| Wild animals are non-domesticated animals and survive in natural environment, which include various mammals, birds, reptiles, amphibians, fish, mollusks, insects and some other animals. Wildlives are the natural host of most pathogens in nature, The great loemia of wild animals are closely related to the public health of humans. Parasitosis is one of the most common diseases in wildlives. Due to various pathogens of parasitosis, with a wide distribution, and large number of infected wild animans, etc. It ofen causes the loss of body weight, the decline of body defense capability, and even leads to death. So, parasitosis is one of the most reasons for causing the number decreasing of wildlife populations.162, 140 and 162 fecal samples were respectively collected from 17 species birds, 18 species herbivores, 23 zoophagous, and examined for understanding the prevalence of parasites infection by Sheather's sugar flotation technique.The overall infection rate of parasites in birds was 19.75%(32/162), the common parasites found were Coccidia and Nematode. And the average infection rate of herbivore was 25.71%(36/140), thereinto, the infection rates of Coccidia, Nematode, Trichuris trichiura, and Lumbricoide were 14.29(20/140), 9.29% (13/140), 7.86%(11/140) and 1.43%(2/140), respectively. The infection rate of Neofelis nebulosa in whipworm was 36.4%(4/11). The infection rates of Coccidia in black panther, Panthera pardusl and tiger were 100%(1/1), 50%(1/2)and 100%(4/4), respectively. However, no eggs or oocysts were discovered in Grand Anglo-Francais Tricolore. 37.8% (34/90) of infection rate was found in the 99 fecal samples from macaque, thereinto, the positive numbers of Coccidia, Trichuris trichiura and lumbricoide were respectively five, 27 and one. Moreove, there were various infection rate in other monkeys, namely, 40%(2/5)of Trichuris trichiura infection in Rhinopithecus roxellanae, and 20%(1/5)of Coccidia infection in Cynomolgi.Survey on the prevalence of Cryptosporidium in partial wild and captive reptiles and amphibians was conducted in Shanghai area. The methods of Sheather's sugar flotation and modified acid-fast staining were used to check the oocysts of Cryptosporidium. The results showed that 45.2%(14/31) and 13.6%(6/44) of infection rates were respectively found in 31 wild snakes and 44 captive snakes by modified acid-fast staining method. Similarly, the infection rates of 30%(3/9), 100%(1/1), 100%(1/1) and 25%(1/4) were also discovered in lizard, tortoise, huge tortoise and python. However, only two positive samples were found in lizards by Sheather's sugar flotation technique, with a average infection rate of 22.2%(2/9). This survey showed that the Cryptosporidium infection rate in wild snakes was significantly higher than that of infection rate in captive snakes in Zoo. Therefore, the results propose a essential warning to persons who have the bad habits that like eating wild snakes.The specific fragment of 18S rRNA gene was amplified by nested-PCR from a Cryptosporidium isolate obtained from snake. PCR product was cloned and the positive clones were selected and sequenced, and the results showed that the length of fragment was 854bp. Moreover, the PCR product was digested by SspI restriction enzyme to determine the Cryptosporidium species and genotype. After digestion, two fragments of 383bp and 414bp were discovered, and based on the profiles, the Cryptosporidium isolate was primarily identified as C. serpentis. In order to determine the Cryptosporidium species/genotype from the aspect of molecular phylogenetic relationship, sequence obtained was searched by Blast and Fasta provided in three nucleotide databases, i.e., NCBI, DDBJ and EMBL. Then, sequence alignment, constructing phylogenetic trees and similarities were respectively performed by using the softwares of Clustal X1.81,Phylip3.64, DNAstar4.0, and etc. According to the results of phylogenetic analyses based on 18S rRNA gene locus, the isolate from snake was identified as C. serpentis.A total of 499 fecal samples were collected from four deer farms in Zhengzhou City and examined for Cryptosporidium by Sheather's sugar flotation technique and modified fast acid staining method. Two Cryptosporidium isolates were obtained. The partial sequences of 18S rRNA, HSP70, actin and COWP genes were amplified from one of the two isolates. Based on the four gene loci, the phylogenetic analyses were conducted to identify the Cryptosporidum species and genotype. Thus to determine the molecular evalution relationship between the cervine isolate and other Cryptosporidium species, and to provide a theory foundation for the prevention and treatment of cryptosporidiosis in deer.Investigation on cryptosporidiosis in deer was conducted in Zhengzhou Zoo, wildlife rescure center, deer farm in Mangshan, etc. Tow positive samples were found from two one-year-old deer and the average infection rate was 0.40%(2/499). The low infection rate suggests that deer studied here are not an important reservior of cryptosporidiosis. Moreover, the farms are all far from the drinking water supply system. Therefore, the deer populations may be not a major source of cryptosporidiosis in humans in Zhengzhou region. The sequences of 18SrRNA,Actin,COWP and HSP70 genes were used to conduct phylogenetic analyses based on neighbour-joining method. The analyses of four genes produced similar topology structure, i.e., the Cryptosporidium isolate shared a identical group with other Cryptosporidium cervine genotypes. The similarities in 18S rRNA, HSP70, actin and COWP genes between this isolate and other Cryptosporidium cervine genotype isolates were respectively 99.1%-99.8%, 99.8%, 99.7% and 100%. All in all, the isolate from sika deer was identified as Cryptosporidium cervine genotype although minor difference was noticed in three of four gene loci studied. |
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