Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria, Australia.
Electrophoresis. 2013 Aug;34(15):2259-67. doi: 10.1002/elps.201300146.
In the present study, we undertook a molecular epidemiological survey of Cryptosporidium and Giardia in calves on three dairy and two beef farms within an open drinking water catchment area (Melbourne, Australia). Faecal samples (n = 474) were collected from calves at two time points (5 months apart) and tested using a PCR-based mutation scanning-targeted sequencing phylogenetic approach, employing regions within the genes of small subunit (SSU) of ribosomal RNA (designated partial SSU), 60 kDa glycoprotein (pgp60) and triose phosphate isomerase (ptpi) as genetic markers. Using partial SSU, the C. bovis, C. parvum, C. ryanae and a new genotype of Cryptosporidium were characterised from totals of 74 (15.6%), 35 (7.3%), 37 (7.8%) and 9 (1.9%) samples, respectively. Using pgp60, C. parvum genotype IIa subgenotype A18G3R1 was detected in 29 samples. Using ptpi, G. duodenalis assemblages A and E were detected in totals of 10 (2.1%) and 130 (27.4%) samples, respectively. The present study showed that a considerable proportion of dairy and beef calves in this open water catchment region excreted Cryptosporidium (i.e. subgenotype IIaA18G3R1) and Giardia (e.g. assemblage A) that are consistent with those infecting humans, inferring that they are of zoonotic importance. Future work should focus on exploring, in a temporal and spatial way, whether these parasites occur in the environment and water of the catchment reservoir.
在本研究中,我们对开放饮用水集水区(澳大利亚墨尔本)内的三个奶牛场和两个肉牛场的犊牛进行了隐孢子虫和贾第鞭毛虫的分子流行病学调查。从犊牛采集粪便样本(n = 474),在两次时间点(相隔 5 个月)采集,并使用基于 PCR 的突变扫描靶向测序系统发育方法进行检测,该方法采用核糖体 RNA 小亚基(SSU)内的基因、60 kDa 糖蛋白(pgp60)和磷酸丙糖异构酶(ptpi)作为遗传标记。使用部分 SSU,从总共 74 份(15.6%)、35 份(7.3%)、37 份(7.8%)和 9 份(1.9%)样本中分别鉴定出 C. bovis、C. parvum、C. ryanae 和一种新型隐孢子虫基因型。使用 pgp60 检测到 29 份样本中的 C. parvum 基因型 IIa 亚基因型 A18G3R1。使用 ptpi,共检测到 G. duodenalis 聚集 A 和 E 分别为 10 份(2.1%)和 130 份(27.4%)。本研究表明,该开放水域集水区的相当一部分奶牛和肉牛犊牛排泄了隐孢子虫(即亚基因型 IIaA18G3R1)和贾第鞭毛虫(如聚集 A),与感染人类的隐孢子虫和贾第鞭毛虫一致,表明它们具有动物源性。未来的工作应集中于以时间和空间的方式探索这些寄生虫是否存在于集水区水库的环境和水中。