Vu Khac Hung, Holoda Emil, Pilipcinec Emil, Blanco Miguel, Blanco Jesús E, Mora Azucena, Dahbi Ghizlane, López Cecilia, González Enrique A, Blanco Jorge
Institute of Microbiology and Immunology, Department of Food Hygiene and Technology, University of Veterinary Medicine, Komenskeho 73, Slovakia.
BMC Vet Res. 2006 Mar 20;2:10. doi: 10.1186/1746-6148-2-10.
Postweaning diarrhoea (PWD) in pigs is usually the main infectious problem of large-scale farms and is responsible for significant losses worldwide. The disease is caused mainly by enterotoxigenic E. coli (ETEC) and Shiga-toxin producing E. coli (STEC). In this study a total of 101 E. coli isolated from pigs with PWD in Slovakia were characterized using phenotypic and genotypic methods.
These 101 isolates belonged to 40 O:H serotypes. However, 57% of the isolates belonged to only six serotypes (O9:H51, O147:H-, O149:H10, O163:H-, ONT:H-, and ONT:H4), including two new serotypes (O163:H- and ONT:H4) not previously found among porcine ETEC and STEC isolated in other countries. Genes for EAST1, STb, STa, LT and Stx2e toxins were identified in 64%, 46%, 26%, 20%, and 5% of isolates, respectively. PCR showed that 35% of isolates carried genes for F18 colonization factor, and further analyzed by restriction endonuclease revealed that all of them were F18ac. Genes for F4 (K88), F6 (P987), F17, F5 (K99), F41, and intimin (eae gene) adhesins were detected in 19 %, 5%, 3%, 0.9%, 0.9%, and 0.9% of the isolates, respectively. The study of genetic diversity, carried out by PFGE of 46 representative ETEC and STEC isolates, revealed 36 distinct restriction profiles clustered in eight groups. Isolates of the same serotype were placed together in the dendrogram, but high degree of polymorphism among certain serotypes was detected.
Seropathotype O149:H10 LT/STb/EAST1/F4 (14 isolates) was the most commonly detected followed by O163:H- EAST1/F18 (six isolates), and ONT:H4 STa/STb/Stx2e/F18 (five isolates). Interestingly, this study shows that two new serotypes (O163:H- and ONT:H4) have emerged as pig pathogens in Slovakia. Furthermore, our results show that there is a high genetic variation mainly among ETEC of O149:H10 serotype.
仔猪断奶后腹泻(PWD)通常是大型养猪场的主要感染性问题,在全球范围内造成重大损失。该疾病主要由产肠毒素大肠杆菌(ETEC)和产志贺毒素大肠杆菌(STEC)引起。在本研究中,使用表型和基因型方法对从斯洛伐克患有PWD的猪中分离出的总共101株大肠杆菌进行了鉴定。
这101株分离株属于40种O:H血清型。然而,57%的分离株仅属于六种血清型(O9:H51、O147:H-、O149:H10、O163:H-、ONT:H-和ONT:H4),包括两种在其他国家分离的猪ETEC和STEC中未发现的新血清型(O163:H-和ONT:H4)。分别在64%、46%、26%、20%和5%的分离株中鉴定出EAST1、STb、STa、LT和Stx2e毒素的基因。PCR显示35%的分离株携带F18定植因子基因,通过限制性内切酶进一步分析表明它们均为F18ac。分别在19%、5%、3%、0.9%、0.9%和0.9%的分离株中检测到F4(K88)、F6(P987)、F17、F5(K99)、F41和紧密黏附素(eae基因)黏附素的基因。对46株代表性ETEC和STEC分离株进行PFGE的遗传多样性研究,揭示了36种不同的限制性图谱,分为八组。同一血清型的分离株在树状图中聚在一起,但在某些血清型中检测到高度的多态性。
血清致病型O149:H10 LT/STb/EAST1/F4(14株)是最常检测到的,其次是O163:H- EAST1/F18(6株)和ONT:H4 STa/STb/Stx2e/F18(5株)。有趣的是,本研究表明两种新血清型(O163:H-和ONT:H4)已在斯洛伐克成为猪病原体。此外,我们的结果表明主要在O149:H10血清型的ETEC之间存在高度的遗传变异。