Kang'ethe Erastus K, Mulinge Erastus K, Skilton Robert A, Njahira Moses, Monda Joseph G, Nyongesa Concepta, Mbae Cecilia K, Kamwati Stanley K
Department of Public Health, Pharmacology and Toxicology, University of Nairobi, PO Box 29053, Nairobi, Kenya.
Trop Anim Health Prod. 2012 Sep;44 Suppl 1:S25-31. doi: 10.1007/s11250-012-0202-5. Epub 2012 Jul 15.
A total of 1,734 cattle faecal samples from 296 dairy-keeping households were collected from urban settings in Nairobi, Kenya. Modified Ziehl-Neelsen staining method and an immunofluorescence assay were used to identify those samples with Cryptosporidium oocyst infection. Oocysts from positive faecal samples were isolated by Sheather's sucrose flotation method and picked from the concentrate using cover slips. Genomic DNA was extracted from 124 of the faecal samples that were positive for Cryptosporidium and was used as template for nested PCR of the 18S rRNA gene. Twenty-five samples (20 %) were PCR-positive for Cryptosporidium, and 24 of the PCR products were successfully cloned and sequenced. Sequence and phylogenetic analysis identified 17 samples (68 %) as Cryptosporidium parvum-like, four samples (16 %) as Cryptosporidium ryanae, three samples (12 %) as Cryptosporidium andersoni and one sample (4 %) as Cryptosporidium hominis. To the best of our knowledge, this is the first genotyping study to report C. parvum-like, C. andersoni and C. hominis in cattle from Kenya. The results of this study show Cryptosporidium infections in calves and cattle may be potential zoonotic reservoirs of the parasite that infects humans.
从肯尼亚内罗毕的城市地区296个奶牛养殖户中总共采集了1734份牛粪样本。采用改良齐-尼氏染色法和免疫荧光分析法鉴定隐孢子虫卵囊感染的样本。通过谢弗氏蔗糖漂浮法从阳性粪便样本中分离出卵囊,并用盖玻片从浓缩物中挑选出来。从124份隐孢子虫阳性粪便样本中提取基因组DNA,并用作18S rRNA基因巢式PCR的模板。25份样本(20%)隐孢子虫PCR检测呈阳性,其中24份PCR产物成功克隆并测序。序列和系统发育分析确定17份样本(68%)为微小隐孢子虫样,4份样本(16%)为瑞氏隐孢子虫,3份样本(12%)为安氏隐孢子虫,1份样本(4%)为人隐孢子虫。据我们所知,这是首次在肯尼亚牛群中对微小隐孢子虫样、安氏隐孢子虫和人隐孢子虫进行基因分型研究。本研究结果表明,犊牛和奶牛中的隐孢子虫感染可能是该寄生虫感染人类的潜在人畜共患病源。