The aim of the study was to compare the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli bovine isolates on a conventional dairy cattle farm with high consumption of parenteral and intramammary cephalosporins (farm A) and on an organic dairy farm with no cephalosporin use (farm B). ESBL-producing E. coli were isolated from rectal swabs and milk filters by selective cultivation on MacConkey agar with cefotaxime (2mg/l). ESBL genes were identified by polymerase chain reaction (PCR) and sequencing, and the genetic diversity of the isolates was determined by XbaI pulsed field gel electrophoresis (PFGE). Conjugative transfer, incompatibility group, and restriction fragment length polymorphism (RFLP) profiles of the ESBL-carrying plasmids were studied. Higher prevalence (39%, n(rectal samples in cows)=309) of CTX-M-1-producing E. coli isolates was found on farm A compared to farm B (<1%, n(rectal samples in cows)=154; 0%, n(rectal samples in calves)=46). Using PFGE, the isolates from farm A were divided into nine pulsotypes. In all ESBL-positive isolates, the bla(CTX-M-1) gene was carried on 40 kb IncN conjugative plasmids of three related HincII restriction profiles. Horizontal gene transfer through transmission of IncN plasmids harboring bla(CTX-M-1) as well as clonal dissemination of a particular clone seems to be involved in dissemination of CTX-M-1-producing E. coli isolates in cows on the farm using cephalosporins in treating bacterial infections. The study demonstrates a possible role of cephalosporin use in the widespread occurrence of CTX-M-1-producing E. coli on the conventional dairy cattle farm compared to the organic farm.
- MeSH
- beta-laktamasy genetika MeSH
- cefalosporiny farmakologie MeSH
- DNA bakterií genetika MeSH
- Escherichia coli klasifikace genetika izolace a purifikace MeSH
- infekce vyvolané Escherichia coli mikrobiologie veterinární MeSH
- nemoci skotu mikrobiologie MeSH
- plazmidy genetika MeSH
- polymerázová řetězová reakce MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- přenos genů horizontální MeSH
- proteiny z Escherichia coli genetika MeSH
- pulzní gelová elektroforéza MeSH
- rezistence k cefalosporinům MeSH
- skot mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- skot mikrobiologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
OBJECTIVES: The aim of this study was to determine the occurrence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli at an equine clinic and a horseback riding centre, and to discuss the impact of antimicrobial treatment on resistance selection. METHODS: Faeces from horses, environmental smears and flies were sampled at both the clinic and riding centre. Staff at the equine clinic were also examined. The samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) to isolate ESBL-producing E. coli. The presence of bla and qnr genes was tested by PCR, and transferability was determined by conjugation. Replicon typing and restriction analysis of plasmids harbouring ESBL and qnr genes were performed. RESULTS: E. coli with the blaCTX-M-1 gene were isolated from horses, staff, environmental smears and flies at the two sites. E. coli isolates from the equine clinic harboured an IncHI1 conjugative 235-285 kb plasmid containing blaCTX-M-1, catA1, strA, sul2 and tet(B) genes. Some of these were positive for qnrS1 and/or qnrB19, and were located on 40 or 45 kb IncN or IncX1 conjugative plasmids. The gene blaCTX-M-1 in isolates from the riding centre was carried by IncN (30 kb) and IncI1 (85 kb) conjugative plasmids. Horizontal gene transfer seems to be involved in disseminating E. coli with ESBL and qnr genes at the clinic and riding centre. CONCLUSIONS: The study illustrates that ESBL-producing E. coli, as well as plasmids carrying ESBL genes of clinical interest, can be easily transferred among horses, humans and flies living in close contact.
- MeSH
- beta-laktamasy genetika MeSH
- Diptera mikrobiologie MeSH
- DNA bakterií chemie genetika MeSH
- Escherichia coli genetika izolace a purifikace MeSH
- feces mikrobiologie MeSH
- koně mikrobiologie MeSH
- konjugace genetická MeSH
- mikrobiologie životního prostředí MeSH
- molekulární sekvence - údaje MeSH
- plazmidy MeSH
- polymerázová řetězová reakce MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- proteiny z Escherichia coli genetika MeSH
- sekvenční analýza DNA MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
AIM: To examine surface water from a pond in the northeastern part of the Czech Republic and young black-headed gulls (Larus ridibundus) nesting on the same pond for the presence of antibiotic-resistant Salmonella and Escherichia coli. METHODS AND RESULTS: A total of 16% (n = 87) of water and 24% (n = 216) of gull samples yielded Salmonella. Salmonella Enteritidis PT8 and PT4 were the most prevalent. Antibiotic resistance was found in 12% (n = 14) of water and 28% (n = 51) of gull salmonellae. Escherichia coli were found in 83 (95%) and 213 (99%) of pond water and gull samples, respectively. Totals of 18% (n = 83) of water and 28% (n = 213) of gull E. coli isolates were resistant to antimicrobial agents tested. Class 1 integrons were found in 21% (n = 14) of water and 15% (n = 60) of gull antibiotic-resistant E. coli isolates. Class 2 integrons and extended-spectrum beta-lactamase-producing E. coli isolates (with bla(CTX-M-1), bla(CTX-M-15)-like, bla(SHV-2) and bla(SHV-12)) were found in 13% (eight positive, n = 60 gull-resistant E. coli isolates) and 3% (seven positive, n = 216 gull E. coli isolates) of gull isolates, respectively. Antibiotic-resistant E. coli isolates with identical pulsed field gel electrophoresis (PFGE) patterns were found in either gulls or water, but not both. Salmonellae of the same serotype and PFGE profile were found in both gulls and water. CONCLUSION: A high prevalence of antibiotic-resistant salmonellae and E. coli were found in both pond water and in sympatric black-headed gulls. SIGNIFICANCE AND IMPACT OF THE STUDY: Intensive contamination of pond surface water by antibiotic-resistant E. coli and salmonellae was documented. Black-headed gulls were identified as important reservoirs of antibiotic-resistant salmonellae and E. coli, including extended-spectrum beta-lactamase-producing isolates.
- MeSH
- antibiotická rezistence MeSH
- beta-laktamasy genetika MeSH
- Charadriiformes mikrobiologie MeSH
- DNA bakterií genetika MeSH
- Escherichia coli účinky léků genetika izolace a purifikace MeSH
- integrony MeSH
- mikrobiální testy citlivosti MeSH
- polymerázová řetězová reakce metody MeSH
- pulzní gelová elektroforéza MeSH
- Salmonella účinky léků genetika izolace a purifikace MeSH
- zdroje nemoci veterinární MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
