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Detection of microcin production by pathogenic Escherichia coli isolates from children with acute diarrhoea.

Production of low molecular weight antibiotic substances was detected among pathogenic Escherichia coli isolates from diarrhoeal children's feces during evaluation of a rotavirus vaccine in Caracas city. One of these products, microcin V627a, was partially purified and characterized. Microcin synthesis and immunity system appears not to be plasmid determined and the antibiotic was produced to a high level in minimal medium during the stationary phase. This microcin has many features in common with the mccA15 family.

Anti-Bacterial Agents↗

Influence of temperature on Pythium splendens--induced root disease on carambola, Averrhoa carambola.

A series of glasshouse and incubator studies were conducted to investigate the role played by Pythium splendens in a decline disorder of carambola, Averrhoa carambola. Plants, 4-6 months old, were grown in native calcareous soil either infested or not infested with the pathogen. Isolates recovered from atemoya, carambola and passion fruit grew optimally at 30 degrees C, and significantly (P < 0.05) increased root necrosis and reduced root, shoot and total biomass of carambola. Temperature had a profound impact on the latter relationships. Two or more times more necrosis developed at 10 and 15 degrees C than at 25 and 30 degrees C. Total biomass accumulations were over four times greater at 30 degrees C than at 10 degrees C, and were always lower in soil infested with P. splendens. When biomass totals from infested and noninfested soil were compared, relative values were lowest at 15 and 20 degrees C and were almost two times greater at 30 degrees C than at 20 degrees C. Root infection by P. splendens was greatest at 15 and 20 degrees C, far below the species' optimum for growth, and at 30 degrees C was over nine times lower than at 15 and 20 degrees C. This is the first detailed report of P. splendens as a pathogen of carambola.

Fruit↗

Genetic structures of the B2 and B1 Escherichia coli strains responsible for extra-intestinal infections.

Escherichia coli strains causing human extra-intestinal infections may be divided into two groups, B1 and B2 according to the electrophoretic patterns of carboxylesterase B. This study compares the restriction fragment length polymorphism (RFLP) of ribosomal DNA (rDNA) for 45 B1 strains and 45 B2 strains to examine the genetic structure of B2 strains and to distinguish them from B1 strains. The isolates were chosen for diversity in their allozymes of esterases, B, A, C and I, their production of virulence factors (alpha-haemolysin, mannose resistant haemagglutinin and cytotoxic necrotizing factor) and certain O antigens, and their pathological and geographical origins. DNA was digested with HindIII and BamHI restriction enzymes and analysed by Southern blotting. The resulting rDNA RFLP patterns of B2 strains were distinct from those of the B1 strains. Moreover, the B2 strains appeared to be less heterogeneous than the B1 strains. The B2 strains gave 13 ribotypes (resulting from the combination of the rDNA RFLP patterns obtained with HindIII and BamHI digestions) while the B1 strains gave 32 ribotypes. Correspondence analysis of the data showed that several clusters of strains were identified in the B2 strains by particular ribotypes, certain associations of esterase B and A electrophoretic variants, O serotypes and virulence factor production. In contrast, these parameters appeared to be unrelated in the B1 strains, reflecting their heterogeneity. These findings, which differentiate two levels of genetic heterogeneity within E. coli pathogenic isolates, indicate that the B2 strains constitute a phylogenetically distinct group within the species.

Carboxylesterase↗

The identification and phylogenetic relationship of pathogenic species of Aspergillus based on the mitochondrial cytochrome b gene.

To study the identification and phylogeny of pathogenic isolates of Aspergillus, we designed primers from known cytochrome b amino acid sequences. Using these primers, 426 bp fragments of a mitochondrial (mt) cytochrome b gene were amplified by polymerase chain reaction (PCR), directly sequenced, and compared among Aspergillus fumigatus, A. flavus, A. niger, A. terreus and Emericella nidulans. Except for E. nidulans, all strains produced the 426 bp fragment by PCR. The E. nidulans strains demonstrated both an intron-presence fragment ( approximately 1500 bp) and intron-absence fragment (426 bp). Species-specific nucleotides were found in each of the five species. Based on sequence analysis, the strains were further divided into several groups within each species. When a 142-amino-acid sequence was estimated from the 426 bp nucleotide sequence using the yeast mt genetic code, the amino acid sequences showed no difference among strains of the individual species. DNA-based phylogenetic and amino acid-based trees were constructed. In conclusion, the DNA sequences of the cytochrome b gene may be of use in identification of pathogenic Aspergillus species and the amino acid-based tree suitable for discussing their phylogenetic relationships.

Amino Acid Sequence↗

Comparative analysis of prevalence, risk factors, and molecular epidemiology of antibiotic-associated diarrhea due to Clostridium difficile, Clostridium perfringens, and Staphylococcus aureus.

We prospectively studied the comparative epidemiology and risk factors for Clostridium difficile, Clostridium perfringens, and Staphylococcus aureus antibiotic-associated diarrhea (AAD). Four thousand six hundred fifty-nine inpatient fecal specimens (11 months) were tested for C. difficile cytotoxin, C. perfringens enterotoxin, and S. aureus by Vero cell assay, enzyme-linked immunosorbent assay, and growth on fresh blood agar, respectively. Two distinct age-, sex-, and location-matched control patient groups were used for multivariate logistic regression risk factor analyses: symptomatic patients who were AAD pathogen negative and asymptomatic patients with histories of recent antimicrobial therapy. All AAD pathogen isolates were DNA fingerprinted. In AAD cases, the prevalences of C. difficile cytotoxin, C. perfringens enterotoxin, and S. aureus were 12.7%, 3.3%, and 0.2%, respectively (15.8% overall). Age of >70 years was a common risk factor. Other risk factors for infective AAD and C. difficile AAD included length of hospital stay and use of feeding tubes (length of stay odds ratios [OR], 1.017 and 1.012; feeding tube OR, 1.864 and 2.808). Female gender and use of antacids were significantly associated with increased risk of C. perfringens AAD (OR, 2.08 and 2.789, respectively), but unlike what was found for C. difficile AAD, specific antibiotic classes were not associated with increased risk. A limited number of genotypes caused the majority of C. difficile and C. perfringens AAD cases. Similar to what was found for C. difficile AAD, there was epidemiological evidence of C. perfringens AAD case clustering and reinfection due to different strains. C. difficile AAD was approximately 4 and 60 times more common than C. perfringens AAD and S. aureus AAD, respectively. Risk factors for these AAD pathogens differed, highlighting the need to define specific control measures. There is evidence of nosocomial transmission in cases of C. perfringens AAD.

Adolescent↗

Double-blind comparative study on the care of the neonatal umbilical cord using 80% ethanol with or without chlorhexidine.

We conducted a double-blind comparative study on the effectiveness of 80% ethanol (EtOH), with or without chlorhexidine (CHD), in the prevention of neonatal umbilical colonization by Staphylococcus aureus, the most frequently isolated pathogen on the neonatal umbilicus in the early period. A total of 100 neonates born at the National Tokyo Medical Centre from March to May 2000 and nursed at a maternity ward were enrolled. Forty-eight were randomly allocated to the group for whom umbilical cord disinfection was performed using 80% EtOH containing 0.5% CHD (CHD group) and 52 to disinfection with 80% EtOH alone (EtOH group). The mothers of the neonates and the nursing staff were unaware as to which disinfectant was being used. Disinfection of the umbilicus and the surrounding area was done immediately after birth and twice daily thereafter, after bathing and in the evening, throughout the hospitalization period. Specimens for bacterial culture were taken from the umbilical cord and surrounding skin on day 4 or 5 after birth. As a disinfectant susceptibility test, we checked the minimum killing concentration (MKC) of CHD and EtOH. There was no statistically significant difference between the two groups with respect to sex, gestation period, birthweight, APGAR score or delivery method. In the CHD group, S. aureus was isolated from 25% of the patients, while it was isolated from 57.7% in the EtOH group (P<0.001). In the CHD group, 50% of the S. aureus strains were MRSA, compared with 73.3% in the EtOH group (non-significant). All the S. aureus strains were killed by the combination of both CHD and EtOH at the concentrations used. In terms of the MKC90, there was no significant difference between the CHD group and the EtOH group. For the daily care of the neonatal umbilicus, disinfection using 80% EtOH containing CHD was found to be more effective than that using 80% EtOH alone in preventing colonization by S. aureus.

Anti-Infective Agents, Local↗

Evaluation of the clinical microbiology profile of moxifloxacin.

Moxifloxacin is a new broad-spectrum antibacterial agent for treatment of respiratory tract infection of pathogens, including the major pathogens isolated in respiratory tract infections. The pharmacokinetic and pharmacodynamic properties of moxifloxacin are: excellent bioavailability, long half-life, and superior tissue penetration. Consequently, the 90% minimum inhibitory concentration (MIC(90)) values exhibited by moxifloxacin are generally lower than the concentrations of moxifloxacin found in circulation and in pulmonary tissues after a standard 400-mg dose given for up to 30 h. The relationship between moxifloxacin MIC(90) values and clinical response was investigated. The results of 13 clinical trials, performed in 30 countries between 1997 and 1998 and comprising 2618 patients treated with moxifloxacin or a comparator drug, were reviewed. Overall, 94% clinical success and 95% bacterial eradication was observed with moxifloxacin. These results were equivalent or superior to results seen with the comparator drugs. Clinical response rates and bacterial eradication rates with moxifloxacin were not significantly affected by bacterial resistance to other antibiotics (i.e., penicillin, clarithromycin, or amoxicillin). The majority (89%-97%) of the different bacterial strains with MICs for moxifloxacin < or =2 mg/L were successfully eradicated. In conclusion, moxifloxacin has potent in vivo bactericidal activity, and pathogen sensitivity to moxifloxacin is in accordance with US Food and Drug Administration and European suggested breakpoint values.

Anti-Infective Agents↗

[Treatment of community-acquired pneumonia by pristinamycin (Pyostacine 500). Results of a non comparative open study].

UNLABELLED: Activity of natural streptogramin (NSG) appears well adapted to pathogens responsible for CAP. The goal of this multicenter pilot study was to bring first data about efficacy of NSG in treatment of CAP. PATIENTS METHOD: Ten days of a NSG (1 gr b.i.d. or t.i.d.) regimen was administered to 46 hospitalized adult patients for CAP defined with fever > 38 degrees C, respiratory symptoms and X-ray opacity. Severely ill patients were excluded. A broncho-pulmonar sample (expectoration or trantracheal aspiration or protected distal sample) was performed in all patients. RESULTS: two patients were excluded because of pulmonary embolism (n = 1) or tuberculosis (n = 1) and 44 patients were analyzed. 50% of them had associated disease, 20% had failure of prior antibiotherapy. At inclusion, mean fever was 39.2 +/- 0.7 degrees C, respiratory rate was 22 +/- 5/mn, PaO2 was 74 +/- 10 mmHg, chest X-ray showed bilateral opacity in 16%, unilateral in 84% and pleural fluid level in 6 cases. Etiological diagnosis was determined in 70% of cases. Streptococcus pneumoniae (n = 14), Haemophilus influenzae (n = 5), Legionella pneumophila (n = 2), Mycoplasma pneumoniae (n = 2) and Chlamydia psittaci (n = 1) were the most frequent isolated pathogens. 40 patients (91%) were cured with NSG and delay to obtain apyrexia was 4.4 +/- 3.9 days. NSG was stopped in 4 patients: 1 clinical and bacteriological failure (Klebsiella pneumoniae), 2 clinical failures (1 pneumococcus with purulent pleurisy, 1 pneumococcus with worsening of respiratory status), 1 patient with resistant H. influenzae strain in spite of favourable clinical evolution. NSG was well tolerated in 86% of patients. CONCLUSION: these data invite to carry on evaluation of first line therapy of CAP with NSG.

Adult↗

Staphylococcus aureus types 5 and 8 capsular polysaccharide-protein conjugate vaccines.

BACKGROUND: Staphylococcus aureus, the first or second most common pathogen isolated from patients, is capsulated; there are at least 12 capsular types, and types 5 and 8 comprise approximately 85% of blood. Types 5 and 8, composed of a trisaccharide repeat unit including a mannose uronic acid and 2 fucoses, are non-immunogenic. As protein conjugates, they induce opsonophagocytic antibodies that confer type-specific active and passive protection in mice. METHODS: A phase II study of patients with end-stage renal disease showed that these conjugates induced approximately one third of the immunoglobulin G antibody of healthy individuals. Increasing the dose to 100 microg of polysaccharide induced levels similar to that in healthy individuals injected with 25 microg. RESULTS: In a double-blinded randomized and controlled study of patients undergoing renal dialysis, the conjugates induced statistically significant protection against bacteremia for as long as 10 months after immunization. The estimated protective level was 80 microg Ab/mL. At re-injection approximately 2 years later, 83 of 83 recipients responded with protective levels. CONCLUSIONS: Conjugate vaccine-induced antibodies to the types 5 and 8 capsular polysaccharide antibodies of S aureus prevent bacteremia caused by this pathogen. The extent and duration of conjugate-induced immunity can be extended by re-immunization approximately 1 year later. Studies of patients undergoing cardiovascular surgery who would be immunized with the staphylococcus conjugates when they are immunologically intact are planned.

Adult↗

Identification of sat, an autotransporter toxin produced by uropathogenic Escherichia coli.

Urinary tract infection (UTI) is a very common extraintestinal infection, and Escherichia coli is by far the most common causative organism. Uropathogenic E. coli possess traits that distinguish them from commensal strains of E. coli, such as secretion systems that allow virulence factors to be targeted to extracytoplasmic compartments. One of at least five characterized secretion mechanisms is the autotransporter system, which involves translocation of a protein across the inner membrane, presumably via the sec system, and across the outer membrane through a beta-barrel porin structure formed by the carboxy-terminus autotransporter domain. We identified a 107 kDa protein that was expressed significantly more often by E. coli strains associated with the clinical syndrome of acute pyelonephritis than by faecal strains (P = 0.029). We isolated the protein from E. coli CFT073, a strain cultured from the blood and urine of a patient with acute pyelonephritis. The N-terminal amino acid sequence showed highest similarity to two known SPATE (serine protease autotransporters of Enterobacteriaceae) proteins, Pet and EspC. Using a 509 bp probe from the 5' region of pet, 10 cosmid clones of an E. coli CFT073 gene library were positive for hybridization. From one cosmid clone, a 7.5 kb EcoRI restriction fragment, which reacted strongly with the probe, was shown to include the entire 3885 bp gene. The predicted 142 kDa protein product possesses the three domains that are typical of SPATE autotransporters: an unusually long signal sequence of 49 amino acids; a 107 kDa passenger domain containing a consensus serine protease active site (GDSGSG); and a C-terminal autotransporter domain of 30 kDa. The protein exhibited serine protease activity and displayed cytopathic activity on VERO primary kidney, HK-2 bladder and HEp-2 cell lines; the name Sat (secreted autotransporter toxin) was derived from these properties. In addition, Sat antibodies were present in the serum of mice infected with E. coli CFT073. Based upon its association with pathogenic isolates, its cytopathic phenotype and its ability to elicit a strong antibody response after infection, we postulate that Sat represents a novel virulence determinant of uropathogenic E. coli.

Amino Acid Sequence↗

Genomic subtraction for the identification of putative new virulence factors of an avian pathogenic Escherichia coli strain of O2 serogroup.

To identify putative new virulence factors of avian pathogenic Escherichia coli (APEC) strains, a genomic subtraction was performed between the APEC strain MT512 and the non-pathogenic E. coli strain of avian origin EC79. Seventeen DNA fragments were cloned that were specific for the APEC strain. Among them, nine were identified that were more frequent among pathogenic than non-pathogenic isolates in a collection of 67 avian E. coli. Chromosome or plasmid location, and the nucleotide sequence of these nine fragments were characterized. Four fragments were plasmid-located. The nucleotide sequence of two of them exhibited identity with the sequence of the RepF1B replicon of E. coli plasmids, and the amino-acid deduced sequences from the two other fragments exhibited similarity to the products of genes sitA of Salmonella Typhimurium and iroD of E. coli, which are involved in iron metabolism. Of the five chromosome-located fragments, three were predicted to encode parts of proteins that were significantly homologous to previously described proteins: TktA (transketolase) of Haemophilus influenzae, a FruA (fructokinase) homologue of Listeria innocua and Gp2 (large terminal subunit) of phage 21. The putative products of the two other chromosome-located fragments were homologous to proteins with unknown functions: Z0255 of E. coli strain EDL933 (EHEC) and RatA of Salmonella Typhimurium strain LT2. Both these chromosomal fragments, whose presence is correlated with serogroups O1 and O2 and to the virulence of APEC strains belonging to these serogroups, are good candidates for being part of novel virulence determinants of APEC. Moreover, several fragments were shown to be located close to tRNA selC, asnT or thrW, which suggests they could be part of pathogenicity islands. Six fragments that were shown to be part of whole ORFs present in the APEC strain MT 512 were also present in extra-intestinal pathogenic E. coli (ExPEC) strains of human and animal origin. Thus, the putative novel virulence factors identified in this study could be shared by ExPEC strains of different origins.

Animals↗

[Isolation and identification of a pathogen of grasshoppers].

A pathogen was isolated from natural dead pests of Ceracris kiangsu in Geleshan farm of Chongqin. Its pathogenecity was confirmed by the law of KOCK. It was identified as Pseudomonas pseudoalcaligenes according to its physiological and biochemical properties as well as the G + C content of DNA (63.73mol%). The results of preliminary bioassay show that the pathogen can infect the grasshoppers and Ceracris kiangsu, and also can infect other pests of grassland in a certain extent.

Animals↗

In vitro antibacterial activity of moxifloxacin against hospital isolates: a multicentre study.

OBJECTIVE: To evaluate the in vitro antibacterial activity of moxifloxacinin in comparison to that of other fluoroquinolones (ciprofloxacin, ofloxacin and trovafloxacin). METHODS: A total of 2,196 strains was collected in 11 French hospitals in 1998. Minimum inhibitory concentrations (MICs) (mg/L) were determined by agar dilution and agar diffusion was performed with 5-microg discs. Internal quality control was carried out with genetically defined strains. RESULTS: MIC50s and MIC90s of moxifloxacin against nalidixic acid (NAL)-susceptible Enterobacteriaceae (n = 663) were 0.12 and 0.5. As for other quinolones, the activity of moxifloxacin (4-32) was reduced against NAL-intermediate and NAL-resistant strains (n = 222). MIC50s and MIC90s of moxifloxacin were 2 and 4 for ciprofloxacin-susceptible P. aeruginosa (n = 128); moxifloxacin had no activity against ciprofloxacin-resistant strains (n = 56). The activity of moxifloxacin was maintained against NAL-susceptible A. baumannii (n = 11; 0.032-0.125), but reduced against NAL-resistant strains (n = 30; 16-32). H. influenzae (n = 97) and M. catarrhalis (n = 40) were inhibited by low concentrations (0.03-0.06 and 0.06-0.25, respectively). Moxifloxacin had better activity (0.06-0.12) than other tested quinolones against methicillin-susceptible S. aureus strains (n = 110); ciprofloxacin-resistant strains (n = 85) (2-8) were usually methicillin-resistant. Moxifloxacin was moderately active against enterococci (n = 149) (E. faecalis: 0.5-16; E. faecium: 2-4). Streptococci (n = 194) and pneumococci (n = 136), including 70 penicillin G-intermediate or G-resistant strains, were inhibited by low concentrations (0.25-0.5 for each species). Based on the regression curve, tentative zone diameter breakpoints could be > or =21 and <18 mm for MIC breakpoints of < or =1 and >2 mg/L, respectively. CONCLUSIONS: While retaining activity against Enterobacteriaceae, moxifloxacin was moderately active against P. aeruginosa. Its activity was inferior to that of ciprofloxacin for these species. This study confirmed the comparatively high in vitro activity of moxifloxacin against Gram-positive cocci and other pathogens isolated from community-acquired respiratory tract infections.

Anti-Infective Agents↗

Simian immunodeficiency virus promoter exchange results in a highly attenuated strain that protects against uncloned challenge virus.

Among the many simian immunodeficiency virus (SIV) immunogens, only live attenuated viral vaccines have afforded strong protection to a natural pathogenic isolate. Since the promoter is crucial to the tempo of viral replication in general, it was reasoned that promoter exchange might confer a novel means of attenuating SIV. The core enhancer and promoter sequences of the SIV macaque 239nefstop strain (NF-kappaB/Sp1 region from -114 bp to mRNA start) have been exchanged for those of the human cytomegalovirus immediate-early promoter (CMV-IE; from -525 bp to mRNA start). During culture of the resulting virus, referred to as SIVmegalo, on CEMx174 or rhesus macaque peripheral blood mononuclear cells, deletions arose in distal regions of the CMV-IE sequences that stabilized after 1 or 2 months of culture. However, when the undeleted form of SIVmegalo was inoculated into rhesus macaques, animals showed highly controlled viremia during primary and persistent infection. Compared to parental virus infection in macaques, primary viremia was reduced by >1,000-fold to undetectable levels, with little sign of an increase of cycling cells in lymph nodes, CD4(+) depletion, or altered T-cell activation markers in peripheral blood. Moreover, in contrast to wild-type infection in most infected animals, the nef stop mutation did not revert to the wild-type codon, indicating yet again that replication was dramatically curtailed. Despite such drastic attenuation, antibody titers and enzyme-linked immunospot reactivity to SIV peptides, although slower to appear, were comparable to those seen in a parental virus infection. When animals were challenged intravenously at 4 or 6 months with the uncloned pathogenic SIVmac251 strain, viremia was curtailed by approximately 1,000-fold at peak height without any sign of hyperactivation in CD4(+)- or CD8(+)-T-cell compartment or increase in lymph node cell cycling. To date, there has been a general inverse correlation between attenuation and protection; however, these findings show that promoter exchange constitutes a novel means to highly attenuate SIV while retaining the capacity to protect against challenge virus.

AIDS Vaccines↗

In-vitro antibacterial activity of levofloxacin against hospital isolates: a multicentre study.

The objective of this study was to evaluate the activity of the fluoroquinolone, levofloxacin, against hospital isolates of bacteria. MICs of levofloxacin were determined for 2154 strains by agar dilution. Breakpoints for susceptibility testing were calculated using the agar diffusion technique with 5 micrograms discs. The activity of levofloxacin against nalidixic acid- and pefloxacin-susceptible Enterobacteriaceae (n = 668) was higher (MIC50/90 0.06-0.12 mg/L) than previously reported for ofloxacin. As seen with other fluoroquinolones, this activity was reduced against nalidixic acid-resistant and pefloxacin-intermediate and -resistant strains (MIC 1-8 mg/L). MICs for Pseudomonas aeruginosa (n = 104) were between 0.12 and 128 mg/L. Levofloxacin had good activity against nalidixic acid- and pefloxacin-susceptible Acinetobacter baumannii (n = 12; MIC 0.06-0.25 mg/L), but the activity was reduced against nalidixic acid- and pefloxacin-resistant strains (n = 80; MIC 1-32 mg/L). Haemophilus influenzae (n = 70), Haemophilus parainfluenzae (n = 47) and Moraxella catarrhalis (n = 64) were inhibited by low concentrations of levofloxacin (MICs 0.016-0.03 mg/L, 0.03-0.12 mg/L) and 0.03-0.12 mg/L, respectively). Clostridium perfringens (n = 23; MIC 0.25-1 mg/L) was more susceptible than Bacteroides fragilis (n = 60; MIC 0.5-4 mg/L). Levofloxacin showed superior activity compared with ofloxacin against methicillin-susceptible staphylococci (n = 107; MIC 0.03-0.5 mg/L); the resistant strains (MICs 2-32 mg/L) were usually also resistant to methicillin. Levofloxacin was less effective against enterococci (n = 105; MIC 1-32 mg/L), but streptococci (n = 192) and pneumococci (n = 129), including 58 penicillin-non-susceptible strains, were inhibited by low concentrations (MICs 0.5-2 mg/L). According to the regression curve, zone diameters were usually 20-22 mm, 17-19 mm and 15-16 mm for MICs of 1, 2 and 4 mg/L, respectively. In conclusion, this study, performed on a large number of strains, confirms the superior anti-bacterial activity of levofloxacin compared with ofloxacin, especially against pathogens isolated from respiratory tract infections.

Acinetobacter↗

Organization of aerobactin, hemolysin, and antibacterial resistance genes in lactose-negative Escherichia coli strains of serotype O4 isolated from children with diarrhea.

Epidemiologically related, non-lactose-fermenting (NLF) Escherichia coli strains of serotype O4 have been isolated at a high frequency from children with diarrhea in Somalia (M. Nicoletti, F. Superti, C. Conti, A. Calconi, and C. Zagaglia, J. Clin. Microbiol. 26:524-529, 1988). In order to define the virulence potential of these strains, we characterized the replication properties of their high-molecular-weight plasmids and studied the genetic locations and organization of the aerobactin (aer) and hemolysin (hly) determinants encoded by 23 NLF O4 E. coli strains. Southern blot hybridizations, mobilization assays of nonconjugative plasmids, and incompatibility-exclusion experiments conducted with a conjugative incompatibility group FI (IncFI) plasmid showed that (i) 20 out of the 23 strains examined harbor a 160- to 180-kb IncFI plasmid that shares homology with the basic replicons RepFIA, RepFIB, and (except for the plasmid of one strain) RepFIC, and 22 strains also contain a 40- to 140-kb IncFII plasmid sharing homology with the RepFIIA replicon; (ii) the IncFI plasmid is nonconjugative and carries antibiotic resistance genes; (iii) the aer system is located on the IncFI plasmids and/or the chromosomes in the three strains not harboring IncFI, and it is found in an inverted orientation; (iv) the hly determinants are located on the chromosome, and their genetic organization is well conserved and closely resembles that of the reference hemolytic plasmid pHly152; and (v) Hly- mutants obtained by transposon insertion mutagenesis are not cytotoxic to HeLa cell monolayers, indicating that hemolysin is responsible for the high cytotoxic activity we have previously reported for these strains. The structural organization of the plasmid-encoded aer operon, together with the finding that those plasmids also carry antibiotic resistance genes, indicates that the IncFI plasmid of the NLF O4 E. coli strains studied more closely resembles aer-encoding virulence IncFI Salmonella R plasmids than E. coli ColV plasmids. The data presented here cannot rule out whether the strains examined are potentially intestinal or extraintestinal pathogens. Nevertheless, the genetic organization of the virulence genes, together with the epidemiological behavior and the wide spectrum of antibiotic resistance of the NLF O4 E. coli strains, indicates that these strains are structured as typical E. coli pathogenic isolates of human origin.

Child↗

Microbiological evaluation of contact lenses and contact lens disinfection solutions in an asymptomatic population and in medical personnel.

PURPOSE: To compare the contamination of contact lenses and contact lens disinfection solutions in a population of contact lenses wearers comprised of medical personnel and non-medical subjects. METHODS: Forty-six medical personnel working in an infectious environment and 35 non-medical subjects were enrolled in the study. Contact lenses and contact lens disinfecting solutions were cultured and potential pathogens were isolated. RESULTS: The frequency of isolation of pathogens from the contact lenses of medical personnel was significantly higher than from non-medical subjects. There was no growth in both the contact lens and the disinfecting solution cultures in 56.5% of the subjects in the medical personnel group, and in 57.1% of non-medical group of subjects. Both the contact lens and disinfecting solution cultures were positive in 21.7% of the medical personnel group, while in the non-medical group, only 2.9% of subjects had positive cultures. CONCLUSIONS: Microbacterial contamination of contact lenses and disinfection solution was more frequent in the medical personnel group compared with non-medical subjects. In the medical personnel group, the most frequently isolated microorganism was Staphylococcus albus (19.8%), which is seen in hospital infections. Because soft and rigid gas permeable contact lens usage was approximately equal, we conclude that medical personnel are more prone to contamination.

Adolescent↗

Antibacterial effect of cyanoacrylate glue.

Enbucrilate (butyl-2-cyanoacrylate; Histoacryl [West Germany], no comparable US product) is a tissue adhesive used in the management of corneal perforations that may have a therapeutic value in suppressing selective pathogens isolated, from corneal infections. In three patients with confirmed corneal infections, varying degrees of inhibition were observed. Bacteriostatic activity was most pronounced against gram-positive microorganisms, and no activity was observed against gram-negative organisms. It appears that this chemical adhesive provided an ancillary means of promoting healing of corneal perforations.

Aged↗