树鼩源大肠杆菌分离鉴定及耶尔森强毒力岛相关基因的检测及序列分析

doi:10.16431/j.cnki.1671-7236.2022.11.038

Abstract

Abstract: 【Objective】 The purpose of this experiment was to study the carriage and drug resistance of Escherichia coli (E.coli) high pathogenicity island (HPI) related genes in artificially bred Tupaia belangeri and to provide some ideas for the breeding and management of Tupaia belangeri and the prevention and treatment of colibacillosis.【Method】 129 samples of Tupaia belangeri anal wipe were collected aseptically.The bacteria were isolated and identified by McConkey medium, LB agar medium, Gram staining and biochemical test.HPI-related genes were detected by PCR.HPI-positive strains were screened after PCR identification.The similarity analysis and phylogenetic tree construction of the main structural genes irp2 and fyuA of the isolates were performed.Disk diffusion method was used for drug sensitivity test of the isolates.【Result】 A total of 123 strains of E.coli were isolated from 129 Tupaia belangeri anal wipe samples, and 95.35% (123/129) were isolated.The results of Gram staining showed that the isolates were microscopically examined as stubby rubrum.The biochemical identification results showed that the isolates were positive for lactose, glucose, maltose, peptone, and mannitol biochemical reactions, and negative for hydrogen sulfide and urease biochemical reactions, all of which conformed to the characteristics of E.coli.The electrophoresis results of PCR products showed that the detection rate of HPI-related genes was 88.62% (109/123), the carrying rates of irp2 and fyuA genes were 34.15% (42/123) and 47.97% (59/123), respectively.The results of sequence analysis of the main structural genes showed that the irp2 and fyuA genes were more than 98.6% and 98.9% similar to the sequences published irp2 and fyuA gene nucleotide sequences in GenBank, respectively, and the phylogenetic tree results showed that the isolates were closely related to Yersinia spp.The results of drug resistance analysis showed that the isolated strains were sensitive to amikacin and florfenicol, and resistant to amoxicillin and oxacillin, the drug resistance rates were 87.80% and 81.30% respectively.【Conclusion】 HPI related genes were widely distributed in E.coli of Tupaia belangeri, which further confirmed that HPI could be horizontally transferred, and the inheritance of the main structural genes irp2 and fyuA were highly conserved.

Key words: Tupaia belangeri; Escherichia coli; high pathogenicity island

CLC Number:

S852.61^＋2

CHEN Bingxun, HAN Dongmei, ZHAO Weiwei, ZHANG Bo, DENG Jing, WAN Quan, WANG Xi, XIAO Peng, LYU Longbao, GAO Hong. Isolation and Identification of Escherichia coli from Tupaia belangeri and Detection and Sequence Analysis of Genes Related to High Pathogenicity Island[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4466-4475.

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Isolation and Identification of Escherichia coli from Tupaia belangeri and Detection and Sequence Analysis of Genes Related to High Pathogenicity Island

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