Preliminary Study on Phenotypic Detection of ESBL and Amp-C β-Lactamase-Producing Escherichia Coli Isolates from Slaughtered Pigs

Marinela Tipișcă, A. Cozma, D. Anita, E. Işan, L. Oslobanu, A. Aniță, G. Savuta
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Abstract

The presence of extended-spectrum beta-lactamase-producing Escherichia coli in food animals is a public health concern. The aim of this study was to phenotypically detect the extended β-lactamase and AmpC β-lactamase-producing E. coli isolated from pig caecum from three slaughterhouses in the North-East of Romania. After collecting the cecal samples, they were phenotypically processed. ESBL and AmpC screening was carried out by cultivation on MacConkey medium with addition of cefotaxime (MC+CTX). The isolates were confirmed as being E. coli based on the biochemical properties using MIU, TSI and API 20E and as ESBL based on synergy tests between clavulanic acid and ceftazidime and/or clavulanic acid and cefotaxime. Phenotypical detection of AmpC beta-lactamase-producing strains was carried out through a stable AmpC cephalosporin (cefepime). Following ESBL/AmpC screening, out of the 128 analysed samples, 51 (39.84%) grew on MC+CTX and they were identified as E. coli. After performing microdilution in broth using EUVSEC2 plates, 78.43% isolates were identified with ESBL phenotype, 9.8% isolates with ESBL/AmpC phenotype, and 11.77% isolates with AmpC phenotype. In this study, although a limited number of strains were analysed, the obtained results phenotypically confirm the presence of ESBL and AmpC enzymes in slaughtered pig populations from the North-East of Romania.
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屠宰猪ESBL和产Amp-C β-内酰胺酶大肠杆菌分离株表型检测的初步研究
食用动物中产生广谱β -内酰胺酶的大肠杆菌的存在是一个公共卫生问题。本研究的目的是表型检测从罗马尼亚东北部三个屠宰场的猪盲肠中分离出的扩展β-内酰胺酶和产生AmpC β-内酰胺酶的大肠杆菌。收集盲肠样本后,对其进行表型处理。在mcconkey培养基上添加头孢噻肟(MC+CTX)进行ESBL和AmpC筛选。根据MIU、TSI和API 20E的生化特性鉴定,分离株为大肠杆菌;根据克拉维酸与头孢他啶和/或克拉维酸与头孢噻肟的协同作用鉴定,分离株为ESBL。采用稳定的AmpC头孢菌素(头孢吡肟)对产AmpC β -内酰胺酶菌株进行表型检测。通过ESBL/AmpC筛选,在128个分析样本中,51个(39.84%)在MC+CTX上生长,鉴定为大肠杆菌。用EUVSEC2板在肉汤中微量稀释后,78.43%分离株为ESBL表型,9.8%分离株为ESBL/AmpC表型,11.77%分离株为AmpC表型。在这项研究中,虽然分析了有限数量的菌株,但获得的结果在表型上证实了罗马尼亚东北部屠宰猪群中存在ESBL和AmpC酶。
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