Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, Banat University of Agricultural Sciences and Veterinary Medicine Timişoara, Calea Aradului nr. 119, 300645 Timişoara, Romania. kalman
Vet Parasitol. 2011 Sep 27;181(2-4):321-4. doi: 10.1016/j.vetpar.2011.04.042. Epub 2011 May 6.
Diarrheic fecal samples from 258 pre-weaned calves (1-30 day-old) from 9 dairy farms located in Banat region, Romania, were microscopically examined for the presence of Cryptosporidium oocysts. Overall, 65 (25%) samples were found positive. A higher percent of infection was recorded in calves aged between 8 and 14 days compared with other age categories (1-7, 8-14, 15-21 and 22-30 days; p<0.05). Genetic characterization was carried out on all Cryptosporidium-positive samples. After DNA extraction, Cryptosporidium species were determined by a nested PCR of the small subunit rRNA gene (18S) followed by RFLP analysis with SspI, VspI and MboII restriction enzymes. The restriction patterns showed that animals were infected with Cryptosporidium parvum. Subsequently, subtyping of 13 C. parvum isolates, based on sequence analysis of the 60 kDa glycoprotein (GP60) gene, showed 2 subtypes (IIaA15G2R1 and IIaA16G1R1) belonging to the subtype family IIa. This is the first molecular study of bovine Cryptosporidium infection in Romania.
从罗马尼亚巴纳特地区 9 个奶牛场的 258 头 1-30 日龄未断奶小牛的腹泻粪便样本中，通过显微镜检查是否存在隐孢子虫卵囊。总体而言，有 65（25%）个样本呈阳性。与其他年龄段（1-7 天、8-14 天、15-21 天和 22-30 天）相比，8-14 天龄的小牛感染率更高（p<0.05）。对所有隐孢子虫阳性样本进行了遗传特征分析。在提取 DNA 后，通过 18S 核糖体 RNA 基因（18S）的嵌套 PCR 确定隐孢子虫种类，然后用 SspI、VspI 和 MboII 限制性内切酶进行 RFLP 分析。限制图谱显示动物感染了微小隐孢子虫。随后，基于 60 kDa 糖蛋白（GP60）基因的序列分析，对 13 株微小隐孢子虫分离株进行了亚分型，显示属于 IIa 亚家族的 2 种亚型（IIaA15G2R1 和 IIaA16G1R1）。这是罗马尼亚首次对牛隐孢子虫感染进行的分子研究。
