Acta Veterinaria et Zootechnica Sinica ›› 2021, Vol. 52 ›› Issue (9): 2569-2578.doi: 10.11843/j.issn.0366-6964.2021.09.020

• PREVENTIVE VETERINARY MEDICINE • Previous Articles     Next Articles

Isolation and Identification of a Strain of Pseudomonas aeruginosa from Forest Musk and Analysis of the Whole Genome Sequence

HE Jian, XING Xiaoyong, WU Xiaochun, WEN Fengqin, ZHANG Yangyang, ZHANG Shengying, LIU Jia, BAO Shijun*   

  1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2021-01-08 Online:2021-09-23 Published:2021-09-26

Abstract: The purpose of this study was to identify the pathogenic bacteria that killed the forest musk deer in a forest musk deer farm in Tianshui, Gansu Province, and to analyze its pathogenicity, drug resistance and the whole genome sequence. The bacterial strains were isolated and purified from the lungs of dead forest musk deer. Then the isolate was identified through biochemical test and 16S rRNA gene sequence analysis. Subsequently, the pathogenicity and drug sensitivity of the isolate were analyzed. And based on the whole genome sequence, the whole genome sequence of the isolate was assembled and annotated. In addition, the phylogenetic analysis of the virulence genes toxA and exoT was completed. The results showed that a strain of Pseudomonas aeruginosa was isolated from the lung of the dead forest musk deer and named TS2019. The LD50 of TS2019 in mice was 2.82×107 CFU·mL-1. The results of drug sensitivity test demonstrated that TS2019 had multiple drug resistance, but it was sensitive to ciprofloxacin, lomefloxacin, etc. The results of genome sequencing showed that the genome size of TS2019 was 6 308 327 bp, including 5 929 genes, of which 1 035 genes were involved in metabolic pathways. There were 875 genes encoding virulence factors which included adhesion proteins, regulatory factors, toxic proteins, etc. There are 5 288 genes related to antibiotic resistance such as tetracycline and aminoglycoside in the genome. The results of the phylogenetic analysis showed that virulent genes toxA and exoT of TS2019 shared over 99% homology with corresponding genes of many P. aeruginosa strains in GenBank. And the phylogenetic analysis of the eoxT gene showed the genetic relationship between TS2019 and P33 strain isolated from human in Hangzhou, was the closest, but it was an independent branch. In this study, a strain of P. aeruginosa was isolated and identified from the lungs of the dead forest musk deer, and the strong pathogenicity and multi-drug resistance of the isolate had been confirmed. The virulence genes toxA and exoT had high homology with the corresponding gene sequences of P. aeruginosa in GenBank. The results provided the theoretical support for the prevention and treatment of P. aeruginosa infection-related diseases of forest musk deer, and also laid a foundation for further study on the pathogenic mechanism and drug resistance mechanism of P. aeruginosa.

Key words: forest musk deer, Pseudomonas aeruginosa, isolation and identification, pathogenic analysis, genome sequence analysis

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