Investigation Of Cryptopsoridum Infection Of Yak And Crab-eating Meacaques And The Identification Of Molecular Characteristics

With the development of society, the contact of human and animal directly or indirectly, more and more. People pay more attention to about the Cryptosporidium infection of wild animals. However, reports on the yak and non-human primates infect Cryptosporidium is limited at home and abroad. Based on the special relationship between human, the author made an detailed investigation of Cryptosporidium infection in wild animals in China. In the result, we get 60 Cryptosporidium isolates of yak and 24 Cryptosporidium isolates of Crab-eating macaques. Specific fragments of 18S rRNA gene and GP60 gene were amplified by nested PCR respectively, and sequenced. The certain types of wildlife animal isolates of Cryptosporidium apecies and genotypes were identified. Phylogenetic relationship of certain wild animals'Cryptosporidium and other Cryptosporidium species is determined. This study provided a rationale for understand the Cryptosporidium relationship with non-human primates and other mammal, and provided a reference for evaluate human and non-human primates close activity security.In order to learn about the infection of yak parasite, We used two methods of modified acid fast staining and Sheater'sugar flotation techniques to detect 456 specemens of yak from Qinghai. In the result with the method of Sheater'sugar flotation techniques,286 fecal samples were positive, with the average infection rate of coccidian62.7%(286/456). The infecting rate of 1-5 moon'day is 53.6%(15/28),46.2%(24/52),62.0%(98/158).66.0%(68/103),70.4%(81/115); 60 fecal samples were positive use the method of modified acid fast staining, with the average infection rate of Cryptosporidium cycst 13.2%(60/456), the infecting rate of 1-5 moon'day is27.8% (13/28),9.1%(12/52),10.1%(16/158),13.6%(14/103),4.3%(5/115) respectively. The investigation result suggest that the Cryptosporidium infection of yak deffer with defferent olds. The coccidian infection rate is fairly high, and to be worth to pay attention to.In order to learn about the infection of Cryptosporidium in yak, Specific fragments of 18S rRNA gene was amplified by nested PCR-RFLP, and sequenced. two isolates were idenfied as Cryptosporidium ryane (FJ463193), homology of 100%。This is the first time that Cryptosporidium ryane be discovered in yak.We used two methods of modified acid fast staining and Sheater'sugar flotation techniques to detect 245 specemens of Crab-eating macaques from Nanning zoon. in the result, it was found that 24 fecal samples were positive, with the average infection rate of Cryptosporidium cycst 9.8% (24/245). The Small-Subunit rRNA(SSU rRNA) specific fragments of 4 Crab-eating macaques Cryptosporidium isolates and GP60 specific fragments of 13 Crab-eating macaques Cryptosporidium isolates were amplified by nested PCR. The PCR products were connection and transformation, cloned and sequenced. Then, Blast or Fasta methods were used to search homological sequences in NCBI, DDBJ and EMBL. After that, homological sequences were alignmented. Phylogenetic tree and homological analysis were made by some biological software such as Clustal X 1.81, and DNAstar 4.0. The result suggest that the Small-Subunit rRNA(SSU rRNA) specific fragments of 4 Crab-eating macaques Cryptosporidium isolates are C. muris, homology of 100% and 99.5%; and GP60 specific fragments of 13 Crab-eating macaques Cryptosporidium isolates are C. homisis, homology of 100% and 99.8%.This is the first time to provide molecular biology evidence for Cryptosporidium muris infection non-human primates, And the first finding of the Cryptosporidium in non-human primates in china and Cryptosporidium ryane can be islated from yak.