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Identification of virulence factors by multiplex PCR in Escherichia coli isolated from calves in Minas Gerais, Brazil

机译:通过多重PCR在巴西米纳斯吉拉斯州牛犊分离的大肠杆菌中鉴定毒力因子

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In this study, multiplex PCR was employed to investigate the virulence factors of Escherichia coli strains isolated from 60-day-old calves. Faecal samples were collected from 54 calves at 12 dairy farms in the state of Minas Gerais, Brazil. A total of 156 isolates were obtained after culture and microbiological isolation and were tested by multiplex PCR for the presence of genes encoding toxins (Stx1, Stx2 and STa) and adherence factors (intimin, F41 and F5). Seventy of 156 isolates were positive for at least one virulence factor: ten (14.3 %) from diarrhoeic animals and 60 (85.7 %) from healthy calves. The virulence markers identified were: Stx1 (82.8 %), eae (24.3 %), F41 (11.4 %), F5 (10 %), STa (4.28 %) and Stx2 (4 %). In diarrhoeic animals, Stx1 (70 %) and F41 (30 %) were identified, while Stx1 (83.3 %), eae (28.3 %), F41 (8.3 %), F5 (11.6 %), STa (5 %) and Stx2 (1.6 %) were detected in isolates from healthy calves. Mixed infections with pathotypes Shiga toxin-producing E. coli (STEC)/enteropathogenic E. coli, STEC/enterohaemorrhagic E. coli and STEC/other (eae/F5, Stx1/STa) were detected in five healthy calves. Pathogenic E. coli were identified in 59.26 % of all calves and on 75 % of the dairy farms studied, not only in diarrhoeic (five of six) but also in healthy calves (27 of 48), which demonstrates the importance of this agent in the aetiology of diarrhoea in calves in the state of Minas Gerais.
机译:在这项研究中,采用多重PCR来研究从60天龄犊牛分离出的大肠杆菌菌株的毒力因子。从巴西米纳斯吉拉斯州12个奶牛场的54头犊牛收集了粪便样品。培养和微生物分离后,共获得156个分离株,并通过多重PCR检测编码毒素(Stx1,Stx2和STa)和粘附因子(内膜素,F41和F5)的基因的存在。 156株分离物中的70株至少有一种毒力因子呈阳性:腹泻动物有10种(14.3%),健康犊牛有60种(85.7%)。鉴定出的毒力标记为:Stx1(82.8%),eae(24.3%),F41(11.4%),F5(10%),STa(4.28%)和Stx2(4%)。在腹泻动物中,鉴定出Stx1(70%)和F41(30%),而Stx1(83.3%),eae(28.3%),F41(8.3%),F5(11.6%),STa(5%)和Stx2在健康小牛的分离物中检出(1.6%)。在五只健康的牛犊中检出了致病性产志贺毒素大肠杆菌(STEC)/致病性大肠杆菌,STEC /肠出血性大肠杆菌和STEC /其他(eae / F5,Stx1 / STa)混合型感染。在所有研究的犊牛中,有59.26%的奶牛和75%的奶牛场中都鉴定出了致病性大肠杆菌,不仅在腹泻(五分之六)中,而且在健康的犊牛中(48个中的27个)都得到了证实,这证明了这种病原在大肠杆菌中的重要性。米纳斯吉莱斯州小牛腹泻的病因。

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