G. Arlet

Rapid detection of the O25b-ST131 clone of Escherichia coli encompassing the CTX-M-15-producing strains

OBJECTIVESnRecently, a CTX-M-15 extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli O25b-ST131 clone, belonging to the B2 phylogenetic group and with a high virulence potential, has been reported all over the world, representing a major public health problem. The present study was carried out to develop a rapid and simple detection assay that identifies members of this clone.nnnMETHODSnA total of 627 E. coli isolates of which 373 produced an ESBL, collected across four continents, were screened using a O25b-ST131 clone allele-specific PCR for the pabB gene.nnnRESULTSnOne hundred and forty-three ESBL isolates were found positive with the assay. These isolates were all of O25b type and, when studied by multilocus sequence typing (25 cases), were all of ST131. The O25b-ST131 clone was found to produce ESBLs other than CTX-M-15, specifically CTX-M-2, -3, -14, -27, -32 and -61 as well as TEM-24. This clone represents 3% of non-ESBL B2 isolates originating from urinary tract infections in Paris.nnnCONCLUSIONSnWe have developed a PCR-based assay that easily identifies a clone with high likelihood of producing ESBLs, including CTX-M-15.

Fecal recovery in humans of viable Bifidobacterium sp ingested in fermented milk

Bifidobacterium sp is a natural component of the dominant colonic microflora that was recently introduced into several fermented dairy products. The aim of the present study was to study the fate of this microorganism in the human gut. On the basis of antibiotic resistance characters, a variant of Bifidobacterium sp that could be distinguished from indigenous bifidobacteria in the fecal flora was selected, and its survival and colonization in the colon was examined. This strain was used to ferment milk, and 125 g of the fermented product obtained was ingested by eight healthy volunteers three times daily for 8 days. Stools were recovered and weighed throughout the study. The results showed that the exogenous Bifidobacterium sp appeared in the stools and reached a mean level of 8.8 +/- 0.1 log colony-forming units per gram. This level was maintained as long as the fermented dairy product was consumed. When its ingestion stopped, the exogenous Bifidobacterium sp gradually decreased and was no longer detectable 8 days after cessation. The mean recovered quantity during the 8-day period of administration of the ingested bifidobacteria excreted in stools was 12.1 +/- 0.1 log colony-forming units per gram, i.e., 29.7% +/- 6% of the ingested bacteria, which was similar to the percentage that reached the colon in previous studies. It is concluded that under physiological conditions, exogenously administered Bifidobacterium sp do not colonize the human colon. However, the high fecal concentrations of exogenous bifidobacteria reached are compatible with metabolic probiotic activities.

Origin and impact of plasmid-mediated extended-spectrum beta-lactamases

Resistance to oxyimino cephalosporins was originally highlighted by the emergence of plasmid-encoded extended-spectrum β-lactamases deriving by mutation from TEM-1, TEM-2 and SHV type enzymes (class A). The broader spectrum of resistance produced by these enzymes is related to more amino acid substitutions, but susceptibility to seven alpha-methoxyimino cephalosporins and carbapenems was preserved until recently. Clavulanate-sensitive extended-spectrum β-lactamases are distributed worldwide, mainly amongKlebsiella pneumoniae isolates. Novel clavulanate-sensitive extended-spectrum β-lactamases deriving from other class A enzymes (e.g. MEN-1 from βlaOXY, OXA-11 inPseudomonas aeruginosa from PSE-2) have been reported. Recently, clavulanate-resistant extended-spectrum β-lactamases (class C) were encountered amongst single isolates, mostlyKlebsiella pneumoniae. These cephalosporinases or cefamycinases (usually chromosomally mediated) have expanded the spectrum of plasmid-encoded resistance to include seven alpha-methoxyimino cephalosporins. Thus far, only two isolates (1Pseudomonas aeruginosa, 1Bacteroides fragilis), both recovered in Japan, with plasmid-mediated resistance to carbapenems have been found.

Clonal Dissemination of a CTX-M-15 β-Lactamase-Producing Escherichia coli Strain in the Paris Area, Tunis, and Bangui

ABSTRACT One hundred twenty CTX-M-15-producing Escherichia coli strains isolated in 10 different hospitals from Paris (France), in the Hospital Charles Nicolle in Tunis (Tunisia), and in the Pasteur Institute in Bangui, Central African Republic (CAR), between 2000 and 2004 were studied. Eighty isolates, recovered from the three countries, were clonally related by repetitive extragenic palindromic PCR and pulsed-field gel electrophoresis. Various resistance profiles were identified among these clonal strains. After conjugation or electroporation of plasmids from E. coli strains representative of each profile and each geographic region, we observed seven resistance profiles in the recipient strains. Incompatibility typing showed that all the plasmids transferred from the clonal strains studied, except one, belonged to the incompatibility group FII. They all shared a multidrug resistance region (MDR) resembling the MDR region located in pC15-1a, a plasmid associated with an outbreak of a CTX-M-15-producing E. coli strain in Canada. They also shared the common backbone of an apparent mosaic plasmid, including several features present in pC15-1a and in pRSB107, a plasmid isolated from a sewage treatment plant. This study suggests that although the plasmid-borne blaCTX-M-15 gene could be transferred horizontally, its dissemination between France, Tunisia, and CAR was due primarily to its residence in an E. coli clone with a strong propensity for dissemination.

Campylobacter Bacteremia: Clinical Features and Factors Associated with Fatal Outcome

BACKGROUNDnCampylobacter bacteremia is uncommon. The influence of underlying conditions and of the impact of antibiotics on infection outcome are not known.nnnMETHODSnFrom January 2000 through December 2004, 183 episodes of Campylobacter bacteremia were identified in 23 hospitals in the Paris, France, area. The medical records were reviewed. Characteristics of bacteremia due to Campylobacter fetus and to other Campylobacter species were compared. Logistic regression analysis was performed to identify risk factors for fatal outcome within 30 days.nnnRESULTSnMost affected patients were elderly or immunocompromised. C. fetus was the most commonly identified species (in 53% of patients). The main underlying conditions were liver disease (39%) and cancer (38%). The main clinical manifestations were diarrhea (33%) and skin infection (16%). Twenty-seven patients (15%) died within 30 days. Compared with patients with bacteremia due to other Campylobacter species, patients with C. fetus bacteremia were older (mean age, 69.5 years vs. 55.6 years; P = .001) and were more likely to have cellulitis (19% vs. 7%; P = .03), endovascular infection (13% vs. 1%; P = .007), or infection associated with a medical device (7% vs. 0%; P = .02). Independent risk factors for death were cancer (odds ratio [OR], 5.1; 95% confidence interval [CI], 1.2-20.8) and asymptomatic infection (OR, 6.7; 95% CI, 1.5-29.4) for C. fetus bacteremia, the absence of prescription of appropriate antibiotics (OR, 12.2; 95% CI, 0.9-157.5), and prescription of third-generation cephalosporins (OR, 10.2; 95% CI, 1.9-53.7) for bacteremia caused by other species.nnnCONCLUSIONSnCampylobacter bacteremia occurs mainly in immunocompromised patients. Clinical features and risk factors of death differ by infection species.

Nosocomial outbreak of acute gastroenteritis in a neonatal intensive care unit in tunisia caused by multiply drug resistantSalmonella wien producing SHV-2 beta-lactamase

In a Tunisian hospital 27 babies, including 12 who were premature, in a single intensive care unit suffered acute gastroenteritis in the period from January to May 1988. The mean age at the onset of gastroenteritis was 8.4 days; nine babies died.Salmonella wien was isolated from stools (all babies) and blood (4 babies). It was also isolated from the stools of one nurse and from a mattress. Twelve of the babies had received cefotaxime, which was successfully replaced by oral colimycin. The outbreak was stopped by the implementation of infection control measures. All isolates ofSalmonella wien were of the same biotype, and had the same antibiotic resistance pattern (third generation cephalosporins, monobactams, aminoglycosides, chloramphenicol, trimethoprim and sulphonamides) and plasmid DNA restriction pattern. The isolates were all susceptible to a combination of cefotaxime and clavulanic acid (a β-lactamase inhibitor), which displayed synergy, suggesting the presence of a β-lactamase (geometric mean MICs 11.24 µg/ml for cefotaxime alone and 0.24 µg/ml in combination with 0.1 µg/ml potassium clavulanate). All isolates produced TEM-1 and SHV-2 β-lactamase which was not transferable toEscherichia coli by conjugation. The presence of the SHV-2 enzyme inSalmonella wien may allow it to adapt to newer β-lactams which is a cause for concern in this hospital.

Dissemination of ESBL and Qnr determinants in Enterobacter cloacae in Algeria

OBJECTIVESnThe aim of this study is to evaluate the prevalence and diversity of extended-spectrum beta-lactamases (ESBLs) in Enterobacter cloacae clinical isolates collected from Algerian hospitals and to verify the association with qnr genes.nnnMETHODSnMICs were determined by Etest for isolates giving positive double-disc synergy tests, and all isolates were screened by PCR and sequenced, respectively, for bla(TEM), bla(CTX-M), bla(SHV) and bla(VEB) genes and for qnr genes (qnrA, qnrB, qnrS), using specific primers.nnnRESULTSnThe prevalence of ESBLs was 25/141 (17.7%) with 11, 9, 4 and 1 isolates testing positive for genes encoding CTX-M-15, CTX-M-3, SHV-12 and VEB-1, respectively. Two SHV-12 producers and one CTX-M-15 producer expressed QnrS1, one isolate produced CTX-M-15 and QnrB1 and one SHV-12 producer co-expressed QnrS1 and QnrB4. qnrA was not detected in our collection, and qnr alleles were not detected in non-ESBL-producing isolates.nnnCONCLUSIONSnSHV-12, QnrS1, QnrB1 and QnrB4 were reported for the first time in Algeria. This study also described a co-expression of qnrS1 and qnrB4 by an SHV-12 producer isolate.

Prevalence and characterization of extended-spectrum β-lactamases in Klebsiella pneumoniae in Algiers hospitals (Algeria)

AIM OF THE STUDYnTo determine the prevalence and the diversity of extended-spectrum beta-lactamases (ESBLs) in 196 Klebsiella pneumoniae clinical isolates collected from three hospitals in Algiers.nnnMATERIALS AND METHODSnAntibiograms were done on Mueller-Hinton agar plates with the disc-diffusion method and MICs were determined by agar-dilution method. Mating experiments were performed in agar medium. Plasmid DNA was extracted by the alcalin-lysis method. Total DNA was extracted with a Qiagen mini kit and screened for bla(TEM) and bla(CTX-M) genes by PCR. Linkage of bla(CTX-M) genes with insertion sequence ISEcp1B and class 1 integrons was investigated by PCR. PCR products were sequenced by the Sanger method. The epidemiological relationships between ESBL-producing K. pneumoniae isolates were analyzed by ERIC-PCR.nnnRESULTSnThirty-nine (19.9%) isolates were found to produce ESBLs belonging to CTX-M-1 group and TEM penicillinases (CTX-M-3, CTX-M-15 and TEM-1). ERIC-PCR analysis showed that the isolates are genetically unrelated. The bla(TEM) and bla(CTX-M) genes as well as aminoglycosides and sulfonamides resistance determinants were found located in self-transferable plasmids of approximately 85 kb. The class 1 integrons and the insertion sequence ISEcp1B were present in the isolates and in their transconjugants. ISEcp1B was found genetically linked to the bla(CTX-M) genes and located 127bp upstream, with the presence of the V and W sequences.nnnCONCLUSIONnThe study revealed a high rate of ESBL-producing K. pneumoniae in Algerian hospitals, resulting from horizontal dissemination of mobile bla(CTX-M) genes.

The CTX-M-15-producing Escherichia coli clone O25b: H4-ST131 has high intestine colonization and urinary tract infection abilities.

Increasing numbers of pyelonephritis-associated uropathogenic Escherichia coli (UPEC) are exhibiting high resistance to antibiotic therapy. They include a particular clonal group, the CTX-M-15-producing O25b:H4-ST131 clone, which has been shown to have a high dissemination potential. Here we show that a representative isolate of this E. coli clone, referred to as TN03, has enhanced metabolic capacities, acts as a potent intestine- colonizing strain, and displays the typical features of UPEC strains. In a modified streptomycin-treated mouse model of intestinal colonization where streptomycin was stopped 5 days before inoculation, we show that TN03 outcompetes the commensal E. coli strains K-12 MG1655, IAI1, and ED1a at days 1 and 7. Using an experimental model of ascending UTI in C3H/HeN mice, we then show that TN03 colonized the urinary tract. One week after the transurethral inoculation of the TN03 isolates, the bacterial loads in the bladder and kidneys were significantly greater than those of two other UPEC strains (CFT073 and HT7) belonging to the same B2 phylogenetic group. The differences in bacterial loads did not seem to be directly linked to differences in the inflammatory response, since the intrarenal expression of chemokines and cytokines and the number of polymorphonuclear neutrophils attracted to the site of inflammation was the same in kidneys colonized by TN03, CFT073, or HT7. Lastly, we show that in vitro TN03 has a high maximum growth rate in both complex (Luria-Bertani and human urine) and minimum media. In conclusion, our findings indicate that TN03 is a potent UPEC strain that colonizes the intestinal tract and may persist in the kidneys of infected hosts.

Genetic Context of Plasmid-Carried blaCMY-2-Like Genes in Enterobacteriaceae

ABSTRACT Analysis of 15 European clinical Enterobacteriaceae isolates showed that differences in the genetic context of blaCMY-2-like genes reflected the replicon type, usually IncA/C or IncI1. These blaCMY-2 loci may originate from the same ISEcp1-mediated mobilization from the Citrobacter freundii chromosome as structures described in earlier studies.