Molecular Characterization of Bubaline Isolate of Cryptosporidium Species from Egypt

Khaled A. Abdelrahma, K. N. Megeed, A. Hammam, G. Morsy, Moshera M.E. Seliem, Dina Aboelsoued
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引用次数: 6

Abstract

Cryptosporidium an apicomplexan parasite has the ability to induce diarrhea in bovines, goats, pigs, dogs and cats worldwide. In this study, buffalo calves fecal samples were examined after staining their smears with Modified Ziehl-Neelsen Stain (MZN). Ileal sections were examined for the detection of pathological changes. Further molecular characterization was done using nested PCR amplification and partial sequence analysis. The detected oocysts were morphologically similar to Cryptosporidium parvum. Light microscopic examination of Cryptosporidium infected ileal Tissue Section (TS) stained with H and E revealed the presence of altered mucosal architecture with congestion of blood vessels, infiltration, sloughing and complete erosion of epithelial cells and shortening, blunting, stunting and atrophy of the intestinal villi. Molecular characterization gave PCR amplicons of 18S SSU rRNA gene products approximately at 823 bp. Sequences proved specified generalized relatedness with 21 species of Cryptosporidium but the nucleotide homogeneity percentage was insufficient to designate species or genotypes. Further bioinformatics analysis showed that resulting Cryptosporidium isolates had the closest match with three isolates. It was implied that the Cryptosporidium isolates is mostly like Cryptosporidium parvum (JX237832.1) previously isolated from buffaloes in Ismailia province.
埃及隐孢子虫Bubaline分离物的分子特征
隐孢子虫是一种顶端复合体寄生虫,能够在全世界的牛、山羊、猪、狗和猫中引起腹泻。本研究采用改良Ziehl-Neelsen染色法(MZN)对水牛粪便进行涂片染色。检查回肠切片,观察病理变化。进一步的分子鉴定采用巢式PCR扩增和部分序列分析。所检卵囊形态与细小隐孢子虫相似。隐孢子虫感染回肠组织切片(TS)的光镜下H和E染色显示粘膜结构改变,血管充血,上皮细胞浸润、脱落和完全糜烂,肠绒毛缩短、变钝、发育迟缓和萎缩。分子鉴定获得了18S SSU rRNA基因产物约823 bp的PCR扩增产物。序列证实与21种隐孢子虫具有特定的广义亲缘关系,但核苷酸同源性百分比不足以确定种或基因型。进一步的生物信息学分析表明,得到的隐孢子虫分离株与三个分离株最接近。该隐孢子虫分离株与先前从伊斯梅里亚省水牛中分离到的小隐孢子虫(JX237832.1)相似。
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