College of Animal Science and Veterinary Medicine, Henan Agricultural University, 15, University Park of Longzihu, Zhengzhou 450046, PR China.
College of Information and Management Science, Henan Agricultural University, 15, University Park of Longzihu, Zhengzhou 450046, PR China.
Infect Genet Evol. 2018 Oct;64:46-51. doi: 10.1016/j.meegid.2018.06.012. Epub 2018 Jun 9.
Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are globally ubiquitous pathogens. However, little is known about the occurrence and distribution of Cryptosporidium spp., G. duodenalis, and E. bieneusi in Tibetan sheep. In the present study, fecal specimens of 177 Tibetan sheep were examined by nested PCR. 4.5% (n = 8), 1.7% (n = 3) and 34.5% (n = 61) of the Tibetan sheep were positive for Cryptosporidium spp., G. duodenalis, and E. bieneusi, respectively. Cryptosporidium ubiquitum was the only species found by small subunit (SSU) rRNA-based PCR, and subtyping of C. ubiquitum (n = 6) by 60-kDa glycoprotein (gp60) revealed that all positive isolates belonged to zoonotic XIIa subtype 2. Multilocus genotyping at the SSU rRNA and β-giardin (bg) genes suggested that three G. duodenalis positive specimens belonged to assemblage E. Sequences analysis of internal transcribed spacer (ITS) gene characterized four E. bieneusi genotypes which all belonged to Group 2, one novel CGS1 (n = 3), and three known: CM7 (n = 34), BEB6 (n = 22), and CHS3 (n = 2). We employed multilocus sequence typing (MLST) using three microsatellites (MS1, MS3 and MS7), one minisatellite (MS4), and sequence analysis of MLST yielded 3, 2, 2 and 2 genotypes at the MS1, MS3, MS4, and MS7 loci, respectively, forming 4 MLGs. Our findings elucidate the occurrence and distribution of three zoonotic pathogens in Tibetan sheep in China. More subsequent and detailed data are required to better understand the transmission of Cryptosporidium spp., G. duodenalis, and E. bieneusi in sheep.
隐孢子虫、十二指肠贾第鞭毛虫和肠微孢子虫是全球普遍存在的病原体。然而，对于藏绵羊中隐孢子虫、十二指肠贾第鞭毛虫和肠微孢子虫的发生和分布知之甚少。本研究采用巢式 PCR 检测了 177 份藏绵羊粪便标本。4.5%（n=8）、1.7%（n=3）和 34.5%（n=61）的藏绵羊分别对隐孢子虫、十二指肠贾第鞭毛虫和肠微孢子虫呈阳性。基于小亚基（SSU）rRNA 的 PCR 仅发现了微小隐孢子虫，对 60kDa 糖蛋白（gp60）的微小隐孢子虫（n=6）进行亚分型显示所有阳性分离株均属于人畜共患的 XIIa 亚型 2。SSU rRNA 和 β-微管蛋白（bg）基因的多位点基因分型表明，3 个十二指肠贾第鞭毛虫阳性标本属于 E 组。内部转录间隔区（ITS）基因序列分析鉴定出 4 种肠微孢子虫基因型，均属于第 2 组，其中 1 种新型 CGS1（n=3）和 3 种已知基因型：CM7（n=34）、BEB6（n=22）和 CHS3（n=2）。我们采用三个微卫星（MS1、MS3 和 MS7）、一个微卫星（MS4）和 MLST 的序列分析，在 MS1、MS3、MS4 和 MS7 基因座上分别获得了 3、2、2 和 2 种基因型，形成了 4 种 MLG。本研究阐明了中国藏绵羊中三种人畜共患病原体的发生和分布。需要更多后续和详细的数据来更好地了解绵羊中隐孢子虫、十二指肠贾第鞭毛虫和肠微孢子虫的传播。
