The prevalence of Cryptosporidium species in calves and heifers with relation to diarrhea from several herds was investigated in this study. Fecal samples were collected from 135 and 120 pre-weaned calves and 79 and 130 heifers raised in the Central Anatolia (CAR) and Mediterranean Regions (MR) of Turkey, respectively. A total of 86 post-weaned calves in CAR were also included in the study. For diagnostic comparison, all samples were examined by microscopic examination, SSU rRNA nested PCR and TaqMan real-time PCR for the presence of oocyst and Cryptosporidium DNA. In total, 102 (34.0 %) and 93 (37.2 %) of the examined samples from CAR and MR were found positive for Cryptosporidium DNA with both nested PCR and real-time PCR analyses, respectively with an overall prevalence of 35.5 %. The diagnostic sensitivity and specificity of microscopic examination were determined as 68.7 % and 100.0 % compared to molecular tools, respectively. RFLP and sequence analyses of the SSU rRNA from the PCR products revealed that 138 (70.8 %) out of 195 positive isolates were C. parvum further confirming the species-specific real-time PCR results. Among the remaining 57 (29.2 %) positive isolates, 30 (15.4 %) and 27 (13.8 %) were characterized as C. ryanae and C. bovis, respectively. C. parvum was the dominant species in pre-weaned calves especially with diarrhea while C. bovis and C. ryanae were mostly found in post-weaned calves and heifers. The sequence analyses of the gp60 gene of C. parvum isolates revealed two subtypes (IIaA13G2R1, IIaA14G1R1) belonging to zoonotic family IIa, with IIaA13G2R1 being the most common in diarrheic calves.

中文翻译：

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土耳其地中海和安纳托利亚中部地区牛隐孢子虫种类的流行和基因分型。

在这项研究中，研究了犊牛和小母牛中隐孢子虫种类的流行与腹泻的关系。分别从土耳其中部安纳托利亚（CAR）和地中海地区（MR）饲养的135和120头断奶小牛以及79和130头小母牛收集粪便样品。该研究中还包括了总共86只断奶后的犊牛。为了进行诊断比较，所有样品均通过显微镜检查，SSU rRNA巢式PCR和TaqMan实时PCR检查是否存在卵囊和隐孢子虫DNA。通过巢式PCR和实时PCR分析，从CAR和MR总共检查出102个（34.0％）和93个（37.2％）隐孢子虫DNA阳性，总体患病率为35.5％。与分子工具相比，显微镜检查的诊断敏感性和特异性分别确定为68.7％和100.0％。RFLP和来自PCR产物的SSU rRNA的序列分析表明，在195株阳性分离株中，有138株（占70.8％）是小球隐孢菌，进一步证实了物种特异性实时PCR结果。在其余的57个（29.2％）阳性分离株中，分别有30个（15.4％）和27个（13.8％）的特征为黑穗假丝酵母和牛分枝杆菌。在断奶前的犊牛中，小腿念珠菌是主要菌种，尤其是腹泻，而在断奶后的犊牛和小母牛中主要发现牛毛弯曲杆菌和黑麦隐孢子虫。细小衣原体分离株gp60基因的序列分析揭示了人畜共患病家族IIa的两个亚型（IIaA13G2R1，IIaA14G1R1），其中IIaA13G2R1在腹泻犊牛中最常见。