versus repeat testing in order to investigate the possibility of refraining from repeat testing . We also assessed the clinical risk of reporting results based on direct testing only . We inoculated Vitek 2 (bioMerieux) directly from 410 positive BACTEC 9240 (BD) blood culture bottles containing gram-negative rods and studied the ID and AST results . In a comparison of direct inoculation with the standard method, a total of 344 isolates of Enterobacteriaceae and Pseudomonas aeruginosa were tested, and 93.0% were correctly identified . Of the 39 (10.2%) samples that contained bacilli not identifiable by Vitek 2, only 1 gave a conclusive, correct result . The overall MIC agreement among 312 isolates was 99.2%, with 0.8% very major and 0.02% major error rates . Of only three (polymicrobial) samples, the direct susceptibility pattern would be reported to the clinician as too sensitive . Vitek 2 results obtained from direct inoculation of blood culture bottles containing gram-negative bacilli are safe enough for immediate reporting, provided that ID and AST are consistent . Repeat testing is not necessary, unless Gram stain or overnight subculture results raise doubt about the purity of the culture.

BMC Biochem . 2004 Jan 12;5(1):1.
Physicochemical characterization of the endotoxins from Coxiella burnetii strain Priscilla in relation to their bioactivities; Toman R et al.; BACKGROUND: Coxiella burnetii is the etiological agent of Q fever found worldwide . The microorganism has like other Gram-negative bacteria a lipopolysaccharide (LPS, endotoxin) in its outer membrane, which is important for the pathogenicity of the bacteria . In order to understand the biological activity of LPS, a detailed physico-chemical analysis of LPS is of utmost importance . RESULTS: The lipid A moiety of LPS is tetraacylated and has longer (C-16) acyl chains than most other lipid A from enterobacterial strains . The two ester-linked 3-OH fatty acids found in the latter are lacking . The acyl chains of the C . burnetii endotoxins exhibit a broad melting range between 5 and 25 degrees C for LPS and 10 and 40 degrees C for lipid A . The lipid A moiety has a cubic inverted aggregate structure, and the inclination angle of the D-glucosamine disaccharide backbone plane of the lipid A part with respect to the membrane normal is around 40 degrees . Furthermore, the endotoxins readily intercalate into phospholipid liposomes mediated by the lipopolysaccharide-binding protein (LBP) . The endotoxin-induced tumor necrosis factor alpha (TNFalpha) production in human mononuclear cells is one order of magnitude lower than that found for endotoxins from enterobacterial strains, whereas the same activity as in the latter compounds is found in the clotting reaction of the Limulus amebocyte lysate assay . CONCLUSIONS: Despite a considerably different chemical primary structure of the C . burnetii lipid A in comparison with enterobacterial lipid A, the data can be well understood by applying the previously presented conformational concept of endotoxicity, a conical shape of the lipid A moiety of LPS and a sufficiently high inclination of the sugar backbone plane with respect to the membrane plane . Importantly, the role of the acyl chain fluidity in modulating endotoxicity now becomes more evident.

Mol Plant Microbe Interact, 2004 Jan, 17(1), 43 - 54
The phytoalexin-inducible multidrug efflux pump AcrAB contributes to virulence in the fire blight pathogen, Erwinia amylovora; Burse A et al.; The enterobacterium Erwinia amylovora causes fire blight on members of the family Rosaceae, with economic importance on apple and pear . During pathogenesis, the bacterium is exposed to a variety of plant-borne antimicrobial compounds . In plants of Rosaceae, many constitutively synthesized isoflavonoids affecting microorganisms were identified . Bacterial multidrug efflux transporters which mediate resistance toward structurally unrelated compounds might confer tolerance to these phytoalexins . To prove this hypothesis, we cloned the acrAB locus from E . amylovora encoding a resistance nodulation division-type transport system . In Escherichia coli, AcrAB of E . amylovora conferred resistance to hydrophobic and amphiphilic toxins . An acrB-deficient E . amylovora mutant was impaired in virulence on apple rootstock MM 106 . Furthermore, it was susceptible toward extracts of leaves of MM 106 as well as to the apple phytoalexins phloretin, naringenin, quercetin, and (+)-catechin . The expression of acrAB was determined using the promoterless reporter gene egfp . The acrAB operon was up-regulated in vitro by the addition of phloretin and naringenin . The promoter activity of acrR, encoding a regulatory protein involved in acrAB expression, was increased by naringenin . In planta, an induction of acrAB was proved by confocal laser scanning microscopy . Our results strongly suggest that the AcrAB transport system plays an important role as a protein complex required for virulence of E . amylovora in resistance toward apple phytoalexins and that it is required for successful colonization of a host plant.

Am J Kidney Dis, 2004 Jan, 43(1), 103 - 11
Peritoneal dialysis catheter removal for acute peritonitis: a retrospective analysis of factors associated with catheter removal and prolonged postoperative hospitalization; Choi P et al.; BACKGROUND: Most patients with acute peritoneal dialysis (PD) peritonitis respond to antibiotic therapy, but a significant minority of patients require surgical catheter removal to eradicate the infection . These patients may experience an adverse postsurgical course . METHODS: We retrospectively analyzed 64 episodes of acute peritonitis requiring PD catheter removal in comparison to 426 episodes treated with antibiotics alone . RESULTS: There were no differences between patients who required PD catheter removal and medically treated patients in sex (62% versus 60% men; P > 0.05), PD modality (31% versus 27% automated PD; P > 0.05), time spent on PD therapy (35 versus 26 months; P > 0.05), or cause of end-stage renal failure . Catheter removal was more likely to occur in elderly (mean age, 61 versus 54 years; P = 0.023) and South Asian patients (38% versus 22%; P = 0.020) and after peritonitis caused by Escherichia coli (16% versus 4%; P = 0.0005), Enterobacter species (5% versus 0.7%; P = 0.031), and Pseudomonas species (5% versus 0.7%; P = 0.031) . The most significant correlation with requirement for surgical catheter removal was duration of peritonitis (mean, 7.5 versus 2.8 days; P = 1.3 x 10(-6)) . Fifty-three percent of catheter removals resulted in postoperative hospitalization longer than 10 days . Delayed discharges were caused by multiple reasons . Compared with discharges within 10 days, prolonged hospitalization was associated with increased age (mean, 64 versus 58 years; P = 0.028) and delay in time to catheter removal (mean, 7.9 versus 5.3 days; P = 0.027) . After catheter removal, only 4% of patients successfully returned to maintenance PD therapy . CONCLUSION: Increased age and duration of peritonitis are associated with both requirement for PD catheter removal and prolonged postoperative hospitalization.

J Antimicrob Chemother, 2004 Feb, 53(2), 290 - 6 Epub 2004 Jan 07.
Antimicrobial usage and resistance trend relationships from the MYSTIC Programme in North America (1999-2001); Mutnick AH et al.; BACKGROUND: The MYSTIC Programme is a global, longitudinal antimicrobial surveillance network of hospitals that frequently utilize carbapenems . One aspect of the programme is the ability to capture antimicrobial consumption data from participating institutions . The current report evaluates these relationships for Enterobacteriaceae and Pseudomonas aeruginosa over the initial 3 year period of the programme in the USA . METHODS: Between 10 and 15 medical centres participated during 1999-2001, each submitting up to 200 isolates/year (7003 strains overall) . Evaluations of the relationship between drug usage and antimicrobial resistance in P . aeruginosa and Enterobacteriaceae for the carbapenems (imipenem and meropenem), cefepime, ceftazidime, ciprofloxacin, gentamicin and piperacillin-tazobactam were determined . Data were analysed based on: (1) aggregate usage results; (2) medical centre-specific usage compared with resistance rates; and (3) medical centre-specific usage results compared with yearly changes in resistance rates (DeltaR) . The parameter of drug usage was the defined daily dose (DDD)/100 patient days calculated from total grams administered, using WHO definitions . RESULTS: Resistance (1999-2001) among Enterobacteriaceae did not change significantly for beta-lactams, but tended to increase slightly for gentamicin (+1.1%) and ciprofloxacin (+3.1%) . P . aeruginosa resistance rates (1999-2001) for gentamicin (+9.0%) and ciprofloxacin (+10.2%) increased, in contrast to a significantly decreased resistance rate for meropenem (-7.7%) . Formulary-use changes were noted: increased meropenem and ciprofloxacin use and decreased consumption for imipenem, aminoglycosides, ceftazidime and cefepime . Aggregate ciprofloxacin DDD/100 days rates were directly related (+3.3 DDD) to Enterobacteriaceae and P . aeruginosa resistance changes, whereas among P . aeruginosa, usage and resistance were inversely correlated for gentamicin (-3.8 DDD; +9.0% resistant) . Medical centre-specific antimicrobial usage calculations did not demonstrate a correlation to rates of resistance (r = -0.38 to 0.61) or yearly changes in resistance rates (r = -0.56 to 0.43) . CONCLUSIONS: The availability of aggregate USA medical centre antimicrobial usage data enabled us to identify several important trends in the incidence of resistance among P . aeruginosa and Enterobacteriaceae: (1) increased use of ciprofloxacin associated with a higher resistance among Enterobacteriaceae; and (2) a correlation between ciprofloxacin categories of resistance and levels of resistance to other antimicrobial classes in P . aeruginosa . Medical centre-specific antimicrobial usage and resistance did not demonstrate direct statistical relationships, and require a continued search for other monitoring methods that can better identify antimicrobial/environmental factors that lead to resistance.

Diagn Microbiol Infect Dis, 2003 Dec, 47(4), 595 - 600
Increasing prevalence of antimicrobial resistance among Enterobacteriaceae uropathogens in Dakar, Senegal: a multicenter study; Dromigny JA et al.; To assess antibiotic susceptibility among Enterobacteriaceae isolated in urine from outpatients in Dakar, Senegal, a prospective multicenter study involving 3 laboratories had been conducted between June and October 2001 . During this period, 300 strains were isolated and susceptibility testing was performed against antibiotics commonly used in treatment of community-acquired urinary tract infections (UTI) . E . coli and K . pneumoniae represented 89% of isolates . The overall resistance rates of ampicillin, amoxicillin-clavulanic acid, nalidixic acid, fluoroquinolones and cotrimoxazole were respectively 77.3%, 34.7%, 14.7%, 13.3%, and 55% . In the light of these results, a re-evaluation of first line therapies and prudent use of fluoroquinolones is advised . At the same time a continued surveillance of antimicrobial resistance should be developed in Senegal in order to control the emergence of multidrug resistant strains and to establish a national therapeutic guideline for treatment of UTI.

J Mol Evol, 2003 Oct, 57(4), 479 - 86
MgtC as a horizontally-acquired virulence factor of intracellular bacterial pathogens: evidence from molecular phylogeny and comparative genomics; Blanc-Potard AB et al.; MgtC is a virulence factor required for intramacrophage survival and growth in low Mg2+ medium in two pathogens that are not phylogenetically related, Salmonella typhimurium and Mycobacterium tuberculosis . In S . typhimurium, mgtC is carried by the SPI-3 pathogenicity island and hybridization studies have suggested that the distribution of mgtC among enterobacteria is limited . In the present study, we searched for the presence of mgtC-like sequences in eubacterial genomes . Analyses of MgtC-like proteins phylogeny and mgtC-like chromosomal context support the hypothesis that mgtC has been acquired by horizontal gene transfer repeatedly throughout bacterial evolution . In addition, the phylogenetic analysis revealed the existence of a subgroup of proteins, that includes the S . typhimurium and M . tuberculosis MgtC proteins, as well as MgtC-related proteins from other pathogens that are able to survive in macrophages, B . melitensis and Y . pestis . We propose that MgtC has a similar function in all these distantly related pathogens, most likely providing the ability to grow in a low Mg2+ environment.

Zhongguo Wei Zhong Bing Ji Jiu Yi Xue, 2004 Jan, 16(1), 2 - 5
{Impact of secondary pancreatic infection on the prognosis of patients with severe acute pancreatitis: an analysis of 60 cases}; Li GK et al.; OBJECTIVE: To investigate the impact of secondary pancreatic infection (SPI) on the prognosis of patients with severe acute pancreatitis (SAP) . METHODS: Clinical data of 60 patients with SAP (January 1, 1980-December 31, 1999), especially the data at the onset of SPI, were retrospectively analyzed . RESULTS: Sixty patients were divided into two groups: SPI group (29 cases) and non-SPI group (31 cases) . There was no significant difference in gender, average age, scores of Ranson and of high risk factors of China Medical Association between two groups, but the average stay days in hospital, payment, acute physiology and chronic health evaluation II (APACHE II) scores, days of fever, prehospital days, days of high white blood cell (WBC) count, the duration of constipation after the onset of SAP and average times of operation were significantly higher in SPI group than that of non-SPI group . Fatality rate of SPI group (7/29, 24.14 percent) was higher than that of non-SPI (1/31, 3.23 percent) . There were 14 cases of infection with single microorganism (48.3 percent) and 15 cases of mixed infection (51.7 percent) in SPI group . Twenty-seven patients were infected with bacteria of Enterobacteriaecae (Escherichia 25, Klebsiella pneumoniae 1, Proteus morgani 1, 8 with bacteria of Pseudomonas (P . aeruginosa 7, P . stanieri 1), 2 with fungus, 5 with other bacteria (Bacillus subtilis 2, Tetracoccus 1, Acetobacteraceae 1, Staphylococcus epidermidis 1) in SPI group . Only in 2 patients with infective pancreatic necrosis the blood culture was positive (Escherichia coli) . CONCLUSION: These results showed that the major source of SPI is entergenous, and SPI would affect the prognosis of SAP patients . With the present therapeutic regimes it was hard to prevent entergenous infections . It is necessary to find a new strategy.

Med Dosw Mikrobiol, 2003, 55(3), 285 - 95
{Susceptibility and frequency of selected groups of bacteria isolated in 2001 from patients at the Holy Cross Cancer Center}; Durnas B et al.; The aim of the presented study was the analysis of microbiological data obtained from patients hospitalized in The Holly Cross Cancer Center in Kielce in 2001 . The frequency of important nosocomial pathogens in selected specimens and their susceptibility to antibiotics were determined . The strains were identified by using commercial tests (bioMerieux) and their antibiotic susceptibility patterns were performed by disc diffusion technique . The most prevalent bacteria were Gram-negative rods of Enterobacteriaceae family (43%), mainly Escherichia coli . Only 2.7% strains of isolated Escherichia coli isolated from clinical specimens collected from hospitalized patients were beta-lactamase--positive (ESBL+) . The second important group of microorganisms were Staphylococci, followed by Enterococcus spp., Pseudomonas aeruginosa and Candida spp . About twenty eight percent of Staphylococcus aureus isolates were resistant to methicillin.

J Bacteriol, 2004 Jan, 186(2), 400 - 10
Complete sequence and evolutionary genomic analysis of the Pseudomonas aeruginosa transposable bacteriophage D3112; Wang PW et al.; Bacteriophage D3112 represents one of two distinct groups of transposable phage found in the clinically relevant, opportunistic pathogen Pseudomonas aeruginosa . To further our understanding of transposable phage in P . aeruginosa, we have sequenced the complete genome of D3112 . The genome is 37,611 bp, with an overall G+C content of 65% . We have identified 53 potential open reading frames, including three genes (the c repressor gene and early genes A and B) that have been previously characterized and sequenced . The organization of the putative coding regions corresponds to published genetic and transcriptional maps and is very similar to that of enterobacteriophage Mu . In contrast, the International Committee on Taxonomy of Viruses has classified D3112 as a lambda-like phage on the basis of its morphology . Similarity-based analyses identified 27 open reading frames with significant matches to proteins in the NCBI databases . Forty-eight percent of these were similar to Mu-like phage and prophage sequences, including proteins responsible for transposition, transcriptional regulation, virion morphogenesis, and capsid formation . The tail proteins were highly similar to prophage sequences in Escherichia coli and phage Phi12 from Staphylococcus aureus, while proteins at the right end were highly similar to proteins in Xylella fastidiosa . We performed phylogenetic analyses to understand the evolutionary relationships of D3112 with respect to Mu-like versus lambda-like bacteriophages . Different results were obtained from similarity-based versus phylogenetic analyses in some instances . Overall, our findings reveal a highly mosaic structure and suggest that extensive horizontal exchange of genetic material played an important role in the evolution of D3112.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 329 - 32
blaVIM-7, an evolutionarily distinct metallo-beta-lactamase gene in a Pseudomonas aeruginosa isolate from the United States; Toleman MA et al.; As part of the CANCER Antimicrobial Surveillance Program in North America, a Pseudomonas aeruginosa isolate, strain 07-406, was shown to possess a metallo-beta-lactamase, designated VIM-7 . bla(VIM-7) is located on a 24-kb plasmid which can be readily transferred into Enterobacteriaceae and other pseudomonads . This is the first report of a mobile metallo-beta-lactamase gene, bla(VIM-7), being detected within the United States.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 305 - 12
Genetic and biochemical characterization of the chromosomal class A beta-lactamases of Raoultella (formerly Klebsiella) planticola and Raoultella ornithinolytica; Walckenaer E et al.; Enterobacterial strains of Raoultella spp . display a penicillinase-related beta-lactam resistance pattern suggesting the presence of a chromosomal bla gene . From whole-cell DNA of Raoultella planticola strain ATCC 33531(T) and Raoultella ornithinolytica strain ATCC 31898(T), bla genes were cloned and expressed into Escherichia coli . Each gene encoded an Ambler class A beta-lactamase, named PLA-1 and ORN-1 for R . planticola and R . ornithinolytica, respectively . These beta-lactamases (291 amino acids), with the same pI value of 7.8, had a shared amino acid identity of 94%, 37 to 47% identity with the majority of the chromosome-encoded class A beta-lactamases previously described for Enterobacteriaceae, and 66 to 69% identity with the two beta-lactamases LEN-1 and SHV-1 from Klebsiella pneumoniae . However, the highest identity percentage (69 to 71%) was found with the plasmid-mediated beta-lactamase TEM-1 . PLA-1, which displayed very strong hydrolytic activity against penicillins, also displayed significant hydrolytic activity against cefepime and, to a lesser extent, against cefotaxime and aztreonam, but there was no hydrolytic activity against ceftazidime . Such a substrate profile suggests that the Raoultella beta-lactamases PLA-1 and ORN-1 should be classified into the group 2be of the beta-lactamase classification of K . Bush, G . A . Jacoby, and A . A . Medeiros (Antimicrob . Agents Chemother . 39:1211-1233, 1995) . The highly homologous regions upstream of the bla(PLA-1A) and bla(ORN-1A) genes comprised a nucleotide sequence identical to the -35 region and another one very close to the -10 region of the bla(LEN-1) gene . From now on, as the bla gene sequences of the most frequent Raoultella and Klebsiella species are available, the bla gene amplification method can be used to differentiate these species from each other, which the biochemical tests currently carried out in the clinical laboratory are unable to do.

Lett Appl Microbiol, 2004, 38(1), 19 - 23
Isolation of endophytic diazotroph Pantoea agglomerans and nondiazotroph Enterobacter asburiae from sweetpotato stem in Japan; Asis CA Jr et al.; AIMS: To isolate and identify diazotrophic endophytes in the stem of Japanese sweetpotato cv . Koganesengan . METHODS AND RESULTS: Surface-sterilized and thinly sliced (1-2 mm) sweetpotato stem samples were incubated in test tubes with semi-solid modified Rennie (MR) medium . The test tubes were assayed for acetylene reduction activity (ARA) 5 days after incubation at 30 degrees C . Twelve isolates were obtained from MR plates inoculated with a loop of semi-solid MR medium from ARA+ tubes . However, ARA test showed that only nine isolates were diazotrophic and three were nondiazotrophic strains . Using the API 20E diagnostic kit, four diazotrophic isolates were identified as strains of Pantoea spp . and five isolates as Klebsiella spp . The nondiazotrophic bacteria were strains of Enterobacter spp . A diazotrophic isolate Pantoea sp . MY1 and nondiazotrophic isolate Enterobacter sp . MY2 were identified to the species level by full sequence analysis of 16S rRNA gene . The results showed that MY1 had 99.2% similarity to Pantoea agglomerans ATCC 27155 and MY2 had 99.5% similarity to Enterobacter asburiae ATCC 35953 . CONCLUSION: The stem of sweetpotato cv . Koganesengan was colonized by diazotrophic endophyte P . agglomerans and nondiazotrophic endophyte E . asburiae . SIGNIFICANCE AND IMPACT OF THE STUDY: This study is an essential step toward understanding the ecology and interaction between endophytic bacteria and sweetpotato.

J Pediatr (Rio J), 1999 Jan-Feb, 75(1), 39 - 44
{Sepsis in childhood: epidemiological profile and microbiologic diagnosis}; Ribeiro AM et al.; OBJECTIVE: To establish an etiological and epidemiological profile of sepsis among children that were hospitalized in the Hospital Infantil Albert Sabin - HIAS (a reference public hospital for all pediatric diseases in the state of Ceara), since this disease has been of high prevalence in this hospital with considerable mortality . METHODS: All children admitted in the hospital from January 1993 to June 1994, who presented sepsis or developed nosocomial sepsis and presented positive blood cultures were studied prospectively . The epidemiologic study was conducted by gathering information in questionnaire form about the following factors: sex, age, reference area, nutritional status, complaints at admission, sepsis origin (nosocomial or not), and outcome for each case . Microbiologic analysis were formed by means of blood culture, concomitant with sensitivity tests of the isolated bacteria to the most commonly used antibiotics . All samples were processed at the Laboratory of Microbiology of the Health's Science Center of the Federal University in Ceara . RESULTS: 205 children were studied of which 17 presented two septic episodes, making a total of 222 episodes in the period . 56.1% of them were male, 81.4% were less than one-year old, 71.1% were malnourished children and 60.5% came from the countryside of the Ceara State . At the moment of their admission, two-thirds of the children presented gastrointestinal problems (diarrhea) or respiratory complaints, whereas 47.7% of the episodes of sepsis were acquired in hospital . The mortality rate was 56.1% . Blood culture test showed a predominance of the following bacteria: Staphylococcus aureus (24.8%), Klebsiella pneumoniae (22.6%), Pseudomonas aeruginosa (15.2%), Enterobacter sp (11.2%), Escherichia coli (7%), and others (19.2%) . CONCLUSIONS: The epidemiological profile of the population in this study was the following: these children were less than one-year old, the majority of them were malnourished and when they were hospitalized, presented with symptoms of diarrhea or respiratory disease . 40% of them had a disease or situation that favored the development of sepsis . The predominant bacterias were Staphylococcus aureus and Klebsiella pneumoniae.

J Bacteriol, 2004 Jan, 186(1), 122 - 35
Growth phase-dependent regulation and stringent control of fis are conserved processes in enteric bacteria and involve a single promoter (fis P) in Escherichia coli; Mallik P et al.; The intracellular concentration of the Escherichia coli factor for inversion stimulation (Fis), a global regulator of transcription and a facilitator of certain site-specific DNA recombination events, varies substantially in response to changes in the nutritional environment and growth phase . Under conditions of nutritional upshift, fis is transiently expressed at very high levels, whereas under induced starvation conditions, fis is repressed by stringent control . We show that both of these regulatory processes operate on the chromosomal fis genes of the enterobacteria Klebsiella pneumoniae, Serratia marcescens, Erwinia carotovora, and Proteus vulgaris, strongly suggesting that the physiological role of Fis is closely tied to its transcriptional regulation in response to the nutritional environment . These transcriptional regulatory processes were previously shown to involve a single promoter (fis P) preceding the fis operon in E . coli . Recent work challenged this notion by presenting evidence from primer extension assays which appeared to indicate that there are multiple promoters upstream of fis P that contribute significantly to the expression and regulation of fis in E . coli . Thus, a rigorous analysis of the fis promoter region was conducted to assess the contribution of such additional promoters . However, our data from primer extension analysis, S1 nuclease mapping, beta-galactosidase assays, and in vitro transcription analysis all indicate that fis P is the sole E . coli fis promoter in vivo and in vitro . We further show how certain conditions used in the primer extension reactions can generate artifacts resulting from secondary annealing events that are the likely source of incorrect assignment of additional fis promoters.

J Appl Microbiol, 2004, 96(1), 96 - 109
Modelling and predicting the simultaneous growth of Listeria monocytogenes and spoilage micro-organisms in cold-smoked salmon; Gimenez B et al.; AIMS: To evaluate and model the simultaneous growth of Listeria monocytogenes and spoilage micro-organisms in cold-smoked salmon . METHODS AND RESULTS: Growth kinetics of L . monocytogenes, lactic acid bacteria (LAB), Enterobacteriaceae, enterococci and Photobacterium phosphoreum were determined in two series of challenge tests with sliced and vacuum-packed cold-smoked salmon (SVP-CSS) . The product contained a high level of smoke components and at 2 degrees C levels of L . monocytogenes increased <100-fold in 193 days . Without the addition of spoilage micro-organisms, L . monocytogenes reached ca 108 CFU g-1 at 5, 10, 17.5 and 25 degrees C . Inoculation with spoilage micro-organisms reduced this level to 102-104 CFU g-1 . LAB dominated the spoilage microfora of SVP-CSS and competition between LAB and L . monocytogenes in SVP-CSS was appropriately described by a simple expansion of the Logistic model . This interaction model aided in predicting the growth of L . monocytogenes in naturally contaminated SVP-CSS when it was used in combination with expanded versions of existing secondary models for L . monocytogenes and LAB . CONCLUSIONS: Temperature, water activity/NaCl, simultaneous growth of LAB, smoke components and to a lesser extent lactate and pH control growth of L . monocytogenes in SVP-CSS . These factors must be included in mathematical models to predict growth of the pathogen in this product . SIGNIFICANCE AND IMPACT OF THE STUDY: The suggested predictive model can be used to support assessment and management of the human health risk due to L . monocytogenes in SVP-CSS.

Dakar Med, 2000, 45(1), 59 - 61
{Urinary tract infections in Dakar: etiologies, therapeutic basis}; Sow AI et al.; This prospective study, performed in Fann University Teaching Hospital from January 1st to December 31st 1998, concern 1446 samples of urine . Enterobacteria (87.56%) were the most frequent aetiology, and Escherichia coli (48.7%) was the leading species in this family . The strains of E . coli present more resistant profil to beta-lactams (70.27%) . Fluoroquinolons are active on more than 80% of the strains responsible of urinary tract infection in Dakar.

Rev Hosp Clin Fac Med Sao Paulo, 2003 Sep-Oct, 58(5), 254 - 9 Epub 2003 Nov 11.
Prevalence rates of infection in intensive care units of a tertiary teaching hospital; Toufen Junior C et al.; OBJECTIVE: To determine the prevalence rates of infections among intensive care unit patients, the predominant infecting organisms, and their resistance patterns . To identify the related factors for intensive care unit-acquired infection and mortality rates . DESIGN: A 1-day point-prevalence study . SETTING:A total of 19 intensive care units at the Hospital das Cl nicas - University of S o Paulo, School of Medicine (HC-FMUSP), a teaching and tertiary hospital, were eligible to participate in the study . PATIENTS: All patients over 16 years old occupying an intensive care unit bed over a 24-hour period . The 19 intensive care unit s provided 126 patient case reports . MAIN OUTCOME MEASURES: Rates of infection, antimicrobial use, microbiological isolates resistance patterns, potential related factors for intensive care unit-acquired infection, and death rates . RESULTS: A total of 126 patients were studied . Eighty-seven patients (69%) received antimicrobials on the day of study, 72 (57%) for treatment, and 15 (12%) for prophylaxis . Community-acquired infection occurred in 15 patients (20.8%), non- intensive care unit nosocomial infection in 24 (33.3%), and intensive care unit-acquired infection in 22 patients (30.6%) . Eleven patients (15.3%) had no defined type . The most frequently reported infections were respiratory (58.5%) . The most frequently isolated bacteria were Enterobacteriaceae (33.8%), Pseudomonas aeruginosa (26.4%), and Staphylococcus aureus (16.9%; {100% resistant to methicillin}) . Multivariate regression analysis revealed 3 risk factors for intensive care unit-acquired infection: age > 60 years (p = 0.007), use of a nasogastric tube (p = 0.017), and postoperative status (p = 0.017) . At the end of 4 weeks, overall mortality was 28.8% . Patients with infection had a mortality rate of 34.7% . There was no difference between mortality rates for infected and noninfected patients (p=0.088) . CONCLUSION: The rate of nosocomial infection is high in intensive care unit patients, especially for respiratory infections . The predominant bacteria were Enterobacteriaceae, Pseudomonas aeruginosa, and Staphylococcus aureus (resistant organisms) . Factors such as nasogastric intubation, postoperative status, and age 60 years were significantly associated with infection . This study documents the clinical impression that prevalence rates of intensive care unit-acquired infections are high and suggests that preventive measures are important for reducing the occurrence of infection in critically ill patients.

Rev Saude Publica, 2003 Dec, 37(6), 775 - 9 Epub 2003 Nov 27.
{Microbiological quality of human milk from a Brazilian milk bank}; Serafini AB et al.; OBJECTIVE: The objectives of the present study were to determine the prevalence of potentially pathogenic microorganisms that indicate the hygienic and sanitary conditions of human milk samples collected at a Human Milk Bank . METHODS: Three hundred and thirty eight (338) samples of human milk collected from a milk bank in a maternity in the municipality of Goiania, in the state of Goias, Brazil were submitted to microbiological analysis . The latter were plated on McConkey agar according to the type of bacteria . Among the total number of samples collected, 194 consisted of raw milk and the remaining 144 were pasteurized milk . RESULTS: The presence of Staphylococcus spp., Streptococcus spp., yeasts and molds, and Enterobacteriaceae was verified in the raw milk samples . Staphylococcus aureus were isolated in 10 (5.2%) samples, Staphylococcus epidermidis in 28 (14.4%) samples, Streptococcus spp . in three (1.6%) samples, yeasts and molds in 43 (22.2%) and Enterobacteriaceae in 49 (25.3%) samples . In a hundred and forty four (144) samples which underwent thermal treatment Staphylococcus aureus was detected in five (3.5%) samples, Staphylococcus epidermidis in 15 (10.4%), Staphylococcus lugdenensis in two (1.4%), Streptococcus spp . in four (2.8%), yeasts and molds in 37 (25.7%), and Enterobacteriaceae in nine (6.3%) . CONCLUSIONS: Analysis indicated a high degree of contamination in raw human milk, and as for the pasteurized milk, despite elimination of the great majority of potentially pathogenic microorganisms, the percentage of yeasts and molds was higher than in raw milk, demonstrating that a lower degree of initial contamination would be necessary for pasteurization to be an efficient means of microbiological control.

J Clin Microbiol, 2003 Dec, 41(12), 5689 - 94
Highly resistant metabolically deficient dwarf mutant of Escherichia coli is the cause of a chronic urinary tract infection; Trulzsch K et al.; We present the case of a 68-year-old diabetic woman who has been suffering from chronic urinary tract infections, recurring over a period of at least 5 years, caused by a slowly growing metabolically deficient dwarf mutant (MDD) of Escherichia coli . This MDD strain was auxotrophic for histidine, was resistant to multiple antibiotics, and showed atypical growth behavior . Colonies were tiny on routine media but were able to revert to normal growth after extended incubation . This strain was identified as E . coli by 16S ribosomal DNA sequencing, and virulence factor profiles were determined by PCR . Seven MDD isolates collected over the 5-year period were grown from midstream urine to significant colony counts and shown to belong to the same clonal group by pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR . These MDDs were repeatedly misidentified by biochemical methods due to their slow growth and atypical colony morphology . This case highlights the importance of recognizing MDDs of Enterobacteriaceae in patients with chronic infections . To our knowledge this is the first report of an MDD of E . coli causing a chronic urinary tract infection.

Appl Environ Microbiol, 2003 Dec, 69(12), 7188 - 93
Inoculum size influences bacterial cross contamination between surfaces; Montville R et al.; Many factors have been shown to influence bacterial transfer between surfaces, including surface type, bacterial species, moisture level, pressure, and friction, but the effect of inoculum size on bacterial transfer has not yet been established . Bacterial cross contamination rates during performance of common food service tasks were previously determined in our laboratory using nalidixic acid-resistant Enterobacter aerogenes . Eight different transfer rates were determined, each involving a minimum of 30 volunteers . The influence of source inoculum level on the percentage of bacteria transferred (percent transfer rates) and log10 CFU per recipient surface was determined using statistical analysis . The effect of inoculum size on transfer rate was highly statistically significant (P < 0.0001) for all transfer rate data combined (352 observations) and for each individual cross contamination rate, except for data on contamination via transfer from chicken to hand through a glove barrier (P = 0.1643) . Where inoculum size on the source was greater, transfer rates were lower, and where inoculum size on the source was less, transfer rates were higher . The negative linear trend was more obvious for activities that had a larger range of inoculum sizes on the source surface . This phenomenon has serious implications for research seeking to determine bacterial cross contamination rates, since the different transfer efficiencies that were previously shown to be associated with certain activities may actually be the result of differing initial inoculum levels . The initial inoculum size on the source and the amount of bacteria transferred must both be considered to accurately determine bacterial transfer rates.

Int J Syst Evol Microbiol, 2003 Nov, 53(Pt 6), 2007 - 11
Staphylococcus nepalensis sp . nov., isolated from goats of the Himalayan region; Spergser J et al.; Four coagulase-negative, novobiocin-resistant cocci, designated CW1(T), PM34, MM3 and RW78, were isolated from the respiratory tract of goats kept in the Himalayan region . The four isolates were assigned to a single species on the basis of almost identical biochemical and physiological traits, protein profiles obtained after SDS-PAGE and identical genomic fingerprints generated after enterobacterial repetitive intergenic consensus (ERIC)-PCR . Strain CW1(T) showed highest 16S rDNA sequence similarities to Staphylococcus cohnii subsp . urealyticus ATCC 49330(T), Staphylococcus saprophyticus subsp . saprophyticus ATCC 15305(T), S . cohnii subsp . cohnii ATCC 29974(T), Staphylococcus arlettae ATCC 43957(T), Staphylococcus gallinarum ATCC 35539(T), Staphylococcus succinus ATCC 700337(T) and Staphylococcus xylosus ATCC 29971(T) (99.0, 98.8, 98.8, 98.4, 98.2, 98.1 and 98.1 %, respectively), indicating its classification within the genus Staphylococcus . The polar lipid composition, fatty acid profiles, quinone systems and diagnostic cell-wall diamino acid were in agreement with the characteristics of the genus Staphylococcus . DNA-DNA hybridization with closely related Staphylococcus species suggested that strain CW1(T) represents an as-yet unrecognized species . Based on these results, a novel species of the genus Staphylococcus is described, Staphylococcus nepalensis sp . nov . The type strain is CW1(T) (=DSM 15150(T)=CCM 7045(T)) and the most dissimilar strain is PM34 (=DSM 15151=CCM 7046).

Hunan Yi Ke Da Xue Xue Bao, 2003 Aug, 28(4), 405 - 8
{Infective pathogens and drug resistance in burned patients}; Zhang X et al.; OBJECTIVE: To find out the changes in the infective pathogens and their drug resistance in burned patients . METHODS: The patients were divided into two groups . The first group was from July 1991 to June 1996, and the second group was from July 1996 to June 2001 . The bacteria of burned body surface and blood were cultured, and the bacteria and their drug sensitivity were analyzed . RESULTS: Gram-negative bacteria were the major bacteria in burn infection, among which Pseudomonas aeruginosa ranked the top . Staphylococcus aureous ranked the first among the Gram-positive bacteria, and the isolation rate of methicillin resistant Staphylococcus aureus increased; The isolation rate of Enterobacter cloacae (10.4%), Escherichia coli (8.3%), Klebsiella pneumoniae (7.3%), and fungus (4.2%) all rose . The antibiotic resistant strains of Pseudomonas aeruginosa and Staphylococcus aureous increased . CONCLUSION: The changes in pathogens of burn infection and bacterial drug resistance are related to the wide use of broad spectrum antibiotics such as cefazidime and imepenem, suggesting that dynamic observation of changes in pathogenic strains and sensitivity of bacteria to antibiotics are useful for clinical prevention and cure of burn infection.

Mol Microbiol, 2004 Jan, 51(1), 215 - 25
Lack of O-antigen is essential for plasminogen activation by Yersinia pestis and Salmonella enterica; Kukkonen M et al.; The O-antigen of lipopolysaccharide (LPS) is a virulence factor in enterobacterial infections, and the advantage of its genetic loss in the lethal pathogen Yersinia pestis has remained unresolved . Y . pestis and Salmonella enterica express beta-barrel surface proteases of the omptin family that activate human plasminogen . Plasminogen activation is central in pathogenesis of plague but has not, however, been found to be important in diarrhoeal disease . We observed that the presence of O-antigen repeats on wild-type or recombinant S . enterica, Yersinia pseudotuberculosis or Escherichia coli prevents plasminogen activation by PgtE of S . enterica and Pla of Y . pestis; the O-antigen did not affect incorporation of the omptins into the bacterial outer membrane . Purified His6-Pla was successfully reconstituted with rough LPS but remained inactive after reconstitution with smooth LPS . Expression of smooth LPS prevented Pla-mediated adhesion of recombinant E . coli to basement membrane as well as invasion into human endothelial cells . Similarly, the presence of an O-antigen prevented PgtE-mediated bacterial adhesion to basement membrane . Substitution of Arg-138 and Arg-171 of the motif for protein binding to lipid A 4'-phosphate abolished proteolytic activity but not membrane translocation of PgtE, indicating dependence of omptin activity on a specific interaction with lipid A . The results suggest that Pla and PgtE require LPS for activity and that the O-antigen sterically prevents recognition of large-molecular-weight substrates . Loss of O-antigen facilitates Pla functions and invasiveness of Y . pestis; on the other hand, smooth LPS renders plasminogen activator cryptic in S . enterica.

J Perinatol, 2003 Dec, 23(8), 620 - 3
Effects of powdered human milk fortifiers on the antibacterial actions of human milk; Chan GM; OBJECTIVES: To evaluate the effects of powdered fortifiers and the addition of iron and medium-chain triglycerides on preterm human milk antibacterial activity . STUDY DESIGN: Human milk samples were obtained from 42 preterm lactating mothers after the first week of postnatal life . Enfamil (EHMF) and Similac (SHMF) Human Milk Fortifiers were evaluated . All mothers were healthy and were on no medications except for vitamins during lactation . The effects of each fortifier against E . coli (E . coli), Staphylococcus (Staph), Enterobacter sakazakii (ES), and Group B Streptococcus (GBS) were measured by the filter paper method and growth of the bacteria with human milk alone as control . The addition of iron and medium-chain triglycerides (MCT) to human milk was also tested . RESULTS: Human milk inhibited the growth of E . coli, Staph, ES, and GBS . Only the SHMF and the addition of MCT had similar antibacterial action as human milk alone . EHMF and the addition of iron to human milk removed the milk's antibacterial action against these four organisms . CONCLUSIONS: Preterm human milk has antimicrobial activity against E . coli, Staph, ES, and GBS . This activity can be affected by the addition of iron and fortifiers that contain iron.

Bioorg Med Chem Lett, 2003 Dec 15, 13(24), 4489 - 92
Novel mechanism of inhibiting beta-lactamases by sulfonylation using beta-sultams; Page MI et al.; Beta-sultams are the sulfonyl analogues of beta-lactams and N-acyl beta-sultams are novel inactivators of the class C beta-lactamase of Enterobacter cloacae P99 . The rates of inactivation show a similar pH-rate dependence as that exhibited by the beta-lactam antibiotics and with ESIMS data it is suggested that beta-sultams sulfonylate the active site serine residue to form a sulfonate ester.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3989 - 93
Prevalence of extended-spectrum beta-lactamase-producing Enterobacter cloacae in the Asia-Pacific region: results from the SENTRY Antimicrobial Surveillance Program, 1998 to 2001; Bell JM et al.; Enterobacter cloacae strains from hospitalized patients with a range of infections were collected by 17 laboratories in the Asia-Pacific region and South Africa . Isolates for which ceftriaxone MICs were above 1 microg/ml and/or ceftazidime MICs were above 2 microg/ml, as well as 46 strains for which ceftriaxone and/or ceftazidime MICs were at or below these values, were screened for levels of extended-spectrum beta-lactamase (ESBL) production through the use of broth microdilution for the detection of clavulanate enhancement of the activity of ceftriaxone, ceftazidime, and cefepime . Of the isolates examined, ceftriaxone and/or ceftazidime had elevated MICs for 44%, of which 36% were ESBL positive . ESBL-positive strains were commonly susceptible to piperacillin-tazobactam and more frequently resistant to several other antimicrobials studied . A cefepime MIC above 0.25 microg/ml had the highest sensitivity (100%) and specificity (74%) for predicting the presence of an ESBL.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3750 - 9
In vitro and in vivo antibacterial activities of DK-507k, a novel fluoroquinolone; Otani T et al.; The antibacterial activities of DK-507k, a novel quinolone, were compared with those of other quinolones: ciprofloxacin, gatifloxacin, levofloxacin, moxifloxacin, sitafloxacin, and garenoxacin (BMS284756) . DK-507k was as active as sitafloxacin and was as active as or up to eightfold more active than gatifloxacin, moxifloxacin, and garenoxacin against Streptococcus pneumoniae, methicillin-susceptible and methicillin-resistant Staphylococcus aureus, and coagulase-negative staphylococci . DK-507k was as active as or 4-fold more active than garenoxacin and 2- to 16-fold more active than gatifloxacin and moxifloxacin against ciprofloxacin-resistant strains of S . pneumoniae, including clinical isolates and in vitro-selected mutants with known mutations . DK-507k inhibited all ciprofloxacin-resistant strains of S . pneumoniae at 1 microg/ml . A time-kill assay with S . pneumoniae showed that DK-507k was more bactericidal than gatifloxacin and moxifloxacin . The activities of DK-507k against most members of the family Enterobacteriaceae were comparable to those of ciprofloxacin and equal to or up to 32-fold higher than those of gatifloxacin, levofloxacin, moxifloxacin, and garenoxacin . DK-507k was fourfold less active than sitafloxacin and ciprofloxacin against Pseudomonas aeruginosa, while it was two to four times more potent than levofloxacin, gatifloxacin, moxifloxacin, and garenoxacin against P . aeruginosa . In vivo, intravenous treatment with DK-507k was more effective than that with gatifloxacin and moxifloxacin against systemic infections caused by S . aureus, S . pneumoniae, and P . aeruginosa in mice . In a mouse model of pneumonia due to penicillin-resistant S . pneumoniae, DK-507k administered subcutaneously showed dose-dependent efficacy and eliminated the bacteria from the lungs, whereas gatifloxacin and moxifloxacin had no significant efficacy . Oral treatment with DK-507k was slightly more effective than that with ciprofloxacin in a rat model of foreign body-associated urinary tract infection caused by a P . aeruginosa isolate for which the MIC of DK-507k was fourfold higher than that of ciprofloxacin . Oral administration of DK-507k to rats achieved higher peak concentrations in serum and higher concentrations in cumulative urine than those achieved with ciprofloxacin . These data indicate the potential advantages of DK-507k over other quinolones for the treatment of a wide range of community-acquired infections.

Antimicrob Agents Chemother, 2003 Dec, 47(12), 3724 - 32
Prevalence and molecular epidemiology of CTX-M extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in Russian hospitals; Edelstein M et al.; A total of 904 consecutive nosocomial isolates of Escherichia coli and Klebsiella pneumoniae collected from 28 Russian hospitals were screened for production of extended-spectrum beta-lactamases (ESBLs) . The ESBL phenotype was detected in 78 (15.8%) E . coli and 248 (60.8%) K . pneumoniae isolates . One hundred fifteen isolates carried the genes for CTX-M-type beta-lactamases, which, as shown by PCR-restriction fragment length polymorphism analysis, were distributed into the two genetic groups of CTX-M-1 (93%)- and CTX-M-2 (7%)-related enzymes . Isolates producing the enzymes of the first group were found in 20 hospitals from geographically distant regions of the country and were characterized by considerable diversity of genetic types, as was demonstrated by enterobacterial repetitive consensus PCR typing . Within this group the CTX-M-3 and the CTX-M-15 beta-lactamases were identified . In contrast, the enzymes of the CTX-M-2 group (namely, CTX-M-5) were detected only in eight clonally related E . coli isolates from a single hospital . Notably, the levels of resistance to ceftazidime were remarkably variable among the CTX-M producers . This study provides further evidence of the global dissemination of CTX-M type ESBLs and emphasizes the need for their epidemiological monitoring.

FEMS Microbiol Lett, 2003 Nov 21, 228(2), 273 - 9
Selenate reduction by Enterobacter cloacae SLD1a-1 is catalysed by a molybdenum-dependent membrane-bound enzyme that is distinct from the membrane-bound nitrate reductase; Watts CA et al.; Enterobacter cloacae SLD1a-1 is capable of reducing selenium oxyanions to elemental selenium under both aerobic and anaerobic conditions . In this study the enzyme that catalyses the initial reduction of selenate (SeO4(2-)) to selenite (SeO3(2-)) has been localised to isolated cytoplasmic membrane fractions . Experiments with intact cells have shown that the putative selenate reductase can accept electrons more readily from membrane-impermeable methyl viologen than membrane-permeable benzyl viologen, suggesting that the location of the catalytic site is towards the periplasmic side of the cytoplasmic membrane . Enzyme activity was enhanced by growing cells in the presence of 1 mM sodium molybdate and significantly reduced in cells grown in the presence of 1 mM sodium tungstate . Non-denaturing polyacrylamide gel electrophoresis (PAGE) gels stained for selenate and nitrate reductase activity have revealed that two distinct membrane-bound enzymes catalyse the reduction of selenate and nitrate . The role of this membrane-bound molybdenum-dependent reductase in relation to selenate detoxification and energy conservation is discussed.

Acta Trop, 2003 Dec, 89(1), 67 - 72
Bacteremia in adults admitted to the Department of Medicine of Bangui Community Hospital (Central African Republic); Kassa-Kelembho E et al.; To determine which pathogens are responsible for bloodstream infections in Bangui and to which antibiotics these pathogens are resistant, we conducted a prospective study of the bacteria isolated from the blood of febrile patients hospitalized in the department of medicine of the Bangui Community Hospital after the failure of antimalarial treatment . One hundred and thirty-one patients were included in this study . Bacteria were identified in 49 blood cultures (37.4%) . Eleven different species were identified . Bacteremia was more frequent in HIV-positive patients than in HIV-negative patients . Salmonella typhimurium, Mycobacterium tuberculosis and Streptococcus pneumoniae were the most frequently isolated pathogens . Eighty percent of enterobacteria were resistant to amoxicillin and 85% to trimethoprim-sulfamethoxazole . Ciprofloxacin and ceftriaxone were the most efficient antibiotics for the enterobacteria, but chloramphenicol and gentamicin were efficient in most cases . Some strains of S . pneumoniae displayed reduced susceptibility to penicillin G, but all strains were susceptible to erythromycin.

Microb Drug Resist, 2003, 9 Suppl 1, S25 - 33
Complex endemic situation regarding extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a hospital in Slovenia; Jutersek B et al.; The first detailed epidemiological study of extended-spectrum beta-lactamase (ESBL)-producing organisms identified in Slovenia was carried out . It was performed on a group of 40 Klebsiella pneumoniae isolates that were randomly selected from all putative ESBL producers of this species recovered in a large hospital in Celje in 1997-2001 . At least three different ESBLs, SHV-2, -5, and -12, were produced by the isolates and these enzymes seem to be common in nosocomial Enterobacteriaceae populations in countries of the region (e.g., Italy, Hungary, Croatia) . The analysis revealed a complex epidemiology of the organisms, illustrated mostly by their high clonal variety but also by the diversity of their beta-lactamase and plasmid content, mating capability, and antimicrobial susceptibility . Although some cases of a 'fresh' dissemination of strains or plasmids could be identified, the overall situation should be described rather as endemic, and its complexity may be in part attributed to the late introduction of the ESBL detection procedure to the hospital.

Lett Appl Microbiol, 2003, 37(5), 410 - 4
Caseinolysis in cheese by Enterobacteriaceae strains of dairy origin; Morales P et al.; AIMS: To study the effect of Enterobacteriaceae strains of dairy origin on caseins under cheese manufacture and ripening conditions . METHODS AND RESULTS: Strains belonging to the genera Enterobacter, Escherichia, Hafnia and Serratia were isolated from fresh raw milk cheeses . Residual caseins in cheeses made from milk individually inoculated with 10 strains of Enterobacteriaceae were determined by capillary electrophoresis . Hierarchical cluster analysis of strains based on data of residual caseins grouped together strains from the same genus, excepting Hafnia strains, which were separated into two groups . Serratia was the most proteolytic genus in our study . Preferences for degradation of casein fractions differed among the four genera studied . CONCLUSIONS: Enterobacteriaceae strains posses proteolytic systems active on all casein fractions under cheese manufacture and ripening conditions . The effects on caseins were similar for strains belonging to the same genus . SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Enterobacteriaceae in cheeses may affect proteolysis during ripening . Assays of Enterobacteriaceae proteolytic activity on milk agar plates may underestimate their caseinolytic activity in cheese.

J Appl Microbiol, 2003, 95(5), 967 - 73
Desiccation and heat tolerance of Enterobacter sakazakii; Breeuwer P et al.; AIMS: Enterobacter sakazakii is an opportunistic pathogen which has been isolated at low levels from powdered infant formulas . This study was performed to demonstrate that Ent . sakazakii is not particularly thermotolerant, but can adapt to osmotic and dry stress . METHODS AND RESULTS: We determined the heat, osmotic and dry stress resistance of Ent . sakazakii . The D-value at 58 degrees C ranged from 0.39 to 0.60 min, which is comparable with that of other Enterobacteriaceae, but much lower than reported previously (Nazarowec-White and Farber 1997, Letters in Applied Microbiology 24: 9-13) . However, stationary phase Ent . sakazakii cells were found to be more resistant to osmotic and dry stress than Escherichia coli, Salmonella and other strains of Enterobacteriaceae tested . Further analysis indicated that the dry resistance is most likely linked to accumulation of trehalose in the cells . CONCLUSIONS: The high tolerance to desiccation provides a competitive advantage for Ent . sakazakii in dry environments, as found in milk powder factories, and thereby increases the risk of postpasteurization contamination of the finished product . SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding of the physiology and survival strategies of Ent . sakazakii is an important step in the efforts to eliminate this bacterium from the critical food production environments.

J Appl Microbiol, 2003, 95(6), 1182 - 90
Seven-hour fluorescence in situ hybridization technique for enumeration of Enterobacteriaceae in food and environmental water sample; Ootsubo M et al.; AIMS: A fluorescent in situ hybridization (FISH) technique using an Enterobacteriaceae-specific probe (probe D) to target 16S rRNA was improved in order to enumerate, within a single working day, Enterobacteriaceae present in food and environmental water samples . METHODS AND RESULTS: In order to minimize the time required for the FISH procedure, each step of FISH with probe D was re-evaluated using cultured Escherichia coli . Five minutes of ethanol treatment for cell fixation and hybridization were sufficient to visualize cultured E . coli, and FISH could be performed within 1 h . Because of the difficulties in detecting low levels of bacterial cells by FISH without cultivation, a FISH technique for detecting microcolonies on membrane filters was investigated to improve the bacterial detection limit . FISH with probe D following 6 h of cultivation to grow microcolonies on a 13 mm diameter membrane filter was performed, and whole Enterobacteriaceae microcolonies on the filter were then detected and enumerated by manual epifluorescence microscopic scanning at magnification of x100 in ca 5 min . The total time for FISH with probe D following cultivation (FISHFC) was reduced to within 7 h . FISHFC can be applied to enumerate cultivable Enterobacteriaceae in food (above 100 cells g-1) and environmental water samples (above 1 cell ml-1) . CONCLUSIONS: Cultivable Enterobacteriaceae in food and water samples were enumerated accurately within 7 h using the FISHFC method . SIGNIFICANCE AND IMPACT OF THE STUDY: A FISHFC method capable of evaluating Enterobacteriaceae contamination in food and environmental water within a single working day was developed.

Antibiot Khimioter, 2003, 48(7), 5 - 11
{Characteristics and clinical value of extended-spectrum beta-lactamases}; Berezin AG et al.; The literature data and the findings of the authors' studies on the role of extended-spectrum bata-lactamases (ESBL) in providing the clinical effect in the treatment of infections due to ESBL-producing organisms were analyzed . The analysis allowed to consider the NCCLS recommendations not sufficiently valid . According to these recommendations the ESBL-producing organisms (among E . coli and Klebsiella spp.) should be regarded as resistant to penicillins, cephalosporins and aztreonam . Susceptibility of 62 Enterobacteriaceae strains that were isolated in 4 therapeutic centres of Tomsk, Nazran and Moscow and proved to be ESBL-producing organisms was tested and it was shown that the 3rd and 4th generation cephalosporins could not be referred to a homogenous group . The same was confirmed by the prospective and retrospective investigations of a multiprofile hospital on the clinical and bacteriological efficacies of the 3rd generation cephalosporins and cefepime in the treatment of hospital-acquired infections due to Enterobacteriaceae strains producing ESBL.

J Food Prot, 2003 Nov, 66(11), 2146 - 50
A new alginate-based rapid method for determining coliforms in milk; Chang SS et al.; A new rapid method for monitoring coliforms was developed on the basis of the instant gelling effects of alginate and calcium . The effectiveness of this new method in the detection of coliforms was evaluated . Tests involving Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, total coliforms in milk, cold-injured coliforms, and total coliforms in raw milk were carried out . The bacterial samples were diluted in 0.2% peptone water containing 90 mM CaCl2 and added into test tubes containing modified purple broth base medium . Coliform concentrations were determined on the basis of the time of color change and gas production in the alginate tubes . All results obtained by the alginate method correlated strongly with those obtained by the conventional violet red bile agar (VRBA) plating method . The alginate method reduced detection time by 12 to 14 h compared with the conventional VRBA plating method . The alginate method can be applied in field studies more easily than melted-agar systems can . The results of this study indicate that the alginate method is an accurate, rapid, simple, and economical way to monitor and estimate concentrations of total coliforms in food.

J Food Prot, 2003 Nov, 66(11), 2005 - 9
Effect of chemical dehairing on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and enterobacteriaceae on carcasses in a commercial beef processing plant; Nou X et al.; The objective of this experiment was to test the hypothesis that cleaning cattle hides by removing hair and extraneous matter before hide removal would result in improved microbiological quality of carcasses in commercial beef processing plants . To test this hypothesis, we examined the effect of chemical dehairing of cattle hides on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and Enterobacteriaceae on carcasses . Samples from 240 control (conventionally processed) and 240 treated (chemically dehaired before hide removal) hides (immediately after stunning but before treatment) and preevisceration carcasses (immediately after hide removal) were obtained from four visits to a commercial beef processing plant . Total aerobic plate counts (APC) and Enterobacteriaceae counts (EBC) were not (P > 0.05) different between cattle designated for chemical dehairing (8.1 and 5.9 log CFU/100 cm2 for APC and EBC, respectively) and cattle designated for conventional processing (8.0 and 5.7 log CFU/100 cm2 for APC and EBC, respectively) . However, E . coli O157:H7 hide prevalence was higher (P < 0.05) for the control group than for the treated group (67% versus 88%) . In contrast to hides, the bacterial levels were lower (P < 0.05) on the treated (3.5 and 1.4 log CFU/100 cm2 for APC and EBC) than the control (5.5 and 3.2 log CFU/100 cm2 for APC and EBC) preevisceration carcasses . Prevalence of E . coli O157:H7 was lower (P > 0.05) on treated than on control preevisceration carcasses (1% versus 50%) . These data indicate that chemical dehairing of cattle hides is an effective intervention to reduce the incidence of hide-to-carcass contamination with pathogens . The data also imply that any effective hide intervention process incorporated into beef processing procedures would significantly reduce carcass contamination by E . coli O157:H7.

Mol Microbiol, 2003 Nov, 50(4), 1391 - 9
Response to culture aeration mediated by the nitrate and nitrite sensor NarQ of Escherichia coli K-12; Stewart V et al.; Respiratory enzyme synthesis in enterobacteria is controlled in response to electron acceptor availability . The iron-sulphur protein Fnr and the sensor-regulator proteins ArcB-ArcA control respiratory gene transcription in response to oxygen and quinone pool redox status respectively . The sensor-regulator proteins NarX-NarL and NarQ-NarP control anaerobic respiratory gene expression in response to nitrate and nitrite . Our laboratory recently engineered the lac operon to replace the primary operator O1-lac with the NarL and NarP protein binding site from the nirB operon . Expression of the lacZ gene from this construct is repressed by nitrate in Nar+ strains . Here, we found that lacZ gene expression was repressed in aerated cultures of narQ+narX null strains . This repression was not observed in narX+narQ+ or narX+narQ null strains . Thus, the NarQ sensor responds to aeration as well as to nitrate and nitrite . The NarX and NarQ sensors are composed of three distinct modules: an amino-terminal sensory module, a carboxyl-terminal transmitter module and a central module of unknown function . Experiments with NarX-NarQ hybrid proteins suggest that the NarQ protein central module is necessary for response to aeration . The physiological significance of this additional sensory role for the NarQ sensor remains obscure.

J Assoc Physicians India, 2003 Jul, 51, 669 - 72
Enterobacter bacteremia; Gupta N et al.; OBJECTIVES: To study the incidence, antimicrobial susceptibility pattern, age and sex distribution, mortality and prognostic factors in cases of Enterobacter bacteremia . METHODS: A total of 18,745 indoor patients with suspected bacteremia were included in study . Enterobacter spp were identified and speciated using standard protocols . Antimicrobial susceptibility testing was done using Stoke's disc diffusion method and minimum inhibitory concentration (MIC) was determined by agar dilution method as recommended by NCCLS . RESULTS: Enterobacter spp ranking sixth was responsible for 4.6% of total group of significant bacteremias . The species most commonly causing bacteremia was E . cloacae (61.8%) . The bacteremia was unimicrobial in 85.1% and part of a polymicrobial bacteremia in 14.8% . Portal of entry in decreasing order of frequency were unknown, respiratory tract, urinary tract and surgical wound . The most common clinical finding was fever (97.2%) . Of Enterobacter isolates 72.9% were multiresistant to three or more antimicrobial agents . Overall mortality was 51.1% . CONCLUSION: Enterobacter spp are becoming increasingly important nosocomial pathogens . To prevent further emergence of multidrug resistance it may beprudent to avoid third generation cephalosporins and instead combination therapy may be used.

Clin Exp Immunol, 2003 Nov, 134(2), 351 - 9
HLA-B27-restricted T cells from patients with ankylosing spondylitis recognize peptides from B*2705 that are similar to bacteria-derived peptides; Frauendorf E et al.; Ankylosing spondylitis (AS) is an inflammatory systemic disease affecting the spine, sacroiliacal and peripheral joints . Although the aetiology of AS remains unknown, the strong association with the HLA-B27 allele might reflect directly a detrimental effect of the HLA-B27 molecule itself, resulting from its potential capability to present 'arthritogenic' peptides to CD8+ T cells . Because some forms of SpA are triggered by enterobacterial infection, such arthritogenic peptides might originate from autologous and/or bacterial proteins triggering cross-reactive CD8+ T cell clones . Intriguingly, two peptides from the second extracellular domain of HLA-B*2705 share sequence homologies with several enterobacterial antigens, exhibit the HLA-B27-binding-motif, and are presented by HLA-B*2705 itself . The objective of this study was to examine the clonal T cell reactivity against these peptides in patients with AS . To this end, we screened peripheral blood lymphocytes (PBL) of 26 patients with AS and 24 healthy donors for TNF-alpha-producing cells using ELISPOT assays . PBL and synovial fluid-derived lymphocytes (SFL) of peptide-responsive patients were then stimulated and cultured with the relevant peptide and control peptides in vitro . Antigen-specific T cell lines (TCL) were identified by standard chromium release assays . Clonal analysis was performed subsequently applying TCRB-CDR3 spectratyping . Among eight peptides tested, only the HLA-B27 168-176 peptide LRRYLENGK was recognized by PBL from B27+ AS patients but not from B27+ healthy controls (P=0.001) . LRRYLENGK-specific T cell clones used preferentially the TCRBV5S1 and the BV14 segment . These results suggest that an HLA-B27-derived peptide with homology to bacterial peptides may play a role in AS.

Clin Microbiol Infect, 2003 Nov, 9(11), 1091 - 103
Evaluation of VITEK 2 for analysis of Enterobacteriaceae using the Advanced Expert System (AES) versus interpretive susceptibility guidelines used at Dynacare Kasper Medical Laboratories, Edmonton, Alberta; Blondel-Hill E et al.; OBJECTIVE: To evaluate the applicability and adaptability of the bioMerieux VITEK 2 Advanced Expert System (AES) to the customized interpretive susceptibility guidelines used at Dynacare Kasper Medical Laboratories (DKML) . METHODS: Three hundred isolates of Enterobacteriaceae (not more than 30% Escherichia coli) were tested on the VITEK 2 system and the API 20E for identification . Susceptibility testing for these isolates was performed on the VITEK 2 system and the Pasco broth microdilution panels . Minimal inhibitory concentrations (MICs) and interpreted results according to the AES and DKML antimicrobial susceptibility guidelines were compared . RESULTS: Of 300 isolates tested for susceptibility, 13 did not give AES interpretations . Of the remaining 287 isolates, interpretations between AES and DKML guidelines were compared for 10 antibiotics . The overall correlation between the AES and DKML interpretations was 96.2% . CONCLUSION: This study demonstrated the benefits and limitations of the bioMerieux AES . Automated knowledge-based systems provide a useful laboratory tool for the interpretation of susceptibility results.

Clin Microbiol Infect, 2003 Sep, 9(9), 912 - 8
Comparative evaluation of three chromogenic agars for detection and rapid identification of aerobic Gram-negative bacteria in the normal intestinal microflora; Filius PM et al.; OBJECTIVE: To compare three different chromogenic agars and MacConkey agar for the detection of aerobic Gram-negative bacteria in the normal intestinal microflora and to assess the accuracy of the chromogenic agars for the direct identification of Escherichia coli . METHODS: A total of 164 Gram-negative clinical isolates (E . coli, Proteus, Klebsiella, Enterobacter, Morganella and Pseudomonas species) and 30 stool specimens were inoculated in parallel on four media: Chromagar E . coli/Coliform, Chromogenic urinary tract infection UTI medium, CHROMagar Orientation and MacConkey agar . All colonies that differed by color and/or morphology were selected for further identification by VITEK 1 and/or API 20E from each medium . RESULTS: On E . coli/Coliform agar five out of 32 (16%) E . coli strains failed to produce the color as described by the manufacturer . No remarkable discrepancies were found for the other clinical isolates . There was no significant difference in detection rate (DR) of aerobic Gram-negative bacteria in stool specimens between the different chromogenic agars and MacConkey agar . The overall DR was about 84%, and varied from 100% for monomicrobial specimens to 33% for polymicrobial specimens . The positive predictive values (PPV) for the direct identification of E . coli on Chromagar E . coli/Coliform, Chromogenic UTI medium and CHROMagar Orientation were 1.00, 0.93 and 0.93, respectively . The negative predictive values (NPV) were 0.53, 0.68 and 0.69, respectively . CONCLUSION: Chromogenic UTI medium and CHROMagar Orientation are the preferred media because of the higher NPV . The high PPV of these agars allows accurate and rapid identification of E . coli.

J Agric Food Chem, 2003 Nov 19, 51(24), 7073 - 8
Factors affecting reduction of selenate to elemental selenium in agricultural drainage water by Enterobacter taylorae; Zhang Y et al.; Microbial reduction of selenate {Se(VI)} to elemental selenium {Se(0)} is a useful technique for removing Se from agricultural drainage water . A series of batch experiments were conducted in the laboratory to determine the effects of yeast extract (50-1000 mg/L), salinity (EC, 5-75 dS/m), and NO(3)(-) (5-100 mg/L) on the removal of Se(VI) (2000 microg/L) from drainage water by Enterobacter taylorae . Results showed that relatively high amounts of yeast extract (500 mg/L) were needed for E . taylorae to effectively reduce Se(VI) to Se(0) . During a 7-day experiment, approximately 95% of added Se(VI) was reduced to Se(0) in the low-salinity drainage water (5 dS/m) with NO(3)(-) values of 5-50 mg/L . In the high-salinity drainage water (50-75 dS/m), reduction of Se(VI) to Se(0) was limited . E . taylorae was also capable of reducing Se(VI) to Se(0) in the San Joaquin Valley drainage water, with a reduction of the added Se(VI) to Se(0) (73.8%) and Se(-II) (20%) . This study suggests that E . taylorae may be used to treat Se(VI)-contaminated drainage water in the field.

J Microbiol Methods, 2003 Dec, 55(3), 717 - 25
Evaluation of parameters affecting quantitative detection of Escherichia coli O157 in enriched water samples using immunomagnetic electrochemiluminescence; Shelton DR et al.; We report here the use of immunomagnetic (IM) electrochemiluminescence (ECL) for quantitative detection of Esherichia coli O157:H7 in water samples following enrichment in minimal lactose broth (MLB) . IM beads prepared in-house with four commercial anti-O157 monoclonal antibodies were compared for efficiency of cell capture . IM-ECL responses for E . coli O157:H7 (strain SEA13B88) were similar for all four commercial anti-O157 LPS monoclonal antibodies . The ECL signal was linearly correlated with E . coli O157:H7 cell concentration, indicating a constant ECL response per cell . Twenty-two strains of E . coli O157:H7 or O157:NM gave comparable ECL signals using IM beads prepared in-house . To assess the potential for interference from background bacteria in MLB-enriched water samples, 10(4) cells of E . coli O157:H7 (strain SEA13B88) were added to enriched samples prior to analysis . There was considerable variability in recovery of E . coli O157:H7 cells; net ECL signals ranged from 1% to 100% of expected values (i.e., percent inhibition from 0% to 99%) . Cultures of Klebsiella pneumoniae, Klebsiella oxytoca, and Enterobacter cloacae, subsequently isolated from MLB-enriched water samples via IM separation (IMS), were observed to interfere with the binding of E . coli O157:H7 cells to IM beads . Recoveries of 10(4) E . coli O157:H7 cells were </=10% in the presence of ca . 10(8) K . pneumoniae, K . oxytoca, or E . cloacae cells . None of these strains gave a positive IM-ECL signal . Although competitive binding decreased sensitivity, there still was a linear correlation between ECL signal and higher E . coli O157:H7 cell concentrations . These studies indicate that IM-ECL in conjunction with MLB enrichment is capable of quantitatively detecting as few as 10(3) to 10(5) E . coli O157:H7 cells ml(-1), depending on percent recoveries, in enriched samples that contain ca . 10(9) total lactose-fermenting bacteria ml(-1) . Assuming comparable growth rates for E . coli O157:H7 and other lactose-fermenting bacteria in MLB, it may be possible to detect as few as one E . coli O157:H7 in 100 ml of raw water containing as many as 10(4) to 10(6) lactose-fermenting bacteria (i.e., total coliforms).

J Clin Microbiol, 2003 Nov, 41(11), 5224 - 6
Evaluation of genotyping large numbers of Escherichia coli isolates by enterobacterial repetitive intergenic consensus-PCR; Meacham KJ et al.; We analyzed a large collection of Escherichia coli isolates typed by enterobacterial repetitive intergenic consensus (ERIC)-PCR with BioNumerics gel analysis software . However, the interexperimental variation of ERIC-PCR caused the computer software to classify repeated isolates as different . ERIC-PCR should not be used as the sole determinant of genetic similarity when typing large numbers of bacterial isolates via computer software.

J Clin Microbiol, 2003 Nov, 41(11), 5167 - 72
Detection of Gallibacterium spp . in chickens by fluorescent 16S rRNA in situ hybridization; Bojesen AM et al.; Gallibacterium has recently been included as a new genus of the family Pasteurellaceae Pohl 1981, which encompasses bacteria previously reported as Pasteurella anatis, "Actinobacillus salpingitidis," and avian Pasteurella haemolytica-like organisms . So far, identification has exclusively relied on phenotypic characterization . We present a method based on a cyanine dye 3.18-labeled in situ hybridization probe targeting 16S rRNA to allow specific detection of bacteria belonging to the genus Gallibacterium . The probe, GAN850, showed no cross-reactivity to 25 other poultry-associated bacterial species, including members of the families Pasteurellaceae, Enterobacteriaceae, and Flavobacteriaceae, when cross-reactivities were evaluated by whole-cell hybridization . The probe was further evaluated by hybridization to formalin-fixed spleen and liver tissues from experimentally infected chickens, in which it proved to be useful for the detection of Gallibacterium . Additionally, determination of the spatial distribution and the host cell affiliation of Gallibacterium at various times during the infection process was possible . In conclusion, the in situ hybridization technique described may be of use as a diagnostic tool as well as for studies to elucidate the pathogenesis of Gallibacterium infections in chickens.

J Clin Microbiol, 2003 Nov, 41(11), 5127 - 33
Application of an rRNA probe matrix for rapid identification of bacteria and fungi from routine blood cultures; Marlowe EM et al.; One of the most important functions of the clinical microbiology laboratory is the identification of the etiology of sepsis . For this study, aliquots from 405 positive blood cultures were tested against a unique array of DNA probes directed against rRNA subsequences of bacteria and fungi for identification . Another 280 samples that were negative after 5 days of incubation were also tested . Blood culture bottles were incubated in a BacT/Alert3D instrument . For the rRNA assay, a 0.4-ml aliquot was removed, and the cells were pelleted by centrifugation . The pellet was washed and frozen at -70 degrees C . Analysis of the pellet involved a lysis step and then the addition of samples to the reaction wells containing the probes in a microtiter plate format . Analysis was performed by using a hybridization protection assay . Results were taken through a series of deductive steps to obtain species, or in some cases genus, identification . Batch sample preparation required approximately 15 min, and sample analysis required another 60 min . Probe results were compared to conventional biochemical identifications . The probe test was negative for the 280 samples that were negative by the BacT/Alert 3D system and for another 21 samples that were false positive (the instrument signaled, but there was no growth) . Microorganisms from the remaining 384 signal-positive samples included 60 Enterobacteriaceae, 10 Pseudomonas aeruginosa, 10 other gram-negative bacteria, 40 Staphylococcus aureus, 152 coagulase-negative staphylococci, 28 streptococci, 22 enterococci, 21 other gram-positive bacteria, 8 anaerobes, and 16 yeast organisms . Seventeen cultures were polymicrobial, and one was gram positive and culture negative . Discordance between probe and conventional identification results was noted for only 12 (1.75%) samples . This novel rapid molecular approach to the identification of bacteria and yeast in blood cultures was highly sensitive (100%) and specific (96%).

J Clin Microbiol, 2003 Nov, 41(11), 5103 - 12
Use of O-antigen gene cluster-specific PCRs for the identification and O-genotyping of Yersinia pseudotuberculosis and Yersinia pestis; Bogdanovich T et al.; Yersinia pestis is a very recently evolved clone of Yersinia pseudotuberculosis serotype O:1b . This close relationship causes potential difficulties in DNA-based diagnostic methods . Analysis of the O-antigen gene clusters in these two organisms identified two regions that were used to specifically identify Y . pestis-Y . pseudotuberculosis as a group or Y . pestis alone . Both PCR assays were found to be 100% specific when tested on a large collection of Yersinia species and other Enterobacteriaceae . Furthermore, advantage was taken of the different setups of the O-antigen gene clusters of the 21 known Y . pseudotuberculosis serotypes to develop a multiplex PCR assay to replace the conventional serotyping method of Y . pseudotuberculosis by O-genotyping . The multiplex PCR assay contained nine sets of specific PCRs in a single tube and when used on Y . pseudotuberculosis reference strains allowed the distinction of 14 individual serotypes and two duplex serotypes (O:4a-O:8 and O:12-O:13) . Serotype O:7, O:9, and O:10 strains required additional PCRs for O-genotyping . Once applied to Y . pseudotuberculosis strains of various origins, a very good correlation between classical serotypes and O-genotypes was observed, although some discrepancies were found . O-genotyping also proved useful to correct misidentification of some strains and to type Y . pseudotuberculosis isolates that had lost the expression of the O-antigen . The PCR-based O-genotyping can easily be applied in conventional laboratories, without the need for tedious preparation of a large set of specific antisera.

Appl Environ Microbiol, 2003 Nov, 69(11), 6848 - 55
Protistan grazing analysis by flow cytometry using prey labeled by in vivo expression of fluorescent proteins; Fu Y et al.; Selective grazing by protists can profoundly influence bacterial community structure, and yet direct, quantitative observation of grazing selectivity has been difficult to achieve . In this investigation, flow cytometry was used to study grazing by the marine heterotrophic flagellate Paraphysomonas imperforata on live bacterial cells genetically modified to express the fluorescent protein markers green fluorescent protein (GFP) and red fluorescent protein (RFP) . Broad-host-range plasmids were constructed that express fluorescent proteins in three bacterial prey species, Escherichia coli, Enterobacter aerogenes, and Pseudomonas putida . Micromonas pusilla, an alga with red autofluorescence, was also used as prey . Predator-prey interactions were quantified by using a FACScan flow cytometer and analyzed by using a Perl program described here . Grazing preference of P . imperforata was influenced by prey type, size, and condition . In competitive feeding trials, P . imperforata consumed algal prey at significantly lower rates than FP (fluorescent protein)-labeled bacteria of similar or different size . Within-species size selection was also observed, but only for P . putida, the largest prey species examined; smaller cells of P . putida were grazed preferentially . No significant difference in clearance rate was observed between GFP- and RFP-labeled strains of the same prey species or between wild-type and GFP-labeled strains . In contrast, the common chemical staining method, 5-(4,6-dichloro-triazin-2-yl)-amino fluorescein hydrochloride, depressed clearance rates for bacterial prey compared to unlabeled or RFP-labeled cells.

Appl Environ Microbiol, 2003 Nov, 69(11), 6520 - 6
Construction of Escherichia coli strains for conversion of nitroacetophenones to ortho-aminophenols; Kadiyala V et al.; The predominant bacterial pathway for nitrobenzene (NB) degradation uses an NB nitroreductase and hydroxylaminobenzene (HAB) mutase to form the ring-fission substrate ortho-aminophenol . We tested the hypothesis that constructed strains might accumulate the aminophenols from nitroacetophenones and other nitroaromatic compounds . We constructed a recombinant plasmid carrying NB nitroreductase (nbzA) and HAB mutase A (habA) genes, both from Pseudomonas pseudoalcaligenes JS45, and expressed the enzymes in Escherichia coli JS995 . IPTG (isopropyl-beta-D-thiogalactopyranoside)-induced cells of strain JS995 rapidly and stoichiometrically converted NB to 2-aminophenol, 2-nitroacetophenone (2NAP) to 2-amino-3-hydroxyacetophenone (2AHAP), and 3-nitroacetophenone (3NAP) to 3-amino-2-hydroxyacetophenone (3AHAP) . We constructed another recombinant plasmid containing the nitroreductase gene (nfs1) from Enterobacter cloacae and habA from strain JS45 and expressed the enzymes in E . coli JS996 . Strain JS996 converted NB to 2-aminophenol, 2-nitrotoluene to 2-amino-3-methylphenol, 3-nitrotoluene to 2-amino-4-methylphenol, 4-nitrobiphenyl ether to 4-amino-5-phenoxyphenol, and 1-nitronaphthalene to 2-amino-1-naphthol . In larger-scale biotransformations catalyzed by strain JS995, 75% of the 2NAP transformed was converted to 2AHAP, whereas 3AHAP was produced stoichiometrically from 3NAP . The final yields of the aminophenols after extraction and recovery were >64% . The biocatalytic synthesis of ortho-aminophenols from nitroacetophenones suggests that strain JS995 may be useful in the biocatalytic production of a variety of substituted ortho-aminophenols from the corresponding nitroaromatic compounds.

Int J Antimicrob Agents, 2003 Nov, 22(5), 526 - 31
Evolution of aminoglycoside resistance phenotypes of four Gram-negative bacteria: an 8-year survey in a University Hospital in Greece; Neonakis I et al.; In order to determine the resistance patterns and evolution trends of four common Enterobacteriaceae (Escherichia coli, Proteus spp., Klebsiella spp . and Enterobacter spp.), aminoglycoside resistance phenotypes of 8917 non-repetitive strains, isolated over an 8-year period, were analysed . Phenotypes were defined by examining the susceptibility of the strains to a panel of aminoglycosides, using disk diffusion method . A large diversity of different resistance phenotypes was encountered . A significant progressive increase in the proportions of wild-type E . coli strains was noted . Among resistant strains of Enterobacter spp . and Klebsiella spp., the incidence of phenotype KTANt (kanamycin, tobramycin, amikacin and netilmicin), indicative of AAC(6')-I production, was very high (66.7 and 46.5%, respectively) . Phenotypes indicative for gentamicin-modifying enzymes as well as broad-spectrum combinations (combinations of gentamicin-modifying enzymes with AAC(6')-I) were infrequent.

J Chemother, 2003 Oct, 15(5), 449 - 53
Inhibition of nitrofurantoin reduction by menthol leads to enhanced antimicrobial activity; Shahverdi AR et al.; Nitrofurantoin is a nitroaromatic compound used for the treatment of urinary tract infections . Nitrofurantoin activity is regulated by a nitroreduction process . It is first reduced by bacterial nitroreductases to active short-life intermediates, which are further converted to non-toxic molecules, which negatively affect its antibacterial activity . In this study we have shown that resistant strains of Klebsiella sp . inactivate the bactericidal activity of nitrofurantoin . Also we demonstrated a synergistic effect between menthol and nitrofurantoin bactericidal activities against nitrofurantoin susceptible Enterobacteriaceae.

J Chemother, 2003 Oct, 15(5), 428 - 41
Natural antibiotic susceptibility of Ewingella americana strains; Stock I et al.; The natural susceptibility of 20 Ewingella americana strains to 72 antibiotics was examined . MIC values were determined using a microdilution procedure in cation-adjusted Mueller-Hinton broth . Evaluation of natural antibiotic susceptibility was performed applying the German standard (where applicable) . Beta-lactamases were examined with a conventional nitrocefin colony testing procedure, activity and induction assays, and SDS-PAGE . Ewingella strains were naturally resistant or of intermediate susceptibility to cefaclor, loracarbef, cefazoline, cefuroxime, cefoxitin, benzylpenicillin, oxacillin, fosfomycin, erythromycin, roxithromycin, clarithromycin, lincosamides, dalfopristin-quinupristin, ketolides, linezolid, glycopeptides, fusidic acid and rifampicin . Uniform natural sensitivity was found with acylureidopenicillins except for azlocillin, ticarcillin, several cephalosporins, carbapenems, aztreonam, tetracyclines, aminoglycosides, quinolones, azithromycin, folate-pathway inhibitors and chloramphenicol . Strains of E . americana were naturally sensitive or of intermediate susceptibility to aminopenicillins (with and without beta-lactamase inhibitors), azlocillin and nitrofurantoin . All ewingellae yielded beta-lactamases; testing of representative strains revealed that these enzymes belong to Ambler class C . Inducibility of beta-lactamase was shown for E . americana ATCC 33852T, CCUG 35675 and CCUG 42782 . The present study describes a database concerning the natural susceptibility of E . americana strains to a range of antibiotics, which can be applied to validate forthcoming antibiotic susceptibility tests of these bacteria . It enlarges the number of Enterobacteriaceae expressing naturally-occurring AmpC beta-lactamases.

Plasmid, 2003 Nov, 50(3), 213 - 29
Peripheral sequences of the Serratia entomophila pADAP virulence-associated region; Hurst MR et al.; Some strains of the Enterobacteriaceae Serratia entomophila and Serratia proteamaculans cause amber disease in the grass grub, Costelytra zealandica (Coleoptera: Scarabaeidae), an important pasture pest in New Zealand . The genes responsible for this disease reside on a large, 155-kb plasmid designated amber disease-associated plasmid (pADAP) . Herein, we report the DNA sequencing of approximately 50 kb upstream and 10 kb downstream of the virulence-encoding region . Based on similarity with proteins in the current databases, and potential ribosome-binding sites, 63 potential ORFs were determined . Eleven of these ORFs belong to a type IV pilus cluster (pilL-V) and a further eight have similarities to the translated products of the plasmid transfer traH-N genes of the plasmid R64 . In addition, a degenerate 785-nt direct repeat flanks a 44.7-kb region with the potential to encode three Bacillus subtilis Yee-type proteins, a fimbrial gene cluster, the sep virulence-associated genes and several remnant IS elements.

Diagn Microbiol Infect Dis, 2003 Nov, 47(3), 547 - 50
Contemporary in vitro synergy rates for aztreonam combined with newer fluoroquinolones and beta-lactams tested against gram-negative bacilli; Sader HS et al.; Aztreonam has been commonly used in various combinations to enhance antimicrobial spectrum of co-drugs and produce potential synergistic activity . Although well studied in vitro over 10 years ago, aztreonam combination testing has been poorly documented with newer or commonly used agents against contemporary isolates . All MIC tests (alone or in combination) used in this experiment were reference broth microdilution methods in checkerboard tray designs . Aztreonam was combined with ciprofloxacin, gatifloxacin, levofloxacin, cefepime, ceftazidime and imipenem at clinically relevant concentrations . Interaction categories were defined by established criteria . Forty strains each of Pseudomonas aeruginosa and Enterobacteriaceae (12 species; aztreonam MIC, 1-16 microg/ml) were tested for each antimicrobial combination (480 total determinations) . No antagonism or indeterminate interactions were identified . The overall rates of synergy or partial synergy for aztreonam with fluoroquinolone combinations was 63.4% versus P . aeruginosa, greatest for aztreonam with gatifloxacin (67.5%) . Interaction categories varied greatly among aztreonam with beta-lactam combinations . Aztreonam with ceftazidime or cefepime versus P . aeruginosa had 75.0 - 85.0% partial or complete synergy rates, but aztreonam with imipenem showed dominant indifference (65.0%) . In contrast, aztreonam with imipenem was more likely to exhibit synergy (32.5%) when tested against Enterobacteriaceae . Aztreonam, often used as an aminoglycoside substitute in antimicrobial combinations, continues to demonstrate enhanced, but variable drug activity interactions for contemporary antimicrobial combinations when tested against recent (2002) clinical isolates.

J Dairy Sci, 2003 Oct, 86(10), 3128 - 37
Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge; Bannerman DD et al.; Several species of gram-negative bacteria, including Escherichia coli, Klebsiella pneumoniae, and various species of Enterobacter, are common mastitis pathogens . All of these bacteria are characterized by the presence of endotoxin or lipopolysaccharide (LPS) in their outer membrane . The bovine mammary gland is highly sensitive to LPS, and LPS has been implicated, in part, in the pathogenesis of gram-negative mastitis . Recognition of LPS is a key event in the innate immune response to gram-negative infection and is mediated by the accessory molecules CD14 and LPS-binding protein (LBP) . The objective of the current study was to determine whether LBP levels increased in the blood and mammary gland following LPS challenge . The left and right quarters of five midlactating Holstein cows were challenged with either saline or LPS (100 microg), respectively, and milk and blood samples collected . Basal levels of plasma and milk LBP were 38 and 6 microg/ml, respectively . Plasma LBP levels increased as early as 8 h post-LPS challenge and reached maximal levels of 138 microg/ ml by 24 h . Analysis of whey samples derived from LPS-treated quarters revealed an increase in milk LBP by 12 h . Similar to plasma, maximal levels of milk LBP (34 microg/ml) were detected 24 h following the initial LPS challenge . Increments in milk LBP levels paralleled a rise in soluble CD14 (sCD14) levels and initial rises in the levels of these proteins were temporally coincident with maximal neutrophil recruitment to the inflamed gland . Because LBP and sCD14 are known to enhance LPS-induced host cell activation and to facilitate detoxification of LPS, these data are consistent with a role for these molecules in mediating mammary gland responses to LPS.

Mol Biol (Mosk), 2003 Sep-Oct, 37(5), 843 - 9
{Computer analysis of regulating metabolism of glycerol-3-phosphate in proteobacteria genome}; Danilova LV et al.; Comparative computer-assisted analysis was used to study putative GlpR-regulons responsible for metabolism of glycerol and glycerol-3-phosphate in genomes of alpha-, beta-, and gamma-proteobacteria . New palindromic GlpR-binding signals were identified in gamma-proteobacteria; consensus sequences being TGTTCGATAACGAACA for Enterobacteriaceae, wTTTTCGTATACGAAAAw for Pseudomonadaceae, and AATGCTCGATCGAGCATT for Vibrionaceae . The signals in alpha- and beta-proteobacteria were also identified: they contained 3-4 direct TTTCGTT repeats separated by 3-4 nucleotide pairs.

Mikrobiyol Bul, 2003 Apr-Jun, 37(2-3), 151 - 5
{Antibacterial effect of nitric oxide}; Coban AY et al.; In this study, the antibacterial effects of nitric oxide (NO) have been investigated against 4 strains of multidrug-resistant Mycobacterium tuberculosis and 8 Klebsiella, 7 Escherichia coli, 7 Staphylococcus, 5 Enterobacter, 7 Pseudomonas and 7 Proteus clinical isolates by using DETA-NO ((Z)-1-{N-(2-aminoethyl)-N-(2-ammonioethyl)amino}diazen-1-ium-1,2-diolate) as the NO donor . Minimum inhibitory concentrations (MICs) were determined by broth microdilution method for all tested microorganisms . The MIC values found in our study were as follows; 0.25 mg/ml for all of the multidrug-resistant Mycobacterium tuberculosis strains, 1 mg/ml for Staphylococcus strains, and approximately 2 mg/ml for the rest of the bacterial strains . These results showed that NO (or NO donor DETA-NO) was more effective on multidrug-resistant M . tuberculosis strains than the other bacterial species . It can be concluded that further studies are needed to explain the clinical use of NO donor DETA-NO, especially, in patients infected with multidrug-resistant M . tuberculosis.

Appl Microbiol Biotechnol, 2004 Feb, 63(6), 691 - 7 Epub 2003 Oct 31.
Production of L-asparaginase in Enterobacter aerogenes expressing Vitreoscilla hemoglobin for efficient oxygen uptake; Geckil H et al.; This study is the first utilizing Vitreoscilla hemoglobin in a heterologous bacterium, Enterobacter aerogenes, to determine the effect of such a highly efficient oxygen-uptake system on the production of l-asparaginase, an enzyme that has attracted considerable attention due to its anti-tumor activity . Here, we show that the Vitreoscilla hemoglobin expressing strain has from 10-fold to more than two orders of magnitude lower l-asparaginase activity than the wild type or the control without the Vitreoscilla hemoglobin gene under different aeration conditions . Aeration and agitation were also determining factors for enzyme production . The enzyme activity was reduced considerably under both full aerobic and anaerobic conditions, while the highest enzyme activity was determined in cultures under low aeration and low agitation . Also, the effect of different concentrations of glucose on enzyme production showed catabolic repression . Glucose at 1% caused almost total inhibition of enzyme activity, while at 0.1% it showed a slightly stimulatory effect on enzyme production, compared with glucose-free medium.

J Gynecol Obstet Biol Reprod (Paris), 2003 Oct, 32(6), 555 - 61
{Bacterial flora in the genital tract the last trimester of pregnancy}; Balaka B et al.; Very widespread in our clinical setting, early-onset sepsis is due to organisms that commonly colonize or infect the maternal genital tract; identifying such organisms would help improve prevention and treatment . OBJECTIVE: To determine the bacterial ecology and the pathological status of the genital organs during the last trimester of pregnancy, in order to evaluate the risk of materno-fetal infections and to improve the present prophylactic measures based on monitoring bacterial carriage during the first trimester . METHOD: Vaginal and endocervical samples, usually taken during the first trimester of pregnancy were delayed and taken during the last trimester of pregnancy, in patients with no signs of sepsis and not taking antibiotics . A macroscopic examination described the aspect of the vagina, the cervix uteri, leukorrhea and possible inflammatory lesions or ulcerations . A microscopic examination searched for parasites, epithelial cells, Clue cells and leukocytes . The appropriate bacteriological cultures were performed after reading the Gram stain and scoring the vaginal flora . The clinical and cytobacteriological aspects were used to identify the bacterial ecology and the pathological genital states . RESULTS: Genital samples were collected from 306 pregnant women . Among them 118 were at 29-32 weeks of gestation, 104 at 33-36 and 84 at 37-40 . The most frequent germs were C . albicans (33.3%), Enterobacteriaceae (20.3%) including E . coli (10.9%), S . aureus (15.4%), Gardnerella (13.6%), and Trichomonas (10.6%), in monomicrobian (79.2%) or polymicrobian carriage (20.8%) . Lower genital tract pathological states such as vaginitis (29.4%), bacterial vaginosis (21.5%) or cervicitis (10.4%) and asymptomatic bacterial carriage (23.5%) and normal genital flora (15%) were identified . CONCLUSION: This is the first report of genital bacterial carriage in African women during the last trimester of pregnancy . Larger studies are required to evaluate the risk of maternofetal infections and to improve current prophylaxis measures.

Infect Control Hosp Epidemiol, 2003 Oct, 24(10), 780 - 2
Meningitis due to Enterobacter aerogenes subsequent to resection of acoustic neuroma and percutaneous endoscopic gastrostomy tube placement: a rare nosocomial event; Poetker DM et al.; We present a case of meningitis after percutaneous endoscopic gastrostomy (PEG) tube placement subsequent to acoustic neuroma resection and cranioplasty . Four days following PEG tube placement, the patient developed Enterobacter aerogenes meningitis, requiring explantation of infected cranioplasty material . His condition subsequently improved . Etiology and future intervention strategies are discussed.

J Antimicrob Chemother, 2003 Dec, 52(6), 1022 - 4 Epub 2003 Oct 29.
Sulphonamide resistance gene sul3 found in Escherichia coli isolates from human sources; Grape M et al.; OBJECTIVES: The aim of this study was to investigate the molecular basis of an observed increasing resistance to trimethoprim and sulphonamides despite a simultaneous decline in co-trimoxazole consumption . The distribution of sulphonamide resistance genes sul1, sul2 and the recently discovered sul3 was studied in a collection of clinical isolates of Enterobacteriaceae . METHODS: PCR with primers specific for sul1, sul2 and sul3 was used to detect the three known sulphonamide resistance genes in the isolate collection . Sequence analysis was used for confirmation of results . Restriction endonuclease digestion and conjugational transfer assays were used for plasmid analysis . RESULTS: In 64 sulphonamide-resistant isolates, 39 sul1 genes and 48 sul2 genes were detected . Twenty-five isolates carried both sul1 and sul2 and two were negative for both genes . With PCR and sequence analysis these two were shown to harbour the new sulphonamide resistance gene sul3, which was carried by different plasmids . CONCLUSIONS: Sulphonamide resistance gene sul3, which is widespread among pigs in Switzerland, has now also been identified in two different clinical isolates of Escherichia coli, located in urinary tract infections in patients in Sweden.

J Microbiol Immunol Infect, 2003 Sep, 36(3), 197 - 202
Bacteremia in hematological and oncological children with febrile neutropenia: experience in a tertiary medical center in Taiwan; Lai HP et al.; A retrospective study of bacteremia in children with febrile neutropenia admitted to a medical center in Taiwan from January 1999 to December 1999 was performed . There were 190 episodes of febrile neutropenia during this period and 46 pathogens were isolated from blood specimens in 38 bacteremic episodes (7 mixed infections) . Pseudomonas aeruginosa (17.4%), Staphylococcus aureus (10.9%), Klebsiella pneumoniae (10.9%), and Enterobacter cloacae (10.9%) were the most common isolates . Three of the 5 isolates of S . aureus were resistant to methicillin . Twenty-three episodes of bacteremia (four mixed infections) were associated with recent antibiotic use . Of the 23 bacteremic episodes with recent antibiotic use, P . aeruginosa (20%), methicillin-resistant S . aureus (10%), K . pneumoniae (10%), Escherichia coli (10%), and E . cloacae (10%) were isolated most often . Relapsed leukemia (odds ratio 3, 95% confidence interval 1.4-6.5) and recent antibiotic therapy (odds ratio 3.4, 95% confidence interval 1.7-7.7) were the independent risk factors of bacteremia . There were 9 mortality cases in patients with bacteremia, including 4 cases with mixed infections, and 5 cases with P . aeruginosa, E . coli, Klebsiella oxytoca, S . aureus, and Streptococcus mitis, respectively . Broad-spectrum antibiotics were necessary in febrile neutropenic children because of the high percentage of mixed infection.

J Endotoxin Res, 2003, 9(5), 271 - 9
Endotoxemia and Gram-negative bacteremia as predictors of outcome in sepsis: a meta-analysis using ROC curves; Hurley JC; The prognostic value of the detection of endotoxin in plasma of patients with suspected Gram-negative sepsis is unclear despite numerous studies . Sixteen studies have reported endotoxemia detection and blood culture results with outcome data for 1937 patients with suspected Gram-negative sepsis . Comparisons of the fatality rates for the subgroups of patients with, respectively, either endotoxemia (group 3), or Gram-negative bacteremia (group 2), or both detected (group 1), versus the fatality rate for the subgroup of patients with these factors absent (group 4), were made based on summary odds ratios (and 95% CI) . Summary odds ratios for the comparison of group 2 versus group 4 and group 3 versus group 4 were non-significant (2.2; 0.9-5.8 and 2.0; 0.8-4.8, respectively) . The summary odds ratio was significant for group 1 versus group 4 (3.6; 2.1-6.3), and more so among studies in which non-Enterobacteriaceae were common among the Gram-negative blood culture isolates (4.8; 2.1-10.7) . Endotoxemia, like Gram-negative bacteremia, is a weak prognostic indicator . Endotoxemia appears to have most prognostic significance when it is detected in the co-presence of Gram-negative bacteremia . The differing interpretations among published studies could be attributed to the variability in types of Gram-negative bacteremia observed.

Ann Dermatol Venereol, 2003 Aug-Sep, 130(8-9 Pt 1), 723 - 8
{Community-acquired cutaneous infections: causal role of some bacteria and sensitivity to antibiotics}; Lorette G et al.; OBJECTIVE: Identify the bacteria responsible for cutaneous infections observed in private practice and test their sensitivity to currently used antibiotics . PATIENTS AND METHODS: The patients were examined by dermatologists . A bacteriological sample was taken and sent to a central laboratory for identification of the germ and antibiograms were performed and the minimal inhibiting concentrations (MIC) determined . RESULTS: Folliculitis, impetigo and furuncles were the three most frequent primary infections . Four hundred and forty-three patients were included and 442 samples were placed in culture . Cultures were positive in 265 cases (a single bacterial strain in 231 cases) . Staphylococcus aureus was isolated in 208 cases, streptococci in 11 and enterobacteria in 3; occasionally several germs were present . Eight strains of staphylococci were meti-R (4 p . 100) . All the strains were sensitive to pristinamycin and mupirocin and 90 p . 100 were sensitive to fusidic acid . CONCLUSION: The occurrence and diffusion of resistant strains is a daily concern in hospitals . In general practice, although care must be taken, the problem rarely occurs and the antibiotics used remain effective.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3506 - 14
Extended-spectrum beta-lactamase-producing Enterobacteriaceae in community and private health care centers; Arpin C et al.; In 1999, 39 of 2,599 isolates of the family Enterobacteriaceae (1.5%) collected by eight private laboratories in the Aquitaine region in France produced an extended-spectrum beta-lactamase (ESBL) . Among these were 19 Enterobacter aerogenes isolates; 8 Klebsiella pneumoniae isolates; 6 Escherichia coli isolates; 3 Proteus mirabilis isolates; and 1 isolate each of Serratia marcescens, Morganella morganii, and Providencia stuartii . ESBL producers were isolated from 38 patients, including 33 residents of 11 clinics or nursing homes and 5 ambulatory patients . Seven different ESBLs were characterized . These mainly consisted of TEM-24 (25 isolates) and TEM-21 (9 isolates), but TEM-15 (2 isolates) and TEM-3, TEM-19, SHV-4, and CTX-M-1 (1 isolate each) were also characterized . Seven strains showed the coexistence of different TEM- and/or SHV-encoding genes, including a new SHV-1 variant, SHV-44, defined by the substitution R205L previously reported for SHV-3 in association with S238G . The epidemiology of the ESBL producers was investigated by random amplification of polymorphic DNA, typing by enterobacterial repetitive intergenic consensus PCR, analysis of resistance cotransferred with the ESBL, and analysis of the restriction profiles of the ESBL-encoding plasmids . Of the TEM-24-expressing strains, 18 were E . aerogenes isolates, including 9 from the same clinic, that were representatives of the epidemic clone disseminating in France . Of the TEM-21-producing strains that belonged to different species of the family Enterobacteriaceae (E . coli, K . pneumoniae, and P . mirabilis), 8 were isolated in the same nursing home . Outbreaks due to strain and/or plasmid dissemination in these clinic and nursing home were demonstrated . The presence of ESBL producers in five ambulatory patients probably resulted from nosocomial acquisition . Our data highlight the serious need to monitor patients for ESBL-producing Enterobacteriaceae in general practice.

Infect Immun, 2003 Nov, 71(11), 6307 - 19
Transfer region of pO113 from enterohemorrhagic Escherichia coli: similarity with R64 and identification of a novel plasmid-encoded autotransporter, EpeA; Leyton DL et al.; Enterohemorrhagic Escherichia coli (EHEC) is a prominent, food-borne cause of diarrhea, bloody diarrhea, and the hemolytic uremic syndrome in industrialized countries . Most strains of EHEC carry the locus for enterocyte effacement (LEE) pathogenicity island, but a proportion of isolates from patients with severe disease do not carry LEE and very little is known about virulence factors in these organisms . LEE-negative strains of EHEC typically express Shiga toxin 2 and carry a large plasmid that encodes the production of EHEC hemolysin . In this study, we determined the nucleotide sequence of the transfer region of pO113, the large hemolysin plasmid from LEE-negative EHEC O113:H21 (EH41) . This 63.9-kb region showed a high degree of similarity with the transfer region of R64, and pO113 was capable of self-transmission at low frequencies . Unlike R64 and the related dot/icm system of Legionella pneumophila, however, pO113 was unable to mobilize RSF1010 . In addition, the pO113 transfer region encoded a novel high-molecular-weight serine protease autotransporter of Enterobacteriaceae (SPATE) protein, termed EpeA . Like other SPATEs, EpeA exhibited protease activity and mucinase activity, but expression was not associated with a cytopathic effect on epithelial cells . Analysis of a second high-molecular-weight secreted protein revealed that pO113 also encodes EspP, a cytopathic SPATE identified previously in EHEC O157:H7 . The nucleotide sequences encoding the predicted beta-domains of espP and epeA were identical and also shared significant homology with a third SPATE protein, EspI . Both espP and epeA were detected in several LEE-negative clinical isolates of EHEC and thus may contribute to the pathogenesis of this subset of EHEC.

J Food Prot, 2003 Oct, 66(10), 1798 - 804
Development of a PCR assay for detection of Enterobacteriaceae in foods; Nakano S et al.; A broad-range PCR assay for the detection of bacteria belonging to the Enterobacteriaceae family was developed in this study . Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay . To determine the specificity of the assay, 72 different bacterial species (of 49 genera), 2 fungi, 3 animals, and 4 plants were tested . Results were positive for every tested Vibrioaceae or Enterobacteriaceae strain except Proteus mirabilis . For all other bacterial strains and eukaryotes tested, results were negative . Bacterial DNA for PCR was prepared by a simple procedure with the use of Chelex 100 resin from culture after growth in brain heart infusion medium . To test this PCR assay for the monitoring of the Enterobacteriaceae family, either Escherichia coli or Salmonella Enteritidis was inoculated into various foods as an indicator . Prior to the PCR, the inoculation of 10 to 40 CFU of bacteria per g of food was followed by a 5-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells . When actual examinations of the contamination of 15 noodle foods with Enterobacteriaceae by this PCR assay were conducted, 33% (5 of 15) of the samples tested positive . These results agreed with those of the Petrifilm Enterobacteriaceae Count Plate assay . Including the enrichment culture step, the entire PCR detection process can be completed within 7 h.

Can J Microbiol, 2003 Jul, 49(7), 479 - 82
A low molecular weight enterotoxic hemolysin from clinical Enterobacter cloacae; Simi S et al.; Seven of 50 Enterobacter cloacae strains from clinical isolates produced small turbid zones of hemolysis in horse and sheep blood agar plates, and the culture supernatants were also positive for hemolytic activity . The hemolysin was partially purified from the culture supernatant of E . cloacae by ultrafiltration (PM-10 membrane) and extraction with acetone . Semipurified hemolysin was stable to heating (100 degrees C, 30 min) and was soluble in organic solvents (acetone, ethanol, and methanol) . The toxin showed no loss of biological activity after treatment with trypsin and was stable to acid treatment at pH 2.0 but not at a pH greater than 7.0 . In the rat intestinal loop assay, the hemolysin caused hemorrhagic fluid accumulation and severe histological alterations . These findings indicate that this hemolysin may be a putative virulence factor in E . cloacae infections.

Pathol Biol (Paris), 2003 Oct, 51(8-9), 516 - 9
{Antibacterial effect of two glycine betaine analogues against Escherichia coli}; Queau M et al.; Strains of enterobacteria that cause urinary tract infections are able to grow in urine with high tonicity . In such conditions bacterias adapt to osmotic forces by incorporation of osmoprotectant compounds including glycine betaine . Accumulation of toxic analogues in bacteria, using inducible betaine transporters, has been previously proposed for development of antibiotics . In this study we report antibacterial effect of two analogues against 82 strains of Escherichia coli isolated form urinary tract infections . Minimal inhibitory concentrations have been measured with and without osmotic stress . The betaine analogues have antibacterial effect against E . coli strains, but only in presence of an osmotic stress.

J Bacteriol, 2003 Nov, 185(21), 6287 - 94
Molecular characterization of the Mg2+-responsive PhoP-PhoQ regulon in Salmonella enterica; Lejona S et al.; The PhoP/PhoQ two-component system controls the extracellular magnesium deprivation response in Salmonella enterica . In addition, several virulence-associated genes that are mainly required for intramacrophage survival during the infection process are under the control of its transcriptional regulation . Despite shared Mg(2+) modulation of the expression of the PhoP-activated genes, no consensus sequence common to all of them could be detected in their promoter regions . We have investigated the transcriptional regulation and the interaction of the response regulator PhoP with the promoter regions of the PhoP-activated loci phoPQ, mgtA, slyB, pmrD, pcgL, phoN, pagC, and mgtCB . A direct repeat of the heptanucleotide sequence (G/T)GTTTA(A/T) was identified as the conserved motif recognized by PhoP to directly control the gene expression of the first five loci, among which the first four are ancestral to enterobacteria . On the other hand, no direct interaction of the response regulator with the promoter of phoN, pagC, or mgtCB was apparent by either in vitro or in vivo assays . These loci are Salmonella specific and were probably acquired by horizontal DNA transfer . Besides, sequence analysis of pag promoters revealed the presence of a conserved PhoP box in 6 out of the 12 genes analyzed . Our results strongly suggest that the expression of a set of Mg(2+)-controlled genes is driven by PhoP via unknown intermediate regulatory mechanisms that could also involve ancillary factors.

Vet Microbiol, 2003 Oct 30, 96(3), 277 - 87
IS900/ERIC-PCR as a tool to distinguish Mycobacterium avium subsp . paratuberculosis from closely related mycobacteria; Englund S; There is an increasing demand for fast and reliable methods to distinguish Mycobacterium avium subsp . paratuberculosis (M . paratuberculosis) from closely related mycobacteria and also a need for rapid strain specific typing of clinical isolates for epidemiological reasons . In the present study, the potential of rep-PCR as a fingerprinting method for M . paratuberculosis was assessed and compared to conventional RFLP . A PCR assay was designed and optimised to obtain reproducible fingerprints of mycobacterial DNA with primers targeting the enterobacterial intergenic consensus (ERIC) sequence and the M . paratuberculosis specific insertion sequence IS900 . Reproducible fingerprints were obtained with 60 strains of M . paratuberculosis, 16 strains of M . avium subsp . avium, 3 strains of M . intracellulare, and 11 other mycobacterial strains . A species-specific band pattern that was clearly distinguishable from that of other mycobacteria was obtained with M . paratuberculosis . The rep-PCR did not detect any differences among M . paratuberculosis strains of different RFLP types, and was therefore not considered as an alternative fingerprinting method . However, the species-specific band pattern make IS900/ERIC-PCR a suitable alternative for distinguishing M . paratuberculosis from other mycobacteria, especially in cases of IS900 PCR positive mycobacteria . The fingerprinting method reported was fast and easy to perform, and produced highly reproducible results.

Nefrologia, 2003 Jul-Aug, 23(4), 333 - 43
{An outbreak of gram-negative bacteremia (GNB), especially enterobacter cloacae, in patients with long-term tunnelled haemodialysis catheters}; Rodriguez Jornet A et al.; Vascular access through a venous catheter for haemodialysis is associated with increased risk of thrombosis, central venous stenosis, short access survival and inadequate dialysis . The most important catheter-related complications, which determine method survival, are infection and dysfunction . In particular, infectious episodes are in some studies the leading cause for untimely catheter removal and for catheter-related morbidity but also for morbidity in dialysis patients . Double-lumen central venous catheters used for haemodialysis, are common causes of septicaemia . Most cases are caused by staphylococci . Episodes of gram-negative bacteriemia have been traced to bacterial contamination of water and/or dialysate, errors in dialyzer reprocessing, and improper setup procedures . In this paper, we describe and outbreak of gram-negative bacteremia, firstly E . cloacae, in an outpatients haemodialysis unit, in the patients with long-term tunnelled haemodialysis catheters . We describe the epidemic investigation that we achieved to identify the source of contaminating bacteria and the route by which bacteria gained access to the bloodstream . We prove the contamination by gram-negative bacterium of the water-distribution lines and haemodialysis machines . Moreover, E . cloacae strains isolated from the lines and machines are genotypically identical to the isolated from the patients . Also, we prove that the hands of health care personnel are unintentional carriers . The outbreak was finished when decontamination of dialysis machines was enhanced and dialyzer-priming fluid was modified.

Parasitol Res, 2003 Dec, 91(6), 520 - 4 Epub 2003 Oct 14.
Effect of native Xenorhabdus on the fitness of their Steinernema hosts: contrasting types of interaction; Sicard M et al.; Steinernema species are entomopathogenic nematodes . They are symbiotically associated with Enterobacteriaceae of the genus Xenorhabdus . These nematode-bacteria symbioses are extremely diversified and constitute an important new model in ecology and evolution to investigate symbioses between microbes and invertebrates . However, no study has so far adequately evaluated either the outcome of the interactions or the obligate nature of interactions in different Steinernema species in the same way . Studying three different species of Steinernema, we showed that symbiotic nematodes are always fitter than aposymbiotic ones . Nevertheless, we revealed contrasting types of interaction in terms of outcome and obligate nature of the interaction . Bacterial analyses showed that nematode species differed dramatically in the number of symbiotic Xenorhabdus they carried . We suggested that when the interaction appeared more facultative for a nematode species, the nematodes carried fewer Xenorhabdus cells than strongly dependent worm species . Thus, the symbiont transmission appeared to become more efficient as the relationship between the nematode and the bacteria became tighter.

J Infect, 2003 Nov, 47(4), 307 - 16
Are infections due to resistant pathogens associated with a worse outcome in critically ill patients?
Peres-Bota D, Rodriguez H, Dimopoulos G, DaRos A, Melot C, Struelens MJ, Vincent JL.
OBJECTIVES: To evaluate the outcome of critically ill patients infected with antimicrobial resistant microorganisms, and to analyse the factors involved in the development of antimicrobial resistance . METHODS: All patients admitted to a 31-bed mixed medico-surgical intensive care unit who developed a nosocomial infection were prospectively followed until discharge or death . RESULTS: Of 949 consecutive patients admitted, 186 developed a nosocomial infection: 79 with an antimicrobial-resistant pathogen and 107 with susceptible strains . The lungs were the main source of infections in both groups . The main resistant microorganisms were Enterobacter aerogenes, methicillin resistant Staphylococcus aureus (MRSA), and Enterobacter cloacae . The main susceptible microorganisms were Enterobacter spp., methicillin susceptible S . aureus (MSSA), and Proteus mirabilis . Patients infected with resistant strains had a longer length of stay prior to infection (9+/-4 vs . 5+/-3 days), longer total length of stay (18+/-16 vs . 11+/-7 days), longer duration of mechanical ventilation (12+/-15 vs . 6+/-7 days), and more severe coagulation, liver, and renal dysfunction (all p<0.05) . The maximum degrees of organ failure during the ICU stay, and the respiratory dysfunction, but not infection with a resistant pathogen, were independent predictors for death . Multivariate logistic regression revealed previous use of multiple antibiotics, duration of length of stay prior to infection, and the degree of liver failure as independent factors for development of infection with resistant organisms . CONCLUSIONS: Infection with antimicrobial resistant microorganisms is not an independent predictor for death . The development of antimicrobial resistance is related to the previous use of multiple antibiotics, the ICU length of stay, and the severity of hepatic dysfunction.

Microbes Infect, 2003 Oct, 5(12), 1057 - 63
Structural requirements for TLR4-mediated LPS signalling: a biological role for LPS modifications; Backhed F et al.; Cells of the mucosal lining are the first to encounter invading bacteria during infection, and as such, they have developed numerous ways of detecting microbial intruders . Recently, we showed that epithelial cells recognize lipopolysaccharide (LPS) through the CD14-Toll-like receptor (TLR)-4 complex . Here, we identify the substructures of LPS that are recognized by the TLR4 receptor complex . In contrast to lipid A, the O-antigen does not mediate an inflammatory response; rather it interferes with the lipid A recognition . An Escherichia coli strain genetically modified to express penta-acylated lipid A not only showed reduced immunogenicity, but was also found to inhibit pro-inflammatory signalling induced by wild-type E . coli (hexa-acylated lipid A) as well as LPS from other bacteria of the Enterobacteriaceae family . Furthermore, penta-acylated LPS from Pseudomonas aeruginosa acted as an antagonist to hexa-acylated E . coli LPS, as did E . coli, as shown by its inhibitory effect on IL-8 production in stimulated cells . Hypo-acylated lipid A, such as that of P . aeruginosa, is found in several species within the gut microflora as well as in several bacteria causing chronic infections . Thus, our results suggest that the composition of the microflora may be important in modulating pro-inflammatory signalling in epithelial cells under normal as well as pathologic conditions.

J Ayub Med Coll Abbottabad, 2003 Apr-Jun, 15(2), 24 - 7
Bacterial aetiology of osteomyelitis cases at four hospitals of Lahore; Malik F; BACKGROUND: The conclusive diagnosis of osteomyelitis requires isolation of pathogen in aspirate from bone lesion, bone debridement and blood culture . The present research was undertaken to study the microbiological pattern of cases of osteomyelitis reporting to four hospitals in Lahore . METHOD: One hundred and fifty patients of osteomyelitis were selected from outpatient departments and Orthopaedic wards of Lahore General Hospital, Sir Ganga Ram Hospital, Services Hospital and Mayo Hospital, Lahore . Specimens of pus from bone, blood and bone debridement were collected . All samples were inoculated onto two Blood Agar and one MacConkey agar plates . One Blood Agar plate was incubated anaerobically for 48 hours and the other two plates aerobically for 24 hours . Smears were made from samples and stained by the Gram's stain . The colonies obtained were processed according to the technique of Mackie and MacCartney . RESULTS: The commonest isolates belonged to the Enterobacteriaceae (32.8%), followed by Staphylococcus aureus (29.5%) . Pseudomonas aeruginosa (15.5%), anaerobes (2.6%) and miscellaneous (19.3%) . Five (2.7%) anaerobic bacteria were isolated . Anaerobic bacteria were peptostreptococci, peptococci and bacteroides either alone or as a mixed infection . CONCLUSION: The present study highlights the importance of microbiological examination of bone in cases of osteomyelitis . Different types of bacteria either alone or as a mixed infection could be the causative agent(s).

Biotechnol Adv, 1994, 12(2), 393 - 448
Structure-function relationships in microbial exopolysaccharides; Sutherland IW; Sufficient well-characterized microbial exopolysaccharides are now available to permit extensive studies on the relationship between their chemical structure and their physical attributes . This is seen even in homopolysaccharides with relatively simple structures but is more marked when greater differences in structure exist, as are found in several heteropolysaccharides . The specific and sometimes unique properties have, in the case of several of these polymers, provided a range of commercial applications . The existence of "families" of structurally related polysaccharides also indicates the specific role played by certain structures and substituents; the characteristics of several of these microbial polysaccharide families will be discussed here . Thus, microbial exopolysaccharides frequently carry acyl groups which may profoundly affect their interactive properties although these groups often have relatively little effect on solution viscosity . Xanthan with or without acylation shows marked differences in synergistic gelling with plant gluco- and galacto-mannans, although the polysaccharides with different acylation patterns show similar viscosity . Similarly "gelrite" from the bacterium originally designated as Auromonas (Pseudomonas)elodea is of greater potential value after deacetylation, when it provides a valuable gelling agent, than it is as a viscosifier in the natural acylated form . The Klebsiella type 54 polysaccharide only forms gels when it, too, has been chemically deacetylated to give a structure equivalent to the Enterobacter XM6 polymer . Both these polysaccharides form gels due to the enhanced interaction with cations following deacylation and to the conformation adopted after removal of the acyl groups . Recent work in our laboratory suggests that deacetylation of certain bacterial alginates also significantly increases ion binding by these polysaccharides, making them more similar in their properties to algal alginates even although the alginates from some Pseudomonas species lack poly-L-guluronic acid sequences . The existence within families of polysaccharides of types in which monosaccharides are altered within a specific structure, or with varying side-chains, also gives an indication of the way in which such substituents affect the physical properties of the polymers in aqueous solution.

J Vet Med B Infect Dis Vet Public Health, 2003 Sep, 50(7), 352 - 6
The effect of commercial steam pasteurization on the levels of Enterobacteriaceae and Escherichia coli on naturally contaminated beef carcasses; Minihan D et al.; The aim of the study was to assess the reduction achieved by steam pasteurization of beef carcasses of Escherichia coli, Enterobacteriaceae and total aerobic mesophilic plate counts (APCs) . In total, 30 carcass halves were exposed to steam pasteurization (90 degrees C, 10 s exposure time) and the 30 corresponding carcass halves remained as untreated controls . The neck, midline and rump were sampled on each carcass half . Significant reductions in E . coli incidence (P < 0.05) and counts, 0.5 log10 CFU 1000 cm(-2) (P < 0.05), were observed on rump sites only . Significant reductions (>0.8 log10 CFU 1000 cm(-2)) of Enterobacteriaceae were observed at all carcass sites sampled (P < 0.05) . Enterobacteriaceae reductions (>2 log10 CFU 1000 cm(-2)) were highly significant at the more contaminated sites (P < 0.001) . Reductions in total APCs were inconsistent . Steam pasteurization significantly reduced the level of E . coli and Enterobacteriaceae at more contaminated sites, but did not result in complete decontamination . Therefore, steam pasteurization should be classed as an aid to hygienic beef processing, but not as a critical control point.

J Clin Microbiol, 2003 Oct, 41(10), 4836 - 8
Dissemination of extended-spectrum beta-lactamase-producing Enterobacteriaceae in pediatric intensive care units; Wu TL et al.; To study the growing trend of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in pediatric intensive care units (PICUs), 88 nonrepetitive ESBL-producing isolates were prospectively collected and analyzed by molecular methods during a 16-month period . The emergence and dissemination of ESBL-producing Enterobacteriaceae in PICUs are the consequence of the clonal dissemination of a few epidemic strains along with the horizontal transmission of resistance gene-carrying plasmids among bacterial organisms.

Cell, 2003 Oct 3, 115(1), 25 - 35
Vesicle-mediated export and assembly of pore-forming oligomers of the enterobacterial ClyA cytotoxin; Wai SN et al.; The ClyA protein is a pore-forming cytotoxin expressed by Escherichia coli and some other enterobacteria . It confers cytotoxic activity toward mammalian cells, but it has remained unknown how ClyA is surface exposed and exported from bacterial cells . Outer-membrane vesicles (OMVs) released from the bacteria were shown to contain ClyA protein . ClyA formed oligomeric pore assemblies in the OMVs, and the cytotoxic activity toward mammalian cells was considerably higher than that of ClyA protein purified from the bacterial periplasm . The redox status of ClyA correlated with its ability to form the oligomeric pore assemblies . In bacterial cells with a defective periplasmic disulphide oxidoreductase system, the ClyA protein was phenotypically expressed in a constitutive manner . The results define a vesicle-mediated transport mechanism in bacteria, and our findings show that the localization of proteins to OMVs directly may contribute to the activation and delivery of pathogenic effector proteins.

Sante, 2003 Apr-Jun, 13(2), 107 - 12
{Enterobacterial susceptibility to antibiotics in northern Lebanon (1998-2001)}; Hamze M et al.; The purpose of this study was to assess the epidemiology of local enterobacterial susceptibility to antibiotics . Between 1 January, 1998, and 31 December, 2001, we studied the sensitivity of 2,238 Enterobacteria to 26 different antibiotic agents in northern Lebanon, in the Microbiology department and Laboratory of the Islami Hospital, Tripoli, Lebanon . We used the diffusion disk method and complied with the guidelines of the French Microbiology Society antibiogram committee . Urinary samples were the most frequent source (67.5%), followed by blood cultures (12.7%) . The dominant species in blood cultures was S . typhi (44.7%) . We found 194 strains that produced extended-spectrum beta lactamases (ESBL), with the highest prevalence in Serratia spp . (44.3%), followed by Klebsiella pneumoniae (23.7%), Escherichia coli (20.7%) and Klebsiella oxytoca (11.3%) . The global susceptibility of these strains to aminopenicillin was 15%; it reached 30% when combined with clavulanic acid . Susceptibility of the ESBL strains to these agents was 0% . The global susceptibility (and that of the ESBL strains, when greater than 0%) to other antibiotics was as follows: ticarcillin 38.5%, piperacillin 38.5%, piperacillin-tazobactam 88% (64%), imipenem 99.4%, (100%), cefalexin 41%, cefoxitin 65% (40.3%), cefuroxime 75%, amikacin 89%, chloramphenicol 30%, gentamicin 78% (42%), tetracycline 28% (16%), minocycline 30% (18.4%), colistin 67% (75%), nitrofuran 40% (45%), cotrimoxazol 40% (13%), nalidixic acid 53% (5.6%), pefloxacin 63% (23%), ciprofloxacin 71% (39%), and levofloxacin 72% (47%).

J Pediatr Hematol Oncol, 2003 Oct, 25(10), 801 - 5
Continuous veno-venous hemofiltration may improve survival from acute respiratory distress syndrome after bone marrow transplantation or chemotherapy; DiCarlo JV et al.; PURPOSE: Acute respiratory distress syndrome (ARDS) may result from immunologic activity triggered by irradiation and/or chemotherapy . Hemofiltration removes plasma water and soluble components below 25 kilodaltons . The authors hypothesized that early hemofiltration might attenuate the inflammatory component of ARDS, resulting in increased survival in immunocompromised children and young adults . METHODS: Ten children (6 bone marrow transplantation, 3 chemotherapy, 1 lymphoma/hemophagocytosis) with ARDS (Pao2/Fio2 94 +/- 37 torr) received early continuous veno-venous hemodiafiltration as adjunctive therapy for respiratory failure, regardless of renal function . Six children had normal urine output and initial serum creatinine (range 0.1-1.2 mg/dL); four had renal insufficiency (initial creatinine 1.7-2.4 mg/dL) . Hemofiltration was instituted coincident with intubation . Respiratory failure was precipitated by Enterobacter sepsis in two patients and by Aspergillus in one . RESULTS: Hemodiafiltration was performed for 13 +/- 9 days . A high rate of clearance was achieved (52 +/- 17 mL/min/1.73 m2) . Duration of mechanical ventilation was 14 +/- 9 days . Nine of the 10 children were successfully extubated; 8 survived . CONCLUSIONS: Early hemofiltration may improve survival from ARDS following bone marrow transplantation or chemotherapy . Possible mechanisms include strict fluid balance, immunomodulation through filtration of inflammatory constituents, and immunomodulation through intensive extracellular water exchange that delivers biochemicals to organs of metabolism as well as the hemofilter.

Int J Antimicrob Agents, 2003 Oct, 22 Suppl 2, 73 - 8
Pivmecillinam--therapy of choice for lower urinary tract infection; Graninger W; Pivmecillinam is the pro-drug of mecillinam, a beta-lactam antibiotic with a novel site of action and with specific and high activity against Gram-negative organisms such as Escherichia coli and other Enterobacteriaceae . Since its introduction, it has been widely used for the treatment of acute lower urinary tract infections (UTI), primarily in the Nordic countries . In contrast to the increasing resistance of urinary pathogens to other beta-lactams particularly ampicillin/amoxycillin and to other UTI antibiotics such as trimethoprim and trimethoprim/sulphamethoxazole (TMP/SMX), the level of resistance has remained on a low level . Less than 2% of E . coli community isolates are resistant to mecillinam . This paper reviews the clinical data on pivmecillinam with a special focus on the safety aspects . A large number of studies from the 70s to 80s have proven the clinical efficacy and safety of pivmecillinam for empirical treatment of acute cystitis . More recent studies confirm short-term treatment with pivmecillinam results in clinical and bacteriological cure rates similar to those obtained with other UTI agents . Both clinical studies in pregnant women with UTI and large epidemiological studies have confirmed the safety of pivmecillinam used in pregnancy . In the Nordic countries pivmecillinam has been the most widely used agent for treatment of UTI in pregnancy for many years . Ecological aspects of antibiotic treatment are important both with regard to adverse effects and development of resistance due to disturbance of the normal micro flora . Studies have shown that pivmecillinam has a very minor impact on the normal oropharyngeal, intestinal and skin microflora . The clinical implications of this are a low frequency of diarrhoea and Candida vaginitis as confirmed in the clinical studies . The high and increasing level of resistance among E . coli to currently recommended first-line agents for acute cystitis requires a re-evaluation of treatment guidelines . With the low resistance, its proven efficacy and favourable safety profile, pivmecillinam is a suitable first-line agent for empirical treatment of acute cystitis.

Mikrobiologiia, 2003 Jul-Aug, 72(4), 528 - 34
{Comparative study of microbial communities from cultured and natural population of the mussel Mytilus trossulus in Peter the Great bay}; Beleneva IA et al.; The 525 strains of heterotrophic bacteria isolated from natural and cultured populations of the mussel Mytilus trossulus and the surrounding seawater were identified to a genus level on the basis of phenotypic analysis and the fatty acid composition of cell lipids . Gram-negative isolates were dominated by six genera of the family Enterobacteriaceae and by the genera Pseudoalteromonas, Vibrio, Photobacterium, Cytophaga/Flavobacterium/Bacteroides, Pseudomonas, and Moraxella, Gram-positive isolates were mainly represented by the genus Streptomyces . The taxonomic compositions of natural and cultured populations of the mussel M . trossulus in Peter the Great Bay were similar.

Enferm Infecc Microbiol Clin, 2003 Oct, 21(8), 404 - 9
{Comparative in vitro activity of garenoxacin (BMS-284756) . Sentry program, Spain (1999-2000)}; Loza E et al.; INTRODUCTION: To evaluate the in vitro activity of the new des-fluoro quinolone, garenoxacin (BMS-284756), compared to activities of ciprofloxacin, levofloxacin, and gatifloxacin in clinical isolates recovered over 1999 and 2000 within the SENTRY antimicrobial surveillance program . METHODS: Quinolone-MICs were performed using the standard NCCLS microdilution technique in 2599 isolates recovered from Hospital Ramon y Cajal (Madrid), Virgen Macarena (Sevilla), and Bellvitge (Barcelona) . RESULTS: The modal MIC range value exhibited by garenoxacin ( < or = 0.03-0.12 mg/L) for Enterobacteriaceae was similar to that of the other quinolones tested . A total of 70% of Pseudomonas aeruginosa isolates were susceptible to garenoxacin and 85% to ciprofloxacin and levofloxacin . Garenoxacin exhibited the highest activity in Staphylococcus aureus, including both methicillin-susceptible and -resistant isolates, with MIC90 values of < or = 0.03 and 2 mg/L, respectively . All Streptococcus pneumoniae isolates were susceptible to garenoxacin, regardless of their penicillin susceptibility status; in terms of MIC90, garenoxacin was 16 times more active than ciprofloxacin and levofloxacin and 4-8 times more active than gatifloxacin . All 6 ciprofloxacin-resistant S . pneumoniae strains showed garenoxacin MIC values ranging from < or = 0.03 to 0.5 mg/L . In Haemophilus influenzae and Moraxella catarrhalis, garenoxacin displayed excellent in vitro activity (MIC < or = 0.06 mg/L), similar to that of the other quinolones tested . CONCLUSIONS: Garenoxacin activity was similar to the activity of other quinolones in Enterobacteriaceae, but was lower in P . aeruginosa . Garenoxacin activity was clearly higher than that of other quinolones in gram-positive isolates, including methicillin-resistant S . aureus and S . pneumoniae with reduced penicillin susceptibility.

Cell Mol Life Sci, 2003 Aug, 60(8), 1764 - 73
Crystal structure of Enterobacter cloacae 908R class C beta-lactamase bound to iodo-acetamido-phenyl boronic acid, a transition-state analogue; Wouters J et al.; The structures of the class C beta-lactamase from Enterobacter cloacae 908R alone and in complex with a boronic acid transition-state analogue were determined by X-ray crystallography at 2.1 and 2.3 A, respectively . The structure of the enzyme resembles those of other class C beta-lactamases . The structure of the complex with the transition-state analogue, iodo-acetamido-phenyl boronic acid, shows that the inhibitor is covalently bound to the active-site serine (Ser64) . Binding of the inhibitor within the active site is compared with previously determined structures of complexes with other class C enzymes . The structure of the boronic acid adduct indicates ways to improve the affinity of this class of inhibitors . This structure of 908R class C beta-lactamase in complex with a transition-state analogue provides further insights into the mechanism of action of these hydrolases.

Prikl Biokhim Mikrobiol, 2003 Jul-Aug, 39(4), 489 - 92
{Protein hydrolysates, obtained from crustaceans by an electrochemical method, as a basis for microbiological culture media}; Nianikova GG et al.; The utility of protein hydrolysates extracted electrochemically from crustaceans (Gammarus pulex and shrimp) as major components of microbiological nutrient media was demonstrated . Saprophytic soil bacteria of the genera Bacillus and Pseudomonas and the family Enterobactericeae displayed a good growth and typical morphology of their colonies on the experimental media in question.

Kansenshogaku Zasshi, 2003 Aug, 77(8), 611 - 7
{Prostate histopathology of NIH category IV prostatitis detected by sextant prostate needle biopsy from the patients with high prostatic specific antigen}; Shimomura T et al.; Asymptomatic prostatitis is classified as category IV in NIH classification of prostatitis syndrome (1999) . No report concerning this category has been present . We investigated this category histopathologically and clinically, in order to clarify the histopathological distribution and its correlation to the clinical features, in this study . Among 785 patients who were suspected prostate cancer because of their high prostatic specific antigen (PSA) values and to have a sextant prostate needle biopsy was performed between January, 1996 and December, 2000, 88 patients (11.2%) were diagnosed as NIH category IV prostatitis (asymptomatic prostatitis) . We observed all pathological specimens stained with Hematoxylin-Eosine, and classified them into subtypes according to the classification criteria for prostatitis defined by True et al . (1999) . We also investigated the relationship between histopathological distribution and clinical features such as PSA values, PSA density, the incidence of pyuria or bacteriuria . In the histopathological study, grade distributions were 12.5% (11/88) in mild, 71.6% (63/88) in moderate, and 15.9% (14/88) in severe . Location distributions were 2.3% (2/88) in glandular, 68.2% (60/88) in periglandular, and 29.5% (26/88) in stromal . No relationship between these subtypes and clinical features was recognized statistically . However, 7 patients (7.95%) were diagnosed as prostate cancers, later . Pyuria was found in 29.1% (23/79) . Bacteriuria was present in 14.3% (11/77) . Isolated bacteria were 4 strains of Enterococcus faccalis, 2 strains of each of Pseudomonas aeruginosa and Staphylococcus aureus, and one strain of each of Escherichia coli, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Staphylococcus haemolyticus, and Staphylococcus epidermidis . Gram positive rod, and Candida sp . No relationship between these subtypes and bacterial species was recognized . These results indicated that the incidence of NIII category IV prostatits was not low without correlation to any clinical features . However, we should pay attention to the presence of prostate cancer, because a small number of the patients were diagnosed as prostate cancer, later.

BMC Infect Dis . 2003 Sep 25;3(1):22.
Dio-Sensimedia: a novel culture medium for rapid detection of extended spectrum beta-lactamases; Cagatay AA et al.; BACKGROUND: Resistance to contemporary broad-spectrum beta-lactams, mediated by extended-spectrum beta-lactamases (ESBL), is an increasing problem worldwide . Many of the emerging antimicrobial resistance problems of this decade have been characterized by difficulty in the recognition of resistance in the laboratory, particularly by rapid susceptibility test methods . The plasmid-encoded ESBL represent such a resistance phenomenon that is difficult to recognize.We compared Dio-Sensimedia-ES (DSM-ES; Diomed, Istanbul, Turkey) and Mueller-Hinton (MH) agar in the double-disk synergy test (DDST) as a novel rapid system for detecting ESBL directly from bacterial culture . METHODS: Sixty ESBL-producing Klebsiella pneumoniae isolates cultured from blood (30), endotracheal aspirates (20), urine (5) and pus (5), as well as 40 Escherichia coli isolates cultured from endotracheal aspirates (15), urine (10), blood (8) and pus (7) were studied . Isolates positive for ESBL by the combined disk tests were tested with the DDST using MH and DSM-ES agar to detect ESBL-mediated resistance in K . pneumoniae and E . coli . DSM-ES agar was also used to determine the susceptibility of Enterobacteriaceae and staphylococci . RESULTS: Among 60 ESBL-producing K . pneumoniae isolates, 59 (98.3%) were identified as ESBL-positive by the DDST using MH, and 58 (96.6%), using DSM-ES agar . Of 40 ESBL-producing E . coli isolates, 38 (95%) were ESBL-positive by the DDST on MH agar, and 37 (92.5%), on DSM-ES agar . The average incubation period required for ESBL detection by the DDST on DSM-ES agar was 4 hours . CONCLUSIONS: Since the DDST results were available within 4 hours when DSM-ES agar was used, the use of this media may significantly lower the length of hospital stay, the total cost for patient care and even the mortality rate by facilitating early treatment against ESBL-producing organismsPublication Types:

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