Cerqueira A M, Guth B E, Joaquim R M, Andrade J R
Departamento de Microbiologia, Imunologia e Parasitologia, UNIFESP, São Paulo, Brazil.
Vet Microbiol. 1999 Oct;70(1-2):111-21. doi: 10.1016/s0378-1135(99)00123-6.
In order to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) strains, 197 fecal samples of healthy cattle from 10 dairy farms, four beef farms and one slaughterhouse at Rio de Janeiro State, Brazil, were examined for Shiga toxin (Stx) gene sequences by polymerase chain reaction (PCR). For presumptive isolation of O157:H7 E. coli, the Cefixime-potassium tellurite-sorbitol MacConkey Agar (CT-SMAC) was used. A high occurrence (71%) of Stx was detected, and was more frequently found among dairy cattle (82% vs. 53% in beef cattle), in which no differences were observed regarding the age of the animals. Dot blot hybridization with stx1 and stx2 probes revealed that the predominant STEC type was one that had the genes for both stx1 and stx2 in dairy cattle and one that had only the stx1 gene for beef cattle. Three (1.5%) O157:H7 E. coli strains were isolated from one beef and two dairy animals by the use of CT-SMAC. To our knowledge, this is the first report of O157:H7 isolation in Brazil. A PCR-based STEC detection protocol led to the isolation of STEC in 12 of 16 randomly selected PCR-positive stool samples. A total of 15 STEC strains belonging to 11 serotypes were isolated, and most of them (60%) had both stx1 and stx2 gene sequences. Cytotoxicity assays with HeLa and Vero cells revealed that all strains except two of serotype O157:H7 expressed Stx. The data point to the high prevalence of STEC in our environment and suggest the need for good control strategies for the prevention of contamination of animal products.
为了评估产志贺毒素大肠杆菌(STEC)菌株的流行情况,对来自巴西里约热内卢州10个奶牛场、4个肉牛场和1个屠宰场的197份健康牛粪便样本进行了聚合酶链反应(PCR)检测,以确定志贺毒素(Stx)基因序列。为了初步分离O157:H7大肠杆菌,使用了头孢克肟-亚碲酸钾-山梨醇麦康凯琼脂(CT-SMAC)。检测发现Stx的发生率很高(71%),且在奶牛中更为常见(82%,肉牛中为53%),不同年龄的动物之间未观察到差异。用stx1和stx2探针进行的斑点杂交显示,奶牛中主要的STEC类型是同时具有stx1和stx2基因的菌株,而肉牛中主要是仅具有stx1基因的菌株。通过使用CT-SMAC,从1头肉牛和2头奶牛中分离出3株(1.5%)O157:H7大肠杆菌。据我们所知,这是巴西首次报告分离出O157:H7。基于PCR的STEC检测方案在16份随机选择的PCR阳性粪便样本中的12份中分离出了STEC。共分离出15株属于11种血清型的STEC菌株;其中大多数(60%)同时具有stx1和stx2基因序列。用HeLa细胞和Vero细胞进行的细胞毒性试验表明,除两株O157:H7血清型菌株外,所有菌株均表达Stx。这些数据表明我们环境中STEC的流行率很高,并表明需要采取良好的控制策略来预防动物产品的污染。