基于粪便样本的野生犬科动物物种鉴别与棘球绦虫感染调查

doi:10.11843/j.issn.0366-6964.2021.012.021

Abstract

Abstract: This study was conducted to explore the methods and its feasibility of identifying Canidae species and to investigate the infection of Echinococcus in wild Canidae based on non-invasive fecal sample analysis. A total of 101 DNA samples extracted from feces of wild Canidae were amplified and sequenced with a pair of universal primers designed for the mitochondrial DNA control region of mammalian species. The species origins of the fecal samples were molecularly identified by the analysis of nucleotide variation, the comparison of sequence similarity, the analysis of genetic distance and the construction of phylogenetic tree. Meanwhile, copro-antigen ELISA was used to investigate the infection of Echinococcus. The test success rate for the species identification of the samples was 73.27%. A total of 20 haplotypes were inferred from the 74 sequences obtained in the study. Each of the haplotype sequences was found to match with one single species in the GenBank database with 98.52%-100% sequence similarity. All the haplotypes can be divided into 4 haplotype sets according to sequence differences. The average number of nucleotide differences among the 4 haplotype sets is 17.83-70.10, and the number of nucleotide differences among the haplotypes within each set is 1-10. The genetic distance among the 4 haplotype sets is 0.068-0.342. The value is much larger than that of the genetic distance among the haplotypes within each set, which is 0.009-0.022. The phylogenetic analysis showed that haplotypes in the same sets clustered into same clades with high bootstrap values. Based on all the analysis results, it can be inferred that the sources of the fecal samples were Canis lupus familiaris, Canis lupus, Vulpes vulpes and Vulpes ferrilata respectively. A total of 11 positive samples were found in the copro-antigen ELISA test. The positive rate of Echinococcus antigen was 10.89%. The analysis of mitochondrial DNA control region sequence can be used for the molecular identification of the Canidae. The detection accuracy rate would be further improved if other molecular markers such as nuclear genes were aslo used. The positive rate of Echinococcus antigen is high in feces of wild Canidae in the study area.

Key words: Canidae, Echinococcus, definitive host, mtDNA, species identification

CLC Number:

SHANG Jingye, ZHANG Guangjia, DANBA Zeli, WANG Qi, YU Wenjie, HE Wei, LIAO Sha, SHEN Yunzhou, HUANG Yan, WANG Qian, ZHONG Bo, LIU Yang. Identification of Wild Canidae Species and Investigation of Echinococcus Infection Based on Fecal Samples[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3546-3556.

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Identification of Wild Canidae Species and Investigation of Echinococcus Infection Based on Fecal Samples

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