1.Isolation and identification of yak-derived Enterococcus from four provinces of Tibetan plateau and detection of virulence resistance genes and biofilms
Jiaojiao XIN ; Guiqin GAO ; Xiaolong ZHAO ; Yongzhi LOU ; Jing LI ; Pan CHANG ; Ren-de SONG ; Xiangying KONG ; Hongmei SHI ; Xiaolin LUO ; Zhuoma GESANG ; Sizhu SUOLANG ; Ga GONG
Chinese Journal of Veterinary Science 2025;45(2):289-297
This study aims to understand the epidemic distribution characteristics,antimicrobial resistance,virulence genes,and biofilm adhesion ability of Enterococcus in yaks on the Tibetan plateau.Three hundred and forty-six fresh yak fecal samples and 311 milk samples were collected from four provinces on the Tibetan plateau(Xizang,Sichuan,Gansu,Qinghai),totaling 657 sam-ples.Bacterial isolation and identification were conducted,followed by 16S rDNA gene detection and the construction of a systematic evolutionary tree.The isolated strains were tested for antimi-crobial resistance and virulence genes using PCR,and sensitivity tests were performed using 18 types of antibiotics.The biofilm adhesion ability of the isolated bacteria was determined using an improved semi-quantitative crystal violet staining method.The results showed that the total isola-tion rate of Enterococcus was 32.27%,with Sichuan having the highest at 60.23%,followed by Gansu,Qinghai,and Tibet autonomous region at 42.70%,23.47%,and 18.31%respectively.In terms of sample types,the isolation rate in fecal samples was 36.71%,and in milk samples,it was 27.33%.Through PCR amplification,bands of approximately 1 400 bp were obtained,and 5 strains were selected for evolutionary analysis,forming a separate cluster.Among the 212 isolated strains,a high resistance to clindamycin,quinupristin-dalfopristin,linezolid,levofloxacin,and erythromycin was observed,with various resistance phenomena,accounting for 60.85%.Only 5 out of 12 resist-ant genes were detected,namely erm(B),tet(L),tet(O),tet(M),and ant(6)-Ia.All 13 virulence genes were detected in Enterococcus,with detection rates in the range of 5.19%to 95.76%,where cpd was 95.75%,gelE was 91.98%,efaA was 86.79%,asal was 86.32%,and the rest ranged from 5.19%to 55.66%.The fsr virulence gene was not detected in Enterococcus from milk sources.Among the isolated strains,3.30%showed medium adhesive ability,48.58%showed weak adhesive ability,and 48.11%showed no adhesive ability.The above research revealed the preva-lence of yak derived Enterococcus,the carrying status of resistance and virulence genes,and the correlation between biofilm phenotypes,laying the foundation for mastering research data on yak-derived Enterococcus in the Tibetan plateau.
2.Isolation and identification of yak-derived Enterococcus from four provinces of Tibetan plateau and detection of virulence resistance genes and biofilms
Jiaojiao XIN ; Guiqin GAO ; Xiaolong ZHAO ; Yongzhi LOU ; Jing LI ; Pan CHANG ; Ren-de SONG ; Xiangying KONG ; Hongmei SHI ; Xiaolin LUO ; Zhuoma GESANG ; Sizhu SUOLANG ; Ga GONG
Chinese Journal of Veterinary Science 2025;45(2):289-297
This study aims to understand the epidemic distribution characteristics,antimicrobial resistance,virulence genes,and biofilm adhesion ability of Enterococcus in yaks on the Tibetan plateau.Three hundred and forty-six fresh yak fecal samples and 311 milk samples were collected from four provinces on the Tibetan plateau(Xizang,Sichuan,Gansu,Qinghai),totaling 657 sam-ples.Bacterial isolation and identification were conducted,followed by 16S rDNA gene detection and the construction of a systematic evolutionary tree.The isolated strains were tested for antimi-crobial resistance and virulence genes using PCR,and sensitivity tests were performed using 18 types of antibiotics.The biofilm adhesion ability of the isolated bacteria was determined using an improved semi-quantitative crystal violet staining method.The results showed that the total isola-tion rate of Enterococcus was 32.27%,with Sichuan having the highest at 60.23%,followed by Gansu,Qinghai,and Tibet autonomous region at 42.70%,23.47%,and 18.31%respectively.In terms of sample types,the isolation rate in fecal samples was 36.71%,and in milk samples,it was 27.33%.Through PCR amplification,bands of approximately 1 400 bp were obtained,and 5 strains were selected for evolutionary analysis,forming a separate cluster.Among the 212 isolated strains,a high resistance to clindamycin,quinupristin-dalfopristin,linezolid,levofloxacin,and erythromycin was observed,with various resistance phenomena,accounting for 60.85%.Only 5 out of 12 resist-ant genes were detected,namely erm(B),tet(L),tet(O),tet(M),and ant(6)-Ia.All 13 virulence genes were detected in Enterococcus,with detection rates in the range of 5.19%to 95.76%,where cpd was 95.75%,gelE was 91.98%,efaA was 86.79%,asal was 86.32%,and the rest ranged from 5.19%to 55.66%.The fsr virulence gene was not detected in Enterococcus from milk sources.Among the isolated strains,3.30%showed medium adhesive ability,48.58%showed weak adhesive ability,and 48.11%showed no adhesive ability.The above research revealed the preva-lence of yak derived Enterococcus,the carrying status of resistance and virulence genes,and the correlation between biofilm phenotypes,laying the foundation for mastering research data on yak-derived Enterococcus in the Tibetan plateau.

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