Taha Shahinaz, Elmalik Khitma, Bangoura Berit, Lendner Matthias, Mossaad Ehab, Daugschies Arwid
Department of Preventive Medicine and Veterinary Public Health, Faculty of Veterinary Medicine, University of Khartoum, P.O. Box 32, Khartoum North, Sudan.
Institute of Parasitology, Leipzig University, An den Tierkliniken 35, 04103, Leipzig, Germany.
Parasitol Res. 2017 Nov;116(11):2971-2979. doi: 10.1007/s00436-017-5606-8. Epub 2017 Sep 13.
Cryptosporidiosis is a common protozoan infection causing morbidity and mortality in young cattle and may be zoonotically transmitted to humans. So far, there is no data available on the presence of Cryptosporidium spp. in the Sudan. The aim of this study was to isolate, identify, and genotype Cryptosporidium oocysts sampled from diarrheic calves housed at different farms in three different municipalities in Khartoum State (Khartoum, Khartoum North, Omdurman). A total of 149 fecal samples were evaluated microscopically for the presence of Cryptosporidium oocysts using the modified Ziehl-Neelsen staining method and 87 (58.3%) samples tested positive. Positive and negative samples were further analyzed by nested PCR targeting the SSU rRNA region. Positive samples were subjected to restriction enzyme analysis of PCR amplicons (PCR-RFLP). Nested PCR identified Cryptosporidium DNA in 53 samples (35.5%); restriction digestion of the PCR products revealed the presence of C. parvum (73.5%), C. ryanae (13.2%), C. andersoni (7.5%), and C. bovis (1.8%). Species distribution was clearly related to age with C. parvum being the predominant species in dysenteric pre-weaned calves. Sequencing of three genes (SSU rRNA, COWP, and GP60) for three C. parvum isolates originating from the three different municipalities showed that all belong to C. parvum subtype family IId. Based on data obtained by GP60, sequencing the two C. parvum isolates from Khartoum and Omdurman represent subtype IIdA18G1, whereas oocysts isolated in Khartoum North belong to subtype IIdA19G1. The observed genotypes are zoonotic and thus C. parvum in calves is potentially a health risk to humans in Khartoum State, Sudan. To the best of our knowledge, this is the first reported attempt to characterize Cryptosporidium isolated from cattle in the Sudan.
隐孢子虫病是一种常见的原生动物感染病，可导致幼牛发病和死亡，并且可能人畜共患传播给人类。到目前为止，苏丹尚无关于隐孢子虫属存在情况的数据。本研究的目的是对从喀土穆州三个不同城市（喀土穆、北喀土穆、恩图曼）不同农场腹泻犊牛采集的隐孢子虫卵囊进行分离、鉴定和基因分型。使用改良的齐尔-尼尔森染色法对总共149份粪便样本进行显微镜检查，以检测隐孢子虫卵囊的存在，87份（58.3%）样本检测呈阳性。对阳性和阴性样本进一步通过靶向小亚基核糖体RNA（SSU rRNA）区域的巢式聚合酶链反应（PCR）进行分析。对阳性样本进行PCR扩增产物的限制性内切酶分析（PCR-RFLP）。巢式PCR在53份样本（35.5%）中鉴定出隐孢子虫DNA；PCR产物的限制性消化显示存在微小隐孢子虫（73.5%）、雷氏隐孢子虫（13.2%）、安氏隐孢子虫（7.5%）和牛隐孢子虫（1.8%）。物种分布与年龄明显相关，微小隐孢子虫是腹泻的断奶前犊牛中的主要物种。对来自三个不同城市的三株微小隐孢子虫分离株的三个基因（SSU rRNA、COWP和GP60）进行测序表明，它们均属于微小隐孢子虫亚型家族IId。根据GP60获得的数据，对来自喀土穆和恩图曼的两株微小隐孢子虫分离株进行测序代表亚型IIdA18G1，而在北喀土穆分离的卵囊属于亚型IIdA19G1。观察到的基因型是人畜共患的，因此犊牛中的微小隐孢子虫对苏丹喀土穆州的人类有潜在健康风险。据我们所知，这是首次报道对从苏丹牛中分离的隐孢子虫进行特征描述的尝试。
