Chen F, Huang K
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
Zoonoses Public Health. 2007;54(9-10):393-400. doi: 10.1111/j.1863-2378.2007.01078.x.
Faecal samples from 1488 pigs were collected from six pig farms in Jiangsu province and Shanghai city, eastern China, stained by the modified acid-fast staining technique (MAFS) and observed by light and phase contrast microscopy to identify Cryptosporidium oocysts. A total of 178 (12%) positive animals were detected. Three randomly selected MAFS-positive samples from each farm were further analysed by reverse-transcription polymerase chain reaction (RT-PCR) using primers designed to amplify DNA fragments from the small subunit ribosomal RNA (SSU rRNA) gene. PCR products from two positive samples were sequenced. Phylogenetic analysis and a distances matrix generated from these sequences showed that the isolates belonged to the species C. parvum, and were most similar to the C. parvum'mouse' genotype. The identification of this C. parvum genotype in pigs has not been reported previously. These findings are discussed in the context of the epidemiology of Cryptosporidium on pig farms.
从中国东部江苏省和上海市的六个养猪场收集了1488份猪粪便样本，采用改良抗酸染色技术（MAFS）进行染色，并通过光学显微镜和相差显微镜观察以鉴定隐孢子虫卵囊。共检测到178头（12%）阳性动物。从每个猪场随机选取3份MAFS阳性样本，使用设计用于从小亚基核糖体RNA（SSU rRNA）基因扩增DNA片段的引物，通过逆转录聚合酶链反应（RT-PCR）进一步分析。对两份阳性样本的PCR产物进行测序。系统发育分析以及由这些序列生成的距离矩阵表明，分离株属于微小隐孢子虫物种，并且与微小隐孢子虫“小鼠”基因型最为相似。此前尚未报道在猪中鉴定出这种微小隐孢子虫基因型。在养猪场隐孢子虫病流行病学的背景下对这些发现进行了讨论。
