| The wild Siberian tiger(Panthera tigris altaica)and Amur Leopard(Panthera pardus orientalis)are precious and endangered animals.Parasitic disease is a common diseases which can harm the health of wildlife animals.In this study,the parasites were identified by microscopic observation.And then the part of ITS-1 gene and 5.8S gene of the Toxascaris leonina and Toxocara cati and the part of 18 S gene of the Strongyloides cati were amplificated by traditional PCR and phylogenetic analyzed.Moreover,intestinal microflora of wild Siberian tiger and Amur leopard which were infected different number of parasites were determinated by macrogeome technique,and compare the features.The results of this study show that wild Siberian tiger infected with eight kinds of parasites,the wild Amur leopard infected seven kinds of parasites.Phylogenetically,wild Siberian tiger's T.cati was distant from felis catu.That T.leonina of wild Siberian tiger sequence resided in a monophyletic group with other tigers indicated that tiger's T.leonina has a close relationship to others.The results of evolution analysis of S.cati show that the S.cati in wild Siberian tiger was close to raccoon dog(Nyctereutes procyonoides).In addition,the infected number and rate of the T.cati(51/68,75%)were the largest in wild Siberian tiger feces,followed by whipworm(16/68,23.5%).The infected number and rate of the T.cati(26/42,61.9%)were the largest in Wild Amur leopard,followed by Spirometra mansoni(21/42,50%).The average infected intensity of T.cati is the highest in the wild Siberian tiger(490.29 n/g),and the average infected intensity of S.mansoni is the highest in the wild Amur leopard(428.21 n/g).The Alpha diversity analysis showed that the bacterial enrichment in intestinal microbes is characterized by high and nonuniformity.For the bacteria phylum,the samples which were infected by large amount of parasite in wild Siberian tiger had 23 and 27 bacteria phylums,which was slightly higher than that of the parasite free samples(23 and 24 phylums).Meanwhile,the samples which were infected by large amount of parasite in Wild Amur leopard had 24 and 17 phylums,which was significantly higher than that of the parasite free samples(12 and 16 phylums).These results indicated that different infection intensity of parasite leading to Wild Siberian tiger and Amur leopard intestinal bacteria class of dominant bacteria species were different.The order,families and genus data also prove this point.The results of principal Component Analysis(PCA)in Beta diversity shows that the wild Siberian tiger all samples?parasitic infection in Amur leopard samples(FPPW1,FPPW2)and captive Siberian tigers(FPTZ1,FPTZ2)were clustered together,which indicated that they had similar bacterial structure.However,Wild Amur leopard,which no parasite infection in the sample,FPPW3 and FPPW4 were scattered with other samples,which indicated that they had different bacterial structure.The results of sample distance matrix based on UniFrac analysis showed that the similarity(0.29)in wild Siberian tiger samples with parasite infection intensity was higher than that of no parasitic infected samples(0.31).The similarity(0.38)in wild Amur Leopard was significantly higher than that no parasitic infected samples(0.59).This study explores the relationship between parasites and intestinal microflora in wild Siberian tiger and wild Amur leopard,providing a basis for further research on the relationship between parasitic diseases and intestinal microbes in endangered feline.And providing the theoretical basis for the prevention of wild Siberian tiger and wild Amur leopard parasitic diseases. |
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