Molecular analysis of the carbapenem and metronidazole resistance mechanisms of Bacteroides strains reported in a Europe-wide antibiotic resistance survey.

Here we examine the carbapenem and metronidazole resistance mechanisms of 640 Bacteroides strains reported in the 2008-2009 European antibiotic susceptibility survey. Of the 22 strains with elevated imipenem minimum inhibitory concentrations (≥4 μg/mL), 10 were cfiA-positive and out of these 5 carried activating insertion sequence (IS) elements in the upstream regions of the cfiA genes. However, resistant strains with cfiA genes but with no activating IS elements were found (n=2) as well as a resistant strain with no cfiA gene. In the former the resistance phenotypes by Etest were heterogeneous, whilst in the latter no carbapenemase production was seen; both mechanisms have been rarely observed, examined and characterised. Interestingly, few (n=3) nim-positive strains were found, including one metronidazole-resistant strain harbouring nimE activated by ISBf6, and two susceptible strains harbouring chromosomally located nim genes

Molecular analysis of the carbapenem and metronidazole resistance mechanisms of Bacteroides strains reported in a Europe-wide antibiotic resistance survey

Here we examine the carbapenem and metronidazole resistance mechanisms of 640 Bacteroides strains reported in the 2008-2009 European antibiotic susceptibility survey. Of the 22 strains with elevated imipenem minimum inhibitory concentrations (≥4 μg/mL), 10 were cfiA-positive and out of these 5 carried activating insertion sequence (IS) elements in the upstream regions of the cfiA genes. However, resistant strains with cfiA genes but with no activating IS elements were found (n = 2) as well as a resistant strain with no cfiA gene. In the former the resistance phenotypes by Etest were heterogeneous, whilst in the latter no carbapenemase production was seen; both mechanisms have been rarely observed, examined and characterised. Interestingly, few (n = 3) nim-positive strains were found, including one metronidazole- resistant strain harbouring nimE activated by ISBf6, and two susceptible strains harbouring chromosomally located nim genes. © 2012 Elsevier B.V. and the International Society of Chemotherapy

Investigation of the MICs of fidaxomicin and other antibiotics against Hungarian Clostridium difficile isolates

The aim of this study was to investigate in vitro activities of fidaxomicin and other antibiotics against 188 Clostridium difficile strains collected from different centers of Hungary. C. difficile isolates showed minimum inhibitory concentration (MIC) range for fidaxomicin of </=0.008-0.5 mg/L, with a MIC90 of 0.125 mg/L. Only four isolates (2.1%) had 0.5 mg/L MIC to fidaxomicin. The obtained MICs showed identical distribution to those found in the EUCAST database for wild-type strains

Analysis of Romanian Bacteroides isolates for antibiotic resistance levels and the corresponding antibiotic resistance genes

As part of an ESCMID Study Group on Anaerobic Infections (ESGAI) project, a study was conducted to measure the antibiotic susceptibilities and corresponding gene contents of 53 Bacteroides fragilis group strains isolated in Romania. The antibiotic resistance data was comparable with the data found for other East-European countries. Here, no resistant isolate was found for imipenem, metronidazole and tigecycline. An increasing role of the cepA, cfxA and cfiA genes was observed in their corresponding antibiotic resistances. Moreover, no isolate was found that harbored the cfiA gene with a possible activating IS element. Clindamycin resistance was low, similarly to that the rate for the ermF gene. However, we did find some isolates with nimB, ermB, msrSA, linA, satG, tetX, tetM and bexA genes. This study was the first to provide antibiotic resistance data for clinical Bacteroides strains from Romania

The prevalence of antibiotic resistance genes in Bacteroides fragilis group strains isolated in different European countries

From the 2008-2009 European Bacteroides antibiotic resistance survey, we selected 161 strains for detection of antibiotic resistance genes (cepA, cfxA, cfiA, nim, ermB, ermF, ermG, linA, mefA, msrSA, tetM, tetQ, tetX, tetX1, tet36 and bexA). To facilitate the throughput, the genes were detected by Real-Time PCR. The presence of the genes was correlated with the known MIC data of the strains for the appropriate antibiotics. For the beta-lactams, the cepA gene was found in 70.8 % of the tested strains (all resistant to ampicillin), but its presence did not correlate with the ampicillin MIC values. The cepA gene occurred at different frequencies among Bacteroides fragilis and non-fragilis Bacteroides strains. The cfxA gene was not a major factor in determining cefoxitin resistance and it was found with higher prevalence in non-fragilis Bacteroides strains than in B. fragilis. Among the five possible clindamycin resistance genes, ermF was the most common and had the highest effect on clindamycin resistance after linA. The ermG-mefA-msrSA combination was found in a set of strains and their linked occurrence implied that they were harbored by the conjugative transposon CTnGERM1. All strains tested were susceptible to metronidazole and none of them harbored nim genes. TetQ was prevalent among both the B. fragilis and non-fragilis Bacteroides strains (78.9 and 84.8%, respectively) and no gene could be clearly linked to tigecycline resistance other than tetQ. BexA, which codes for the fluoroquinolone efflux pump, was found in 7.5% of strains and occurred at different frequencies among B. fragilis and non-fragilis Bacteroides strains, but was represented only in a minor proportion of moxifloxacin-resistant strains

Development of EUCAST disk diffusion method for susceptibility testing of the Bacteroides fragilis group isolates.

With the emergence of antibiotic resistance among Bacteroides fragilis group isolates the need of susceptibility testing in routine laboratories is increasing. The aims of the present study were to evaluate the disk diffusion method for susceptibility testing in case of different clinical isolates of Bacteroides spp by comparing zone diameter results with MICs obtained earlier during an Europe-wide antibiotic susceptibility surveillance, and to propose zone diameter breakpoints, which correlate for the EUCAST MIC breakpoints. We tested 381 clinical isolates of the B. fragilis group to amoxicillin/clavulanic acid, cefoxitin, clindamycin, imipenem, metronidazole, moxifloxacin, piperacillin/tazobactam, tigecycline by agar dilution method previously. The inhibition zones of the same antibiotics including meropenem disc were determined by the disc diffusion on Brucella blood agar supplemented with haemin and vitamin K1. Plates were incubated at 37 degrees C in an anaerobic atmosphere for 24 h. The zone diameters were read at 100% inhibition. In case of discrepant results MICs were determined by gradient test and compared with the inhibition zones on the same plate. We found a good agreement between the inhibition zone diameters and the MICs for imipenem, metronidazole, moxifloxacin and tigecyclin. The inhibition zone diameters of meropenem also separated clearly the isolates, which can be considered wild-type isolates. In case of amoxicillin/clavulanic acid and piperacillin/tazobactam intermediate and susceptible isolates according to the MIC determination, overlap during the zone diameter determination. Isolates with an inhibition zone <23 mm for amoxicillin/clavulanic acid and <25 mm for piperacillin/tazobactam should be retested by a MIC determination method. The 10 mug clindamycin disc clearly separated the resistant and the susceptible population of B. fragilis group strains. In the case of cefoxitin only resistant population could be separated with an inhibition zone <17 mm, intermediate and susceptible isolates overlap. In conclusion, we suggest that disk diffusion can be an option for susceptibility testing of B. fragilis group isolates for most relevant antibiotics in routine laboratories

Occurrence and analysis of rare cfiA-bft doubly positive Bacteroides fragilis strains

We detected four cfiA-bft1 doubly positive Bacteroides fragilis strains out of 486 B. fragilis isolates analyzed for antibiotic susceptibilities and antibiotic resistance genes from a recent pan-European survey. The prevalence of the enterotoxin bft genes was roughly equal among cfiA-negative and -positive B. fragilis strains. We also demonstrated that the cfiA-bft doubly positive strains had the most common B. fragilis genomic pattern (I.1.). Thus we concluded that the bft-carrying CTn86 conjugative transposons are mobile accounting for this unexpected simultaneous occurrence of the cfiA and bft genes

Occurrence and analysis of rare cfiA–bft doubly positive Bacteroides fragilis strains

We detected four cfiAebft1 doubly positive Bacteroides fragilis strains out of 486 B. fragilis isolates analyzed for antibiotic susceptibilities and antibiotic resistance genes from a recent pan-European sur- vey. The prevalence of the enterotoxin bft genes was roughly equal among cfiA-negative and -positive B. fragilis strains. We also demonstrated that the cfiAebft doubly positive strains had the most common B. fragilis genomic pattern (I.1.). Thus we concluded that the bft-carrying CTn86 conjugative transposons are mobile accounting for this unexpected simultaneous occurrence of the cfiA and bft genes