RESUMO
OBJECTIVES: To characterize ESBL-encoding Escherichia coli cultured from pigs and their plasmids carrying these genes following conjugation into recipient strains. METHODS: Six ESBL-producing E. coli were recovered from faecal samples taken from pigs along with a further isolate from the environment of a farrowing house on three pig farms in Ireland. These isolates were characterized by phylogenetic grouping, MLST and ESBL genotype analyses. Conjugation experiments were carried out in broth mating assays. S1-nuclease PFGE was used to determine the plasmid profiles. Whole-genome sequences of the seven E. coli were determined and subsequently analysed. RESULTS: Phylogenetic groups and the corresponding MLST STs identified among the seven tested E. coli isolates included A/ST10, A/ST34, C/ST23 and C/ST1629. All seven isolates carried one or more high-molecular-weight plasmids and demonstrated the ability to transfer their cefotaxime resistance phenotype at high frequencies. Five transmissible plasmid replicon types were detected, including IncK/B (nâ=â3), IncI1 (nâ=â2), IncFIA (nâ=â1), IncFIB (nâ=â1) and IncN (nâ=â1). ESBL-encoding genes, including blaCTX-M-14, blaCTX-M-15 and blaTEM-20, were identified. CONCLUSIONS: As the first report from pig sources in Ireland, characterization of these ESBL-encoding isolates and their transmissible plasmids extends our understanding on these resistance markers from porcine E. coli.
Assuntos
Infecções por Escherichia coli/veterinária , Escherichia coli/enzimologia , Suínos/microbiologia , beta-Lactamases/metabolismo , Animais , Análise por Conglomerados , Conjugação Genética , Microbiologia Ambiental , Escherichia coli/classificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Fazendas , Fezes/microbiologia , Variação Genética , Irlanda , Tipagem Molecular , Filogenia , Plasmídeos/análise , Análise de Sequência de DNARESUMO
Neonatal enteritis is a common condition of young calves and can be caused by pathogenic strains of Escherichia coli. We hypothesised that on-farm antimicrobial use would result in an increased frequency of resistance in these strains during the calving season. We also sought to determine if the frequency of resistance reflected on-farm antimicrobial use. Faecal samples were collected from cases of calf enteritis on 14 spring-calving dairy farms during two 3 week periods: Period 1 - February 11th through March 2nd 2008 and Period 2 - April 14th through May 5th 2008. E. coli were cultured from these samples, pathogenic strains were identified and antimicrobial susceptibility testing was carried out on these pathogenic isolates. Antimicrobial prescribing data were collected from each farm for the previous 12 months as an indicator of antimicrobial use. The correlation between antimicrobial use and resistance was assessed using Spearman's correlation coefficient. Logistic regression analysis was used to investigate the relationship between resistance, sampling period and pathotype. Penicillins and aminopenicillins, streptomycin, and tetracyclines were the most frequently prescribed antimicrobials and the greatest frequencies of resistance were detected to these 3 antimicrobial classes. A strong correlation (ρ=0.879) was observed between overall antimicrobial use and frequencies of antimicrobial resistance on farms. Sampling period was significant in the regression model for ampicillin resistance while pathotype was significant in the models for streptomycin, tetracycline and trimethoprim/sulphamethoxazole resistance. The frequencies of resistance observed have implications for veterinary therapeutics and prudent antimicrobial use. Resistance did not increase during the calving season and factors other than antimicrobial use, such as calf age and bacterial pathotype, may influence the occurrence of resistance in pathogenic E. coli.