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MacConkey Agar (macconkey + agar)
Selected AbstractsAntimicrobial-resistant faecal Escherichia coli in wild mammals in central Europe: multiresistant Escherichia coli producing extended-spectrum beta-lactamases in wild boarsJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2010I. Literak Abstract Aims:, To determine the presence of antibiotic-resistant faecal Escherichia coli in populations of wild mammals in the Czech Republic and Slovakia. Methods and Results:, Rectal swabs or faeces collected during 2006,2008 from wild mammals were spread on MacConkey agar and MacConkey agar containing 2 mg l,1 of cefotaxime. From plates with positive growth, one isolate was recovered and identified as E. coli. Susceptibility to 12 antibiotics was tested using the disk diffusion method. Resistance genes, class 1 and 2 integrons and gene cassettes were detected in resistant isolates by polymerase chain reaction (PCR). Extended-spectrum beta-lactamases (ESBL) were further characterized by DNA sequencing, macrorestriction profiling and determination of plasmid sizes. Plasmid DNA was subjected to EcoRV digestion, transferability by conjugation and incompatibility grouping by multiplex PCR. The prevalence of resistant isolates was 2% in small terrestrial mammals (rodents and insectivores, nE. coli = 242), 12% in wild ruminants and foxes (nE. coli = 42), while no resistant isolates were detected in brown bears (nE. coli = 16). In wild boars (Sus scrofa) (nE. coli = 290), the prevalence of resistant isolates was 6%. Class 1 and 2 integrons with various gene cassettes were recorded in resistant isolates. From wild boars, five (2%, nrectal smears = 293) multiresistant isolates producing ESBL were recovered: one isolate with blaCTX-M-1 + blaTEM-1, three with blaCTX-M-1 and one with blaTEM-52b. The blaCTX-M-1 genes were carried on approx. 90 kb IncI1 conjugative plasmids. Conclusions:, Antibiotic-resistant E. coli occured in populations of wild mammals in various prevalences. Significance and Impact of the Study:, Wild mammals are reservoirs of antibiotic-resistant E. coli including ESBL-producing strains which were found in wild boars. [source] Prevalence of enterohaemorrhagic Escherichia coli from serotype O157 and other attaching and effacing Escherichia coli on bovine carcasses in AlgeriaJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2006A. Chahed Abstract Aims:, Bovine meat is the principal source of human contamination of attaching and effacing Escherichia coli, including enterohaemorrhagic E. coli O157. The aim was to study the prevalence of these strains on bovine carcasses in Algeria. Methods and Results:, Two-hundred and thirty carcasses were swabbed and analysed by classical microbiological methods for total E. coli counts and for the presence of pathogenic E. coli. The E. coli counts were high, with a 75th percentile of 444·75 CFUs cm,2. For pathogenic E. coli, more than 7% of the tested carcasses were positive for E. coli O157. Eighteen E. coli O157 strains were isolated and typed by multiplex PCR. The main isolated pathotype (78%) was eae+ stx2+ ehxA+. In addition to E. coli O157, other attaching and effacing E. coli (AEEC) were also detected from carcasses by colony hybridization after pre-enrichment and plating on sorbitol MacConkey agar using eae, stx1 and stx2 probes. Thirty carcasses (13%) on the 230 analysed harboured at least one colony positive for one of the tested probes. These positive carcasses were different from those positive for E. coli O157. Sixty-six colonies (2·9%) positive by colony hybridization were isolated. The majority (60·6%) of the positive strains harboured an enteropathogenic E. coli -like pathotype (eae+ stx,). Only three enterohaemorrhagic E. coli (EHEC)-like (eae+ stx1+) colonies were isolated from the same carcass. These strains did not belong to classical EHEC serotypes. Conclusions:, In this study, the global hygiene of the slaughterhouse was low, as indicated by the high level of E. coli count. The prevalence of both E. coli O157 and other AEEC was also high, representing a real hazard for consumers. Significance and Impact of the Study:, This is the first study of this type in Algeria, which indicates that the general hygiene of the slaughterhouse must be improved. [source] Comparison of the sensitivity of manual and automated immunomagnetic separation methods for detection of Shiga toxin-producing Escherichia coli O157:H7 in milkJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2002R.D. Reinders Aim:,To determine the sensitivity of methods for detection of injured and uninjured Escherichia coli O157:H7 (E. coli O157) in raw and pasteurized milk. Methods and Results:,Raw milk, pasteurized milk with 1·5% fat content and pasteurized milk with 3·5% fat content were spiked with E. coli O157 at low levels. The samples were enriched in modified tryptone soya broth with novobiocin (mTSBn) at 37°C. Aliquots of the enriched culture were analysed either by manual immunomagnetic separation (MIMS) and culturing on sorbitol MacConkey agar with or without cefixime and potassium tellurite (SMACct or SMAC), or by automated immunomagnetic separation and integrated ELISA (EiaFossÔ). Uninjured E. coli O157 organisms were detected in milk by both methods at 1 cfu 10 ml,1 sample). Injured organisms were detected at levels of about 4 cfu 10 ml,1 sample. Direct enrichment in mTSBn (22 h incubation) showed better sensitivity for injured cells than enrichment in buffered peptone water (BPW, 22 h incubation), or in a two-step enrichment consisting of BPW (6 h, 37°C) and mTSBn (16 h, 37°C), successively. Conclusions:,The methods showed equal sensitivity in that they were both able to detect 1 cfu 10 ml,1 milk sample. Injured organisms can be detected and isolated at a level almost as low as this. A resuscitation step is not recommended for the detection and isolation of injured and non-injured E. coli O157 from milk. Significance and Impact of the Study:,Due to the dilution of contamination in the bulk tank, analysis of milk for the presence of E. coli O157 requires a very sensitive method. Both methods described here are useful for such analysis. [source] Cefixime,tellurite rhamnose MacConkey agar for isolation of Vero cytotoxin-producing Escherichia coli serogroup O26 from Scottish cattle and sheep faecesLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2008J. Evans Abstract Aims:, To compare rhamnose MacConkey agar supplemented with cefixime and tellurite (CT-RMac) and tryptone bile X-glucuronide (TBX) agars as isolation media for Vero cytotoxin-producing Escherichia coli (VTEC) serogroup O26 from animal faeces. Methods and Results:, Nine VTEC O26 were isolated from sheep faeces; out of which six were isolated only on CT-RMac and one was isolated only on TBX. One hundred and twelve VTEC O26 were isolated from calf faeces; out of which 97% were from CT-RMac and 52% were from TBX. In a study of E. coli O26 strains, 84% of VT-positive O26 did not ferment rhamnose when compared with 16% of VT-negative O26. VT-positive (19%) and VT-negative (39%) E. coli O26 strains did not grow on CT-RMac agar. Conclusions:, It is important to consider that VTEC O26 strains either may ferment rhamnose or may be sensitive to the CT supplement of CT-RMac agar. Significance and Impact of the Study:, This work compares CT-RMac and TBX agars as isolation medium for VTEC O26 from Scottish animal faeces and highlights that VTEC O26 may be missed if only CT-RMac agar is used. [source] Prevalence of Escherichia coli O157:H7 in industrial minced beefLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2002C. Vernozy-Rozand Aims: ,The lack of baseline data on the prevalence of Escherichia coli O157:H7 in retail minced beef in France prompted this survey of industrial minced beef production. Methods and Results: ,An automated enzyme-linked fluorescence immunoassay (ELFA), the VIDAS E. coli O157 method, was used to detect E. coli O157 in industrial minced beef samples. Confirmation of samples positive according to the ELFA was performed using an automated immunoconcentration (ICE) system, VIDAS ICE, which allows the selective capture and release of target organisms. The ICE was followed by culture on cefixime tellurite sorbitol MacConkey agar and a chromogenic medium, O157:H7 ID. Of the 3450 minced beef samples tested, 175 samples were positive with the ELFA method and, of these, four were confirmed by the ICE method. They were identified as sorbitol-negative, O157-positive, H7-positive, mobile, verotoxin-producing E. coli . Conclusions: ,The prevalence of E. coli O157:H7 in industrial French minced beef was 0·12%, consistent with many other reports. Significance and Impact of the Study: ,The low infective dose of E. coli O157:H7 presents a major threat. The main means of combating this organism are thermal destruction and good food hygiene covering activities on-farm, in the abattoir and in minced beef industries. [source] |