ABSTRACT
Objective Based on sequencing the full-length genomes of two Chinese Ferret-Badger, we analyzed the properties of rabies viruses genetic variation in molecular level to get information on prevalence and variation of rabies viruses in Zhejiang,and to enrich the genome database of rabies viruses street swains isolated from Chinese wildlife.Methods Overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses of the N genes from Chinese Ferret-Badger,sika deer,vole,dog.Vaccine strains were then determined.Results The two full-length genomes were completely sequenced to find out that they had the same genetic structure with 11 923 nts including 58 nts-Leader, 1353 nts-NP,894 nts-PP,609 nts-MP,1575 nts-GP,6386 nts-LP, and 2,5,5 nts-intergenic regions (IORs),423 nts-Pseudogene-like sequence(ψ),70 nts-Trailer.Conclusion The two full-length genomes were in accordance with the propertms of Rhabdoviridae Lyssa virus by blast and multi-sequence alignment.Th nucleotide and amino acid sequences among Chinese strains had the highest similarity,especially among animals of the same species.Of the two full-length genomes,the similarity in amino acid level was dramatically higher than that in nucleotide level, so that the nucleotide mutations happened in these two genomes were most probably as synonymous mutations.Compared to the referenced rabies viruses,the lengths of the five protein coding regions did not show any changes or recombination,but only with a few-point mutations.It was evident that the five proteins appeared to be stable.The variation sites and types of the two ferret badgers genomes were similar to the referenced vaccine or street strains.The two strains were genotype 1 according to the multi-sequence and phyiogenetic analyses,which possessing the distinct geographyphic characteristics of China.All the evidence suggested a cue that these two ferret badgers rabies viruses were likely to be street virus that already circulating in wildlife.
ABSTRACT
Objective To understand the prevalence of rabies among wild animals and the animal species in rabies epidemic areas of Zhejiang province. Methods One hundred and sixty samples were collected from the brain tissues of cats, stoats, Apodemus agrarius, Moschus chinensis, and Sus scrofa in Lishui and Chunan cities of Zhejiang province. Each sample was divided into four parts: cerebrum,mesencephal, cerebellum and cyrus hippocampi which were used to determine the positive samples by detection of rabies virus specific antigens and nucleotides, using DFA and RT-PCR methods. Results Positive slides in the tests contained a glaring, apple green brilliance fluorescence using rabies virus specific monoclonal antibody against nucleoprotein. Using Nested-PCR method targeted at part of N gene,five positive samples were identified which consisting of four positive samples from stoats with positive ratio as 8.33% (4/48) and one positive sample from Apodemns agrarius with positive ratio as 1.75% (1/57). However, no positive result was found from cats, Moschus chinensis, and Sus scrofa samples. Conclusion Rabies virus positive samples were identified from stoats and Apodemus agrarins in the mountain areas with biological diversity in Lishui and Chunan cities of Zhejiang province,indicating that stoats and Apodemus agrarius might have played a role in human rabies and acted as host of rabies virus. In order to effectively prevent and control rabies virus under these complicated geographical and ecological environment, we must understand and evaluate the infection situation among animals in these regions.