Inglis G D, Kalischuk L D, Busz H W
Agriculture and Agri-Food Canada Research Centre, Lethbridge, AB, Canada.
Can J Microbiol. 2003 Nov;49(11):655-61. doi: 10.1139/w03-087.
A polymerase chain reaction (PCR)-based survey of campylobacters associated with faeces collected from 382 beef cattle was undertaken. To ensure the removal of PCR inhibitors present in faeces and determine if adequate extraction was achieved, faeces were seeded with internal control DNA (i.e., DNA designed to amplify with the Campylobacter genus primer set, but provide a smaller amplicon) before the extraction procedure. In only two samples (0.5%) were the internal control or Campylobacter genus amplicons not detected. In the remaining 380 faecal samples, Campylobacter DNA was detected in 83% of the faecal samples (80% of the faecal samples were positive for Campylobacter genus DNA, and 3% of the samples were negative for Campylobacter genus DNA but positive for DNA of individual species). The most frequently detected species was Campylobacter lanienae (49%), a species only recently connected to livestock hosts. Campylobacter jejuni DNA was detected in 38% of the faecal samples, and Campylobacter hyointestinalis and Campylobacter coli DNA were detected in 8% and 0.5% of the samples, respectively. Campylobacter fetus DNA was not detected. Twenty-four percent of the faecal samples contained DNA of at least two species of Campylobacter. Of these samples, the majority (81%) contained DNA of C. jejuni and C. lanienae. The results of this study indicate that beef cattle commonly release a variety of Campylobacter species into the environment and may contribute to the high prevalence of campylobacteriosis in humans inhabiting areas of intensive cattle production, such as southern Alberta. Furthermore, this study demonstrates the utility of using PCR as a rapid and accurate method for simultaneously detecting the DNA of a diverse number of Campylobacter species associated with bovine faeces.
对382头肉牛粪便中弯曲杆菌进行了基于聚合酶链反应(PCR)的调查。为确保去除粪便中存在的PCR抑制剂并确定是否实现了充分提取,在提取程序之前,向粪便中接种了内部对照DNA(即设计用于与弯曲杆菌属引物组一起扩增,但产生较小扩增子的DNA)。仅在两个样本(0.5%)中未检测到内部对照或弯曲杆菌属扩增子。在其余380份粪便样本中,83%的粪便样本检测到弯曲杆菌DNA(80%的粪便样本弯曲杆菌属DNA呈阳性,3%的样本弯曲杆菌属DNA呈阴性,但个别物种的DNA呈阳性)。最常检测到的物种是拉内弯曲杆菌(49%),这是一种最近才与家畜宿主相关的物种。空肠弯曲杆菌DNA在38%的粪便样本中被检测到,猪肠弯曲杆菌和结肠弯曲杆菌DNA分别在8%和0.5%的样本中被检测到。胎儿弯曲杆菌DNA未被检测到。24%的粪便样本含有至少两种弯曲杆菌的DNA。在这些样本中,大多数(81%)含有空肠弯曲杆菌和拉内弯曲杆菌的DNA。本研究结果表明,肉牛通常会向环境中释放多种弯曲杆菌物种,这可能导致居住在艾伯塔省南部等肉牛密集养殖地区的人类弯曲杆菌病高发率。此外,本研究证明了使用PCR作为一种快速准确的方法,可同时检测与牛粪相关的多种弯曲杆菌物种的DNA的实用性。