Diagn Microbiol Infect Dis, 2001 Feb, 39(2), 85 - 93 Identification by 16S ribosomal RNA gene sequencing of an Enterobacteriaceae species with ambiguous biochemical profile from a renal transplant recipient; Woo PC et al.; Traditional ways of identification of bacteria by phenotypic characteristics cannot be used for non-cultivable organisms and organisms with unusual biochemical profiles . In this study, an Enterobacteriaceae was isolated in pure growth from the mid-stream urine of a 67-year old renal transplant recipient with urinary tract infection . Conventional biochemical tests did not reveal a pattern resembling any known member of the Enterobacteriaceae family . The Vitek system (GNI+) showed that it was 18% Leclercia adecarboxylata and 55% Klebsiella ozaenae; whereas the API system (20E) showed that it was 99.8% Rahnella aquatilis . 16S ribosomal RNA gene sequencing showed that there was 7 base differences between the isolate and Enterobacter cloacae, 18 base differences between the isolate and Enterobacter asburiae, 17 base differences between the isolate and Enterobacter cancerogenus, 35 base differences between the isolate and K . ozaenae, 27 base differences between the isolate and L . adecarboxylata, and 72 base differences between the isolate and R . aquatilis, indicating that the isolate most closely resembled a strain of E . cloacae . Identification of the organism in this study is important, as the choice of antibiotics would be radically different . In this case, cephalosporins should be avoided regardless of in-vitro susceptibility as cephalosporins are well-known to select for AmpC derepressed mutants in Enterobacter, and previous administration of third-generation cephalosporins is more likely to be associated with multidrug resistant Enterobacter isolates than is administration of antibiotics that do not include a third-generation cephalosporin. Biochim Biophys Acta, 2001 May 1, 1505(1), 15 - 27 Sodium ion-translocating decarboxylases; Buckel W; The review is concerned with three Na(+)-dependent biotin-containing decarboxylases, which catalyse the substitution of CO(2) by H(+) with retention of configuration (DeltaG degrees '=-30 kJ/mol): oxaloacetate decarboxylase from enterobacteria, methylmalonyl-CoA decarboxylase from Veillonella parvula and Propiogenium modestum, and glutaconyl-CoA decarboxylase from Acidaminococcus fermentans . The enzymes represent complexes of four functional domains or subunits, a carboxytransferase, a mobile alanine- and proline-rich biotin carrier, a 9-11 membrane-spanning helix-containing Na(+)-dependent carboxybiotin decarboxylase and a membrane anchor . In the first catalytic step the carboxyl group of the substrate is converted to a kinetically activated carboxylate in N-carboxybiotin . After swing-over to the decarboxylase, an electrochemical Na(+) gradient is generated; the free energy of the decarboxylation is used to translocate 1-2 Na(+) from the inside to the outside, whereas the proton comes from the outside . At high {Na(+)}, however, the decarboxylases appear to catalyse a mere Na(+)/Na(+) exchange . This finding has implications for the life of P . modestum in sea water, which relies on the synthesis of ATP via Delta(mu)Na(+) generated by decarboxylation . In many sequenced genomes from Bacteria and Archaea homologues of the carboxybiotin decarboxylase from A . fermentans with up to 80% sequence identity have been detected. Mikrobiol Z, 2000 Nov-Dec, 62(6), 7 - 14 {Use of PCR for the identification of representatives of Lactobacillus and Streptococcus}; Kovalenko NK et al.; REP- and ERIC-PCR genome analysis of lactic acid bacteria of genera Lactobacillus and Streptococcus have shown the presence of REP- and ERIC-repetitive sequences with high degree of homology . Amplification products which separation in agar gel results in formation of a specific fingerprint are obtained under REP- and ERIC-PCR . It is shown that REP- and ERIC-specific primers can be used for PCR identification of both enterobacteria and lactic acid bacteria isolated from different ecological niches. Mikrobiol Z, 2000 Sep-Oct, 62(5), 23 - 8 {Ecological diagnosis of pyoseptic diseases in newborns using the "Diastaph" disc}; Flegontova VV et al.; The etiological spectrum of pyoseptic diseases was studied in 113 newborns . It was established, that between 187 strains of microorganisms, belonging to 19 species, 73.3% were staphylococci and streptococci, 25.1%--Enterobacteriaceae, 1.6%--Candida albicans . Mixed infection, caused by association of 2-5 Gram-positive and (or) Gram-negative microorganisms, was marked in 51.3% of newborns . In cases of mono-infection (42.5%) staphylococci and streptococci were leading etiological agents . The possibility of generic identification of staphylococci using the diagnostic disks "Diastaph" which contain a new antibiotic batumin has been studied . All the isolated staphylococci strains were highly sensitive to batumin in contrast to other Gram-positive and Gram-negative microorganisms that caused pyodermia in newborns. Clin Exp Rheumatol, 2001 Jan-Feb, 19(1), 47 - 52 The modulation of chlamydial replication by HLA-B27 depends on the cytoplasmic domain of HLA-B27; Kuipers JG et al.; OBJECTIVE: The intracellular persistence of viable Chlamydia trachomatis (CT) within the joint is thought to initiate and maintain the inflammatory process in CT-induced arthritis . CT-induced arthritis is associated with HLA-B27 . Recently it was shown that HLA-B27, besides being a T-cell restriction element, can directly influence the invasion and/or replication of enterobacteriae and alters salmonella-induced signal transduction . It was the aim of this study to analyze the effect of HLA-B27 on CT-invasion and replication in human host cells . METHODS: Human Hela cells and Hela cells transfected with either HLA-B27 cDNA or controls (HLA-A1 cDNA; HLA-B27 mutant = HLA-B27 without cytoplasmic tail; B27Q10 = HLA-B27 Exon 1-4 linked to Exon 5 of murine Q10) were infected with CT . By direct immunofluorescence chlamydial invasion was determined 4 hours post infection (p.i.), chlamydial replication 2 days and 4 days p.i . The number of infective CT in the different cell lines was determined by titration of the cell lysates on Hep-2 cells with subsequent immunoperoxidase staining . RESULTS: Invasion was not affected by HLA-B27 . However, formation of chlamydial inclusion bodies and replication was suppressed by HLA-B27 . Genetically engineered mutants of HLA-B27 (HLA-B27 mutant, B27Q10) lacking the cytoplasmic tail of HLA-B27 did not affect replication . CONCLUSION: The reduction of chlamydial replication by HLA-B27 depends on the cytoplasmic domain of HLA-B27, thus providing a new hypothesis for chlamydial persistence in HLA-B27 positive reactive arthritis. Clin Immunol, 2001 Mar, 98(3), 364 - 9 Spontaneous inflammatory disease in HLA-B27 transgenic mice does not require transporter of antigenic peptides; Khare SD et al.; HLA-B27 is strongly linked with a group of human diseases called spondyloarthropathies . Even though HLA-B27 as an MHC class I molecule would be expected to present endogenously processed peptides such as cytosolic or viral proteins, many of the B27-linked diseases begin after an infection with an enterobacteria, an exogenous antigen . In our previous studies, we have described development of spontaneous inflammatory disease in HLA-B27 transgenic mice expressing beta(2)m free heavy chains on the cell surface . In order to address the role of endogenous versus exogenous antigens and a role for Tap genes in the development of spontaneous diseases, mice lacking Tap-1 (knockout) were mated to HLA-B27/human beta(2)m transgenic mice . B27(+)/human beta(2)m(+) double-transgenic mice (without mouse beta(2)m) lacking the Tap-1 gene developed spontaneous inflammatory disease similar to wild-type Tap-1 gene-expressing counterparts . Our data demonstrate that peptide transporters (Tap) were not involved in the development of spontaneous inflammatory disease in B27(+)/human beta(2)m transgenic animals . Environ Microbiol, 2000 Aug, 2(4), 417 - 27 Detection of abundant sulphate-reducing bacteria in marine oxic sediment layers by a combined cultivation and molecular approach; Wieringa EB et al.; The depth distribution and diversity of sulphate-reducing bacteria (SRB) was analysed in the upper intertidal zone of a sandy marine sediment of the Dutch island Schiermonnikoog . The upper centimetre of the sediment included the oxic-anoxic interface and was cut into five slices . With each slice, most probable number (MPN) dilution series were set up in microtitre plates using five different substrates . In the deeper sediment layers, up to 1 x 10(8) cm(-3) lactate-utilizing SRB were counted, corresponding to 23% of the total bacterial count . From the highest positive dilutions of the MPN series, 27 strains of SRB were isolated in pure culture . Sequencing of a 580 bp fragment of the 16S rDNA revealed that 21 isolates had identical sequences, also identical with that of the previously described species Desulfomicrobium apsheronum . However, the diversity of the isolates was higher with respect to their physiological properties: a total of 11 different phenotypes could be distinguished . Genomic fingerprinting by enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR) revealed an even higher diversity of 22 different genotypes . A culture-independent analysis by PCR and denaturing-gradient gel electrophoresis (DGGE) revealed that the partial 16S rDNA sequence of the isolated D . apsheronum strains constituted a significant fraction of the Desulfovibrionaceae . The high subspecies diversity suggests that this abundant aggregate-forming species may have evolved adaptations to different ecological niches in the oxic sediment layers. Therapie, 2000 Nov-Dec, 55(6), 691 - 7 {Antibacterial activity of urine after administration of ofloxacin for 5 days}; Drugeon H et al.; The antibacterial activity of ofloxacin was evaluated in urine over a period of 96 h after oral administration for 5 days of 200 mg twice a day in 12 healthy female volunteers . Bacteriostatic and bactericidal activity of urines were studied for five strains of enterobacterias recovered from urinary infections: two strains of Escherichia Coli Nal-S and Nal-R, two strains of Proteus mirabilis Nal-S and Nal-R, and one strain of Klebsiella pneumoniae Nal-S . Mean urinary concentrations of ofloxacin were very high during the first 12 h following last intake . They were still above 7 mg/l till the 48th hour and above 1.6 mg/l till the 72nd hour . Bactericidal activity of urine was present for 72 h in respect of four strains studied at that time; urine was not bactericidal as regards E . coli Nal-R . After 5 days of oral treatment with ofloxacin (200 mg b.i.d.), urine retains a bactericidal activity for at least 72 h against bacterial strains of urinary tract infections. Mol Biol (Mosk), 2001 Jan-Feb, 35(1), 157 - 62 {Expression of the NS1 gene of tick-borne encephalitis virus in gram-negative bacteria from the mouse nasopharynx}; Morozova OV et al.; Bacteria were isolated from the nasopharynx of BALB/c mice and electroporated with pUR290(NS1)2 containing two copies of tick-borne encephalitis virus (TBEV) strain Sofjin NS1 under the control of the lac promoter . The plasmid persisted in transformants for at least ten passages . The NS1 gene expression was detected in Gram-negative enterobacteria via immunoblotting with monoclonal antibodies against TBEV nonstructural glycoprotein NS1 . Recombinant NS1 was detected in bacterial cells and in the culture medium . Intranasal immunization with recombinant bacteria activated production of antibodies against NS1 in serum of BALB/c mice . The humoral immune response to NS1 failed to protect immunized mice from a TBEV challenge. Acta Microbiol Immunol Hung, 2001, 48(2), 129 - 41 Detection and toxin production of Staphylococcus aureus in sudden infant death cases in Hungary; Csukas Z et al.; The potential role of microbial agents was investigated in 13 cases of Sudden Infant Death Syndrome and in 9 non-SIDS cases in Budapest between September 1996 and May 1998 . Autopsy, histological examination and microbiological tests were performed on samples of blood, cerebrospinal fluid, pharyngeal samples and lung tissue from infants under one year died suddenly, without previous diseases . The multifactorial pathomechanism of SIDS was suggested by the isolation of toxin producing Staphylococcus aureus-, Enterobacteriaceae and Candida albicans strains in large number and by the detection of Parainfluenza Type 2 virus antigen . S . aureus proved the predominant bacteria in the SIDS cases . Nasopharyngeal microbial flora and S . aureus carrier of 100 age matched healthy infants were tested during the same period . S . aureus was isolated from 54% of SIDS cases and 37% from healthy infants /OR = 1.986 (95% Confidence interval = 0.55-7.33), p = 0243/ . The enterotoxin and TSST-1 toxin producing activity of S . aureus showed the characteristic difference . The toxigenic S . aureus was detected in 46% of SIDS cases and 16% of healthy infants /OR = 4.5 (95% CI = 1.15-17.72), p = 0.010/ . The distribution of toxigenic and nontoxigenic isolates was 86% in SIDS cases and 43% in healthy infants /OR = 7.875 (CI = 0.78-191.89), p = 0.041/. Folia Biol (Praha), 2001, 47(1), 11 - 3 Cytotoxic effects of colicins E1 and E3 on v-myb-transformed chicken monoblasts; Smarda J et al.; Colicins show a considerable cytostatic activity, which is much less known and understood than their killing activity targeting bacteria of the Enterobacteriaceae family . In this communication, the cytotoxic effects of colicins E1 and E3 on v-myb-transformed chicken monoblasts BM2 are presented . We detected clear reduction of the viable cell number induced by colicins E1 and E3, occurring without apparent changes in cell cycle profiles . The level of inhibition was proportional to the colicin concentration within the limits of 0.5-1.25 microg/ml . This result documents that colicins produced by Enterobacteriaceae exert their cytotoxic effects on leukemic cells. J Med Microbiol, 2001 Mar, 50(3), 208 - 14 Characterisation of Hafnia alvei isolates from human clinical extra-intestinal specimens: haemagglutinins, serum resistance and siderophore synthesis; Podschun R et al.; Extra-intestinal Hafnia alvei isolates are rarely considered to be pathogenic . To investigate whether such strains are able to produce virulence factors, a total of 70 clinical H . alvei isolates was compared with clinical extra-intestinal isolates of other members of the enterobacterial tribe Klebsiellae (Kiebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens) . Whereas mannose-sensitive haemagglutination (MSHA) was less common in H . alvei (59%) than in K . pneumoniae (86%) and E . cloacae (89%) isolates, the incidences of mannose-resistant haemagglutination indicative of type 3 pili (MR/K-HA) and of serum resistance properties were not lower . All H . alvei strains secreted siderophores but, unlike the other enterobacterial species examined, the siderophore type was neither enterobactin nor aerobactin . Although the low pathogenicity of H . alvei isolates could not be attributed to any of the factors investigated, the mean number of factors expressed by each H . alvei isolate was significantly lower than that expressed by K . pneumoniae and E . cloacae isolates but did not differ significantly from that of S . marcescens . Based on these findings, the low pathogenicity of H . alvei appears to be due to its low frequency of expression of virulence factors as compared with clinically significant species such as K . pneumoniae and E . cloacae. J Clin Microbiol, 2001 Mar, 39(3), 1190 - 4 Isolation and characterization of a Salmonella enterica serotype Typhi variant and its clinical and public health implications; Woo PC et al.; We report the isolation and characterization of a member of the family Enterobacteriaceae isolated from the gallbladder pus of a food handler . Conventional biochemical tests suggested Salmonella enterica serotype Typhi, but the isolate agglutinated with poly(O), 2O, 9O, and Vi Salmonella antisera but not with poly(H) or any individual H Salmonella antisera . 16S rRNA gene sequencing showed that there were two base differences between the isolate and Salmonella enterica serotype Montevideo, four base differences between the isolate and serotype Typhi, five base differences between the isolate and Salmonella enterica serotype Typhimurium, and six base differences between the isolate and Salmonella enterica serotype Dublin, indicating that the isolate was a strain of S . enterica . Electron microscopy confirmed that the isolate was aflagellated . The flagellin gene sequence of the isolate was 100% identical to that of the H1-d flagellin gene of serotype Typhi . Sequencing of the rfbE gene, which encoded the CDP-tyvelose epimerase of the isolate, showed that there was a point mutation at position +694 (G-->T), leading to an amino acid substitution (Gly-->Cys) . This may have resulted in a protein of reduced catalytic activity and hence the presence of both 2O and 9O antigens . We therefore concluded that the isolate was a variant of serotype Typhi . Besides antibiotic therapy and cholecystectomy, removal of all stones in the biliary tree was performed for eradication of the carrier state. J Clin Microbiol, 2001 Mar, 39(3), 983 - 9 Controlled clinical comparison of BACTEC plus anaerobic/F to standard anaerobic/F as the anaerobic companion bottle to plus aerobic/F medium for culturing blood from adults; Wilson ML et al.; To determine the optimal anaerobic companion bottle to pair with BACTEC Plus Aerobic/F medium for recovery of pathogenic microorganisms from adult patients with bacteremia and fungemia, we compared Plus Anaerobic/F bottles with Standard Anaerobic/F bottles, each of which was filled with 4 to 6 ml of blood . The two bottles were paired with a Plus Aerobic/F bottle filled with 8 to 12 ml of blood . A total of 14,011 blood culture sets were obtained . Of these, 11,583 sets were received with all three bottles filled adequately and 12,257 were received with both anaerobic bottles filled adequately . Of 818 clinically important isolates detected in one or both adequately filled anaerobic bottles, significantly more staphylococci (P < 0.001), streptococci (P < 0.005), Escherichia coli isolates (P < 0.02), Klebsiella pneumoniae isolates (P < 0.005), and all microorganisms combined (P < 0.001) were detected in Plus Anaerobic/F bottles . In contrast, significantly more anaerobic gram-negative bacilli were detected in Standard Anaerobic/F bottles (P < 0.05) . Of 397 unimicrobial episodes of septicemia, 354 were detected with both pairs, 30 were detected with Plus Aerobic/F-Plus Anaerobic/F pairs only, and 13 were detected with Plus Aerobic/F-Standard Anaerobic/F pairs only (P < 0.05) . Significantly more episodes of bacteremia caused by members of the family Enterobacteriaceae (P < 0.05) and aerobic and facultative gram-positive bacteria (P < 0.025) were detected with Plus Anaerobic/F bottles only . In a paired-bottle analysis, 810 of 950 isolates were recovered from both pairs, 90 were recovered from Plus Aerobic/F-Plus Anaerobic/F pairs only, and 50 were recovered from Plus Aerobic/F-Standard Anaerobic/F pairs only (P < 0.001) . Paired Plus Aerobic/F-Plus Anaerobic/F bottles yielded significantly more staphylococci (P < 0.001), streptococci (P < 0.05), and members of the family Enterobacteriaceae (P <0.001) . We conclude that Plus Anaerobic/F bottles detect more microorganisms and episodes of bacteremia and fungemia than Standard Anaerobic/F bottles as companion bottles to Plus Aerobic/F bottles in the BACTEC 9240 blood culture system. J Clin Microbiol, 2001 Mar, 39(3), 889 - 96 National epidemiologic surveys of Enterobacter aerogenes in Belgian hospitals from 1996 to 1998; De Gheldre Y et al.; Two national surveys were conducted to describe the incidence and prevalence of Enterobacter aerogenes in 21 Belgian hospitals in 1996 and 1997 and to characterize the genotypic diversity and the antimicrobial resistance profiles of clinical strains of E . aerogenes isolated from hospitalized patients in Belgium in 1997 and 1998 . Twenty-nine hospitals collected 10 isolates of E . aerogenes, which were typed by arbitrarily primed PCR (AP-PCR) using two primers and pulsed-field gel electrophoresis . MICs of 10 antimicrobial agents were determined by the agar dilution method . Beta-lactamases were detected by the double-disk diffusion test and characterized by isoelectric point . The median incidence of E . aerogenes colonization or infection increased from 3.3 per 1,000 admissions in 1996 to 4.2 per 1000 admissions in the first half of 1997 (P < 0.01) . E . aerogenes strains (n = 260) clustered in 25 AP-PCR types . Two major types, BE1 and BE2, included 36 and 38% of strains and were found in 21 and 25 hospitals, respectively . The BE1 type was indistinguishable from a previously described epidemic strain in France . Half of the strains produced an extended-spectrum beta-lactamase, either TEM-24 (in 86% of the strains) or TEM-3 (in 14% of the strains) . Over 75% of the isolates were resistant to ceftazidime, piperacillin-tazobactam, and ciprofloxacin . Over 90% of the strains were susceptible to cefepime, carbapenems, and aminoglycosides . In conclusion, these data suggest a nationwide dissemination of two epidemic multiresistant E . aerogenes strains in Belgian hospitals . TEM-24 beta-lactamase was frequently harbored by one of these epidemic strains, which appeared to be genotypically related to a TEM-24-producing epidemic strain from France, suggesting international dissemination. Antibiot Khimioter, 2001, 46(1), 15 - 7 {Comparative antibiotic susceptibility in vitro of microbial isolates from patients with complications after gastrectomy for stomach carcinoma and from contents of their large intestine}; Levanov AV; Antibiotic susceptibility of the main isolates from the large intestine contents and pathological substrates of 12 patients after gastroectomy for stomach carcinoma was studied . Three of them developed esophagoenterostomy incompetence, 4 had intraabdominal abscesses and in 5 infection of the operation wound was stated . In all, 30 isolates of enterobacteria, 28 isolates of enterococci and 38 isolates of bacteroides were tested . Antibiotic susceptibility of the isolates from various sources was practically identical, which showed that before surgical operations for stomach carcinoma it was necessary that data on the patients intestine microbiocenosis and antibioticograms of the main isolates should be available to correct severe dysbiotic disorders. Pediatr Infect Dis J, 2001 Feb, 20(2), 134 - 40 Increase of Enterobacter in neonatal sepsis: a twenty-two-year study; Hervas JA et al.; BACKGROUND: Data on the incidence of Enterobacter infections in neonates over prolonged periods of time are scant . We determined the epidemiology of Enterobacter sepsis and/or meningitis and the trends of infection in a neonatal unit . METHODS: Retrospective review of sepsis and/or meningitis in inborn neonates admitted to Son Dureta University Hospital during a 22-year period . Molecular study by ribotyping of the Enterobacter strains isolated from 1995 to 1997 . RESULTS: There were 513 cases of culture-proved sepsis and/or meningitis in neonates . In late onset infections Klebsiella pneumoniae and Staphylococcus epidermidis were the most frequent isolates in the period 1977 through 1991 . Enterobacter was the most common isolate in the period 1992 through 1998 . During this latter period Candida infections also increased, and the resistance rate of Enterobacter to cefotaxime was higher (59.2%) . Decrease in early onset infections and increase in late onsets (4.6/1,000 live births) were observed in the second period . From 1977 to 1998, 45 episodes of sepsis and/or meningitis by Enterobacter species were identified in 44 patients (8.7% of all neonatal bacteremias) . Three patients with Enterobacter bacteremia died (6.6%, 0.03/1,000 live births) . During 1995 through 1997 5 different clones causing sepsis were identified and 3 were predominant . In 1997 there was an outbreak of Enterobacter disease . After cleaning, cohort nursing and hygiene reinforcement, Enterobacter was not isolated in the next 2 years . No change in the antibiotic policy was made . CONCLUSIONS: We observed a resurgence of Enterobacter infections in our neonatal intensive care unit . The sudden disappearance of this microorganism after reinforcement of hygienic measures, without withdrawing cefotaxime, confirms the importance of patient-to-patient transmission of this nosocomial infection . Further studies are needed to establish the role of antibiotics in the emergence of microorganisms in neonatal intensive care units. J Bacteriol, 2001 Mar, 183(6), 1870 - 80 Molecular characterization of global regulatory RNA species that control pathogenicity factors in Erwinia amylovora and Erwinia herbicola pv . gypsophilae; Ma W et al.; rsmB(Ecc) specifies a nontranslatable RNA regulator that controls exoprotein production and pathogenicity in soft rot-causing Erwinia carotovora subsp . carotovora . This effect of rsmB(Ecc) RNA is mediated mostly by neutralizing the function of RsmA(Ecc), an RNA-binding protein of E . carotovora subsp . carotovora, which acts as a global negative regulator . To determine the occurrence of functional homologs of rsmB(Ecc) in non-soft-rot-causing Erwinia species, we cloned the rsmB genes of E . amylovora (rsmB(Ea)) and E . herbicola pv . gypsophilae (rsmB(Ehg)) . We show that rsmB(Ea) in E . amylovora positively regulates extracellular polysaccharide (EPS) production, motility, and pathogenicity . In E . herbicola pv . gypsophilae, rsmB(Ehg) elevates the levels of transcripts of a cytokinin (etz) gene and stimulates the production of EPS and yellow pigment as well as motility . RsmA(Ea) and RsmA(Ehg) have more than 93% identity to RsmA(Ecc) and, like the latter, function as negative regulators by affecting the transcript stability of the target gene . The rsmB genes reverse the negative effects of RsmA(Ea), RsmA(Ehg), and RsmA(Ecc), but the extent of reversal is highest with homologous combinations of rsm genes . These observations and findings that rsmB(Ea) and rsmB(Ehg) RNA bind RsmA(Ecc) indicate that the rsmB effect is channeled via RsmA . Additional support for this conclusion comes from the observation that the rsmB genes are much more effective as positive regulators in a RsmA(+) strain of E . carotovora subsp . carotovora than in its RsmA(-) derivative . E . herbicola pv . gypsophilae produces a 290-base rsmB transcript that is not subject to processing . By contrast, E . amylovora produces 430- and 300-base rsmB transcripts, the latter presumably derived by processing of the primary transcript as previously noted with the transcripts of rsmB(Ecc) . Southern blot hybridizations revealed the presence of rsmB homologs in E . carotovora, E . chrysanthemi, E . amylovora, E . herbicola, E . stewartii and E . rhapontici, as well as in other enterobacteria such as Escherichia coli, Salmonella enterica serovar Typhimurium, Serratia marcescens, Shigella flexneri, Enterobacter aerogenes, Klebsiella pneumoniae, Yersinia enterocolitica, and Y . pseudotuberculosis . A comparison of rsmB sequences from several of these enterobacterial species revealed a highly conserved 34-mer region which is predicted to play a role in positive regulation by rsmB RNA. J Antimicrob Chemother, 2001 Mar, 47(3), 271 - 5 Antimicrobial activity of fluoroquinolone photodegradation products determined by parallel-line bioassay and high performance liquid chromatography; Sunderland J et al.; The fluoroquinolones produce multiple photodegradation products . Little is known about these products, particularly whether any possess antimicrobial activity . To investigate this, we used the parallel-line bioassay to investigate discrepancies in zone of inhibition size in conjunction with high performance liquid chromatography (HPLC) analysis . A continuous flow photochemical reaction unit ('Beam-Boost') was used to partially photodegrade the fluoroquinolones ofloxacin, levofloxacin, ciprofloxacin and moxifloxacin (0.02 mM) by between 15 and 89%, as confirmed by HPLC . The concentration of residual parent fluoroquinolone in each irradiated sample was measured by HPLC and a non-irradiated control solution was prepared at the same concentration . These were compared by parallel-line bioassays using Escherichia coli, Enterobacter cloacae and Klebsiella oxytoca . With ofloxacin and levofloxacin, the zone size for the control solution was significantly less than that of the irradiated solutions, with >15% photodegradation in at least two of the indicator organisms, indicating that the photodegradation products possess antimicrobial activity . No difference was seen with ciprofloxacin at any level of photodegradation with any of the indicator organisms, nor with moxifloxacin at 30 and 54% photodegradation . A significant difference was observed with E . cloacae only, at 83% photodegradation. CLAO J, 2001 Jan, 27(1), 47 - 52 Microbiological profile of a shipboard environment and the flora on contact lenses of seamen; Theng JT et al.; PURPOSE: The purpose of this study was to culture and identify the spectrum of organisms (and their sensitivities) that contaminate the extended wear contact lenses of seamen in their working environment . A secondary aim was to identify the most appropriate first-line antibiotic regimen to be used on seamen who sustain contact lens-related corneal keratitis on board ship . METHODS: Twenty pairs of contact lenses of 20 seamen in one group and 24 pairs in another group wearing contact lenses were collected at the end of 1 week of extended wear . Groups one and two differed only in the way lenses were stored prior to culturing . All contact lenses were then brought to the microbiological lab within 24 hours for culture and sensitivity testing . RESULTS: The most common contaminants on the contact lenses in this study were Staphylococcus epidermidis and Staphylococcus aureus, organisms resident on the normal eyelids . Enterobacterand Pseudomonas species, present in a shipboard environment, were also identified as contaminants on the contact lenses . The organisms cultured from the contact lenses correlated well with those of normal lid flora as well as from the ship environment and are probably derived from these sources . Storage in saline-containing preservatives yielded significantly less positive bacterial cultures from the contact lenses in our study . All bacterial isolates were sensitive to ciprofloxacin whereas several bacteria resistant to cefazolin and gentamicin were identified . CONCLUSION: The most common contaminants on the contact lenses in this study were Staphylococcus epidermidis and Staphylococcus aureus, organisms resident on the normal eyelids . Enterobacter and Pseudomonas species, which are normally present in a shipboard environment, were also identified as contaminants on the contact lenses . Ciprofloxacin is effective against all organisms identified as contaminants on the contact lenses in this study . Of all the antibiotics tested, it is probably the most suitable agent against contact lensrelated keratitis under such shipboard circumstances and is thus recommended in these situations. Environ Microbiol, 2000 Dec, 2(6), 620 - 31 Variations in antibiotic resistance profile in Enterobacteriaceae isolated from wild Australian mammals; Sherley M et al.; We carried out a retrospective analysis of 946 strains of Enterobacteriaceae isolated from wild Australian mammals between 1993 and 1997 . The prevalence of resistance to fixed concentrations of 32 antimicrobial agents was determined, and the respective roles that taxonomic family of the host, state of origin and bacterial species play in defining prevalence and range of resistance were investigated . Our results demonstrated a low but widespread prevalence of antimicrobial resistance in wild isolates . Only amikacin, ciprofloxacin, meropenem and gentamicin inhibited growth in all 946 samples . There was extensive variation in the combination of antibiotics to which isolates were resistant, and multiple antibiotic resistance was common . Geographical location and host group significantly influenced the antibiotic resistance profile of an isolate, whereas bacterial species influenced both the resistance profile of an isolate and the number of antibiotics it was resistant to . The role of these factors in determining observed antibiotic resistance profiles suggests that any study measuring resistance in wild isolates should include the broadest possible range of bacterial species, host species and sampling locations . As such, this study provides an important new baseline for future measurements of antibiotic resistance in the Australian environment. J Med Liban, 2000 Jul-Aug, 48(4), 278 - 82 Management of urinary tract infections; Nassar NT; Urinary tract infections (UTIs) are commonly encountered in medical practice and range from asymptomatic bacteruria to acute pyelonephritis . Enterobacteriaceae with E . coli being the most prevalent, are responsible for most commonly acquired uncomplicated UTIs and usually respond promptly to oral antibiotics . In contradistinction, more resistant pathogens cause nosocomially acquired infections which often require parenteral antibiotic therapy . Patients with acute bacterial prostatitis, usually caused by Enterobacteriaceae present with a tender prostate gland and respond promptly to antibiotic therapy . Chronic bacterial prostatitis on the other hand, is a subacute infection characterized by recurrent episodes of bacterial UTI where the patient presents with vague symptoms of pelvic pain and voiding problems . Treatment is protracted and may be frustrating . Nonbacterial prostatitis and chronic pelvic pain syndrome produce symptoms similar to those of chronic bacterial prostatitis . Treatment is not well defined due to their uncertain etiologies . Most episodes of catheter associated bacteruria are asymptomatic, where less than 5% will be complicated by bacteremia . The use of systemic antibiotics for treatment or prevention of bacteruria is not recommended, particularly in the geriatric age group, since it helps select for resistant organisms . Prevention thus remains the best option to control it . Few patients without catheters who have asymptomatic bacteruria develop serious complications and therefore routine antimicrobial therapy is not justified with only two exceptions : before urologic surgery and during pregnancy. J Med Liban, 2000 Jul-Aug, 48(4), 227 - 32 Current status and changing trends of antimicrobial resistance in Saudi Arabia; Akhter J et al.; Due to modern travel and ease of spread of infections, it is desirable to widen knowledge of susceptibility of common bacterial isolates from different parts of the world for optimal clinical management and control programs . Over the past decades, antimicrobial resistance has emerged in all kinds of micro-organisms worldwide including Saudi Arabia . This phenomenon is primarily due to increasing antibiotic use and misuse in humans, animals and agriculture . Additionally, the presence of a large expatriate population and a significant number of visitors to the Kingdom annually for pilgrimage and/or work from all over the world may have also facilitated the importation to Saudi Arabia of drug resistant micro-organisms from other countries . Saudi Arabia has witnessed an increase of drug resistant Mycobacterium tuberculosis, Streptococcus pneumoniae, Staphylococcus aureus and some Enterobacteriaceae in the last decade . We describe the status of antimicrobial resistance in Saudi Arabia which is an important focus of antimicrobial resistance for the Gulf Region. Cell Mol Life Sci, 1999 Nov 30, 56(9-10), 771 - 8 Bacterial antibiotic efflux systems of medical importance; Kohler T et al.; Multidrug efflux systems endow on bacterial cells the ability to limit the access of antimicrobial agents to their targets . By actively pumping out antibiotic molecules, these systems prevent the intracellular accumulation necessary for antibiotics to exert their lethal activity . Drug efflux appears to be one of the most widespread antibiotic resistance mechanisms among microorganisms, since it has been demonstrated to occur in many Gram-positive and Gram-negative bacteria including medically important species like staphylococci, streptococci, enterobacteria and opportunistic pathogens like Pseudomonas aeruginosa . Efflux pumps can be specific for only one substrate or accommodate a more or less wide range of noxious products . Export of structurally unrelated compounds confers a multidrug-resistance phenotype on bacterial cells . Therapeutically critical levels of resistance can be achieved by overexpression of efflux systems, especially in those species such as P . aeruginosa which possess a low outer membrane permeability . It is suspected that the dual physiological function of active efflux systems is both the secretion of intracellular metabolites and the protection against a variety of harmful substances that the microorganism may encounter in its natural environment. J Hepatol, 2001 Jan, 34(1), 32 - 7 Bacterial translocation of enteric organisms in patients with cirrhosis; Cirera I et al.; BACKGROUND/AIMS: The aim of the study was to investigate the prevalence and associated risk factors for bacterial translocation in patients with cirrhosis, a mechanism involved in the pathogenesis of bacterial infections in experimental cirrhosis . METHODS: Mesenteric lymph nodes were obtained for microbiological culture from 101 patients with cirrhosis and from 35 non-cirrhotic patients . RESULTS: Enteric organisms were grown from mesenteric lymph nodes in 8.6% of non-cirrhotic patients . In the 79 cirrhotic patients without selective intestinal decontamination, the prevalence of bacterial translocation significantly increased according to the Child-Pugh classification: 3.4% in Child A, 8.1% in Child B and 30.8% in Child C patients (chi2 = 6.106, P < 0.05) . However, translocation by Enterobacteriaceae, the organisms commonly responsible for spontaneous bacteremia and peritonitis in cirrhosis, was only observed in 25% of the cases . The prevalence of bacterial translocation in the 22 cirrhotic patients undergoing selective intestinal decontamination, all Child-Pugh class B and C, was 4.5% . The Child-Pugh score was the only independent predictive factor for bacterial translocation (odds ratio 2.22, P = 0.02) . CONCLUSIONS: Translocation of enteric organisms to mesenteric lymph nodes is increased in patients with advanced cirrhosis and is reduced to the level found in non-cirrhotic patients by selective intestinal decontamination. Antibiot Khimioter, 2000, 45(11), 6 - 8 {Non-sporulating anaerobic bacteria and anastomosis failure in patients with gastric carcinoma}; Levanov AV et al.; Cases of anastomosis suture failure within the period from 1977 to 1987 and from 1988 to 1998 in 139 patients after various surgical operations for gastric carcinoma were analyzed . Infection in the cases of the anastomosis sUture failure at the early terms was mainly due to representatives of Enterobacteriaceae and at the later terms the failure was mainly due to non-sporulating anaerobes belonging to Bacteroidaceae . The data are indicative of the fact that the use of antimicrobials requires a differential approach. An R Acad Nac Med (Madr), 2000, 117(1), 163 - 78; discussion 178-83 {Current interest of antipneumococcal vaccination}; del Rey Calero J; Pneumonia acquired in Community (CAP) may be a primary disease occurring in healthy individuals or secondary to predisposing factors or comorbidity . Prevalence of CAP is 2.6 to 5% for all ages, in USA 12%, for over 65 years 30% . Streptococcus pneumoniae is the commonest pathogen 30-50%, H . influenzae in COPD, the atypical pneumonia Mycoplasma pn., M . catharralis, Legionella pn., Enterobacteria, anaerobics often in hospital survey . In children is different RSV, Parainfluenzae type 3, Rhinovirus in the first 2 years old . Others are S . pneumoniae, H . influenzae, Chlamydia sp., etc . Appropriate empiric antibiotic therapy choices are based in guidelines . The most common pathogen is S . pneumoniae, isolates raised resistance rates to Penicillin to 20-50%, 40% in our country and also to Macrolides, with potential clinical failure (21-40%) . Specially in elderly people and with the comorbidity are recommended the 23 valent polysaccharide vaccine, effective in bacteremic pneumonia 70-80% . Is not effective in children under 2 years, for that is important conjugated vaccine Hib (toxoids T, D, CRM197, OMP Nm) to prevent carriers, otitis media and reduce exacerbation of these respiratory infections. Pathol Biol (Paris), 2000 Dec, 48(10), 933 - 9 {Comparative antibacterial activity of cefotaxime, ceftazidime and cefepime with regard to different strains of Enterobacter aerogenes selected for their resistance to third generation cephalosporins}; Husson MO et al.; After being confronted with the isolation in our laboratory of numerous antibiotic-multiresistant Enterobacter aerogenes strains, we studied the in vitro antimicrobial activity of cefotaxime, ceftazidime, and cefepime alone or in association with sulbactam . For that, we selected 67 isolates according to their low level of susceptibility to cefotaxime . First, we deduced from a synergy test in presence of clavulanic acid and cloxacillin the production of an extended spectrum beta-lactamase (ESBL) and/or an overproduction of a chromosomal cephalosporinase . Three groups of strains were thus defined: one group of ESBL strains, another group of overproducing strains of chromosomal cephalosporinase, and a last group that produced the two types of enzymes . Minimal inhibitory concentrations (MICs) of each cephalosporin alone or in presence of 8 mg/L of sulbactam, gentamicin or amikacin were measured . Our results demonstrated the best activity of cefepime: MICs were low with a value inferior to 4 mg/L independently of the type of beta-lactamase . They were lower than 0.5 mg/L in presence of sulbactam against ESBL-producing strains . The cephalosporins could be used in association with aminoglycosides according to their susceptibility. Z Naturforsch {C}, 2000 Nov-Dec, 55(11-12), 886 - 9 Antimicrobial activities of Ferulago essential oils; Demirci F et al.; Essential oils from Ferulago asparagifolia Boiss., F . galbanifera (Miller) W . Koch, F . humilis Boiss . (Endemic), F . trachycarpa Boiss . growing in Turkey were evaluated against 15 microorganisms for their antifungal and antibacterial activity using an agar tube dilution and microdilution broth susceptibility assay, respectively . The essential oil compositions were investigated by GC/MS . Inhibitory effects against Escherichia coli, Enterobacter aerogenes, Candida albicans, Gaeumannomyces graminis var . tritici, Sclerotium rolfsii and Fusarium moniliforme were remarkable . Results are discussed in comparison with the chemical composition of the essential oils. Scand J Infect Dis, 2000, 32(6), 615 - 21 Increased incidence of bacteraemia due to viridans streptococci in an unselected population of patients with acute myeloid leukaemia; Persson L et al.; The aetiology, clinical characteristics and outcome of bacteraemia in patients with acute myeloid leukaemia were studied . All positive blood cultures collected at a haematological ward during 2 7-y periods were evaluated . Altogether, 274 episodes of bacteraemia in 152 patients were recorded, 80 episodes during 1980-86 and 194 during 1990-96 . During the 2 periods, trimethoprim-sulfamethoxazol in combination with amikacin was the first-line empirical therapy in patients with neutropaenia and fever . In 1990, antimicrobial prophylaxis with ciprofloxacin and fluconazole was introduced . The incidence of bacteraemia due to viridans streptococci or coagulase-negative staphylococci increased from the first period to the second, whereas the incidence of Enterobacteriaceae decreased . In granulocytopaenic patients during 1990-96, viridans streptococci accounted for 21% of the isolates and in patients treated prophylactically with fluoroquinolone, viridans streptococci accounted for 31% . All viridans streptococci were sensitive to penicillin . At the time of the positive blood cultures, the patients of the second period were granulocytopaenic in 83% of the episodes . The mortality related to septicaemia during the later period was 13% and only 1 of 33 (3%) of the patients with viridans streptococci died . Eight patients (9%) died in relation to septicaemia following curative antileukaemic therapy. J Mol Microbiol Biotechnol, 2001 Jan, 3(1), 103 - 12 Phyloproteomics: species identification of Enterobacteriaceae using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; Conway GC et al.; To evaluate matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) as a tool for rapid identification of common clinical bacterial isolates, we analyzed 25 carefully selected isolates of pathogenic Escherichia coli (E . coli) and additional Enterobacteriaceae members . Organisms were prepared according to clinical microbiological protocols and analyzed with minimal additional processing . Spectra were reproducible from preparation to preparation and comprised 40-100 peaks primarily representing intracellular proteins with masses up to 25 kDa . Spectra of 14 genetically diverse bacteremic isolates of E . coli were compared with isolates representing other genera within the Enterobacteriaceae family . Using a new spectrum comparison algorithm, E . coli isolates were closely related to each other and were readily distinguishable from other Enterobacteriaceae, including Salmonella and Shigella . Presently, the methodology permits the analysis of 40 unknown isolates per hour per instrument . These results suggest that MALDI-ToF MS offers a rapid and reliable approach for performing phyloproteomics i.e., identification of unknown bacterial isolates based on similarities within protein biomarker databases. J Basic Microbiol, 2000, 40(5-6), 343 - 9 Bacterial counts associated with poultry processing at different sampling times; Geornaras I et al.; Aerobic plate counts, Enterobacteriaceae counts and Pseudomonas counts were performed on neck skin samples from six processing steps in a poultry abattoir at three different sampling times . Sampling time 1 was shortly after start-up of processing operations, time 2 after a tea break which was preceded by a cold water rinse-down of equipment surfaces, and time 3 before shut-down . No significant differences (P > 0.05) in microbial numbers of neck skin samples were observed between the three sampling times at the six sampling sites . At this particular processing plant, therefore, sampling at any time of the processing shift would thus not lead to significantly different bacterial counts of neck skins . The lowest aerobic plate counts, over all three sampling times, were obtained for neck skins sampled after spray washing, and the highest for neck skins sampled after packaging . This indicated the efficacy of the washing step in reducing microbial contamination but subsequent re-contamination of carcasses . Despite the Pseudomonas counts of neck skins being lower than the Enterobacteriaceae counts at the beginning of processing, packaging of carcasses resulted in Pseudomonas counts that were higher than the Enterobacteriaceae counts. J Food Prot, 2001 Jan, 64(1), 72 - 80 Quantification and variability analysis of bacterial cross-contamination rates in common food service tasks; Chen Y et al.; This study investigated bacterial transfer rates between hands and other common surfaces involved in food preparation in the kitchen . Nalidixic acid-resistant Enterobacter aerogenes B199A was used as a surrogate microorganism to follow the cross-contamination events . Samples from at least 30 different participants were collected to determine the statistical distribution of each cross-contamination rate and to quantify the natural variability associated with that rate . The transfer rates among hands, foods, and kitchen surfaces were highly variable, being as low as 0.0005% and as high as 100% . A normal distribution was used to describe the variability in the logarithm of the transfer rates . The mean +/- SD of the normal distributions were, in log percent transfer rate, chicken to hand (0.94 +/- 0.68), cutting board to lettuce (0.90 +/- 0.59), spigot to hand (0.36 +/- 0.90), hand to lettuce (-0.12 +/- 1.07), prewashed hand to postwashed hand (i.e., hand washing efficiency) (-0.20 +/- 1.42), and hand to spigot (-0.80 +/- 1.09) . Quantifying the cross-contamination risk associated with various steps in the food preparation process can provide a scientific basis for risk management efforts in both home and food service kitchens. Enferm Infecc Microbiol Clin, 2000 Dec, 18(10), 500 - 5 {False resistance to imipenem in gram negative bacilli with and automatized system}; Fernandez F et al.; BACKGROUND: The aim of this study was to evaluate the reliability of MIC values of imipenem against gramnegative rods obtained with the automated system WalkAway-98 (MicroScan, Dade, USA) . MATERIAL AND METHODS: One-hundred and seventy three consecutive clinical isolates of Gram-negative rods for which the MIC of imipenem were > or = 4 mg/l (Urine-Combo 6I panels, U6I) or > or = 8 mg/l (Neg-Combo 6I panels, N6I) were evaluated, including 104 non-fermenting gram-negative rods (NFGNR) and 69 enterobacteria . Panels were inoculated and read according to manufacturer's instructions . Microdilution, according to NCCLS guidelines, was used as the method of reference . MIC of imipenem determined by WalllAway-96 and microdilution differing > or = 2 dilution steps from those obtained with mirodilution were considered as discrepant results . Discrepancies in clinical categories were also evaluated by calculating three types of errors: very major (false susceptibility), major (false resistance) and minor (either susceptible or resistant by one method and intermediate by the other one) . RESULTS: The percentages of discrepancies in the MIC of imipenem determined with U6I panels were 74% and 84% for NFGNR and enterobacteria, respectively . No very major errors were detected . Major errors were observed for 6% and 12% of the strains with U6I panels in NFGNR and enterobacteria, respectively, and in 12% (NFGNR) and 50% (enterobacteria) with N61 panels . With U61 panels minor errors were observed in 11% and 25% of NFGNR and enterobacteria, respectively, while with N61 panels minor error were observed in 39% and 45% of both groups, respectively . CONCLUSIONS: MIC of imipenem > or = 4 mg/l obtained with the WalkAway-96 system against gramnegative rods, particularly in the case of enterobacteria, should be confirmed with a reference susceptibility method. Mol Plant Microbe Interact, 2001 Jan, 14(1), 10 - 20 Role of the nucleoid-associated protein H-NS in the synthesis of virulence factors in the phytopathogenic bacterium Erwinia chrysanthemi; Nasser W et al.; The ability of the enterobacterium Erwinia chrysanthemi to induce pathogenesis in plant tissue is strongly related to the massive production of plant-cell-wall-degrading enzymes (pectinases, cellulases, and proteases) . Additional factors, including flagellar proteins and exopolysaccharides (EPS), also are required for the efficient colonization of plants . Production of these virulence factors, particularly pectate lyases, the main virulence determinant, is tightly regulated by environmental conditions . The possible involvement of the protein H-NS in this process was investigated . The E . chrysanthemi hns gene was cloned by complementation of an Escherichia coli hns mutation . Its nucleotide sequence contains a 405-bp open reading frame that codes for a protein with 85% identity to the E . coli H-NS protein . An E . chrysanthemi hns mutant was constructed by reverse genetics . This mutant displays a reduced growth rate and motility but an increased EPS synthesis and sensitivity toward high osmolarity . Furthermore, pectate lyase production is dramatically reduced in this mutant . The hns mutation acts on at least two conditions affecting pectate lyase synthesis: induction of pectate lyase synthesis at low temperatures (25 degrees C) is no longer observed in the hns mutant and induction of pectate lyase production occurs in the late stationary growth phase in the hns background, instead of in the late exponential growth phase as it does in the parental strain . Moreover, the E . chrysanthemi hns mutant displays reduced virulence on plants . Taken together, these data suggest that H-NS plays a crucial role in the expression of the virulence genes and in the pathogenicity of E . chrysanthemi. Aust Vet J, 2000 Dec, 78(12), 846 - 9 Application of ERIC-PCR for the comparison of isolates of Haemophilus parasuis; Rafiee M et al.; OBJECTIVE: To validate a polymerase chain reaction (PCR) based method . Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), for the fingerprinting of Haemophilus parasuis strains and to use that method to differentiate isolates from apparently related outbreaks of Glasser's disease on three pig farms . DESIGN: ERIC-PCR was evaluated by comparing 15 different strains that represented all 15 recognised serovars in the Kielstein-Rapp-Gabrielson (KRG) scheme for serotyping H parasuis . Next, ERIC-PCR was used to examine 14 Australian field isolates of H parasuis; 12 collected from three farms suffering apparently related outbreaks of Glasser's disease and two from two other farms with no known connection . RESULTS: The 15 serovar reference strains all gave unique, reproducible ERIC-PCR fingerprints . The 12 isolates from the three apparently related outbreaks all gave a single fingerprint, which was distinct from any seen in the 15 serovar reference strains and the two other Australian field isolates in the studied farms . The confirmation that all 12 isolates were the same strain allowed the development of a prevention and control program that has prevented the emergence of any further outbreaks of Glasser's disease on the three farms . CONCLUSION: ERIC-PCR is a suitable technique for the differentiation of unrelated strains of H parasuis . The finding that the 12 field isolates of H parasuis all shared the same fingerprint is strong evidence that there was a common source of infection on all three farms . This study has shown, for the first time, that ERIC-PCR is a suitable technique for the sub-typing of H parasuis and useful for studying the epidemiology of outbreaks of Glasser's disease. World J Surg, 2000 Dec, 24(12), 1537 - 41 Remaining small bowel length: association with catheter sepsis in patients receiving home total parenteral nutrition: evidence of bacterial translocation; Terra RM et al.; Patients with short bowel syndrome (SBS) receiving total parenteral nutrition (TPN) have a high incidence of catheter-related sepsis, one of its major complications . The aim of this study was to correlate the length of remaining small bowel (RSB) with septic episodes related to the central venous catheter in a group of patients with severe SBS with home TPN . The length of the RSB (<50 cm or > or = 50 cm) was related to the frequency of catheter sepsis, time until the first episode, and the agents responsible in eight SBS patients receiving home TPN . There were 13 episodes of catheter infection (0.88 per patient-year) . The group with a shorter RSB length (five patients) presented 1.3 to 2.76 infections/year and 2 to 9 months until the first episode, compared to 0 to 0.75 infections/ year (p = 0.0357) and 11 to 65 months until the first episode (p = 0.0332) in the group with the longer RSB . In the first group, the agents isolated were Enterobacteriae (Enterobacter sp., Klebsiella sp., Pseudomonas sp., and Proteus sp.) in eight episodes and Candida sp . in one . In the latter sepsis was caused by Staphylococcus sp . in three episodes and Pseudomonas sp . in one . Therefore patients with remaining small bowel shorter than 50 cm have a higher frequency of catheter-related sepsis, particularly by enteric microorganisms . This might be an evidence of the occurrence of bacterial translocation and its role in the pathogenesis of catheter-related sepsis in patients with an extremely short RSB receiving home TPN. J Med Microbiol, 2001 Jan, 50(1), 29 - 34 Molecular analysis of clinical isolates of Providencia alcalifaciens; Sobreira M et al.; In an attempt to elucidate the virulence factors and the pathogenic mechanisms of Providencia alcalifaciens, 36 isolates identified in 1994-1995 in Recife city, Brazil were analysed by PCR to investigate the presence of DNA sequences homologous to virulence genes described in other invasive enterobacteria, as well as their ability to invade HeLa cells, their plasmid profiles and antibiotic resistance patterns . The genetic diversity of the isolates was also analysed by RAPD-PCR . No homologous sequences of virulence genes were observed with any of the P . alcalifaciens isolates studied . Ten isolates had no plasmid and 26 harboured one-to-five plasmids of 147-<6.9 kb . Invasion of HeLa cells was observed in only 10 isolates . No correlation between the plasmid content of the strains, their invasion of HeLa cells or their resistance to antimicrobial drugs could be established . The isolates could be distributed into 10 genotypic groups by RAPD-PCR . Considering the genotypic profile and ability to invade HeLa cells, 7 of the 10 invasive isolates belonged to the same genotypic group . The presence of invasive isolates in the same or a related genotypic group suggests the existence of a clonal lineage responsible for the invasiveness. Akush Ginekol (Sofiia), 2000, 39(3), 19 - 22 {Nosocomial enterobacter - sepsis in neonatal intensive care unit in Pleven}; Rosmanova R et al.; At the recent years considerably increased the role of Enterobacter spp as causes of nosocomial sepsis in different intensive units . It is determined of their possibilities to change quick antibiotic resistance especially to third generation cephalosporins and wide insemination of the environment . During the period from 29.10.1997 to 30.12.1997 in Neonatal intensive care unit--Pleven have been registered 9 cases of nosocomial sepsis caused by Enterobacter aerogenes . The clinic picture run with shock, temperature instability, hyperbilirubinemia, mottling respiratory insufficiency, I/T ratio > 0.2 . Thrombocytopenia . Detection of the pathogen organisms present wide antibiotics resistance to cephalosporins . The microbiological control of the environment have been isolated Enterobacter spp from neonatal intensive unit, delivery room, unit for healthy newborn . The persistence of Enterobacter infections is a result of low supply of household linen, detergents, single-used products and widely used of cephalosporins, insufficient staff and overpopulation with patients. Urologiia, 2000 Mar-Apr, (2), 8 - 15 {Practical approaches of antibiotics choice in uncomplicated urinary tract infections}; Strachunskii LS et al.; Infections of the urinary tract (IUT) belong to the most prevalent infectious diseases . Acute cystitis is the most frequent symptom of uncomplicated IUT . The main agents of IUT are gram-negative enterobacteria, mainly Escherichia coli (80%) . The agents of uncomplicated IUT are the least resistant (5%) to fluoroquinolones (norfloxacin and ciprofloxacin) . The duration of antibiotic therapy for acute cystitis is determined predominantly by risk factors: a 7-day course is recommended for cases with risk factors and a 3-day one for cases without risk factors . In acute pyelonephritis antibiotic therapy should be longer (10-14 days) . Preventive therapy is recommended for patients with frequent relapses of IUT. Int J Antimicrob Agents, 2000 Sep, 16(1), 5 - 15 The fluoroquinolone antibacterials: past, present and future perspectives; Appelbaum PC et al.; The history of the development of the quinolones is described from the first quinolone, nalidixic acid, via the first 6-fluorinated quinolone norfloxacin, to the latest extended-spectrum fluoroquinolones . The structural modifications made to the basic quinolone and naphthyridone nucleus and to the side chains have allowed improvements to be made such that the next group of fluoroquinolones after norfloxacin, exemplified by ciprofloxacin, had high activity against gram-negative species and a number of atypical pathogens, good-to-moderate activity against gram-positive species and were well absorbed and distributed . These compounds have been successfully used in the clinic for a decade and the size of the market has risen in recent years to only a little less than that for penicillins and macrolides . Notwithstanding the broad spectrum of these compounds, defects became evident . The growth in understanding of structure activity relationships with fluoroquinolones has enabled the development of even better compounds . The targets in fluoroquinolone research during the last few years include: improvements in pharmacokinetic properties, greater activity against gram-positive cocci and anaerobes, activity against fluoroquinolone-resistant strains, and improvements in activity against non-fermentative gram-negative species . The compounds developed in the recent years have fulfilled some but not all of these goals; improved bioavailability is one target achieved with most of the more recent compounds allowing for once-daily dosing . Gatifloxacin, moxifoxacin and trovafloxacin have all greatly improved the activity against gram-positive cocci, particularly pneumococci, and against anaerobes . They are not quite as active as ciprofloxacin against Enterobacteriaceae, and show no substantial improvements in activity against non-fermentative species . Clinafloxacin, gemifloxacin and sitafloxacin have even better activity against gram-positive cocci and are as active as ciprofloxacin against most gram-negatives, though gemifloxacin is less active than the other new compounds against gram-negative anaerobes . These three compounds do retain some activity against a number of ciprofloxacin-resistant species (gram-positive and gram-negative), but whether this activity will be adequate for clinical use is at present unclear . Both clinafloxacin and sitafloxacin contain a chloro substituent at position 8 of the quinolone nucleus . A halogen at this position in a number of compounds, though giving good activity, has also been associated with phototoxicity . Several fluoroquinolones have had to be withdrawn or strictly limited in their use post-marketing and in some cases no obvious relationship can be seen between the adverse effects and structural features, making this an area for urgent research. Med Sci Monit, 2000 Mar-Apr, 6(2), 285 - 90 Comparison of bacterial flora found in the peritoneal cavity and drains after intraabdominal surgery; Chylak J et al.; The aim of this study was to evaluate bacterial flora infecting during the surgical procedures the peritoneal cavity and drains which were used after the surgery in 40 patients who underwent surgery for colon and rectum tumors . Smears from the peritoneal cavity, liquid removed from the drain taken at 3-4 days after the surgery and smears from the drain taken at the end of drainage of each patient were examined for bacterial content . The comparison of bacterial flora found in the peritoneal cavity with bacteria found in drains showed that the frequency of isolation of anaerobes decreased in contrast to aerobes which were more often cultured over the time of drainage (p < 0.05) . Bacteroides spp, enterococci, and strains of Enterobacteriaceae were most often isolated from the peritoneal cavity, but over the time of drainage the frequency of isolation of coagulase-negative staphylococci including MRCNS increased as well as the number of patients infected with enterococci, whereas anaerobes and strains of Enterobacteriaceae were rarely isolated. Antimicrob Agents Chemother, 2001 Mar, 45(3), 883 - 92 Phenotypic resistance of Staphylococcus aureus, selected Enterobacteriaceae, and Pseudomonas aeruginosa after single and multiple in vitro exposures to ciprofloxacin, levofloxacin, and trovafloxacin; Gilbert DN et al.; The phenotypic resistance of selected organisms to ciprofloxacin, levofloxacin, and trovafloxacin was defined as a MIC of > or =4 microg/ml . The dynamics of resistance were studied after single and sequential drug exposures: clinical isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MSSA and MRSA), Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens, and Pseudomonas aeruginosa were utilized . After a single 48-h exposure of a large inoculum to four times the initial MIC for the organism, the frequency of selection of resistant mutants of MSSA was greater for trovafloxacin than levofloxacin (P = 0.008); for E . cloacae, the frequency was highest for ciprofloxacin and lowest for levofloxacin and trovafloxacin; for S . marcescens, the frequency was highest for trovafloxacin and lowest for ciprofloxacin (P = 0.003) . The results of serial passage experiments were analyzed both by the Kaplan-Meier product-limited method as well as by analysis of variance of mean inhibitory values . By both methods, MSSA and MRSA expressed mutants resistant to ciprofloxacin after fewer passages than were required for either levofloxacin or trovafloxacin . For the aerobic gram-negative bacilli, two general patterns emerged . Mutants resistant to trovafloxacin appeared sooner and reached higher mean MICs than did mutants resistant to levofloxacin or ciprofloxacin . Mutants resistant to ciprofloxacin appeared later and reached mean MICs lower than the MICs of the other two drugs studied . Even though individual strain variation occurred, the mean MICs were reproduced when the serial passage experiment was repeated using an identical panel of E . coli isolates . In summary, the dynamic selection of fluoroquinolone-resistant bacteria can be demonstrated in experiments that employ serial passage of bacteria in vitro. Antimicrob Agents Chemother, 2001 Mar, 45(3), 825 - 36 In vitro and in vivo properties of Ro 63-9141, a novel broad-spectrum cephalosporin with activity against methicillin-resistant staphylococci; Hebeisen P et al.; Ro 63-9141 is a new member of the pyrrolidinone-3-ylidenemethyl cephem series of cephalosporins . Its antibacterial spectrum was evaluated against significant gram-positive and gram-negative pathogens in comparison with those of reference drugs, including cefotaxime, cefepime, meropenem, and ciprofloxacin . Ro 63-9141 showed high antibacterial in vitro activity against gram-positive bacteria except ampicillin-resistant enterococci, particularly vancomycin-resistant strains of Enterococcus faecium . Its MIC at which 90% of the isolates tested were inhibited (MIC(90)) for methicillin-resistant Staphylococcus aureus (MRSA) was 4 microg/ml . Ro 63-9141 was bactericidal against MRSA . Development of resistance to the new compound in MRSA was not observed . Ro 63-9141 was more potent than cefotaxime against penicillin-resistant Streptococcus pneumoniae (MIC(90) = 2 microg/ml) . It was active against ceftazidime-susceptible strains of Pseudomonas aeruginosa and against Enterobacteriaceae except Proteus vulgaris and some isolates producing extended-spectrum beta-lactamases . The basis for the antibacterial spectrum of Ro 63-9141 lies in its affinity to essential penicillin-binding proteins, including PBP 2' of MRSA, and its stability towards beta-lactamases . The in vivo findings were in accordance with the in vitro susceptibilities of the pathogens . These data suggest the potential utility of Ro 63-9141 for the therapy of infections caused by susceptible pathogens, including MRSA . Since insufficient solubility of Ro 63-9141 itself precludes parenteral administration in humans, a water-soluble prodrug, Ro 65-5788, is considered for development. Antimicrob Agents Chemother, 2001 Mar, 45(3), 723 - 6 Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics; Goldstein C et al.; Many pathogenic and commensal organisms are multidrug resistant due to exposure to various antibiotics . Often, this antimicrobial resistance is encoded by integrons that occur on plasmids or that are integrated into the bacterial chromosome . Integrons are commonly associated with bacterial genera in the family Enterobacteriaceae . We determined that class 1 integrases were present in approximately 46% of the isolates from the family Enterobacteriaceae; class 2 integrases were present only among Escherichia coli and Salmonella isolates . Seven percent of veterinary isolates were positive for class 3 integrase by DNA-DNA hybridization but could not be confirmed to be positive by PCR . None of the veterinary isolates possessed the class 4 integrase gene . The distribution of these integrase genes was variable within the members of the family Enterobacteriaceae when some or all integrase classes were absent from a particular genus . There was also considerable variability in the distribution of these integrases within a species, depending on the animal host . Unlike the class 1 integrases, the other integrase class, intI2, appears to be more restricted in its distribution among the members of the family Enterobacteriaceae . There is also considerable variability in the distribution of the class 1 integrases within E . coli strains isolated from different food animals . The class 1 integrases are the most widely disseminated of the four classes among the members of the family Enterobacteriaceae from both the clinical and normal flora of animals . This is the first report to closely examine the distribution of class 2 integrases in members of the family Enterobacteriaceae isolated in the United States. Luminescence, 2001 Jan-Feb, 16(1), 33 - 8 Enterobacter cloacae leukotoxin: modulation of reactive oxidant species generated by neutrophils; Barnes AI et al.; A leukotoxin purified from Enterobacter cloacae culture by saline precipitation, gel chromatography and HPLC was studied as a modulator of reactive oxidant species (ROS) produced by human neutrophils . Chemiluminescence showed that stimulation of ROS was achieved at a low leukotoxin concentration, but ROS production decreased when the toxin was applied at concentrations above 30 microg/mL . Also, the addition of 100 microg toxin/mL significantly reduced the activating effect of phorbol myristate acetate (PMA) and low doses of toxin did not produce an opposite effect toward the stimulation produced by PMA . Normal neutrophils showed a linear correlation between the inverse of ROS production and time, but the kinetic reaction changed when toxins were added to the cells and the ROS formation increased directly with time . FEMS Microbiol Lett, 2001 Feb 20, 195(2), 185 - 90 Extension of resistance to cefepime and cefpirome associated to a six amino acid deletion in the H-10 helix of the cephalosporinase of an Enterobacter cloacae clinical isolate; Barnaud G et al.; Enterobacter cloacae CHE, a clinical strain with overproduced cephalosporinase was found to be highly resistant to the new cephalosporins, cefepime and cefpirome (MICs> or =128 microg ml(-1)) . The strain was isolated from a child previously treated with cefepime . The catalytic efficiency of the purified enzyme with the third-generation cephalosporins, cefepime and cefpirome, was 10 times higher than that with the E . cloacae P99 enzyme . This was mostly due to a decrease in K(m) for these beta-lactams . The clinical isolate produced large amounts of the cephalosporinase because introduction of the ampD gene decreased ampC expression and partially restored the wild-type phenotype . Indeed, MICs of cefepime and cefpirome remained 10 times higher than those for a stable derepressed clinical isolate (OUDhyp) transformed with an ampD gene . Sequencing of the ampC gene showed that 18 nucleotides had been deleted, corresponding to the six amino acids SKVALA (residues 289--294) . According to the crystal structure of P99 beta-lactamase, this deletion was located in the H-10 helix . The ampR-ampC genes from the clinical isolates CHE and OUDhyp were cloned and expressed in Escherichia coli JM101 . The MICs of cefpirome and cefepime of E . coli harboring ampC and ampR genes from CHE were 100--200 times higher than those of E . coli harboring ampC and ampR genes from OUDhyp . This suggests that the deletion, confirmed by sequencing of the ampC gene, is involved in resistance to cefepime and cefpirome . However, the high level of resistance to cefepime and cefpirome observed in the E . cloacae clinical isolate was due to a combination of hyperproduction of the AmpC beta-lactamase and structural modification of the enzyme . This is the first example of an AmpC variant conferring resistance to cefepime and cefpirome, isolated as a clinical strain. Infect Immun, 2001 Mar, 69(3), 1477 - 82 Signaling by toll-like receptor 2 and 4 agonists results in differential gene expression in murine macrophages; Hirschfeld M et al.; Lipopolysaccharide (LPS) derived from the periodontal pathogen Porphyromonas gingivalis has been reported to differ structurally and functionally from enterobacterial LPS . These studies demonstrate that in contrast to protein-free enterobacterial LPS, a similarly purified preparation of P . gingivalis LPS exhibited potent Toll-like receptor 2 (TLR2), rather than TLR4, agonist activity to elicit gene expression and cytokine secretion in murine macrophages and transfectants . More importantly, TLR2 stimulation by this P . gingivalis LPS preparation resulted in differential expression of a panel of genes that are normally induced in murine macrophages by Escherichia coli LPS . These data suggest that (i) P . gingivalis LPS does not signal through TLR4 and (ii) signaling through TLR2 and through TLR4 differs quantitatively and qualitatively . Our data support the hypothesis that the shared signaling pathways elicited by TLR2 and by TLR4 agonists must diverge in order to account for the distinct patterns of inflammatory gene expression. Curr Microbiol, 2001 Apr, 42(4), 252 - 6 Microbiological and biochemical study of coffee fermentation; Avallone S et al.; The coffee fermentation microflora were rich and mainly constituted of aerobic Gram-negative bacilli, with Erwinia and Klebsiella genuses at the highest frequencies . The best population increase was observed with lactic acid bacteria and yeasts, whereas those microorganisms that counted on a pectin medium remained constant during the fermentation step . Qualitatively, lactic acid bacteria belonged mainly to Leuconostoc mesenteroides species but the others microflora were relatively heterogeneous . The microorganisms isolated on pectin medium were Enterobacteriaceae, identified as Erwinia herbicola and Klebsiella pneumoniae, not reported as strong pectolytic strains . Throughout coffee fermentation, 60% of the simple sugars were degraded by the total microflora and not specifically by pectolytic microorganisms. Pediatr Infect Dis J, 2001 Jan, 20(1), 102 - 3; discussion 120-2 In vitro and in vivo antimicrobial activity of topical ofloxacin and other ototopical agents; Klein JO; BACKGROUND: Ototopical agents are extensively used for otitis externa (OE), acute otitis media identified by otorrhea in patients who have tympanostomy tubes (AOM-TT) and chronic suppurative otitis media (CSOM) . The quinolones have particular value as ototopical agents because of the broad spectrum of antibacterial activity of importance in otic diseases and the high concentrations of antibacterial activity at the site of infection . METHODS: A survey of literature on in vitro activity and microbiologic efficacy in clinical trials of quinolone otic products for OE, AOM-TT and CSOM . RESULTS: OE: Floxin otic and Cortisporin TC otic suspension were equally effective in eradicating the three major pathogens Pseudomonas aeruginosa, Proteus mirabilis and Staphylococcus aureus . CSOM: Ofloxacin otic was effective in an open label trial in uniform eradication of S . aureus, P . aeruginosa, Proteus mirabilis and Enterobacter spp . AOM-TT: Ofloxacin otic and amoxicillin/clavulanate (by mouth) were equivalent clinically; rates of eradication of initial pathogens were similar for Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, but ofloxacin otic was superior in eradication of S . aureus and P . aeruginosa CONCLUSIONS: In each of the studies of OE, CSOM and AOM-TT, ofloxacin otic solution was effective in eradicating the bacterial pathogen from the site of infection: equivalent to Cortisporin for children with OE; superior to amoxicillin/clavulanate for patients with AOM-TT who had acute drainage; and effective in eradicating bacterial pathogens from the external canal of patients with CSOM. Proc Natl Acad Sci U S A, 2001 Feb 13, 98(4), 1883 - 8 Epub 2001 Jan 30. The insect endosymbiont Sodalis glossinidius utilizes a type III secretion system for cell invasion; Dale C et al.; Sodalis glossinidius is a maternally transmitted secondary endosymbiont residing intracellularly in tissues of the tsetse flies, Glossina spp . In this study, we have used Tn5 mutagenesis and a negative selection procedure to derive a S . glossinidius mutant that is incapable of invading insect cells in vitro and is aposymbiotic when microinjected into tsetse . This mutant strain harbors Tn5 integrated into a chromosomal gene sharing high sequence identity with a type III secretion system invasion gene (invC) previously identified in Salmonella enterica . With the use of degenerate PCR, we have amplified a further six Sodalis inv/spa genes sharing high sequence identity with type III secretion system genes encoded by Salmonella pathogenicity island 1 . Phylogenetic reconstructions based on the inv/spa genes of Sodalis and other members of the family Enterobacteriaceae have consistently identified a well-supported clade containing Sodalis and the enteric pathogens Shigella and Salmonella . These results suggest that Sodalis may have evolved from an ancestor with a parasitic intracellular lifestyle, possibly a latter-day entomopathogen . These observations lend credence to a hypothesis suggesting that vertically transmitted mutualistic endosymbionts evolve from horizontally transmitted parasites through a parasitism-mutualism continuum. J Hosp Infect, 2001 Feb, 47(2), 116 - 24 Evaluation of the contribution of isolation precautions in prevention and control of multi-resistant bacteria in a teaching hospital; Eveillard M et al.; From February 1999 to January 2000, a control programme to prevent the spread multi-resistant bacteria (MRB) was implemented in a French teaching hospital . This programme focused on methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended-spectrum beta-lactamases (ESBL), and was based on the application of barrier precautions (washing hands with antiseptic soaps, wearing disposable gloves and gowns, identifying MRB carriers) . No changes in antibiotic policy occurred during the year . Our aim was to conduct an evaluation of this programme by measuring incidence rates . Concurrently, the effect of barrier precautions was estimated in an indirect way, by documenting the availability of barrier precautions in MRB carriers' rooms and by analysing the monthly correlation between the supply of such material and the theoretical cumulated length of MRB carriers' isolation in six randomized wards . All MRB isolated in hospitalized patients were recorded, and differentiated between acquisition in our hospital or from elsewhere . For the analysis of trends, the year was divided in three periods of four months . Over the year, the global MRB incidence was 1.26 per 1000 patient-days (PD) {95% confidence interval (95%CI)=1.16-1.36} . The MRSA incidence was 0.89 per 1000 PD (95%CI=0.81- 0.97) and the ESBL incidence was 0.38 per 1000 PD (95% CI=0.33-0.43) . The MRB incidence decreased significantly in all types of specialties except for surgical wards . The incidence decreased by 17.9% for MRSA, 54.9% for ESBL and 34.8% for both MRB . Concurrently, the proportion of strains acquired in our hospital decreased for MRSA (P for trend > or = 0.05) and ESBL (P for trend > or = 0.01), whereas the incidence of imported strains increased slightly . The proportion of multiresistant strains in S . aureus (36.8%) and Enterobacter aerogenes (37.0%) remained similar throughout the year . Thus, the decrease of the incidence concerned both resistant and susceptible strains . The availability of antiseptic soaps increased significantly (P for trend > or = 0.01) . The amount of antiseptic soap ordered and the theoretical lengths of isolation were correlated on a monthly basis (Spearman coefficient = 0.72; P > or = 0.02) . These results shows the efficacy of such a programme of MRB containment in a large hospital, provided barrier nursing is instigated, together with the availability of such material as antiseptic soap, to allow implementation . Mol Microbiol, 2001 Feb, 39(3), 781 - 91 Substrate complexes and domain organization of the Salmonella flagellar export chaperones FlgN and FliT; Bennett JC et al.; The flagellar proteins FlgN and FliT have been proposed to act as substrate-specific export chaperones, facilitating incorporation of the enterobacterial hook-associated axial proteins (HAPs) FlgK/FlgL and FliD into the growing flagellum . In Salmonella typhimurium flgN and fliT mutants, the export of target HAPs was reduced, concomitant with loss of unincorporated flagellin into the surrounding medium . Gel filtration chromatography of wild-type S . typhimurium cell extracts identified stable pools of FlgN and FliT homodimers in the cytosol, but no chaperone-substrate complexes were evident . Nevertheless, stable unique complexes were assembled efficiently in vitro by co-incubation of FlgN and FliT with target HAPs purified from recombinant Escherichia coli . The sizes of the chaperone-substrate complexes indicated that, in each case, a chaperone homodimer binds to a substrate monomer . FlgN prevented in vitro aggregation of FlgK monomers, generating a soluble form of the HAP . Recombinant polypeptides spanning the potentially amphipathic C-terminal regions of FlgN or FliT could not complement in trans the chaperone deficiency of the respective flgN and fliT mutants, but efficient flagellar assembly was restored by homodimeric translational fusions of these domains to glutathione S-transferase, which bound FlgK and FlgL like the wild-type FlgN . These data provide further evidence for the substrate-specific chaperone function of FlgN and FliT and indicate that these chaperones comprise common N- and C-terminal domains mediating homodimerization and HAP substrate binding respectively . In support of this view, the flgN mutation was specifically complemented by a hybrid chaperone comprising the N-terminal half of FliT and the C-terminal half of FlgN. Clin Microbiol Infect, 2000 Sep, 6(9), 460 - 3 Recommendation for treatment of severe infections caused by Enterobacteriaceae producing extended-spectrum beta-lactamases (ESBLs); Paterson DL; Extended-spectrum beta-lactamase (ESBL)-producing organisms are a global problem . No randomized controlled trials have ever been performed to guide optimal treatment . However, in vitro studies and observational studies strongly suggest that carbapenems (imipenem or meropenem) should be regarded as drugs of choice for serious infections due to ESBL-producing organisms . Other beta-lactam antibiotics (cefepime, beta-lactam/beta-lactamase inhibitor combinations) are not suitable as first-line therapy . The increasing frequency of the association between quinolone resistance and ESBL production have greatly limited the role of this class of antibiotic against ESBL producers. Clin Microbiol Infect, 2000 Jul, 6(7), 376 - 84 Clinical and molecular epidemiology of chromosome-mediated resistance to third-generation cephalosporins in Enterobacter isolates in eastern France; Talon D et al.; OBJECTIVE: To determine both the prevalence of group I beta-lactamase-related resistance and the clinical setting in which resistance to expanded-spectrum cephalosporins occurs . METHODS: Isolates of Enterobacter spp . were sensitivity tested to a range of antibiotics, and selected isolates were DNA fingerprinted by pulsed-field gel electrophoresis . The medical records of all patients with positive cultures for Enterobacter spp . were reviewed to determine the effect of previous antibiotic treatment on the susceptibility profile of these organisms . RESULTS: The crude incidence of colonization/infection (n = 315) was 0.51 per 100 patients and 0.73 per 1000 days of hospitalization . The 4-day and 7-day Kaplan-Meier rates of colonization/infection with Enterobacter were estimated to be 7.57% (standard deviation (SD = 3.26%) and 4.16% (SD = 2.88%)), respectively . The time lag to colonization/infection with isolates producing large amounts of Bush group 1 beta-lactamase (HLBL) (27.35 +/- 27.30 days) was significantly different from that to colonization/infection with wild-type isolates (13.59 +/- 17.93 days) (P = 0.036) . Ninety-six isolates (30.5%) demonstrated acquired resistance to expanded-spectrum cephalosporins: 34 isolates (10.8%) produced extended-spectrum beta-lactamase, and 62 isolates (19.7%) produced HLBL . The 89 Enterobacter isolates susceptible to third-generation cephalosporins yielded 84 major DNA patterns, and the 45 HLBL isolates yielded 38 major DNA patterns . The risk of colonization/infection with HLBL-producing Enterobacter was higher in cases of antimicrobial treatment with third-generation cephalosporins or a fluoroquinolone, and in cases of urinary tract colonization/infection . CONCLUSIONS: The judicious use in hospitals of both expanded-spectrum cephalosporins and other antibiotics such as fluoroquinolones is necessary to curtail the emergence of resistance in Enterobacter spp. Clin Microbiol Infect, 2000 Jun, 6(6), 316 - 23 TEM-24 extended-spectrum beta-lactamase-producing Enterobacter aerogenes: long-term clonal dissemination in French hospitals; Galdbart JO et al.; OBJECTIVE: To investigate interstrain relatedness of TEM-24-producing Enterobacter aerogenes clinical strains isolated between 1993 and 1998 in 10 French hospitals from nine areas by pulsed-field gel electrophoresis (PFGE) and plasmid patterns . METHODS: Fifteen TEM-24-producing strains and a set of 16 control strains having various other antibiotic resistance phenotypes were genotyped by PFGE . Plasmid DNA from TEM-24-producing strains and transconjugants was analyzed . RESULTS: Analysis of XbaI macrorestriction patterns revealed only minor variations, and showed that all 15 TEM-24-producing strains were closely related . Some isolates originating from distant areas had indistinguishable patterns . According to their clustering correlation coefficients, they were also genomically distant from the control strains . Two plasmid patterns were observed in TEM-24-producing strains, one of them in 13 of the strains . Large plasmids of 85 kb encoding TEM-24 beta-lactamase were present in all isolates and, in all except one strain, could be transferred with high frequency by conjugation . CONCLUSIONS: These results confirm that the spread of the TEM-24 extended-spectrum beta-lactamase in France was essentially due to the dissemination of a single clone. Clin Microbiol Infect, 2000 Jun, 6(6), 294 - 302 Meropenem versus imipenem/cilastatin as empirical monotherapy for serious bacterial infections in the intensive care unit; Verwaest C; Belgian Multicenter Study Group; OBJECTIVE: To compare the efficacy and tolerability of meropenem and imipenem/cilastatin as empirical monotherapy in intensive care unit (ICU) patients with serious bacterial infections . METHODS: A multicenter, open-label, randomized, parallel-group trial was conducted in Belgium, evaluating empirical monotherapy with meropenem or imipenem/cilastatin (both 1 g/8 h intravenously) in ICU patients with one or more of the following infections caused by sensitive pathogens: lower respiratory tract infection (LRTI) in ventilated patients, intra-abdominal infection or sepsis . RESULTS: The overall satisfactory clinical response rate at the end of randomized treatment was 77.0% (67/87) with meropenem and 68.1% (62/91) with imipenem/cilastatin (difference 8.9%; 95% confidence interval -4.2% to 21.9%; P = 0.185) . The two drugs produced similar satisfactory clinical response rates against LRTIs: 68.3% (41/60) with meropenem versus 68.6% (35/51) with imipenem/cilastatin . Meropenem appeared to be slightly more effective against intra-abdominal infections: 95.5% (21/22) versus 76.7% (23/30), respectively . All five meropenem recipients with sepsis had a satisfactory clinical response, compared to 40.0% (4/10) of those who received imipenem/cilastatin . The overall satisfactory bacteriologic response rate was 67.1% (49/73) with meropenem and 60.3% (44/73) with imipenem/cilastatin (difference 6.9%; 95% confidence interval -8.7% to 22.4%; P = 0.389) . The predominant pathogens were Escherichia coli, Enterobacter spp . and Pseudomonas aeruginosa . No incidences of drug-related nausea and vomiting were reported, but one probable drug-related seizure occurred in the imipenem/cilastatin group . CONCLUSIONS: Meropenem is at least as efficacious (clinically and bacteriologically) as imipenem/cilastatin for the empirical monotherapy of serious bacterial infections in ICU patients, and it can therefore be considered a useful option in this setting . Moreover, meropenem is well tolerated and offers several potential advantages, including greater in vitro activity against Gram-negative pathogens and the option of bolus administration. Int J Antimicrob Agents, 2001 Feb, 17(2), 123 - 9 Intra- and inter-generic plasmid-mediated spread of cephalosporin and aminoglycoside resistance amongst Klebsiella aerogenes K41 and other enterobacteria; Verma A et al.; Klebsiella aerogenes K41, resistant to third generation cephalosporins and aminoglycosides, was isolated from clinical samples of 153 in-patients . Blood cultures accounted for 24 (15.7%) of isolates . The MIC(90) of ceftazidime for the isolates of 84 patients was >512 mg/l and was reduced to 2.0 by 4 mg/l of clavulanic acid, but only to 64 by 4 mg/l of sulbactam . Isolates of K . aerogenes K41 produced extended-spectrum beta-lactamase (ESBL) SHV-5 and TEM-1, identified by isoelectric focusing . Plasmid profiles showed that co-dissemination of cephalosporin and aminoglycoside resistance, plus ESBL production, coincided with the acquisition of a 116-kb plasmid . This plasmid was transferable in vitro from K . aerogenes K41 to other serotypes and genera of the Enterobacteriaceae. Microbiology, 2001 Jan, 147(Pt 1), 203 - 13 Cloning and characterization of the gene encoding periplasmic 2',3'-cyclic phosphodiesterase of Yersinia enterocolitica O:8; Trulzsch K et al.; The gene encoding periplasmic 2',3'-cyclic phosphodiesterase in Yersinia enterocolitica O:8 (designated cpdB), was cloned and expressed in Escherichia coli . This enzyme enables Y . enterocolitica to grow on 2',3'-cAMP as a sole source of carbon and energy . Sequencing and analysis of a 3 kb ECO:RI fragment containing the cpdB gene revealed an open reading frame of 1179 bp, corresponding to a protein with a molecular mass of 71 kDa . The first 25 amino acid residues show features of a typical prokaryotic signal sequence . The predicted molecular mass of the mature peptide is therefore in agreement with the molecular mass estimated by SDS gel electrophoresis (68 kDa) . The putative cpdB promoter region contains two possible -10 and -35 regions . Furthermore, the 5' untranslated region contains sequences with significant homology to the cyclic AMP-cyclic AMP receptor protein binding site and the sigma(28) consensus . This region is interrupted by an enterobacterial repetitive intergenic consensus (ERIC) sequence . Deletion of the ERIC element from the cpdB promoter region had no effect on cpdB expression . In the 3' untranslated region, a possible rho-independent transcriptional terminator was identified . The deduced amino acid sequence of the Y . enterocolitica CpdB protein shows 76% identity with CpdB of Salmonella typhimurium and E . coli . CpdB of Y . enterocolitica is exported to the periplasmic space . An isogenic Y . enterocolitica cpdB mutant strain, constructed by allelic exchange, was no longer able to grow on 2',3'-cAMP as sole source of carbon and energy . The CpdB mutant showed no significant change in virulence in an oral and intravenous mouse infection model. J Immunol, 2001 Feb 15, 166(4), 2342 - 7 Bacterial lipopolysaccharide activates HIV long terminal repeat through Toll-like receptor 4; Equils O et al.; In HIV-infected patients, concurrent infections with bacteria and viruses are known to induce HIV replication as assessed by increases in plasma HIV RNA levels . In the present study, we determined the cell surface receptor and molecular mechanisms of enterobacterial LPS-induced HIV transcription . Human dermal microvessel endothelial cells (HMEC) were transfected with an HIV-long terminal repeat (LTR)-luciferase construct and subsequently stimulated with purified bacterial LPS . Our studies demonstrate that human Toll-like receptor 4 (TLR4) mediates LPS-induced NF-kappaB and HIV-LTR activation in HMEC through IL-1 signaling molecules, namely myeloid differentiation protein, IL-1R-associated kinase, TNFR-associated factor, and NF-kappaB-inducing kinase . Cotransfection of HMEC with HIV-LTR-luciferase and TLR4 cDNA from LPS-hyporesponsive C3H/HeJ mice abrogates LPS-induced HIV transcription as does the use of dominant-negative mutants of the IL-1 signaling molecules . Transfection of HMEC with an HIV-LTR-mutant that lacks the NF-kappaB binding site or pretreatment of cells with chemical inhibitors of the NF-kappaB pathway also blocked LPS-induced HIV-LTR transactivation . These data support the conclusion that TLR4 mediates enterobacterial LPS-induced HIV transcription via IL-1 signaling molecules and NF-kappaB activation plays an important role in HIV-LTR transactivation. J Immunol, 2001 Feb 1, 166(3), 1871 - 6 Conventional alpha beta T cells are sufficient for innate and adaptive immunity against enteric Listeria monocytogenes; Bregenholt S et al.; We have begun to dissect the cellular requirements for generation of immunity against enteric infection by Listeria monocytogenes using a novel T(-) B(-) NK(-) mouse strain (mice double deficient for the common cytokine receptor gamma-chain (gamma(c)) and the recombinase-activating gene-2 (RAG2/gamma(c) mice) . Initial experiments showed that C57BL/6 mice and alymphoid RAG2/gamma(c) mice had similar kinetics of bacterial accumulation in the spleen, liver, and brain early after intragastric L . monocytogenes infection (up to day 3), calling into question the physiologic role of gut-associated lymphoid cells during the passage of this enterobacterium into the host . However, in contrast to C57BL/6 mice, RAG2/gamma(c) mice rapidly succumbed to disseminated infection by day 7 . Polyclonal lymph node CD4(+) and CD8(+) alphabeta T cells were able to confer RAG2/gamma(c) mice with long-lasting protection against enteric L . monocytogenes infection in the absence of gammadelta T, NK, and NK-T cells . Moreover, these alphabeta T-reconstituted RAG2/gamma(c) mice produced IFN-gamma at levels comparable to C57BL/6 mice in response to L . monocytogenes both in vitro and in vivo . Protection was IFN-gamma dependent, as RAG2/gamma(c) mice reconstituted with IFN-gamma-deficient alphabeta T cells were unable to control enteric L . monocytogenes infection . Furthermore, alphabeta T cell-reconstituted RAG2/gamma(c) mice were able to mount memory responses when challenged with lethal doses of L . monocytogenes . These data suggest that NK, NK-T, gammadelta T, and B cells are functionally redundant in the immunity against oral L . monocytogenes infection, and that in their absence alphabeta T cells are able to mediate the early IFN-gamma production required for both innate and adaptive immunity. Antimicrob Agents Chemother, 2001 Feb, 45(2), 532 - 9 Antimicrobial properties and mode of action of the pyrrothine holomycin; Oliva B et al.; Holomycin, a member of the pyrrothine class of antibiotics, displayed broad-spectrum antibacterial activity, inhibiting a variety of gram-positive and gram-negative bacteria, with the exception of Enterobacter cloacae, Morganella morganii, and Pseudomonas aeruginosa . The antibiotic lacked activity against the eukaryotic microorganisms Saccharomyces cerevisiae and Candida kefyr . Holomycin exhibited a bacteriostatic response against Escherichia coli that was associated with rapid inhibition of RNA synthesis in whole cells . Inhibition of RNA synthesis could have been a secondary consequence of inhibiting tRNA aminoacylation, thereby inducing the stringent response . However, the levels of inhibition of RNA synthesis by holomycin were similar in a stringent and relaxed pair of E . coli strains that were isogenic except for the deletion of the relA gene . This suggests that inhibition of RNA synthesis by holomycin could reflect direct inhibition of DNA-dependent RNA polymerase . Examination of the effects of holomycin on the kinetics of the appearance of beta-galactosidase in induced E . coli cells was also consistent with inhibition of RNA polymerase at the level of RNA chain elongation . However, holomycin only weakly inhibited E . coli RNA polymerase in assays using synthetic poly(dA-dT) and plasmid templates . Furthermore, inhibition of RNA polymerase was observed only at holomycin concentrations in excess of those required to inhibit the growth of E . coli . It is possible that holomycin is a prodrug, requiring conversion in the cell to an active species that inhibits RNA polymerase. Pediatrics . 2001 Feb;107(2):E26. Two cases of diskitis attributable to anaerobic bacteria in children; Brook I; Diskitis, an inflammation of the intervertebral disk, is generally attributable to Staphylococcus aureus and rarely Staphylococcus epidermidis, Kingella kingae, Enterobacteriaciae, and Streptococcus pneumoniae . In many cases, no bacterial growth is obtained from infected intervertebral discs . Although anaerobic bacteria were recovered from adults with spondylodiscitis, these organisms were not reported before from children . The recovery of anaerobic bacteria in 2 children with diskitis is reported . Patient 1 . A 10-year-old male presented with 6 weeks of low back pain and 2 weeks of low-grade fever and abdominal pain . Physical examination was normal except for tenderness to percussion over the spine between thoracic vertebra 11 and lumbar vertebra 2 . The patient had a temperature of 104 degrees F . Laboratory tests were within normal limits, except for erythrocyte sedimentation rate (ESR), which was 58 mm/hour . Blood culture showed no growth . Magnetic resonance imaging with gadolinium contrast revealed minimal inflammatory changes in the 12th thoracic vertebra/first lumbar vertebra disk . There was no other abnormality . A computed tomography (CT)-guided aspiration of the disk space yielded bloody material, which was sent for aerobic and anaerobic cultures . Gram stain showed numerous white blood cells and Gram-positive cocci in chains . Cultures for anaerobic bacteria yielded heavy growth of Peptostreptococcus magnus, which was susceptible to penicillin, clindamycin, and vancomycin . The patient was treated with intravenous penicillin 600 000 units every 6 hours for 3 weeks, and then oral amoxicillin, 500 mg every 6 hours for 3 weeks . The back pain resolved within 2 weeks, and the ESR returned to normal at the end of therapy . Follow-up for 3 years showed complete resolution of the infection . Patient 2 . An 8-year-old boy presented with low back pain and low-grade fever, irritability, and general malaise for 10 days . He had had an upper respiratory tract infection with sore throat 27 days earlier, for which he received no therapy . The patient had a temperature of 102 degrees F, and physical examination was normal except for tenderness to percussion over the spine between the second and fourth lumbar vertebrae . Laboratory tests were normal, except for the ESR (42 mm/hour) . Radiographs of the spine showed narrowing of the third to fourth lumbar vertebra disk space and irregularity of the margins of the vertebral endplates . A CT scan revealed a lytic bone lesion at lumbar vertebra 4, and bone scan showed an increase uptake of (99m)technetium at the third to fourth lumbar vertebra disk space . CT-guided aspiration of the disk space yielded cloudy nonfoul-smelling material, which was sent for aerobic and anaerobic cultures . Gram stain showed numerous white blood cells and fusiform Gram-negative bacilli . Anaerobic culture grew light growth of Fusobacterium nucleatum . The organism produced beta-lactamase and was susceptible to ticarcillin-clavulanate, clindamycin, metronidazole, and imipenem . Therapy with clindamycin 450 mg every 8 hours was given parenterally for 3 weeks and orally for 3 weeks . Back pain resolved within 2 weeks . A 2-year follow-up showed complete resolution and no recurrence . This report describes, for the first time, the isolation of anaerobic bacteria from children with diskitis . The lack of their recovery in previous reports and the absence of bacterial growth in over two third of these studies may be caused by the use of improper methods for their collection, transportation, and cultivation . Proper choice of antimicrobial therapy for diskitis can be accomplished only by identification of the causative organisms and its antimicrobial susceptibility . This is of particular importance in infections caused by anaerobic bacteria that are often resistant to antimicrobials used to empirically treat diskitis . This was the case in our second patient, who was infected by F nucleatum, which was resistant to beta-lactam antibiotics . The origin of the anaerobic bacteria causing the infection in our patient is probably of endogenous nature . The presence of abdominal pain in the first child may have been attributable to a subclinical abdominal pathothology . The preceding pharyngitis in the second patient may have been associated with a potential hematogenous spread of F nucleatum . P magnus has been associated with bone and joint infections . This report highlights the importance of obtaining disk space culture for aerobic and anaerobic bacteria from all children with diskitis . Future prospective studies are warranted to elucidate the role of anaerobic bacteria in diskitis in children. J Clin Microbiol, 2001 Feb, 39(2), 430 - 7 Use of rpoB gene analysis for detection and identification of Bartonella species; Renesto P et al.; Identification of Bartonella species is of increasing importance as the number of infections in which these bacteria are involved increases . To date, these gram-negative bacilli have been identified by various serological, biochemical, and genotypic methods . However, the development of alternative tools is required, principally to circumvent a major risk of contamination during sample manipulation . The aim of our study was to investigate the possible identification of various Bartonella species by comparison of RNA polymerase beta-subunit gene (rpoB) sequences . This approach has previously been shown to be useful for the identification of members of the family Enterobacteriaceae (C . M . Mollet, M . Drancourt, and D . Raoult, Mol . Microbiol . 26:1005-1011, 1997) . Following PCR amplification with specific oligonucleotides, a 825-bp region of the rpoB gene was sequenced from 13 distinct Bartonella strains . Analysis of these sequences allowed selection of three restriction enzymes (ApoI, AluI, and AflIII) useful for discerning the different strains by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis . To confirm the potential value of such an approach for identification of Bartonella, the rpoB PCR was then applied to 94 clinical samples, and the results obtained were identical to those obtained by our reference PCR method . Twenty-four isolates were also adequately identified by PCR-RFLP analysis . In all cases, our results were in accordance with those of the reference method . Moreover, conserved regions of DNA were chosen as suitable primer targets for PCR amplification of a 439-bp fragment which can be easily sequenced. Appl Environ Microbiol, 2001 Feb, 67(2), 986 - 90 Characterization of acetic acid bacteria in traditional acetic acid fermentation of rice vinegar (komesu) and unpolished rice vinegar (kurosu) produced in Japan; Nanda K et al.; Bacterial strains were isolated from samples of Japanese rice vinegar (komesu) and unpolished rice vinegar (kurosu) fermented by the traditional static method . Fermentations have never been inoculated with a pure culture since they were started in 1907 . A total of 178 isolates were divided into groups A and B on the basis of enterobacterial repetitive intergenic consensus-PCR and random amplified polymorphic DNA fingerprinting analyses . The 16S ribosomal DNA sequences of strains belonging to each group showed similarities of more than 99% with Acetobacter pasteurianus . Group A strains overwhelmingly dominated all stages of fermentation of both types of vinegar . Our results indicate that appropriate strains of acetic acid bacteria have spontaneously established almost pure cultures during nearly a century of komesu and kurosu fermentation. Appl Environ Microbiol, 2001 Feb, 67(2), 910 - 21 Molecular analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 strains: clonal relationships between clinical and environmental isolates; Singh DV et al.; A total of 26 strains of Vibrio cholerae, including members of the O1, O139, and non-O1, non-O139 serogroups from both clinical and environmental sources, were examined for the presence of genes encoding cholera toxin (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), hemolysin (hlyA), NAG-specific heat-stable toxin (st), toxin-coregulated pilus (tcpA), and outer membrane protein (ompU), for genomic organization, and for the presence of the regulatory protein genes tcpI and toxR in order to determine relationships between epidemic serotypes and sources of isolation . While 22 of the 26 strains were hemolytic on 5% sheep blood nutrient agar, all strains were PCR positive for hlyA, the hemolysin gene . When multiplex PCR was used, all serogroup O1 and O139 strains were positive for tcpA, ompU, and tcpI . All O1 and O139 strains except one O1 strain and one O139 strain were positive for the ctxA, zot, and ace genes . Also, O1 strain VO3 was negative for the zot gene . All of the non-O1, non-O139 strains were negative for the ctxA, zot, ace, tcpA, and tcpI genes, and all of the non-O1, non-O139 strains except strain VO26 were negative for ompU . All of the strains except non-O1, non-O139 strain VO22 were PCR positive for the gene encoding the central regulatory protein, toxR . All V . cholerae strains were negative for the NAG-specific st gene . Of the nine non-ctx-producing strains of V . cholerae, only one, non-O1, non-O139 strain VO24, caused fluid accumulation in the rabbit ileal loop assay . The other eight strains, including an O1 strain, an O139 strain, and six non-O1, non-O139 strains, regardless of the source of isolation, caused fluid accumulation after two to five serial passages through the rabbit gut . Culture filtrates of all non-cholera-toxigenic strains grown in AKI media also caused fluid accumulation, suggesting that a new toxin was produced in AKI medium by these strains . Studies of clonality performed by using enterobacterial repetitive intergenic consensus sequence PCR, Box element PCR, amplified fragment length polymorphism (AFLP), and pulsed-field gel electrophoresis (PFGE) collectively indicated that the V . cholerae O1 and O139 strains had a clonal origin, whereas the non-O1, non-O139 strains belonged to different clones . The clinical isolates closely resembled environmental isolates in their genomic patterns . Overall, there was an excellent correlation among the results of the PCR, AFLP, and PFGE analyses, and individual strains derived from clinical and environmental sources produced similar fingerprint patterns . From the results of this study, we concluded that the non-cholera-toxin-producing strains of V . cholerae, whether of clinical or environmental origin, possess the ability to produce a new secretogenic toxin that is entirely different from the toxin produced by toxigenic V . cholerae O1 and O139 strains . We also concluded that the aquatic environment is a reservoir for V . cholerae O1, O139, non-O1, and non-O139 serogroup strains. Appl Environ Microbiol, 2001 Feb, 67(2), 895 - 903 Genetic diversity of Pierce's disease strains and other pathotypes of Xylella fastidiosa; Hendson M et al.; Strains of Xylella fastidiosa isolated from grape, almond, maple, and oleander were characterized by enterobacterial repetitive intergenic consensus sequence-, repetitive extragenic palindromic element (REP)-, and random amplified polymorphic DNA (RAPD)-PCR; contour-clamped homogeneous electric field (CHEF) gel electrophoresis; plasmid content; and sequencing of the 16S-23S rRNA spacer region . Combining methods gave greater resolution of strain groupings than any single method . Strains isolated from grape with Pierce's disease (PD) from California, Florida, and Georgia showed greater than previously reported genetic variability, including plasmid contents, but formed a cluster based on analysis of RAPD-PCR products, NotI and SpeI genomic DNA fingerprints, and 16S-23S rRNA spacer region sequence . Two groupings of almond leaf scorch (ALS) strains were distinguished by RAPD-PCR and CHEF gel electrophoresis, but some ALS isolates were clustered within the PD group . RAPD-PCR, CHEF gel electrophoresis, and 16S-23S rRNA sequence analysis produced the same groupings of strains, with RAPD-PCR resolving the greatest genetic differences . Oleander strains, phony peach disease (PP), and oak leaf scorch (OLS) strains were distinct from other strains . DNA profiles constructed by REP-PCR analysis were the same or very similar among all grape strains and most almond strains but different among some almond strains and all other strains tested . Eight of 12 ALS strains and 4 of 14 PD strains of X . fastidiosa isolated in California contained plasmids . All oleander strains carried the same-sized plasmid; all OLS strains carried the same-sized plasmid . A plum leaf scald strain contained three plasmids, two of which were the same sizes as those found in PP strains . These findings support a division of X . fastidiosa at the subspecies or pathovar level. Folia Biol (Krakow), 1999, 47(3-4), 135 - 41 Exoproteinases of the type A in pathogenesis of insect bacterial diseases; Andrejko M; Immune inhibitors produced in infected larvae of Galleria mellonella by such entomopathogens as Pseudomonas aeruginosa, Serratia marcescens and Heterorhabditis bacteriophora effectively blocked in vitro bactericidal activity of insect haemolymph against Escherichia coli D31, both in Galleria mellonella and Pieris brassicae pupae previously vaccinated with Enterobacter cloacae . Even at a trace concentration, the extracellular proteinases, by proteolytic degradation, totally destroyed the activity of cecropin peptides from Galleria and cecropin-like and attacin-family proteins from Pieris, but no ability to destroy antibacterial activity was shown by extracts obtained from Galleria larvae killed by massive doses of bacterial saprophytes . It is suggested that by blocking antibacterial immune response of the host, the proteinases help the bacteria to multiply in the haemolymph, thus they could be considered an important factor in the pathogenesis of bacterial diseases of insects. Rev Med Chil, 1999 Sep, 127(9), 1049 - 55 {Microbiological study of gallbladder bile in a high risk zone for gallbladder cancer}; Roa I et al.; BACKGROUND: Gallbladder cancer is frequent in Chile and there is sparse information about the association between this type of cancer and the presence of bacteria in the gallbladder bile . AIM: To determine the presence of aerobic bacteria in gallbladder bile in patients with and without gallbladder cancer . MATERIAL AND METHODS: A microbiological analysis of bile and pathological study was performed in 608 gallbladders, obtained during to cholecystectomies performed to 513 women and 95 men aged 44 years old as a mean . RESULTS: Pathological study showed a chronic cholecystitis in 468 cases (77%), an acute cholecystitis in 140 (33%), cancer in 24 (3.9%) and dysplasia in 5 cases (0.8%) . A positive culture was obtained in 22.5% of women and 28.5% of males . Twenty seven percent of women over 30 years old had positive cultures compared with 10% of younger women (p < 0.001) . Thirty two percent of acute cholecystitis had positive cultures, compared with 24% of chronic cholecystitis (p = 0.03) . E Coli was isolated in 51% of positive cases, Streptococci-Enterococci in 24%, Enterobacter sp in 9%, Klebsiella and Proteus in lower proportions . Salmonella sp was isolated in 4 cases, being all women with chronic cholecystitis . Thirteen of 29 cases with cancer or dysplasia had positive cultures (45%), compared with 25% of patients with inflammatory gallbladder diseases (p = 0.02) . Streptococci-Enterococci were isolated in 7 cases and Enterobacter sp in three . CONCLUSIONS: The presence of Salmonella sp in gallbladder bile was not frequent in the studied patients . Its ro