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Herd prevalence of Enterobacteriaceae producing CTX‐M‐type and CMY‐2 β‐lactamases among Japanese dairy farms
Authors:M Ohnishi  AT Okatani  H Esaki  K Harada  T Sawada  M Murakami  K Marumo  Y Kato  R Sato  K Shimura  N Hatanaka  T Takahashi
Institution:1. Veterinary Clinical Laboratory, Nemuro District Agricultural Mutual Aid Association, , Nakashibetsu, Hokkaido, Japan;2. Laboratory of Veterinary Public Health II, School of Veterinary Medicine, Azabu University, , Sagamihara, Kanagawa, Japan;3. Department of Biotechnology, Research Institute for Animal Science in Biochemistry and Toxicology, , Sagamihara, Kanagawa, Japan;4. Department of Veterinary Internal Medicine, Tottori University, , Tottori‐shi, Tottori, Japan;5. Laboratory of Veterinary Microbiology, School of Veterinary Medicine, Nippon Veterinary and Life Science University, , Musashino, Tokyo, Japan;6. Laboratory of Molecular Biology, School of Veterinary Medicine, Azabu University, , Sagamihara, Kanagawa, Japan;7. Department of Microbiology and Immunology, Showa University School of Medicine, , Shinagawa, Tokyo, Japan;8. Laboratory of Gynecology and Obstetrics, School of Veterinary Medicine, Azabu University, , Sagamihara, Kanagawa, Japan
Abstract:

Aims

To determine the herd prevalence of Enterobacteriaceae producing CTX‐M‐type extended‐spectrum β‐lactamases (ESBLs) among 381 dairy farms in Japan.

Methods and Results

Between 2007 and 2009, we screened 897 faecal samples using BTB lactose agar plates containing cefotaxime (2 μg ml?1). Positive isolates were tested using ESBL confirmatory tests, PCR and sequencing for CTX‐M, AmpC, TEM and SHV. The incidence of Enterobacteriaceae producing CTX‐M‐15 (= 7), CTX‐M‐2 (= 12), CTX‐M‐14 (= 3), CMY‐2 (= 2) or CTX‐M‐15/2/14 and CMY‐2 (= 4) in bovine faeces was 28/897 (3·1%) faecal samples. These genes had spread to Escherichia coli (= 23) and three genera of Enterobacteriaceae (= 5). Herd prevalence was found to be 20/381 (5·2%) dairy farms. The 23 E. coli isolates showed clonal diversity, as assessed by multilocus sequence typing and pulsed‐field gel electrophoresis. The pandemic E. coli strain ST131 producing CTX‐M‐15 or CTX‐M‐27 was not detected.

Conclusions

Three clusters of CTX‐M (CTX‐M‐15, CTX‐M‐2, CTX‐M‐14) had spread among Japanese dairy farms.

Significance and Impact of the Study

This is the first report on the prevalence of multidrug‐resistant CTX‐M‐15–producing E. coli among Japanese dairy farms.
Keywords:CMY‐2 β  ‐lactamase  CTX‐M type ESBLs  dairy cattle     Escherichia coli     multilocus sequence typing  pulsed‐field gel electrophoresis
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