J AOAC Int, 2003 Jul-Aug, 86(4), 802 - 14 3M Petrifilm enterobacteriaceae count plate method for enumeration of enterobacteriaceae in selected foods: collaborative study; Silbernagel KM et al.; The practice of detecting and enumerating all oxidase-negative, glucose-fermenting-Gram-negative rods (i.e., the family Enterobacteriaceae) is used to indicate unsanitary or inadequate food processing conditions . The objective of this interlaboratory collaborative study was to evaluate and compare the methods described in Standard Methods for the Examination of Dairy Products (SMEDP) and the Compendium of Methods for the Microbiological Examination of Foods (Compendium) with a commercial product, the 3M Petrifilm Enterobacteriaceae Count Plate, for the recovery of Enterobacteriaceae in foods . Six foods--cheddar cheese, milk, flour, frozen prepared meals, frozen broccoli, and nut pieces--were analyzed for Enterobacteriaceae by 12 collaborating laboratories . For each food tested, the collaborators received 8 blind test portions consisting of a control test portion and 3 levels of inoculated test portion, each in duplicate . Each test portion was tested by the Petrifilm Enterobacteriaceae Count Plate method as well as the SMEDP or Compendium methods . The precision estimates (repeatability or within-laboratory variation, and reproducibility or between-laboratory variation) were calculated with standard statistical techniques. Curr Microbiol, 2003 Aug, 47(2), 87 - 92 2-ketogluconic acid production and phosphate solubilization by Enterobacter intermedium; Hwangbo H et al.; Enterobacter intermedium, isolated from grass rhizosphere, exhibited a strong ability to solubilize insoluble phosphate . This bacterium oxidized glucose to gluconic acid and sequentially to 2-ketogluconic acid (2-KGA), which was identified using HPLC and GC-MS . The ability of E . intermedium to solubilize phosphate and produce 2-KGA produce in broth medium containing different components was monitored with air and without air supply . With an air supply, the production of 2-KGA markedly increased to about 110 g/l at day 10 in media containing 0.2 M gluconic acid, while it was about 65 g/l without gluconic acid addition . With an air supply, the concentration of soluble phosphate significantly decreased to 200-250 mg/l in media containing 1% CaCO3, whereas it was about 1000 mg/l without CaCO3 addition . Without an air supply, the concentration of 2-KGA and phosphate were negligible throughout the culture period. Antimicrob Agents Chemother, 2003 Oct, 47(10), 3089 - 98 In vitro susceptibilities of gram-negative bacteria isolated from hospitalized patients in four European countries, Canada, and the United States in 2000-2001 to expanded-spectrum cephalosporins and comparator antimicrobials: implications for therapy; Wenzel RP et al.; Access to current antimicrobial agent surveillance data is an important prerequisite for the optimal management of patients with hospital-acquired infections . The present study used data collected in 2000 to 2001 from 670 laboratories in Europe (France, Germany, Italy, and Spain), Canada, and the United States to report on the in vitro activities of ceftriaxone, cefotaxime, and comparative agents against >125,000 isolates of gram-negative bacteria from hospitalized patients . All but two isolates of Enterobacteriaceae (one isolate of Proteus mirabilis from France and one isolate of Morganella morganii from Canada) were susceptible to imipenem . The susceptibility of Escherichia coli to ceftriaxone or cefotaxime was > or = 97% in each country, and for P . mirabilis, susceptibility was 99% in each country except Italy . In contrast, susceptibility of E . coli to ciprofloxacin varied from 80.5% (Spain) to 94.0% (France); levofloxacin susceptibility ranged from 75.2% (Spain) to 91.6% (United States) . Among Klebsiella pneumoniae and Klebsiella oxytoca isolates, ceftriaxone and cefotaxime susceptibilities ranged from 86.6 to 98.7% and 83.5 to 99.7%, respectively, depending upon the country . Considerable geographic variation in the susceptibilities (generally 85 to 95% susceptible) of Serratia marcescens and M . morganii to ceftriaxone and cefotaxime were observed . For S . marcescens, susceptibility to piperacillin-tazobactam varied from 81.5% (France) to 94.1% (Italy) and susceptibility to ciprofloxacin ranged from 66.2% (Germany) to 90.7% (Spain) . Enterobacter cloacae and Enterobacter aerogenes were less susceptible to ceftriaxone and cefotaxime than were the other species of Enterobacteriaceae studied . The present study demonstrated that established parenteral expanded-spectrum cephalosporin antimicrobial agents retain significant in vitro activity against many clinically important gram-negative pathogens. Toxicon, 2003 Sep 15, 42(4), 359 - 66 Identification of Enterobacter bacteria as saxitoxin producers in cattle's rumen and surface water from Venezuelan Savannahs; Sevcik C et al.; We have previously shown that a paralytic toxin able to block sodium channels in nerve is associated with a cattle disease known as bovine paraplegic syndrome (BPS) {Toxicon . 31 (1993) 1581} . We have now identified this as saxitoxin (STX) using HPLC by either the methods of {Toxicon . 31 (1993) 1581}, or {Toxicon . 25 (1987) 1105} . In recent experiments we were able to collect and cultivate facultative anaerobic bacteria growing on rumen, grass and ponds of corrals with high incidence of BPS; the cultured bacteria produce compounds indistinguishable from STX under both HPLC procedures described above . Two species of the Enterobacter genus (E . asburiae and E . cloacae) and a strain of Klebsiella pneumoniae, were identified using standard biochemical criteria as well as gas chromatography of bacterial lipids . All these bacteria produced STX in aerobic cultures. Chemotherapy, 2003 Sep, 49(5), 229 - 36 Beta-lactamase stability of faropenem; Dalhoff A et al.; Faropenem (FAR) is an orally available member of the penem class unique among carbapenems and other available beta-lactams . This study compared FAR to cephalosporins and imipenem with respect to beta-lactamase (BLA) stability and emergence of resistance to Staphylococcus aureus and Escherichia coli . BLA stability was studied using enzyme preparations from sonicated/centrifuged 24-hour cultures of E . coli, Enterobacter cloacae, Proteus vulgaris, Providencia rettgeri, Klebsiella pneumoniae, S . aureus, and Bacteroides fragilis grown in the presence of 20 mg/l ampicillin or cephaloridine to induce penicillinase or cephalosporinase, respectively . Substrate hydrolysis was quantitated spectrophotometrically . Multistep acquisition of resistance was promoted by growing bacteria in broth containing 2-fold dilutions of antibiotic over 10 cycles . Aliquots from test tubes with visible growth provided the inoculum for the next series of dilutions . FAR as well as other cephalosporins tested were highly stable to penicillinase derived from S . aureus and E . coli . However, E . coli- and P . vulgaris-derived cephalosporinase hydrolyzed cephaloridine, cefaclor and cefotiam considerably, whereas FAR was highly stable . FAR was highly stable against hydrolysis by various BLAs prepared from four B . fragilis strains and the rate of FAR hydrolysis by metallo-BLA was 5 times lower than that for imipenem . Additionally, the acquisition of resistant S . aureus strains was less pronounced for FAR compared to other agents tested . MICs rose 8-fold after the 10th sub-MIC exposure, while MICs rose 16-, 31- and 512-fold for cefixime, cefazolin and cefaclor, respectively . E . coli shifts in MICs were moderate for all the agents tested . In conclusion, FAR is characterized by pronounced BLA stability compared to other cephalosporins and imipenem . Furthermore, a lower propensity for resistance development with FAR as compared to cephalosporins was observed . J Food Prot, 2003 Sep, 66(9), 1716 - 9 Effect of microorganism characteristics on leak size critical to predicting package sterility; Keller S et al.; The effects of microorganism size and motility on the leak size critical to the sterility of a package, along with the imposed pressure required to initiate liquid flow for the critical leak size, were measured . Pseudomonas fragi Lacy-1052, Bacillus atrophaeus ATCC 49337, and Enterobacter aerogenes ATCC 29007 were employed to assess package sterility . One hundred twenty-six 7-mm-long microtubes with interior diameters of 5, 10, and 20 microm were used to simulate package defects . Forty-two solid microtubes were used as controls . No significant differences were found between sizes or motility statuses of test organisms with respect to loss of sterility as a result of microbial ingress into test cells with microtube interior diameters of 5, 10, and 20 microm (P > 0.05) . Interactions between the initiation of liquid flow as a result of applied threshold pressures and sterility loss for test cells were significant (P < 0.05). Ann Hematol, 2003 Oct, 82 Suppl 2, S167 - 74 Epub 2003 Sep 09. Antimicrobial therapy of febrile complications after high-dose chemo-/radiotherapy and autologous hematopoietic stem cell transplantation--guidelines of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO); Bertz H et al.; Infectious complications occur in 60-100% of patients following high-dose chemotherapy (HDC) and autologous hematopoietic stem cell transplantation (HSCT), and are commonly caused by Gram-negative aerobic bacteria (such as Pseudomonas aeruginosa and enterobacteriacea e) and Gram-positive cocci (such as enterococci, streptococci and staphylococci), which should be covered by empiric first-line antibiotic therapy . Less frequently, infections are caused by fungi and anaerobic bacteria, and initial therapy does not necessarily have to cover coagulase-negative staphylococci, oxacillin-resistant S . aureus (MRSA), anaerobic bacteria and fungi . Patients who already receive antibiotics and develop pulmonary infiltrates should immediately be treated with systemic antifungals . Patients with fever and diarrhea or other signs and symptoms of gastrointestinal or perianal infection should be treated with antibiotics covering anaerobic bacteria and enterococci . Clinically stable patients with skin infections or central venous catheter-related infections can be treated with standard empiric antibiotic therapy including a beta-lactam active against Pseudomonas aeruginosa with or without an aminoglycoside, and should only receive glycopeptides if they do not respond to first-line therapy within 72 hours, become clinically unstable, have severe mucositis, or when resistance against the empiric antibiotics is demonstrated . Recombinant hematopoietic growth factors should not be added routinely but may be considered in life-threatening situations such as invasive pulmonary mycoses or sepsis. Int J Antimicrob Agents, 2003 Sep, 22(3), 270 - 3 4-alkoxy and 4-thioalkoxyquinoline derivatives as chemosensitizers for the chloramphenicol-resistant clinical Enterobacter aerogenes 27 strain; Gallo S et al.; Enterobacter aerogenes is a Gram-negative bacteria frequently responsible for nosocomial respiratory tract infections . Strains resistant to chloramphenicol are frequently isolated . Alkoxy and thio-alkoxyquinolines have a potential to act as chemosensitizers that would render multi-drug-resistant (MDR) bacterial infections susceptible to antibiotics to which they were originally resistant . Several new quinoline derivatives have been prepared, characterized and studied for their ability to increase chloramphenicol sensitivity of E . aerogenes 27, a clinical strain that exhibits the MDR phenotype . Drugs investigated were either quinoline ethers or quinoline thio-ethers . Thio-ethers are much more efficient in increasing chloramphenicol sensitivity than other corresponding ethers . In particular, 4-piperidinoethylthio-quinoline increases the strain sensitivity to chloramphenicol by about 20 times at 2 mM concentration . Similarly, sensitivity to quinolone antibiotics dramatically increases . Because these quinoline derivatives act as inhibitors of the drug efflux pump responsible for bacterial resistance to chloramphenicol, they may serve as adjunct to conventional therapy of E . aerogenes infections. Int J Antimicrob Agents, 2003 Sep, 22(3), 265 - 9 Thanatin activity on multidrug resistant clinical isolates of Enterobacter aerogenes and Klebsiella pneumoniae; Pages JM et al.; Efflux pumps protect bacterial cells by ejecting intracellular toxic molecules such as antibiotics, detergents and defensins that have penetrated the cell envelope . Some of these efflux pumps recognise structurally unrelated compounds (mdr pump) and account for the resistance of some organisms to two or more agents . It would be of interest to identify molecules that are able to circumvent the problems created by multidrug resistance phenotypes during antibiotic therapy . We have studied the activity of thanatin, a 21-residue cationic antimicrobial peptide produced by an insect, against three bacterial species . The antibacterial effect depended on the size of lipopolysaccharide side chains . In clinically resistant isolates of Enterobacter aerogenes and Klebsiella pneumoniae, the biological activity of thanatin is independent of the membrane permeability, possibly controlled by one or more porins, and/or the expression of drug efflux pumps, two mechanisms which confer high level antibiotic resistance . In addition, thanatin was able to improve the activity of structurally unrelated antibiotics (norfloxacin, chloramphenicol, tetracycline) on a multidrug- resistant E . aerogenes clinical isolate. Eur J Intern Med, 2003 Aug, 14(5), 341 - 343 Infections in diabetic foot ulcers; Candel Gonzalez FJ et al.; Foot ulcers and infections are common in diabetic patients . A 30-month-long descriptive study was conducted in our hospital in which we analyzed microbiological isolates of all patients admitted with diabetic foot infections . The predominant flora identified were Staphylococcus aureus and coagulase-negative Staphylococcus, followed by Enterococcus spp., Streptococcus spp., and enterobacteriaceaes . In 27 positive cultures (42%) polymicrobial flora were found . There were only 5% anaerobic bacteria . There appears to be a relationship between the sample collection system and microbiological isolates. J Biol Chem, 2003 Dec 5, 278(49), 49215 - 22 Epub 2003 Sep 16. A new view of the mechanisms of UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) and 5-enolpyruvylshikimate-3-phosphate synthase (AroA) derived from X-ray structures of their tetrahedral reaction intermediate states; Eschenburg S et al.; UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) and 5-enolpyruvylshikimate-3-phosphate synthase (AroA) constitute the small enzyme family of enolpyruvyl transferases, which catalyze the chemically unusual reaction of enolpyruvyl transfer . MurA catalyzes the first step in the biosynthesis of the bacterial cell wall; AroA is the sixth enzyme of the shikimate pathway leading to the synthesis of aromatic compounds in numerous microorganisms and plants . Because both metabolic pathways are absent from mammals but essential for the growth of microorganisms, MurA and AroA are attractive targets for the development of novel antimicrobial drugs . We have determined the x-ray structures of the D305A mutant of Enterobacter cloacae MurA and the D313A mutant of Escherichia coli AroA, both of which crystallized in the presence of their substrates . The structures depict the tetrahedral reaction intermediate states of the enzymes and prove that, without the aspartate side chain, the overall addition-elimination reaction in both enzymes is halted after the addition step . The presented structures lead to a new view of the catalytic mechanism and, moreover, provide an ideal starting point for the rational design of potent inhibitors of MurA and AroA. Rev Esp Quimioter, 2003 Jun, 16(2), 233 - 8 {Prevalence and susceptibility patterns of extended-spectrum betalactamase-producing Escherichia coli and Klebsiella pneumoniae in a general university hospital in Beirut, Lebanon}; Daoud Z et al.; Extended-spectrum betalactamases (ESBLs) are recognized worldwide as a problem in hospitalized patients . Their prevalence among clinical isolates of Enterobacteriaceae varies between countries and institutions . We studied the evolution of ESBL production by clinical isolates of Escherichia coli and Klebsiella pneumoniae and analyzed the patterns of susceptibility of these isolates to different antimicrobial agents in a general university hospital in Beirut . Of the 4299 isolates of E . coli and 1248 isolates of K . pneumoniae tested over the five years, 2.0% of the E . coli and 20.0% of K . pneumoniae were ESBL producing . A clear decrease in the susceptibility to all antibiotics was observed between 1999 and 2001, and no resistance to imipenem was detected . The isolates were distributed between the Intensive Care Unit (ICU), medical wards, outpatients, and other origins . The highest numbers were found in the ICU (E . coli 28.1% and K . pneumoniae 34.8%) . Three phenotypes of resistance to cefotaxime and ceftazidime were observed on the basis of microbiological results . The present study was the first to assess the occurrence and susceptibility patterns of extended-spectrum betalactamase-producing Enterobacteriaceae in Lebanon. J Antimicrob Chemother, 2003 Oct, 52(4), 591 - 7 Epub 2003 Sep 12. Evaluation of Oxoid combination discs for detection of extended-spectrum beta-lactamases; De Gheldre Y et al.; OBJECTIVES: We evaluated the reliability of cefpirome/clavulanate (CD04) compared with ceftazidime/clavulanate (CD02) and cefotaxime/clavulanate (CD03) Oxoid combination discs for the detection of extended-spectrum beta-lactamases (ESBL) in several Enterobacteriaceae isolates, including Enterobacter spp . METHODS: Overall, a total of 105 ESBL-positive {positive double-disc synergy test (DDST)} and 94 ESBL-negative (negative DDST) Gram-negative isolates were evaluated . Ninety-eight isolates were confirmed as ESBL-positive on the basis of the sequence alignments of the blaTEM and/or blaSHV gene amplification products, which matched with previously identified ESBLs . The phenotypic detection of ESBLs was performed by the three combination discs according to the NCCLS and BSAC methods . The CD04 disc was evaluated with the manufacturer's recommended zone size difference breakpoint of > or =4 mm . RESULTS: In Escherichia coli and Klebsiella spp., the sensitivities (%)/specificities (%) of CD02, CD03 and CD04 discs, and the combination of CD02 or CD04 discs, were, respectively, 88/92, 90/92, 95/84 and 100/82, while the corresponding figures were 94/100, 4/100, 94/100 and 100/100 in Enterobacter aerogenes . NCCLS and BSAC methods yielded concordant results in 99% of the isolates . CONCLUSIONS: CD04 and CD02 discs were the best combination for detection of ESBLs in our collection of Enterobacteriaceae isolates, including E . aerogenes. Lett Appl Microbiol, 2003, 37(4), 275 - 80 The Yersinia HPI is present in Serratia liquefaciens isolated from meat; Olsson C et al.; AIMS: The aim of the study was to screen the Enterobacteriaceae flora of meat for the presence of bacteria harbouring the Yersinia high-pathogenicity island (HPI) . METHODS AND RESULTS: Bacteria from 29 meat and 29 liver samples were isolated on violet-red bile glucose agar . A total of 197 isolates were screened for the presence of the irp2 gene, encoded within the HPI, by PCR . One isolate that was positive for irp2 gene was also positive for the fyuA, irp1, ybtP/ybtQ, ybtX/ybtS and int/asn tRNA genes by PCR . The presence of fyuA, irp1 and irp2 genes was confirmed by Southern hybridization . CONCLUSIONS: The isolate was identified as Serratia liquefaciens by sequencing of the 16S rRNA gene and by ribotyping . SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a Serratia harbouring the Yersinia HPI . Serratia is a frequently occurring Enterobacteriaceae genus in chill-stored meat. J Appl Microbiol, 2003, 95(4), 744 - 52 Characterization of blaCMY-10 a novel, plasmid-encoded AmpC-type beta-lactamase gene in a clinical isolate of Enterobacter aerogenes; Lee SH et al.; AIMS: We report the description of a novel plasmid-encoded AmpC beta-lactamase gene (blaCMY-10) from Enterobacter aerogenes K9911729 that was isolated from a patient suffering from pneumonia in South Korea . METHODS AND RESULTS: Using antibiotic susceptibility testing, plasmid analysis, transconjugation and Southern blot analysis, the cefoxitin resistance phenotype reflects the presence of a large plasmid {pYMG-1 (130 kb)} in Ent . aerogenes K9911729 . One beta-lactamase with the pI of 8.0 from transconjugant of Ent . aerogenes K9911729 was identified by isoelectric focusing on a gel . A 1475 bp DNA fragment containing the blaCMY-10 gene, identified on pYMG-1 of Ent . aerogenes K9911729, was sequenced and an open reading frame coding for 382 amino acid, CMY-10, was found . The 37 class C beta-lactamases were subclassified into 1a to 1j and CMY-10 into 1a by phylogenetic analysis . A sequence identical to the common regions in In6, In7 and a novel integron from pSAL-1 was found upstream from blaCMY-10 gene at nucleotide 1-71 . CONCLUSIONS: These results clearly show that blaCMY-10 gene belongs to the group of ampC-related bla genes . Homology analysis among AmpC enzymes or ampC genes implied that integration of the chromosomal ampC gene into a large resident plasmid, followed by transconjugation, was involved in the evolution of blaCMY-10 gene . SIGNIFICANCE AND IMPACT OF THE STUDY: The first identification of the blaCMY-10 gene is of concern as chromosomal beta-lactamases may cause serious therapeutic problems if their genes are translocated onto plasmids. J Appl Microbiol, 2003, 95(4), 664 - 76 Investigation of antibiotic and antibacterial agent cross-resistance in target bacteria from homes of antibacterial product users and nonusers; Cole EC et al.; AIM: To describe the relationship between antibiotic and antibacterial resistance in environmental and clinical bacteria from home environments across geographical locations, relative to the use or nonuse of antibacterial products, with a focus on target organisms recognized as potential human pathogens . METHODS AND RESULTS: In a randomized study, environmental and clinical samples were collected from the homes of antibacterial product users (n=30) and nonusers (n=30) for the isolation of target bacteria for antibiotic and antibacterial testing in three geographical areas (in USA and UK) . Isolates were tested for antibiotic susceptibility, with selected antibiotic-resistant and antibiotic-susceptible isolates tested against four common antibacterial agents (triclosan, para-chloro-meta-xylenol, pine oil and quaternary ammonium compound) . Prequalified users and nonusers at each location were randomly selected after meeting exclusionary criteria . Of 1238 isolates, more target bacteria were recovered from nonuser than user homes . Of Staphylococcus aureus isolates (n=33), none showed resistance to oxacillin or vancomycin; for Enterococcus sp . (n=149), none were resistant to ampicillin or vancomycin; and for Klebsiella pneumoniae (n=54)and Escherichia coli (n=24), none were resistant to third generation cephalosporins . Antibiotic resistance to one or more of the standard test panel drugs for Gram-positive and Gram-negative target bacteria was comparable between nonuser and user homes for both environmental and clinical isolates {e.g . resistance of environmental coagulase-negative (CN) Staphylococcus sp . was 73.8% (124/168) from nonuser homes and 73.0% (111/152) from user homes, and Enterobacteriaceae other than E . coli, 75.9% (186/245) from nonuser homes compared with 78.0% from user homes} . Of 524 Gram-negatives tested against preferred/alternative drugs, 97.1% (509/524) were susceptible to all antibiotics, across both groups . Isolates of S . aureus, Enterococcus sp . and CN Staphylococcus sp . susceptible to all preferred treatment drugs showed comparable antibacterial minimum inhibitory concentration (MIC) results between nonuser and user home isolates . For Gram-positives resistant to one or more preferred drugs, greatest resistance to antibacterial active ingredients was found in the nonuser group . For Gram-negatives, the antibacterial MIC data were comparable for isolates that were fully susceptible and resistant to one or more preferred/alternative treatment antibiotics . CONCLUSIONS: The results showed a lack of antibiotic and antibacterial agent cross-resistance in target bacteria from the homes of antibacterial product users and nonusers, as well as increased prevalence of potential pathogens in nonuser homes . SIGNIFICANCE AND IMPACT OF THE STUDY: It refutes widely publicized, yet unsupported, hypotheses that use of antibacterial products facilitates the development of antibiotic resistance in bacteria from the home environment. J Anim Sci, 2003 Sep, 81(9), 2270 - 8 Nutritional evaluation of egg byproducts in diets for early-weaned pigs; Schmidt LS et al.; A total of 272 Cotswold pigs (17 +/- 1 d) were utilized in three experiments to evaluate the nutritive value of spray-dried egg proteins for early-weaned pigs . In all experiments, pigs were stratified by sex and initial BW and then assigned randomly to experimental diets . In Exp . 1, four corn-soybean meal-based diets containing 7% of either spray-dried porcine plasma (SDPP), spray-dried technical albumen (SDTA), SDTA stored at 70 degrees C for 3 d (SDTA-ht), or spray-dried whole egg (SDWE) were assigned to five pens each with four pigs for a 3-wk study period . Average daily gain, ADFI, and gain:feed ratio (G:F) were determined . At the end of wk 3, five pigs per treatment were killed to determine ileal AA and energy digestibilities, as well as Enterobacteriaceae counts . Compared with the SDPP diet, ADG and G:F were lower (P < 0.05) for SDTA-, SDTA-ht- and SDWE-containing diets . Apparent ileal digestibilities of cystine, histidine, isoleucine, methionine, and threonine in the SDPP diet were lower (P < 0.05) than in diets containing spray-dried egg products . Ileal digestible energy content did not differ (P > 0.05) in all diets (3.1 to 3.2 Mcal/kg) . Enterobacteriaceae counts were lower in the SDTA-ht diet than in either the SDTA or SDWE diets (P < 0.05) . In Exp . 2, the effect of substituting SDPP with varying levels of SDTA was investigated . Diets were randomly assigned to five pens (except for the 100% SDTA diet, which had four pens), each with four pigs . Average daily gain, ADFI, and G:F decreased linearly as the level of SDTA was increased in the diet (P < 0.05) . Replacing SDPP with SDTA at 25 or 50% had no effect on pig performance (P > 0.10) . In Exp . 3, phase I diets containing 0, 25, or 50% SDTA in place of SDPP (7% of the diet) were each assigned at random to eight pens each with four pigs for a 14-d period, after which all pigs were switched to a common phase II diet lacking both SDPP and SDTA for another 14 d . Average daily feed intake and ADG did not differ among all diets in phase I and II and overall (d 0 to 28) . Pigs fed the diet containing 50% SDTA in phase I had lower (P < 0.05) G:F than those fed the SDPP diet . The results indicate that technical albumen can replace 25 to 50% of SDPP in early-weaned pig diets without compromising performance, and further suggest that heat-treated SDTA may affect intestinal microbial population in pigs. Diagn Microbiol Infect Dis, 2003 Sep, 47(1), 377 - 83 Contemporary re-evaluation of the activity and spectrum of grepafloxacin tested against isolates in the United States; Gordon KA et al.; Grepafloxacin potency and spectrum of activity were re-evaluated against contemporary pathogens collected from clinical infections in 2001-2002 . A total of 995 isolates were tested for grepafloxacin by the reference agar dilution method and these results were compared to those of 25 other antimicrobial agents . Grepafloxacin activity remained comparable to that of ciprofloxacin, levofloxacin and gatifloxacin against Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae (MIC(90), 0.03-2 microg/ml; 0.0-7.7% resistance rates) . For Pseudomonas aeruginosa, grepafloxacin was active against ciprofloxacin-susceptible (MIC(90), 2 microg/ml), but not against ciprofloxacin-resistant (MIC(90), >8 microg/ml) isolates . Against methicillin-susceptible Staphylococcus aureus, grepafloxacin susceptibility rate was 91.4%, equal to that of levofloxacin . None of the fluoroquinolones showed reasonable activity against methicillin-resistant staphylococci . Gatifloxacin and grepafloxacin had the same MIC(90) against beta-hemolytic streptococci (0.25 microg/ml) and penicillin-susceptible Streptococcus pneumoniae (0.25 microg/ml) . Grepafloxacin and other fluoroquinolone activities were not influenced by penicillin resistance in S . pneumoniae . Grepafloxacin was very active against Haemophilus influenzae (MIC(90), 0.03 microg/ml), Moraxella catarrhalis (MIC(90), 0.03 microg/ml) and Legionella spp . (MIC(90), 0.5 microg/ml) . These results on recently isolated organisms indicate that grepafloxacin has a sustained potency and spectrum against most clinically important and indicated pathogens. Diagn Microbiol Infect Dis, 2003 Sep, 47(1), 361 - 4 Global patterns of susceptibility for 21 commonly utilized antimicrobial agents tested against 48,440 Enterobacteriaceae in the SENTRY Antimicrobial Surveillance Program (1997-2001); Sader HS et al.; A total of 48,440 Enterobacteriaceae isolates collected consecutively from patients hospitalized in participant SENTRY Antimicrobial Surveillance Program sites in four international regions (Asia-Pacific, Europe, Latin America, and North America) were tested by reference broth microdilution method against the most commonly used antimicrobial agents . The most active compounds could be divided in 3 groups based on their spectrum of activity . The first group included meropenem and imipenem, with 99.9% susceptibilty (S) rates for the Enterobacteriaceae . The second group includes amikacin (97.3% S) and cefepime (97.2% S); and a third active group had a rank order of susceptibility of: gatifloxacin = levofloxacin (91.7% S) > ceftazidime (91.4% S) > ceftriaxone (91.2% S) > aztreonam (91.1% S) > gentamicin (90.6% S) > piperacillin/tazobactam = ciprofloxacin (90.5% susceptibility) . These latter antimicrobial agents presented susceptibility rates of approximately 90% (89.8%-91.7%) . Continued resistance surveillance by various programs remain necessary to monitor the in vitro effectiveness of antimicrobial agents currently used in clinical practice. Emerg Infect Dis, 2003 Aug, 9(8), 999 - 1002 NmcA carbapenem-hydrolyzing enzyme in Enterobacter cloacae in North America; Pottumarthy S et al.; An imipenem-resistant Enterobacter cloacae isolate was recovered from the blood of a patient with a hematologic malignancy . Analytical isoelectric focusing, inhibitor studies, hydrolysis, induction assays, and molecular sequencing methods confirmed the presence of a NmcA carbapenem-hydrolyzing enzyme . This first report of NmcA detected in North America warrants further investigation into its distribution and clinical impact. Pharmazie, 2003 Aug, 58(8), 590 - 2 Stimulation of furanochromone accumulation in callus cultures of Ammi visnaga L . by addition of elicitors; Krolicka A et al.; In order to check the possibility of producing secondary metabolites, in vitro cultures of A . visnaga callus were established . The best growth of A . visnaga callus was obtained on Murashige and Skoog medium (MS) containing 6-benzyladenine (BA) and alpha-naphtaleneacetic acid (NAA) . The study was concentrated on the induction of production of secondary metabolites by exposing callus to abiotic elicitors: benzo(1,2,3)-thiadiazole-7-carbothionic acid S-methyl ester (BION) and a suspension of silica (SiO2) and biotic elicitors: autoclaved lysates of Enterobacter sakazaki and scleroglucan . GC analysis indicated that not-elicited callus of A . visnaga grown in darkness accumulated 2 times more visnagin than the one which was grown under a 16-h photoperiod . The highest accumulation of visnagin was observed in the callus culture elicited with scleroglucan or BION . Scleroglucan induced also the accumulation of khellin in A . visnaga callus . The presented work shows that biosynthesis of pharmacologically important secondary metabolites in A . visnaga cultures could be stimulated by application of elicitors. Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 78 - 81 {Effect of gastrointestinal tract secretions on the anti-lysozyme activity of enterobacteria}; Tarasevich AV et al.; The effect of the duodenal contents and its components on the antilysozyme activity (ALA) of enterobacteria was studied . Under the action of native bile and the preparation of dried bile the stimulation of the expression of ALA factor in 86.6-93.3% of enterobacteria was observed . Pancreatin increased the level of the secretion of ALA factor by 50% in comparison with the control . The preparation of egg lysozyme produced no changes in the frequency and level of the expression of ALA factor . The study revealed the dose-dependant character of the action of duodenal contents: in the native form, as well as at concentrations of 50% and 25%, duodenal contents stimulated the expression of the ALA factor of enterobacteria; at concentrations of 12.5% and lower, its effect could be stimulating, inhibitory or neutral. Zh Mikrobiol Epidemiol Immunobiol, 2003 Jul-Aug, (4), 23 - 7 {Molecular mechanisms of relationships between the host organism and pathogenic enterobacteria}; Tuigunov MM et al.; Modern data on the molecular mechanisms of relationships between the host organism and the pathogenic representatives of the family Enterobacteriaceae in the host-parasite system are presented . The process of cytokine and eicosanoid regulation of the immune process of the host in the norm and pathology states are analyzed . The examples of the mechanisms of immune suppression, false antigenic stimulation and the mimicry of pathogens are given. Braz J Infect Dis, 2003 Apr, 7(2), 129 - 34 Plasmid-mediated extended-spectrum beta-lactamase-producing strains of Enterobacteriaceae isolated from diabetes foot infections in a Brazilian diabetic center; Motta RN et al.; We bacteriologically analyzed 156 species of Enterobacteriaceae, isolated from 138 patients with community-acquired diabetic foot ulcers, in a prospective study made at a diabetic center and at the Federal University of Ceara, Brazil, from March, 2000, to November, 2001.The samples were cultured using selective media, and identification, susceptibility tests and detection of plasmid-mediated-extended-spectrum-beta-lactamase (ESBL) producing strains were made with conventional and automated methods . The most frequently occurring pathogens were K . pneumoniae (21.2%), Morganella morganii (19.9%) and E . coli (15.4%) . High resistance rates were noted for ampicillin, first generation cephalosporin, trimethoprim/sulfamethoxazole, tetracycline, amoxicillin-clavulanic acid and chloramphenicol . ESBL-producing strains were detected in 6% of the patients . Resistance among gram-negative bacteria has become increasingly common, even in community-acquired infections. Microb Drug Resist, 2003 Fall, 9(3), 299 - 305 Outbreak of TEM-24-producing Enterobacter aerogenes in a Spanish hospital; Salso S et al.; Organisms encoding multidrug resistance genes are becoming increasingly prevalent . During a 2-month period (December, 2000, to January, 2001), 83 consecutive isolates of Enterobacter spp . were collected in our microbiology department . Antibiotic susceptibility was determined using the Vitek II automatic system . We selected strains with decreased susceptibility to extended-spectrum cephalosporins . The double-disk potentiation test was positive in 10 of these strains, indicating the presence of extended-spectrum beta-lactamases (ESBLs) . Polymerase chain reaction (PCR), isoelectric focusing (IEF), and sequencing identified TEM 24 beta-lactamase in the 10 selected E . aerogenes . Random amplification of polymorphic DNA (RAPD-PCR) revealed the same clonal origin for all the strains tested and strongly suggest an outbreak of multidrug-resistant E . aerogenes . To follow up the trends in ESBLs-producing Enterobacter infections in the hospital over time, we repeated the study 1 year later (December, 2001, to February, 2002) . Only three ESBLs-producing Enterobacter were found . All of them corresponded to the previously characterized clone. Pneumonol Alergol Pol, 2003, 71(1-2), 59 - 67 {Etiologic factors of pleural empyema as a potential source of nosocomial infections}; Jezierska-Anczukow A et al.; Pleural empyema still comprises an important therapeutic problem despite the availability of effective antibiotic therapy . This disorder is characterised by 20% mortality rate . Moreover, the involvement of multi-drug resistant bacterial strains may pose a risk of a nosocomial spread to other hospitalised patients . In the present study we have analysed 184 bacterial strains isolated from 63 patients with pleural empyema . A predominance of aerobic bacteria was detected, both Gram-positive cocci and Gram-negative bacilli . Staphylococci isolated from the clinical samples were characterised by a high percentage of strains resistant to gentamicin (86.3%) and methicillin (38.2%) . The other important etiological agents were Pseudomonas aeruginosa and Gram-negative enteric rods of the Enterobacteriaceae family . Only 1 strain of anaerobic bacteria was detected . The enteric bacilli were characterised by a low percentage of isolates susceptible to most of the tested antibiotics (< 60%), with an exception of ciprofloxacin (68%) and imipenem (100%) . These multi-drug resistant strains may spread nosocomially to other patients. J Clin Microbiol, 2003 Sep, 41(9), 4264 - 9 Multiple CTX-M-type extended-spectrum beta-lactamases in nosocomial isolates of Enterobacteriaceae from a hospital in northern Italy; Pagani L et al.; Twelve isolates of Enterobacteriaceae (1 of Klebsiella pneumoniae, 8 of Escherichia coli, 1 of Proteus mirabilis, and 2 of Proteus vulgaris) classified as extended-spectrum beta-lactamase (ESBL) producers according to the ESBL screen flow application of the BD-Phoenix automatic system and for which the cefotaxime MICs were higher than those of ceftazidime were collected between January 2001 and July 2002 at the Laboratory of Clinical Microbiology of the San Matteo University Hospital of Pavia (northern Italy) . By PCR and sequencing, a CTX-M-type determinant was detected in six isolates, including three of E . coli (carrying bla(CTX-M-1)), two of P . vulgaris (carrying bla(CTX-M-2)), and one of K . pneumoniae (carrying bla(CTX-M-15)) . The three CTX-M-1-producing E . coli isolates were from different wards, and genotyping by pulsed-field gel electrophoresis (PFGE) revealed that they were clonally unrelated to each other . The two CTX-M-2-producing P . vulgaris isolates were from the same ward (although isolated several months apart), and PFGE analysis revealed probable clonal relatedness . The bla(CTX-M-1) and bla(CTX-M-2) determinants were transferable to E . coli by conjugation, while conjugative transfer of the bla(CTX-M-15) determinant from K . pneumoniae was not detectable . Present findings indicate that CTX-M enzymes of various types are present also in Italy and underscore that different CTX-M determinants can be found in a single hospital and can show different dissemination patterns . This is also the first report of CTX-M-2 in P . vulgaris. Appl Environ Microbiol, 2003 Sep, 69(9), 5364 - 71 Morphological, host range, and genetic characterization of two coliphages; Goodridge L et al.; Two coliphages, AR1 and LG1, were characterized based on their morphological, host range, and genetic properties . Transmission electron microscopy showed that both phages belonged to the Myoviridae; phage particles of LG1 were smaller than those of AR1 and had an isometric head 68 nm in diameter and a complex contractile tail 111 nm in length . Transmission electron micrographs of AR1 showed phage particles consisting of an elongated isometric head of 103 by 74 nm and a complex contractile tail 116 nm in length . Both phages were extensively tested on many strains of Escherichia coli and other enterobacteria . The results showed that both phages could infect many serotypes of E . coli . Among the enterobacteria, Proteus mirabilis, Shigella dysenteriae, and two Salmonella strains were lysed by the phages . The genetic material of AR1 and LG1 was characterized . Phage LG1 had a genome size of 49.5 kb compared to 150 kb for AR1 . Restriction endonuclease analysis showed that several restriction enzymes could degrade DNA from both phages . The morphological, genome size, and restriction endonuclease similarities between AR1 and phage T4 were striking . Southern hybridizations showed that AR1 and T4 are genetically related . The wide host ranges of phages AR1 and LG1 suggest that they may be useful as biocontrol, therapeutic, or diagnostic agents to control and detect the prevalence of E . coli in animals and food. Appl Environ Microbiol, 2003 Sep, 69(9), 5306 - 18 Population genetics of the nomenspecies Enterobacter cloacae; Hoffmann H et al.; The genetic heterogeneity of the nomenspecies Enterobacter cloacae is well known . Enterobacter asburiae, Enterobacter cancerogenus, Enterobacter dissolvens, Enterobacter hormaechei, Enterobacter kobei, and Enterobacter nimipressuralis are closely related to it and are subsumed in the so-called E . cloacae complex . DNA-DNA hybridization studies performed previously identified at least five DNA-relatedness groups of this complex . In order to analyze the genetic structure and the phylogenetic relationships between the clusters of the nomenspecies E . cloacae, 206 strains collected from 22 hospitals, a veterinarian, and an agricultural center in 11 countries plus all 13 type strains of the genus and reference strain CDC 1347-71(R) were examined with a combination of sequence and PCR-restriction fragment length polymorphism (PCR-RFLP) analyses of the three housekeeping genes hsp60, rpoB, and hemB as well as ampC, the gene of a class C beta-lactamase . Based on the neighbor-joining tree of the hsp60 sequences, 12 genetic clusters (I to XII) and an unstable sequence crowd (xiii) were identified . The robustness of the genetic clusters was confirmed by analyses of rpoB and hemB sequences and ampC PCR-RFLPs . Sequence crowd xiii split into two groups after rpoB analysis . Only three strains formed a cluster with the type strain of E . cloacae, indicating that the minority of isolates identified as E . cloacae truly belong to the species; 13% of strains grouped with other type strains of the genus, suggesting that the phenotypes of these species seem to be more heterogeneous than so far believed . Three clusters represented 70% of strains, but none of them included a type or reference strain . The genetic clustering presented in this study might serve as a framework for future studies dealing with taxonomic, evolutionary, epidemiological, or pathogenetic characteristics of bacteria belonging to the E . cloacae complex. Lancet Infect Dis, 2003 Sep, 3(9), 537 - 46 Fluoroquinolones in paediatrics: a risk for the patient or for the community? Gendrel D, Chalumeau M, Moulin F, Raymond J. Fluoroquinolones are an important group of antibiotics widely used in adult patients because of their excellent tissue penetration and their bactericidal activity . They are not authorised for paediatric use (except the limited indication of pseudomonas infections in cystic fibrosis), however, because of the potential for joint toxicity reported from experiments with young animals . Despite the absence of official approval, fluoroquinolones are widely used in paediatrics as second-line antibiotics when all other treatments have failed . Most of the information available about paediatric use concerns ciprofloxacin, which is used in children much more often than the other members of this class . The published paediatric series have shown that frequency of articular side-effects varies according to age: all the surveys have reported frequencies of around 0.1% in adults and 2-3% in children . Outside of cystic fibrosis and severe infections in which no other treatment is possible, the only paediatric situations where fluoroquinolones are superior to standard treatments for children, in speed of recovery and comfort as well as in efficacy, are typhoid fever, severe shigella dysenteries, and enterobacteria meningitis . Should the use of new fluoroquinolones active against pneumococci be authorised for upper respiratory infections (including recurrent otitis) in children, the potential emergence and dissemination of pneumococci strains in which multidrug resistance includes fluoroquinolones would create a real risk in the community . It is, therefore, important to continue the policy of second-line use in children, only after failure of an earlier treatment, and when other antibiotics approved for paediatric use cannot be used. Eur J Epidemiol, 2003, 18(7), 703 - 10 Comparative study of three different DNA fingerprint techniques for molecular typing of Shigella flexneri strains isolated in Romania; Surdeanu M et al.; In this study, 97 epidemiologically unrelated Shigella flexneri strains isolated during 1994 (69 isolates) and 1997 (28 isolates) were characterised by ribotyping, enterobacterial repetitive intergenic consensus sequence-based PCR typing, and pulsed-field gel electrophoresis . Number of strains belonging to each of the six serotypes is selected equal to their distribution in Romania . The isolates comprise 24 ribotypes based on combination of two restriction patterns obtained with HindlII and PstI, respectively, 7 enterobacterial repetitive intergenic consensus (ERIC)-PCR types, and 92 XbaI pulsed-field gel electrophoresis (PFGE) patterns grouped in 31 pulsotypes at Dice coefficients of 85% similarity . We find no significant difference in the distribution of isolates collected during the two periods . Macrorestriction analysis by PFGE offers maximal discrimination . There seems to be little genetic variability among circulating S . flexneri strains of serotype 2a, suggesting that even a combination of several molecular techniques, including PFGE, could not easily differentiate an outbreak strain from temporally associated independent isolates. Exp Clin Endocrinol Diabetes, 2003 Aug, 111(5), 288 - 93 Lupus-like panniculitis in a patient with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED); Fuchtenbusch M et al.; Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autosomal recessive disorder, characterised by a loss of self-tolerance to endocrine tissues, chronic candidiasis and ectodermal disorders . APECED is associated with mutations of a single gene, designated autoimmune regulator (AIRE) . We describe a 31-year-old APECED patient with non-traumatic, cutaneous ulcers on both forearms with features of a lupus-like panniculitis . On admission to the ICU in September 2001, the patient suffered from a ketoacidotic, hyperglycemic coma and adrenal crisis due to an Enterobacter-cloacae sepsis, originating from multiple, necrotising deep cutaneous ulcers . These ulcers spontaneously developed on both forearms, some of which were just emerging, full blown or healing with scars . Histological examination showed signs of a scarring panniculitis and vasculitis . Immunohistochemistry and direct immunofluorescence with characterisation of immunoglobulin and complement-factor binding pattern revealed features of a lupus-like panniculitis . Sequence analysis of all 14 exons of the AIRE gene revealed a R257 X mutation in exon 6 resulting in a nonsense mutation at codon 257 confirming the diagnosis of APECED . Oral treatment with 60 mg/day corticosteroids for two weeks led to complete resolution of all ulcers . In conclusion, mutations in the AIRE gene may provide the genetic background against which additional factors can initiate an autoimmune process . Here, autoimmune panniculitis appears to be an associated feature of the APECED syndrome . Our findings support the use of immunosuppressive therapy for autoimmune disease components of the APECED syndrome. FEMS Microbiol Lett, 2003 Aug 29, 225(2), 257 - 62 Genetic evidence for the existence of two pathways for the biosynthesis of methionine in the Leptospira spp; Picardeau M et al.; There are two major pathways for methionine biosynthesis: the enterobacterial type transsulfuration pathway and the sulfhydrylation pathway as previously identified in the spirochete Leptospira meyeri . Sequence analysis of the L . meyeri metYX locus allows the identification of a third gene, called metW, which encodes a protein exhibiting similarities with homologs in many organisms belonging to the alpha-, beta-, and gamma-subdivisions of proteobacteria . The metW, metX and metY genes of L . meyeri were disrupted by a resistance cassette by homologous recombination . While the L . meyeri metX mutant shows methionine auxotrophy, the metY mutant (as well as the metW and metYmetW mutants) conserves methionine prototrophy, suggesting the presence of additional route(s) which may bypass the direct sulfhydrylation pathway . In addition, a L . interrogans gene, called metZ, was found to complement an Escherichia coli metB mutant, further suggesting that the transsulfuration pathway is also present in Leptospira spp. FEMS Microbiol Lett, 2003 Aug 29, 225(2), 201 - 6 GASP phenotype: presence in enterobacteria and independence of sigmaS in its acquisition; Martinez-Garcia E et al.; The appearance of growth advantage in stationary phase or GASP was originally detected in Escherichia coli . The presence of this phenotype in other enterobacteria such as Enterobacter cloacae, Salmonella typhimurium, Providencia stuartii and Shigella dysenteriae is described in this work . E . cloacae GASP strains presented lower levels of RpoS than the parental strain, although no mutation in the gene or its promoter was detected . This work offers evidence of GASP rpoS-independent pathways as GASP was also acquired in knock-out rpoS E . cloacae and E . coli strains. Biopolymers, 2003, 72(5), 391 - 8 Optical properties of Erwinia herbicola bacteria at 0.190-2.50 microm; Arakawa ET et al.; We measure the complex index of refraction of Erwina herbicola (also known as Enterobacter agglomerans or Pantoea agglomerans) bacteria (ATTC 33243) over the spectral region from 0.190 to 2.50 microm (4000-52,632 cm(-1)) . Transmission measurements are made on solid films of E . herbicola and on suspensions of the bacteria in water . These measurements, combined with spectral reflectance and Kramers-Kronig analysis, allow the determination of the real and imaginary parts over the entire wavelength interval . Accurate and consistent results are obtained for this complex and difficult to measure material . This is part of a continuing series of measurements of the optical constants of representative biological materials that are applicable to the development of methods for detection of airborne biological contaminants, where the material under study is used as a surrogate for a pathogenic agent . Microbiology, 2003 Sep, 149(Pt 9), 2557 - 70 The colicin G, H and X determinants encode microcins M and H47, which might utilize the catecholate siderophore receptors FepA, Cir, Fiu and IroN; Patzer SI et al.; The colicin G producer Escherichia coli CA46, the colicin H producer E . coli CA58 and E . coli Nissle 1917 (DSM 6601) were shown to produce microcin H47 and the newly described microcin M . Both microcins were exported like colicin V by an RND-type export system, including TolC . The gene cluster encoding microcins H47 and M in strains CA46 and CA58 is nearly identical to that in strain DSM 6601, except that two additional genes are included . A Fur box identified in front of the microcin-encoding genes explained the observed iron regulation of microcin production . The catecholate siderophore receptors Fiu, Cir and FepA from E . coli and IroN, Cir and FepA from Salmonella were identified as receptors for microcins M, H47 and E492 . IroN takes up the glucose-containing catecholate siderophore salmochelin, whose synthesis is encoded in the iro gene cluster found in Salmonella and certain, often uropathogenic, E . coli strains . A gene in this iro cluster, iroB, which encodes a putative glycosyltransferase, was also found in the microcin H47/M and microcin E492 gene clusters . These microcins could aid the producing strain in competing against enterobacteria that utilize catecholate siderophores. Mol Biol Evol, 2004 Jan, 21(1), 1 - 13 Epub 2003 Aug 29. Site-specific recombination links the evolution of P2-like coliphages and pathogenic enterobacteria; Nilsson AS et al.; The genome of the tailed temperate coliphage P2 (Myoviridae) contains some genes that probably are horizontally transferred additions to the genome . One of these genes, the Z/fun gene, was recently found intact in the genome of Neisseria meningitidis . We have investigated the presence of P2-like phages, and the genetic variation at the position corresponding to the phage P2 Z/fun locus, in the Escherichia coli reference collection (ECOR) . P2-like phages are common in E . coli since they are present in about 30% of the ECOR strains . Hybridizations and PCR amplifications indicate that the overall variation among these phages is small . Amplification of the region corresponding to the phage P2 Z/fun locus in 11 prophages revealed that this is a multivariable locus . Sequencing of the region resulted in 10 completely different sequences but with a similar high AT-content as the Z/fun gene . All sequences contained at least one open reading frame with good transcription and translation signals . All sequences were also surrounded by a highly similar, previously undiscovered, inverted repeat (IR) . We also found this IR in genetically unstable regions in pathogenic enterobacteria . This demonstrates that P2-like phages are important factors in the evolution of bacteria, not only because they carry a diversity of lysogenic conversion genes but also because they can act as vectors for single genes . The genes found between the IRs have unknown functions, and only a few clearly similar genes have been found in other bacteria. J Med Entomol, 2003 May, 40(3), 371 - 4 Bacteria in midguts of field-collected Anopheles albimanus block Plasmodium vivax sporogonic development; Gonzalez-Ceron L et al.; Bacterial infections were investigated in midguts of insectary and field-collected Anopheles albimanus Weidemann from southern Mexico . Serratia marcescens, Enterobacter cloacae and Enterobacter amnigenus 2, Enterobacter sp., and Serratia sp . were isolated in field samples obtained in 1998, but only Enterobacter sp . was recovered in field samples of 1997 and no bacteria were isolated from insectary specimens . These bacteria were offered along with Plasmodium vivax infected blood to aseptic insectary An . albimanus, and the number of infected mosquitoes as well as the oocyst densities assessed after 7d . Plasmodium vivax infections in mosquitoes co-infected with En . amnigenus 2, En . cloacae, and S . marcensces were 53, 17, and 210 times, respectively, lower than in control mosquitoes, and the mean oocyst density in mosquitoes co-infected with En . cloacae was 2.5 times lower than in controls . Mortality was 13 times higher in S . marcensces-infected mosquitoes compared with controls . The overall midgut bacterial infection in mosquito field populations may influence P . vivax transmission, and could contribute to explain the annual variations in malaria incidence observed in the area. Jpn J Antibiot, 2003 Jun, 56(3), 171 - 9 {Susceptibility of major pathogens of acute pharyngitis and tonsillitis to levofloxacin and other oral antimicrobial drugs}; Matsuzaki K et al.; A total of 2865 strains of the causative organisms isolated from the patients with acute pharyngitis and tonsillitis at the primary medical institutions were used in this study . The MICs of levofloxacin (LVFX) and other oral antimicrobial drugs were determined and evaluated by the NCCLS guideline . LVFX, cefditoren (CDTR) and cefcapene (CFPN) were potently active against 773 isolates of Hemophilus influenzae, the MIC50S of LVFX being < or = 0.06 microgram/mL and also the same as the MIC90S of LVFX . LVFX was the most active against 496 isolates of Enterobacteriaceae . The MIC50S of LVFX were < or = 0.06 microgram/mL and were lower than those of CDTR, cefdinir (CFDN) and cefpodoxime (CPDX) (MIC50S: 0.5 microgram/mL) . The MIC90S of these cephems were markedly higher than the respective MIC50S, whereas MIC50 of LVFX was 0.12 microgram/mL, only twice the MIC50 . Against the majority of Streptococcus pyogenes (555 isolates) and Streptococcus spp . (495 isolates), CDTR, CFDN, CPDX and CFPN were highly active (MICs: < or = 0.06 microgram/mL), and clarithromycin (CAM) and azithromycin (AZM) were also active against these organisms (MICs: 0.12 to 0.25 microgram/mL) . Against S . pneumoniae (92 isolates), CDTR and CFDN were active (MIC50S: 0.12 and 0.25 microgram/mL, respectively) . However, the MIC90S of these drugs were 4-8 times the MIC50S . Against Moraxella (Branhamella) catarrhalis (454 isolates), LVFX was potently active, the MIC90 of LVFX being < or = 0.06 microgram/mL and MIC90S of the other cephems being 0.5 microgram/mL or more . When the susceptibility of these strains to LVFX was evaluated by the NCCLS guideline, about 3% of other Streptococcus spp . were resistant to the drug but no test strains resistant to LVFX were detected in H . influenzae, S . pyogenes or Enterobacteriaceae . On the other hand, the percentages of strains susceptible to the cephems tested were 60-90%, which were quite different according to kinds of drugs and species used . Furthermore, the strains of S . pneumoniae resistant to CFDN and CPDX, and those to CAM and AZM were 21-25% and 50% or more, respectively, whereas no LVFX-resistant strains were detected . The major pathogens isolated from patients with pharyngitis and tonsillitis in the primary institutions were highly susceptible to LVFX . These results suggest that LVFX is a useful drug which is potently active against the strains resistant to oral cephem and macrolide antibiotics. J Eur Acad Dermatol Venereol, 2003 Sep, 17(5), 575 - 7 Cutaneous infection caused by Salmonella typhi; Marzano AV et al.; Salmonella typhi is a flagellated, gram-negative bacillus belonging to the family Enterobacteriaceae responsible for typhoid fever, which is a prolonged bacteraemic, systemic illness with minimal, at least initially, diarrhoea . Localized infection is a not uncommon complication of salmonella septicaemia, particularly occurring in immunocompromised patients . However, salmonella localization to the skin presenting as cutaneous ulceration is regarded as a rare event . We report a unique case of such a presentation as the sole clinical manifestation of infection with S . typhi and highlight the possible salmonella aetiology for unusual cutaneous lesions in individuals coming from endemic areas. Infect Control Hosp Epidemiol, 2003 Aug, 24(8), 607 - 12 Resistance in Enterobacteriaceae: results of a multicenter surveillance study, 1995-2000; Friedland I et al.; OBJECTIVES: To assess changes over time in susceptibility of Enterobacteriaceae from patients in ICUs, compare susceptibility rates of isolates from patients in ICUs with those from inpatients outside ICUs, and explore phenotypic patterns of cross-resistance and co-resistance . DESIGN: From 1995 to 2000, centers participating in the ICU Surveillance Study tested 100 to 200 consecutive nosocomial gram-negative bacilli by broth microdilution . SETTING: Each year, 42 to 97 U.S . hospitals tested isolates . RESULTS: In all years, imipenem was the most potent agent tested, followed by amikacin and ertapenem . Extended-spectrum beta-lactam and monobactam agents had good activity against Escherichia coli and Klebsiella species, but limited activity against Enterobacter species . Susceptibility to imipenem and amikacin did not fluctuate during the analysis period, whereas susceptibility to ceftazidime, ceftriaxone, and ciprofloxacin decreased 2% to 5% . The decline was most pronounced for susceptibility of Escherichia coli to ciprofloxacin: 98.7% of ICU isolates were susceptible in 1995 versus 93.2% in 2000 . Susceptibility of ICU isolates was lower than that of non-ICU isolates, except for ciprofloxacin, for which the reverse was true . Cross-resistance was common among extended-spectrum cephalosporins and penicillins, but uncommon between imipenem and ertapenem . Only imipenem and ertapenem remained highly active against Enterobacteriaceae with a phenotype suggesting possible production of an extended-spectrum beta-lactamase and those with a phenotype suggesting possible Amp C hyperproduction . CONCLUSIONS: Imipenem was the most active agent against nosocomial Enterobacteriaceae . Susceptibility to ciprofloxacin decreased from 1995 to 2000, particularly in Escherichia coli, and, in contrast to other agents, was lower among non-ICU isolates. Antimicrob Agents Chemother, 2003 Sep, 47(9), 2864 - 7 Extended-spectrum beta-lactamases in enterobacteriaceae in Buenos Aires, Argentina, public hospitals; Quinteros M et al.; Resistance to extended-spectrum cephalosporins is often associated with plasmid encoded extended spectrum beta-lactamases (ESBL) . In order to evaluate the prevalence and diversity of ESBLs in enterobacteria in our city, a 1-month-period survey was carried out from April to May 2000 . Extended-spectrum-cephalosporin-resistant strains, isolated from inpatient clinical specimens other than stools, were collected among 17 participating hospitals . From a total of 427 enterobacterial strains that were collected during this period, 39 were extended-spectrum cephalosporin resistant . The National Committee for Clinical Laboratory Standards' Screening and Confirmatory Tests for ESBL production were performed using cefotaxime and ceftazidime; cefepime and cefepime-clavulanic acid-containing disks were included . beta-Lactamases were characterized by isoelectric focusing and PCR amplification using specific primers . Three different ESBLs were detected: SHV-related (4 isolates), PER-2-type (9 isolates), and CTX-M-2-related (26 isolates) . Sequencing of the corresponding genes confirmed CTX-M-2 in 19 of 21 and CTX-M-31 (an allelic variant) in the remaining 2 of 21 . CTX-M-2 (or its variant) was detected in all Escherichia coli, Enterobacter aerogenes, Serratia marcescens, Proteus mirabilis, and Providencia stuartii strains, while PER-2 was detected in Enterobacter cloacae, E . aerogenes, and Klebsiella pneumoniae; SHV-related ESBL were found only in K . pneumoniae . These results clearly show that CTX-M-2 is the most prevalent ESBL produced by enterobacterial species isolated from public hospitals in Buenos Aires. Chin Med J (Engl), 2003 Aug, 116(8), 1244 - 7 The status of drug resistance and ampC gene expression in Enterobacter cloacae; Zhou Z et al.; OBJECTIVE: To investigate the status of the drug resistance and the ampC gene expression of Enterobacter cloacae . METHODS: Disk diffusion tests were made for detecting the susceptibility of antimicrobial agents against Enterobacter cloacae . AmpC gene was amplified by polymerase chain reaction (PCR) and verified by DNA sequencing . AmpC gene expression was analyzed according to antimicrobial agent sensitive phenotype . RESULTS: The sensitivity rates of 144 strains to imipenam, cefepime and cefoperazone/sulbactam were 98.61%, 65.97% and 63.89%, respectively . The sensitivity rates of 144 strains to other antimicrobial agents were lower . Among the 144 strains 120 were found to be positive by PCR for ampC . The PCR product showed high homology to the GenBank ampC sequence . Stably derepressed strains, hyperinducible strains and unexpressing or lower level expressing strains accounted for 30.0% (36/120), 37.5% (45/120), and 32.5% (39/120), respectively . Fifty-six out of 120 strains (46.67%) also produced extended spectrum beta-lactamases (ESBLs) . The hyperinducible strains were highly sensitive to all the antimicrobial agents except amoxicillin/clavulanic acid and cefuroxime, while the stably derepressed strains were only sensitive to imipenam and cefepime . However, sensitivity to cefepime decreased if the strains also produced ESBLs . CONCLUSIONS: The drug resistant status of Enterobacter cloacae is severe . Clearing out the expressive status of ampC gene will be helpful in selection of antimicrobial agents in the treatment of clinical infection. J Endotoxin Res, 2003, 9(4), 244 - 9 Molecular insights into the assembly and diversity of the outer core oligosaccharide in lipopolysaccharides from Escherichia coli and Salmonella; Whitfield C et al.; In the Enterobacteriaceae, the core oligosaccharide provides the junction between the highly conserved lipid A and the remarkably diverse polysaccharide O antigen . The basic structure of the inner (lipid A-proximal) core is well conserved, perhaps reflecting constraints imposed by its involvement in the structural integrity of the outer membrane . However, non-stoichiometric modifications do create some structural variants . The outer core may show more variation . Efforts to develop immunoprophylactic strategies based on the core oligosaccharide require a detailed understanding of core immunochemistry, the accessibility of specific epitopes in the LPS, and the distribution of specific structures within natural populations . The availability of sequences for the waa (core biosynthesis) loci and functional data for the gene products provide a molecular basis for the known structural diversity in Escherichia coli and Salmonella core oligosaccharide . Surveys of waa-locus organization have established the distribution of these core types in natural populations and have identified genetic variants that provide candidates for additional novel structures. Tidsskr Nor Laegeforen, 2003 Aug 14, 123(15), 2021 - 2 {Urinary tract pathogens in uncomplicated lower urinary tract infections in women in Norway}; Jureen R et al.; BACKGROUND: We wanted to investigate the antimicrobial susceptibility of urinary tract pathogens in uncomplicated lower urinary tract infections in adult women in Norway . MATERIAL AND METHODS: Urine samples from 312 adult women with symptoms of uncomplicated urinary tract infections from eight general practices were included . RESULTS: Significant bacteriuria was found in 187 samples (60%) . E coli was isolated from 153 (82%) of these samples . Other isolated uropathogens were S saprophyticus 18 (10%), Proteus spp 6 (3%), Klebsiella spp 4 (2%), Enterobacter spp 2 (1%), enterococci 1 (0.5%) and other Gram-positive bacteria 3 (1,5%) . No fungi were isolated . Of the E coli isolates, 1 %, 1 % and 9 % were resistant to nitrofurantoin, mecillinam and trimetoprim respectively . All S saprophyticus isolates were sensitive to nitrofurantoin and trimetoprim . INTERPRETATION: Antibiotic resistance of urinary tract pathogens causing uncomplicated urinary tract infections in adult women in general practice is still low in Norway. Infect Immun, 2003 Sep, 71(9), 5087 - 96 A novel pathogenicity island integrated adjacent to the thrW tRNA gene of avian pathogenic Escherichia coli encodes a vacuolating autotransporter toxin; Parreira VR et al.; We report the complete nucleotide sequence and genetic organization of the Vat-encoding pathogenicity island (PAI) of avian pathogenic Escherichia coli strain Ec222 . The 22,139-bp PAI is situated adjacent to the 3' terminus of the thrW tRNA gene, has a G+C content of 41.2%, and includes a bacteriophage SfII integrase gene, mobile genetic elements, two open reading frames with products exhibiting sequence similarity to known proteins, and several other open reading frames of unknown function . The PAI encodes an autotransporter protein, Vat (vacuolating autotransporter toxin), which induces the formation of intracellular vacuoles resulting in cytotoxic effects similar to those caused by the VacA toxin from Helicobacter pylori . The predicted 148.3-kDa protein product possesses the three domains that are typical of serine protease autotransporters of Enterobacteriaceae: an N-terminal signal sequence of 55 amino acids, a 111.8-kDa passenger domain containing a modified serine protease site (ATSGSG), and a C-terminal outer membrane translocator of 30.5 kDa . Vat has 75% protein homology with the hemagglutinin Tsh, an autotransporter of avian pathogenic E . coli . A vat deletion mutant of Ec222 showed no virulence in respiratory and cellulitis infection models of disease in broiler chickens . We conclude that the newly described PAI and Vat may be involved in the pathogenicity of avian septicemic E . coli strain Ec222 and other avian pathogenic E . coli strains. Oral Microbiol Immunol, 2003 Oct, 18(5), 329 - 32 Sensitivity of genera Porphyromonas and Prevotella to the bactericidal action of C-terminal domain of human CAP18 and its analogues; Isogai E et al.; This paper reports the effect of the synthesized 27-amino acid sequence in the C-terminal domain of human CAP18 (hCAP18), a human cationic antibacterial protein or cathelicidin, on certain strains belonging to the genera Porophyromonas and Prevotella . The domain binds lipopolysaccharides (LPS) from Porophyromonas gingivalis and Porophyromonas circumdentaria as well as enterobacterial LPS . Two analogues of hCAP18, designated LL/CAP18 and FF/CAP18, were also tested to determine whether additional activity was obtained . The analogue peptides replaced with hydrophobic and cationic amino acid residues showed more potent bactericidal and LPS-binding activities than the original one. J Food Prot, 2003 Aug, 66(8), 1486 - 9 Survey of shell egg processing plant sanitation programs: effects on egg contact surfaces; Jones DR et al.; Sanitation standard operating procedures (SSOPs) are an integral component of process control and are often the first step in the implementation of food safety regulations . The objective of this study was to assess and compare the efficacies of sanitation programs used in a variety of shell egg processing facilities . In-line, off-line, and mixed operations were evaluated . Sixteen direct or indirect egg contact surfaces were sampled in various shell egg processing facilities in the southeast United States . Samples were collected at the end of a processing day (POST) and again the next morning before operations began (PRE) . Total aerobic plate counts (APCs) were obtained and Enterobacteriacae were enumerated . No significant differences (P > 0.05) between POST and PRE bacterial counts were found for the 16 sampling sites . In general, high APCs were found on the wall of the recirculating water tank both POST and PRE . The APCs for the rewash belt were considerably high for all plants sampled . APCs were also high for the vacuum loaders . APCs for washers and washer brushes were relatively low for most plants sampled . PRE and POST levels of plant sanitation, as determined by direct microbial plating, did not differ significantly . At this point, it is difficult to draw definitive conclusions about how rigid SSOPs should be for the shell egg processing industry. Int J Antimicrob Agents, 2003 Aug, 22(2), 147 - 54 In vitro susceptibility of 4903 bacterial isolates to gemifloxacin--an advanced fluoroquinolone; Blondeau JM et al.; The in vitro activity of gemifloxacin against over 4900 bacterial isolates was determined by microbroth dilution with interpretation in accordance with NCCLS guidelines . Susceptibility results were compared with those for ciprofloxacin, gatifloxacin, levofloxacin and moxifloxacin . Gemifloxacin and the other fluoroquinolones were not affected by either beta-lactamase production or penicillin-resistance in Streptococcus pneumoniae . The MIC90 values for gemifloxacin were: S . pneumoniae 0.063 mg/l; Haemophilus influenzae 0.016 mg/l; Moraxella catarrhalis 0.008 mg/l, methicillin-susceptible Staphylococcus aureus 0.063 mg/l; methicillin-susceptible Streptococcus pyogenes 0.031 mg/l; Enterobacteriaceae 0.031-0.16 mg/l; Pseudomonas aeruginosa 4 mg/l; Neisseria meningitidis 0.008 mg/l . The MIC90 for gemifloxacin was lower than those for the other quinolones tested against S . pneumoniae (ciprofloxacin 2-4 mg/l, gatifloxacin 0.5 mg/l, levofloxacin 1-2 mg/l, moxifloxacin 0.25 mg/l) . This study confirms the enhanced potent activity of gemifloxacin against Gram-positive pathogens, its broad-spectrum, Gram-negative activity and indicates that gemifloxacin is likely to have an important role in treating patients with Gram-positive and/or Gram-negative infections. Int J Antimicrob Agents, 2003 Aug, 22(2), 128 - 33 Molecular epidemiology of Enterobacteriaceae producing extended-spectrum beta-lactamase in a French university-affiliated hospital; Bertrand X et al.; We conducted a retrospective study to investigate the epidemiology of Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE) in our hospital . We determined the occurrence of extended-spectrum beta-lactamase (ESBL) in Enterobacteriaceae over a 2-year period . We also characterised ESBLs by isoelectric focusing (IEF) and investigated the epidemiological relatedness of EBLSE by pulsed field gel electrophoresis (PFGE) . During this period, 70 patients were colonised/infected with one or several strains of EBLSE, giving a crude incidence of 0.095 per 1000 patient-days . We found that ESBL-producing Enterobacter aerogenes were the main source of ESBLE dissemination . Indeed, 59.5% of ESBLE were E . aerogenes and 21.9% of the other ESBLE resulted from a plasmid transfer originating from E . aerogenes . IEF and PFGE analysis demonstrated that the dissemination of ESBL from E . aerogenes in our hospital was due to a single clone that always harbours TEM-24 . This emphasises the importance of standard contact isolation precautions and the early detection of ESBLE-colonised patients in high risk departments like intensive care units. J Anim Sci, 2003 Aug, 81(8), 2019 - 31 Fermented and nonfermented liquid feed to growing pigs: effect on aspects of gastrointestinal ecology and growth performance; Canibe N et al.; The effect of feeding dry feed (DF), nonfermented liquid feed (NFLF), and fermented liquid feed (FLF) to growing pigs on aspects of gastrointestinal ecology and on performance was investigated . Nonfermented liquid feed was prepared by mixing feed and water at a ratio of 1:2.5 immediately before feeding . Fermented liquid feed was prepared by mixing feed and water in the same ratio as NFLF, and stored in a tank at 20 degrees C for 4 d, after which half the volume was removed twice daily at each feeding and replaced with the same volume of feed and water mixture . A total of 60 pigs (initial BW of 30.7 kg) from 20 litters was used . Twenty pigs, housed individually, were allotted to each of the diets and fed restrictively . Five pigs from each diet were sacrificed at an average BW of 112 kg and digesta from the gastrointestinal tract (GI-tract) was obtained to examine variables describing some aspects of the gastrointestinal ecology . Fermented liquid feed contained high levels of lactic acid bacteria (9.4 log cfu/g) and lactic acid (approximately 169 mmol/kg), low levels of enterobacteria (<3.2 log cfu/g), and had a low pH (4.4) . Nonfermented liquid feed contained 7.2 log cfu/g of lactic acid bacteria, and 6.2 log cfu/g of enterobacteria, which indicated that fermentation had started in the feed . The pigs fed FLF had the lowest levels of enterobacteria along the GI-tract (<3.2 to 5.0 log cfu/g), and those fed NFLF the highest levels (5.7 to 6.6 log cfu/g; P < or = 0.02) . Fermented liquid feed caused a decrease in gastric pH from 4.4 and 4.6 for DF and NLF, to 4.0 (P = 0.003), and increased numerically the gastric concentration of lactic acid (P = 0.17) from 50 to 60 mmol/kg in the DF and NFLF treatments to 113 mmol/kg in the FLF treatment . The animals fed NFLF showed the highest weight gain (995 g/d) and feed intake (2.14 kg/d), and those fed FLF the lowest values (weight gain, 931 g/d; feed intake, 1.96 kg/d; P = 0.003 for weight gain, and P < 0.001 for feed intake) . The results from the present study indicate that feeding FLF as prepared here may be a valid feeding strategy to decrease the levels of enterobacteria in the GI-tract of growing pigs, whereas feeding liquid feed that has started to ferment (high levels of enterobacteria and high pH as with NFLF) increases the presence of these undesirable bacteria . Nonetheless, higher daily feed intake and body weight gain are obtained when feeding NFLF compared with feeding FLF or DF. J Bone Joint Surg Am, 2003 Aug, 85-A(8), 1454 - 60 Necrotizing fasciitis: clinical presentation, microbiology, and determinants of mortality; Wong CH et al.; BACKGROUND: Necrotizing fasciitis is a life-threatening soft-tissue infection primarily involving the superficial fascia . The present report describes the clinical presentation and microbiological characteristics of this condition as well as the determinants of mortality associated with this uncommon surgical emergency . METHODS: The medical records of eighty-nine consecutive patients who had been admitted to our institution for necrotizing fasciitis from January 1997 to August 2002 were reviewed retrospectively . RESULTS: The paucity of cutaneous findings early in the course of the disease makes the diagnosis difficult, and only thirteen of the eighty-nine patients had a diagnosis of necrotizing fasciitis at the time of admission . Preadmission treatment with antibiotics modified the initial clinical picture and often masked the severity of the underlying infection . Polymicrobial synergistic infection was the most common cause (forty-eight patients; 53.9%), with streptococci and enterobacteriaceae being the most common isolates . Group-A streptococcus was the most common cause of monomicrobial necrotizing fasciitis . The most common associated comorbidity was diabetes mellitus (sixty-three patients; 70.8%) . Advanced age, two or more associated comorbidities, and a delay in surgery of more than twenty-four hours adversely affected the outcome . Multivariate analysis showed that only a delay in surgery of more than twenty-four hours was correlated with increased mortality (p < 0.05; relative risk = 9.4) . CONCLUSIONS: Early operative debridement was demonstrated to reduce mortality among patients with this condition . A high index of suspicion is important in view of the paucity of specific cutaneous findings early in the course of the disease. J Bacteriol, 2003 Sep, 185(17), 5328 - 32 Role for Salmonella enterica enterobacterial common antigen in bile resistance and virulence; Ramos-Morales F et al.; Passage through the digestive tract exposes Salmonella enterica to high concentrations of bile salts, powerful detergents that disrupt biological membranes . Mutations in the wecD or wecA gene, both of which are involved in the synthesis of enterobacterial common antigen (ECA), render S . enterica serovar Typhimurium sensitive to the bile salt deoxycholate . Competitive infectivity analysis of wecD and wecA mutants in the mouse model indicates that ECA is an important virulence factor for oral infection . In contrast, lack of ECA causes only a slight decrease in Salmonella virulence during intraperitoneal infection . A tentative interpretation is that ECA may contribute to Salmonella virulence by protecting the pathogen from bile salts. J Bacteriol, 2003 Sep, 185(17), 5314 - 9 IS1999 increases expression of the extended-spectrum beta-lactamase VEB-1 in Pseudomonas aeruginosa; Aubert D et al.; The integron-borne bla(VEB-1) gene encodes an extended-spectrum beta-lactamase . This gene was associated mostly with IS1999 and rarely with an additional IS2000 element in Pseudomonas aeruginosa isolates from Thailand, whereas IS1999 was only very rarely associated with bla(VEB-1) in Enterobacteriaceae . Expression experiments and promoter study identified promoter sequences in IS1999 that increased the expression of VEB-1 in P . aeruginosa. Arch Biochem Biophys, 2003 Sep 1, 417(1), 12 - 7 Antioxidative flavonoid quercetin: implication of its intestinal absorption and metabolism; Murota K et al.; Quercetin is a typical flavonoid ubiquitously present in fruits and vegetables, and its antioxidant effect is implied to be helpful for human health . The bioavailability of quercetin glycosides should be clarified, because dietary quercetin is mostly present as its glycoside form . Although quercetin glycosides are subject to deglycosidation by enterobacteria for the absorption at large intestine, small intestine acts as an effective absorption site for glucose-bound glycosides (quercertin glucosides) . This is because small intestinal cells possess a glucoside-hydrolyzing activity and their glucose transport system is capable of participating in the glucoside absorption . A study using a cultured cell model for intestinal absorption explains that the hydrolysis of the glucosides accelerates their absorption in the small intestine . Small intestine is also recognized as the site for metabolic conversion of quercetin and other flavonoids as it possesses enzymatic activity of glucuronidation and sulfation . Modulation of the intestinal absorption and metabolism may be beneficial for regulating the biological effects of dietary quercetin. Drugs, 2003, 63(17), 1855 - 78 Ertapenem: a review of its use in the management of bacterial infections; Curran M et al.; Ertapenem, a carbapenem antibacterial, has in vitro activity against many Gram- negative (including Enterobacteriaceae) and Gram-positive aerobic and anaerobic bacteria that are commonly associated with various infections.Once-daily parenteral (intravenous or intramuscular) ertapenem 1g was as effective as comparator antimicrobial agents (piperacillin/tazobactam or ceftriaxone +/- metronidazole) in patients with bacterial infections in randomised, double-blind, multicentre clinical trials . Response rates with ertapenem were 84% and 87% (combined microbiological and clinical) in patients with complicated intra-abdominal infections (CIAI), 82% (clinical) in patients with complicated skin and skin structure infections (CSSSI), 86% and 92% (microbiological) in patients with complicated urinary tract infections (CUTI), 92% (clinical) in patients with community-acquired pneumonia (CAP) associated with typical pathogens and 94% (clinical) in patients with acute pelvic infection . Respective response rates were statistically equivalent to those with comparators (81-94%) . The efficacy of ertapenem was equivalent to that of piperacillin/tazobactam in patients infected with Enterobacteriaceae or anaerobes and to ceftriaxone in patients infected with Enterobacteriaceae . Ertapenem was generally well tolerated by patients with bacterial infections, with most adverse events being mild to moderate in severity . The most common ertapenem-associated adverse events were diarrhoea, infused vein complication, nausea, headache, vaginitis in females, phlebitis and/or thrombophlebitis and vomiting . CONCLUSION: Ertapenem is a broad-spectrum parenteral antibiotic with activity against many Gram-negative (including Enterobacteriaceae) and Gram-positive aerobic and anaerobic bacteria and is suitable for once-daily administration . Ertapenem has a role in the treatment of CAP associated with typical respiratory pathogens and is of particular value in the treatment of polymicrobial infections (such as CIAI, CSSSI, CUTI and acute pelvic infections), especially where Enterobacteriaceae and anaerobic bacteria are involved. J Antimicrob Chemother, 2003 Sep, 52(3), 389 - 96 Epub 2003 Aug 13. Comparison of microbial adherence to antiseptic and antibiotic central venous catheters using a novel agar subcutaneous infection model; Gaonkar TA et al.; An agar subcutaneous infection model (agar model), which simulates the rat subcutaneous infection model (rat model), was developed to assess the ability of antimicrobial catheters to resist microbial colonization . The catheters were implanted in the agar and rat models and the insertion sites were infected immediately or on day 7, 14 or 21 post-implantation . The catheters implanted in the agar model were transferred to fresh media one day before infection on day 7, 14 or 21 . The efficacy of chlorhexidine and silver sulfadiazine impregnated (CS) catheters, CS catheters with higher levels of chlorhexidine (CS+ catheters), minocycline-rifampicin (MR) catheters and silver catheters against Staphylococcus aureus and rifampicin-resistant Staphylococcus epidermidis RIF-r2 was compared in the agar and rat models . No significant difference in the adherence or the drug release was found between the in vitro and in vivo models . In both models, CS+ and MR catheters were effective against S . aureus even when infected on day 14, whereas CS catheters were colonized when challenged on day 7 . CS+ catheters were effective against S . epidermidis RIF-r2, whereas MR catheters showed adherence when infected on day 7 . CS+ catheters prevented colonization of all the organisms including, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Candida albicans in the agar model, whereas MR catheters were effective only against S . aureus and S . epidermidis strains . Silver catheters were ineffective against all the organisms . The agar model may be used to predict the in vivo efficacy of antimicrobial catheters against various pathogens. J Antimicrob Chemother, 2003 Sep, 52(3), 331 - 44 Epub 2003 Aug 13. Properties and potential of ertapenem; Livermore DM et al.; Ertapenem is a carbapenem that shares the activity of imipenem and meropenem against most species, but is less active against non-fermenters . Activity is retained against most strains with AmpC and extended-spectrum beta-lactamases, although resistance can arise if these enzymes are combined with extreme impermeability . Resistance can also be caused by IMP, VIM, KPC and NMC carbapenemases, but again, co-requires impermeability . Although the spread of carbapenemases in the future is a concern, they are currently very rare . Given as a 1 g intravenous (iv) infusion once daily, ertapenem has a plasma half-life of approximately 4 h in healthy volunteers, and a Cmax of 155 mg/L and 13 mg/L for total and free drug, respectively . Excretion is largely renal, divided equally between native drug and an open-ring derivative . Trials show equivalence to piperacillin/tazobactam or ceftriaxone in (a) intra-abdominal infections, (b) community-acquired pneumonia, (c) acute pelvic infections, (d) skin and skin structure infections and (e) complicated urinary tract infections . The USA licence grants all these five indications; the EU licence grants the first three . Further potential uses include home iv therapy, directed therapy against Enterobacteriaceae with AmpC or extended-spectrum cephalosporinases, and tentatively, surgical prophylaxis . Widening the usage of carbapenems raises public health concerns, somewhat allayed by the continued rarity of carbapenemases after 17 years of imipenem use, and by the fact that carbapenemases occur mostly in non-fermenters outside the spectrum of ertapenem, and co-require impermeability to confer resistance in Enterobacteriaceae . Nevertheless, if ertapenem is to be used widely, its effects on the resistance ecology need to be monitored carefully. Acta Microbiol Pol, 2003, 52(1), 81 - 6 Siderophore-mediated strategies of iron acquisition by extraintestinal isolates of Enterobacter spp; Mokracka J et al.; A total of 89 examined Enterobacter isolates belonging to three species: E . cloaceae, E . aerogenes and E . sakazakii, produced iron chelators detected in universal CAS assay . In chemical assays the strains were shown to excrete mostly catecholate (88 strains) and hydroxamate (42 strains) type of siderophores . Forty-one strains produced both catecholate and hydroxamate siderophores whereas one isolate produced only hydroxamate . Besides, the isolates were screened for genes coding for another siderophore: yersiniabactin . The genes for biosynthesis and uptake of yersiniabactin are located on the high-pathogenicity island (HPI) of Yersinia spp . The presence of three marker genes irp1, irp2 and fyuA was estimated by polymerase chain reaction . Two strains: E . aerogenes and E . cloaceae possessed irp1, irp2 and fyuA genes . PCR products of irp1, irp2 and fyuA were of 240, 280 and 780 bp, respectively. Antibiot Khimioter, 2003, 48(3), 26 - 9 {Sulperazone in the treatment of severe infections in patients with cancer}; Mitrokhin SD; For the evaluation of rational sulperazone position in oncological hospital and elaboration of its implementation criteria investigation was performed . The trial included 193 patients (88 patients with injuries, 66 with hospital pneumonia, 39 with urinary track infection) . From different clinical material of the patients were isolated 136 straines of Gram-negative bacteria . Suseptibile to sulperazone were 67 per cent of P . aeruginosa, 78-86 per cent of P . vulgaris, Klebsiella and E . coli isolates, 66 {symbol: see text} 60 per cent of Enterobacter and Serratia isolates subsequently . Comparative results of treatment with sulperazone as monotherapy and for combination with aminoglycosides (gentamycin or amicacin) in the case of P . aeruginosa infection demonstrated high sulperazone efficacy not lower than imipenem/cilsatatin and cefepime efficacy in the case of hospital infections treatment at critical care units . Low price of the sulperazone treatment when compared to imipenem/cilastatine is emphasized . Another advantage of sulperazone when compared to cefepime is anaerobic infection treatment--cefepime is administered in combination with metronidazole while sulperazone is used in monotherapy. J Appl Microbiol, 2003, 95(3), 428 - 36 Transfer of antibiotic resistance between commensal and pathogenic members of the Enterobacteriaceae under ileal conditions; Blake DP et al.; AIM: To determine the rate of antibiotic resistance transmission between commensal and pathogenic representatives of the Enterobacteriaceae . METHODS AND RESULTS: Through the use of a validated in vitro simulation of the porcine ileum, the transmission of antibiotic resistance was detected between commensal Escherichia coli, E . coli O157 and Salmonella spp . Countable transconjugant populations arose readily and, in one example, proved capable of indefinite persistence . CONCLUSIONS: Genetic material conferring antibiotic resistance is readily transmissible between members of the Enterobacteriaceae under ileal conditions . Recipient phenotype influences the persistence of multi-resistant transconjugants . SIGNIFICANCE AND IMPACT OF THE STUDY: The observation that the conjugal transmission of antibiotic resistance is commonplace under ileal conditions impacts primarily on the risk of food contamination by multi-resistant bacteria . The establishment of a multi-resistant transconjugant population as a dominant member of the microflora maintains a genetic reservoir of antimicrobial resistance. Appl Microbiol Biotechnol, 2004 Feb, 63(6), 666 - 71 Epub 2003 Aug 09. Synthesis of FimH receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from Penicillium citrinum, Aspergillus phoenicis and almond; Maitin V et al.; Recombinant Penicillium citrinum alpha-1,2-mannosidase, expressed in Aspergillus oryzae, was employed to carry out regioselective synthesis of alpha- d-mannopyranosyl-(1-->2)- d-mannose . Yields (w/w) of 16.68% disaccharide, 3.07% trisaccharide and 0.48% tetrasaccharide were obtained, with alpha1-->2 linkages present at 98.5% of the total linkages formed . Non-specific alpha-mannosidase from almond was highly efficient in reverse hydrolysis and oligosaccharide yields of 45-50% were achieved . The products of the almond mannosidase were a mixture of disaccharides (30.75%, w/w), trisaccharides (12.26%, w/w) and tetrasaccharides (1.89%, w/w) with 1-->2, 1-->3 and 1-->6 isomers . alpha-1,2-linkage specific mannosidase from P . citrinum and alpha-1,6-linkage-specific mannosidase from Aspergillus phoenicis were used in combination to hydrolyse the respective linkages from the mixture of isomers, resulting in alpha- d-mannopyranosyl-(1-->3)- d-mannose in 86.4% purity . The synthesised oligosaccharides can potentially inhibit the adhesion of pathogens by acting as "decoys" of receptors of type-1 fimbriae carried by enterobacteria. J Clin Microbiol, 2003 Aug, 41(8), 3790 - 800 Rapid identification of bacteria from positive blood cultures by terminal restriction fragment length polymorphism profile analysis of the 16S rRNA gene; Christensen JE et al.; Bacteremia results in significant morbidity and mortality, especially among patient populations that are immunocompromised . Broad-spectrum antibiotics are administered to patients suspected to have bloodstream infections that are awaiting diagnosis that depends on blood culture analysis . Significant delays in identification of pathogens can result, primarily due to the dependence on growth-based identification systems . To address these limitations, we took advantage of terminal restriction fragment (TRF) length polymorphisms (T-RFLP) due to 16S ribosomal DNA (rDNA) sequence diversity to rapidly identify bacterial pathogens directly from positive blood culture . TRF profiles for each organism were determined by sizing fragments from restriction digests of PCR products derived from two sets of 16S rDNA-specific fluorescent dye-labeled primers . In addition, we created a TRF profile database (TRFPD) with 5899 predicted TRF profiles from sequence information representing 2860 different bacterial species . TRF profiles were experimentally determined for 69 reference organisms and 32 clinical isolates and then compared against the predicted profiles in the TRFPD . The predictive value of the profiles was found to be accurate to the species level with most organisms tested . In addition, identification of 10 different genera was possible with profiles comprising two or three TRFs . Although it was possible to identify Enterobacteriaceae by using a profile of three TRFs, the similarity of the TRF profiles of these organisms makes differentiation of species less reliable with the current method . The ability to rapidly (i.e., within approximately 8 h) identify bacteria from blood cultures has potential for reducing unnecessary use of broad-spectrum antibiotics and promoting more timely prescription of appropriate antibiotics. Nucleic Acids Symp Ser, 2000, (44), 25 - 6 Study on highly diastereoselective synthesis of (2'R)-2'-deoxy{2'-2H}guanosine; Kawashima E et al.; To develop an efficient method for the synthesis of a highly diasteroselective (2'R)-2'-deoxy{2'-2H}guanosine (1), studies of organic chemical conversion from 2'-bromo-2'-deoxy-N2-Isobutyryl-3',5'-O-TIPDS-guanosine (2) to 1 and a biological transdeoxyribofuranosylation of (2'R > 98% de)-2'-deoxy{2'-2H}uridine (4) were carried out . As the results, a highly diastereoselective synthesis of 1 was achieved by a biological transdeoxyribofuranosylation between 2,6-diaminopurine and 4 by the use of Enterobacter aerogenes AJ-11125, followed by treatment with adenosine deaminase . The results will be described in detail. Appl Environ Microbiol, 2003 Aug, 69(8), 4915 - 26 Colonization of Arabidopsis thaliana with Salmonella enterica and enterohemorrhagic Escherichia coli O157:H7 and competition by Enterobacter asburiae; Cooley MB et al.; Enteric pathogens, such as Salmonella enterica and Escherichia coli O157:H7, have been shown to contaminate fresh produce . Under appropriate conditions, these bacteria will grow on and invade the plant tissue . We have developed Arabidopsis thaliana (thale cress) as a model system with the intention of studying plant responses to human pathogens . Under sterile conditions and at 100% humidity, S . enterica serovar Newport and E . coli O157:H7 grew to 10(9) CFU g(-1) on A . thaliana roots and to 2 x 10(7) CFU g(-1) on shoots . Furthermore, root inoculation led to contamination of the entire plant, indicating that the pathogens are capable of moving on or within the plant in the absence of competition . Inoculation with green fluorescent protein-labeled S . enterica and E . coli O157:H7 showed invasion of the roots at lateral root junctions . Movement was eliminated and invasion decreased when nonmotile mutants of S . enterica were used . Survival of S . enterica serovar Newport and E . coli O157:H7 on soil-grown plants declined as the plants matured, but both pathogens were detectable for at least 21 days . Survival of the pathogen was reduced in unautoclaved soil and amended soil, suggesting competition from indigenous epiphytes from the soil . Enterobacter asburiae was isolated from soil-grown A . thaliana and shown to be effective at suppressing epiphytic growth of both pathogens under gnotobiotic conditions . Seed and chaff harvested from contaminated plants were occasionally contaminated . The rate of recovery of S . enterica and E . coli O157:H7 from seed varied from undetectable to 19% of the seed pools tested, depending on the method of inoculation . Seed contamination by these pathogens was undetectable in the presence of the competitor, Enterobacter asburiae . Sampling of 74 pools of chaff indicated a strong correlation between contamination of the chaff and seed (P = 0.025) . This suggested that contamination of the seed occurred directly from contaminated chaff or by invasion of the flower or silique . However, contaminated seeds were not sanitized by extensive washing and chlorine treatment, indicating that some of the bacteria reside in a protected niche on the seed surface or under the seed coat. Appl Environ Microbiol, 2003 Aug, 69(8), 4697 - 705 Rapid detection of meat spoilage by measuring volatile organic compounds by using proton transfer reaction mass spectrometry; Mayr D et al.; The evolution of the microbial spoilage population for air- and vacuum-packaged meat (beef and pork) stored at 4 degrees C was investigated over 11 days . We monitored the viable counts (mesophilic total aerobic bacteria, Pseudomonas spp., Enterobacteriaceae, lactic acid bacteria, and Enterococcus spp.) by the microbiological standard technique and by measuring the emission of volatile organic compounds (VOCs) with the recently developed proton transfer reaction mass spectrometry system . Storage time, packaging type, and meat type had statistically significant (P < 0.05) effects on the development of the bacterial numbers . The concentrations of many of the measured VOCs, e.g., sulfur compounds, largely increased over the storage time . We also observed a large difference in the emissions between vacuum- and air-packaged meat . We found statistically significant strong correlations (up to 99%) between some of the VOCs and the bacterial contamination . The concentrations of these VOCs increased linearly with the bacterial numbers . This study is a first step toward replacing the time-consuming plate counting by fast headspace air measurements, where the bacterial spoilage can be determined within minutes instead of days. Appl Environ Microbiol, 2003 Aug, 69(8), 4583 - 94 Microbial quality and direct PCR identification of lactic acid bacteria and nonpathogenic Staphylococci from artisanal low-acid sausages; Aymerich T et al.; Detection of six species of lactic acid bacteria and six species of gram-positive catalase-positive cocci from low-acid fermented sausages (fuets and chorizos) was assessed by species-specific PCR . Without enrichment, Lactobacillus sakei and Lactobacillus curvatus were detected in 11.8% of the samples, and Lactobacillus plantarum and Staphylococcus xylosus were detected in 17.6% . Enriched samples allowed the detection of L . sakei and S . xylosus in all of the samples (100%) and of Enterococcus faecium in 11.8% of the sausages . The percentages of L . curvatus, L . plantarum, Staphylococcus carnosus, and Staphylococcus epidermidis varied depending on the sausage type . L . curvatus was detected in 80% of fuets and in 57% of chorizos . L . plantarum was found in 50% of fuets and 100% of chorizos . S . epidermidis was detected in only 11.8% of fuets, and S . carnosus was detected in only 5.9% of chorizos . Lactococcus lactis, Staphylococcus warneri, and Staphylococcus simulans were not detected in any sausage type . From a microbiological point of view, 70.6% of the samples could be considered of high quality, as they had low counts of Enterobacteriaceae and did not contain any of the food-borne pathogens assayed. New Microbiol, 2003 Jul, 26(3), 257 - 62 Prevalence of extended-spectrum beta-lactamases produced by nosocomial isolates of Enterobacteriaceae in Trakya University Hospital, Turkey; Akata F et al.; The prevalence of extended-spectrum beta-lactamase (ESBL) production by 194 nosocomial isolates of Enterobacteriacea recovered from 1995 to 1999 was investigated . The ESBL production was determined by the double-disk synergy test and was confirmed by the E-test ESBL strip . Twenty-three isolates (21 Klebsiella pneumoniae, one Escherichia coli, one Providencia rettgeri) were found as ESBL-producers (11.8%) . These isolates were also usually resistant to non-betalactam antibiotics . Most of them contained a beta-lactamase with a pI of 7.6 . All the strains conjugally transferred their ESBLs to recipient E . coli . Contrary to others, ESBL-producing K . pneumoniae strains isolated in 1999 were resistant to ciprofloxacin, and had the identical plasmid profiles suggestive of an outbreak . Ciprofloxacin resistance in these strains could not be transferred . In conclusion, K . pneumoniae was the main ESBL-producing species among nosocomial isolates of Enterobacteriacae in our hospital. Am J Kidney Dis, 2003 Aug, 42(2), 342 - 9 Levofloxacin pharmacokinetics in ESRD and removal by the cellulose acetate high performance-210 hemodialyzer; Sowinski KM et al.; BACKGROUND: No published data are available describing the pharmacokinetics of intravenous levofloxacin in patients with end-stage renal disease (ESRD) . Objectives of this study are to determine the pharmacokinetics and dialytic clearance of levofloxacin and develop dosing strategies in these patients . METHODS: Eight noninfected subjects receiving long-term thrice-weekly hemodialysis, with no measurable residual renal function, were administered intravenous levofloxacin, 250 mg, over 1 hour after a scheduled hemodialysis session . Blood samples were collected serially during the interdialytic period, during the next intradialytic period, and immediately after the next hemodialysis session . Serum concentrations of levofloxacin were determined by high-performance liquid chromatography . Differential equations describing a 2-compartment open-infusion pharmacokinetic model were fit to each individual subject's serum concentration-time data by iterative nonlinear weighted least-squares regression analysis using Adapt II (Biomedical Simulations Resource, University of Southern California, Los Angeles, CA) . Ratios of maximum serum concentration (C(max)) to minimum inhibitory concentration (MIC) were calculated for common respiratory pathogens by using MIC for 90% of isolates (MIC90) data from published studies . RESULTS: All subjects completed the study, and no adverse events were reported . Median systemic clearance, volume of distribution at steady state, elimination half-life, and C(max) were 37.0 mL/min (range, 12.8 to 42.7 mL/min), 103.3 L (range, 39.8 to 139.3 L), 34.4 hours (range, 28.4 to 39.3 hours), and 5.2 microg/mL (range, 4.1 to 11.3 microg/mL), respectively . Median dialytic clearance and levofloxacin reduction ratios were 84.4 mL/min (range, 61.8 to 107.6 mL/min) and 0.244 (range, 0.181 to 0.412), respectively . Median C(max)-MIC90 ratios were 10 or greater for Haemophilus influenzae, Moraxella catarrhalis, Enterobacter cloacae, and Klebsiella pneumoniae, approximately 5 for Streptococcus pneumoniae, and less than 1 for Pseudomonas aeruginosa . CONCLUSION: The administration of levofloxacin to patients with ESRD as 500 mg initially, followed by 250 mg every 48 hours, will provide adequate C(max)-MIC ratios after the first and subsequent doses for most patients with respiratory tract infections caused by organisms with levofloxacin MICs of 1 microg/mL or less. Eur J Biochem, 2003 Aug, 270(16), 3469 - 76 Cause of mortality in insects under severe stress; Matsumoto H et al.; Mortality in the host armyworm larvae Pseudaletia separata parasitized by the parasitic wasp Cotesia kariyai was dramatically increased when they were simultaneously infected by the entomopathogen Serratia marcescens . Previous studies have shown that this strong insecticidal effect is due to a metalloprotease-like insecticide (MPLI) released from S . marcescens enterobacter . This study was conducted to elucidate the exact cause of the mortality resulting from MPLI . Injection of MPLI caused a sharp increase in hemolymph dopamine concentration followed by elevated levels of brain dopamine in armyworm larvae . {3H}Dopamine injected into the hemocoel, was incorporated into the brains of MPLI-injected larvae to a level eight times greater than in BSA-injected control larvae . Transmission electron microscopy showed an obvious decrease in thickness and density of the brain sheath in insects injected with MPLI . This was probably due to the MPLI-induced elevation of hemocyte metalloprotease activities . Further, electron microscopic and TUNEL staining analyses showed a significant increase in apoptotic cells in the brain 12 h after the injection . Injection of 3-iodotyrosine (a tyrosine hydroxylase inhibitor) before MPLI completely prevented the increase in hemolymph dopamine in test larvae and their following death . From these observations, we conclude that MPLI-injected larvae may have suffered mortal damage through increased apoptosis of brain cells caused by an influx of dopamine from the hemolymph. Bioresour Technol, 2003 Nov, 90(2), 139 - 43 Synergetic effect of gamma irradiation and moisture content on decontamination of sewage sludge; Al-Bachir M et al.; Samples of concentrated municipal sewage sludge, stored for 2, 4 and 6 months, with moisture contents of 2%, 20%, 40%, 60%, and 80% were exposed to doses of 0, 1, 2, 3, 4 and 5 kilogray (kGy) of gamma irradiation . Immediately after irradiation, total microbial count and bacterial pathogens in sewage sludge were determined . The results indicated that in all tested sewage sludge samples, bacterial pathogens including Enterobacter sp., Klebsiella sp., Salmonella sp., and Escherichia coli were initially detected . All doses of gamma irradiation reduced the total counts of microorganisms . D(10) of total count decreased with increase in the moisture content of the sewage sludge . The lowest lethal dose for tested bacterial pathogens was 5 kGy in air dried sewage sludge . In addition for wet sewage sludge having more than 40% moisture, the lethal dose was 1 kGy, for samples taken at different storage periods 2, 4 and 6 months, and therefore the cost per unit could be decreased to half when wet sewage sludge (about 50% moisture) was used. Res Microbiol, 2003 Jul-Aug, 154(6), 425 - 31 Genetic engineering of Enterobacter aerogenes with the Vitreoscilla hemoglobin gene: cell growth, survival, and antioxidant enzyme status under oxidative stress; Geckil H et al.; Hemoglobins in unicellular organisms, like the one here in the bacterium Vitreoscilla, have greater chemical reactivity than their homologues in multicellular organisms . They can catalyze redox reactions and may protect cells against oxidative stress . The ability of Vitreoscilla hemoglobin to complement deficiencies of terminal cytochrome oxidases in Escherichia coli also suggests that this hemoglobin can receive electrons during respiration . In this study, a recombinant strain of Enterobacter aerogenes engineered to produce the Vitreoscilla Hb was investigated with regard to its susceptibility to oxidative stress . The culture response to oxidative stress produced by exogenously applied hydrogen peroxide was characterized in terms of cell growth, survival and the activities of two key antioxidant enzymes (catalase and superoxide dismutase) . The influence of the physiological state of the cells and different media upon these culture dynamics was determined . Results showed that the hemoglobin-expressing strain is quite distinct in terms of growth/survival properties and activity of antioxidant enzymes from that of non-hemoglobin counterparts. Am J Health Syst Pharm, 2003 Jul 15, 60(14), 1440 - 6 Enterobacter cloacae bloodstream infections in pediatric patients traced to a hospital pharmacy; Selenic D et al.; The sources of an outbreak of Enterobacter cloacae bloodstream infections in a pediatric hospital were investigated, as were the risk factors for acquiring the infection: Two retrospective case-control studies were conducted . The study sample included all patients admitted to the general pediatric wards from February 5 through March 30, 2001, who had a positive blood culture for E . cloacae . Pediatric ward and pharmacy infection-control practices were reviewed, personnel and environmental cultures were obtained, and pulsed-field gel electrophoresis (PFGE) molecular typing of the bloodstream isolates was conducted . Four subjects were identified . These infants were more likely than control patients to receive i.v . ranitidine (p < 0.01) . Among patients receiving i.v . ranitidine, subjects were more likely than controls to receive i.v . ranitidine prepared by a pharmacist . No environmental or personnel cultures yielded E . cloacae . Patients' E . cloacae isolates had four different PFGE patterns, suggesting environmental rather than point-source contamination . Ranitidine multidose vials were kept connected to an automatic compounding machine for up to 48 hours at room temperature after the first dose was drawn, contrary to manufacturer recommendations . Further, preparation of ranitidine infusions was not conducted in accordance with recommendations for risk level 2 sterile i.v . products . The use of contaminated ranitidine multidose vials was the most likely cause of an outbreak of E . cloacae . However, a combination of other factors such as inadequate hand-washing techniques, presence of E . cloacae in the environment, noncompliance with guidelines for the preparation of sterile infusions and medications, and a susceptible population may have contributed to the infections. Mol Microbiol, 2003 Aug, 49(4), 1031 - 41 Microcin E492 antibacterial activity: evidence for a TonB-dependent inner membrane permeabilization on Escherichia coli; Destoumieux-Garzon D et al.; The mechanism of action of microcin E492 (MccE492) was investigated for the first time in live bacteria . MccE492 was expressed and purified to homogeneity through an optimized large-scale procedure . Highly purified MccE492 showed potent antibacterial activity at minimal inhibitory concentrations in the range of 0.02-1.2 microM . The microcin bactericidal spectrum of activity was found to be restricted to Enterobacteriaceae and specifically directed against Escherichia and Salmonella species . Isogenic bacteria that possessed mutations in membrane proteins, particularly of the TonB-ExbB-ExbD complex, were assayed . The microcin bactericidal activity was shown to be TonB- and energy-dependent, supporting the hypothesis that the mechanism of action is receptor mediated . In addition, MccE492 depolarized and permeabilized the E . coli cytoplasmic membrane . The membrane depolarization was TonB dependent . From this study, we propose that MccE492 is recognized by iron-siderophore receptors, including FepA, which promote its import across the outer membrane via a TonB- and energy-dependent pathway . MccE492 then inserts into the inner membrane, whereupon the potential becomes destabilized by pore formation . Because cytoplasmic membrane permeabilization of MccE492 occurs beneath the threshold of the bactericidal concentration and does not result in cell lysis, the cytoplasmic membrane is not hypothesized to be the sole target of MccE492. Am J Infect Control, 2003 Aug, 31(5), 280 - 7 Spectrum of microbes and antimicrobial resistance in a surgical intensive care unit, Barbados; Hariharan S et al.; BACKGROUND: To survey the epidemiologic findings of infections and antibiotic resistance patterns in the surgical intensive care unit (ICU) of a tertiary care university teaching hospital . METHODS: The microbiologic culture-sensitivity reports of patients admitted to a surgical ICU were prospectively studied for 6 months each of 3 consecutive years . The antibiotic usage for these patients also was studied concurrently . Reports from general surgical wards for 6 months of 1 year also were analyzed for comparison . The common specimens assayed microbiologically were tracheal aspirate, urine, blood, wound swabs, invasive catheter tips, and screening swabs for methicillin-resistant Staphylococcus aureus . RESULTS: The organisms reported were Enterobacteriaceae, Pseudomonas species, S aureus, and enterococci . Organisms were highly resistant to amoxicillin and first-generation cep