西藏牦牛粪便大肠杆菌毒力特性、耐药性与Ⅰ类整合子分子特征相关性分析

doi:10.16431/j.cnki.1671-7236.2023.10.035

Abstract

Abstract: 【Objective】 Yaks,as one of the important hosts of zoonotic and multi-drug resistant pathogens in Tibet Autonomous Region,the abuse of antibiotics during the breeding process is the main reason for the development and spread of multi-drug resistance in yaks.The aim of this study was to conduct correlation analysis on the virulence characteristics,drug resistance,integrase and biofilm-forming phenotype of Escherichia coli (E.coli) strains from Tibetan yaks.【Method】 200 samples of yaks diarrhea were collected from farmers in Lhasa,Linzhi,and Nagqu in Tibet.Use bacteriological methods to isolate and purify E.coli by MacConkey Agar medium and Eosin-Methylene Blue Agar medium.PCR amplification and sequencing of the suspected E.coli were performed with 16S rDNA universal primers.The obtained sequence were BLAST aligned in NCBI database.7 classes of 16 antimicrobial drug susceptibility tests were performed on E.coli.Using PCR amplification to detect 4 categories of 10 virulence genes,20 common drug resistance genes,and 2 class Ⅰ integrator that were closely related to pathogenicity.The modified semi-quantitative crystalline violet staining method was performed on the isolated E.coli to determine their biofilm-forming phenotype,and conducted correlation analysis.【Result】 91 strains of E.coli from yaks were isolated and identified.The antibiotic drug sensitivity test of 16 antibiotics in 7 categories showed that E.coli had the strongest drug resistance to clindamycin (87.91%).Meanwhile,there was a phenomenon of multiple drug resistance,with some strains being resistant to up to 13 antibiotics.4 types of virulence genes,STEC,ETEC,EPEC and NTEC,were found to be positive in virulence test,among these detected virulence genes,F17 gene accounted for 59.34% (54/91) and stx1 gene accounted for 49.45% (45/91).Drug resistance gene testing showed that the most common among the 22 drug resistance genes was bla_TEM gene,followed by tetracycline tetA and sul1 genes.Class Ⅰ integrator analysis detected intⅠ1 and intⅠ2 integrase genes,with separately rates of 29.67% (27/91) and 1.10% (1/91).The formation of biofilm was one of the main mechanisms for bacteria to survive in harsh environments.The detection of biofilm showed that 48.35% (44/91) of 91 E.coli strains showed weak adhesion ability,and the data analysis showed that the detection rate of STEC and ETEC of E.coli was positively correlated with the detection rate of drug resistant genes,class Ⅰ integrator and biofilm.【Conclusion】 A variety of virulence genes and drug resistance genes were detected from 91 strains of E.coli isolated from yaks,and the data showed that there was a certain correlation between the diversity of its drug resistance genes and class Ⅰ integrator and the ability of biofilm formation.The results provided a theoretical basis for the study of drug resistance mechanism of E.coli from yaks and the reasonable use of antibiotics.

Key words: Tibetan yaks; Escherichia coli; virulence genes; drug resistance; integrase; biofilm

CLC Number:

S852.61⁺2

XIN Jiaojiao, SUONAN Dunzhu, LOU Yongzhi, AWANG Kezhu, WANG Xin, GANG Haojie, YAN Jiahui, CHANG Pan, CHEN Shuyu, ZHOU Hongting, YIXI Cuomu, SUOLANG Sizhu, GONG Ga. Correlation Analysis on Virulence Characteristics,Drug Resistance and Molecular Features of Class Ⅰ Integrator of Escherichia coli Isolated from Feces in Tibetan Yak[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4210-4222.

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Correlation Analysis on Virulence Characteristics,Drug Resistance and Molecular Features of Class Ⅰ Integrator of Escherichia coli Isolated from Feces in Tibetan Yak

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