榆林部分地区湖羊乳房炎病原菌的分离鉴定及耐药性和毒力基因检测

doi:10.16431/j.cnki.1671-7236.2024.04.042

Abstract

Abstract: 【Objective】 The purpose of this experiment was to clarify the distribution of the main pathogenic bacteria,their resistance and virulence genes,and guide the rational use of drugs in the clinical treatment of mastitis in Hu sheep farms.【Method】 43 clinical mastitis milk samples were collected from 6 large-scale sheep farms.Bacterial morphology identification,biochemical identification and 16S rDNA sequence analysis were used to determine the main pathogenic bacteria causing mastitis in Hu sheep.Drug resistance was detected using paper diffusion method,and related virulence genes were detected using PCR method.【Result】 The Gram staining results showed that Gram positive cocci and Gram negative bacilli were obtained. On the Baird Parker agar plate,Gram positive cocci exhibited rounded edges and black colonies in the middle.Biochemical tests showed that the isolated strains were positive for mannitol,hydrogen peroxide and rabbit plasma coagulation tests.After 16S rDNA sequencing comparison results showed that a total of 31 strains were highly similar to Staphylococcus aureus.Gram negative bacilli presented black purple colonies with metallic luster on the Eosin Methyl Blue agar plate,while on the MacConkey agar plate,pink colonies appeared.Biochemical tests showed that the isolated strains were positive for indole and methyl red tests,and negative for VP and citrate utilization tests.After 16S rDNA sequencing comparison,a total of 12 strains were highly similar to Escherichia coli.The drug sensitivity test results showed that the isolates of Staphylococcus aureus were sensitive to drugs such as ciprofloxacin,levofloxacin and cefotaxime,but resistant to penicillin,erythromycin and streptomycin.The isolates of Escherichia coli were sensitive to drugs such as ciprofloxacin and levofloxacin,and were resistant to penicillin and erythromycin.Seven virulence genes were detected in Staphylococcus aureus,namely PVL, Hlb, seb, clfA, clfB, fnbB and icaD genes,but Hla, sea and fnbA genes were not detected.Six virulence genes were detected in Escherichia coli,namely hlyA, etrA, fasA, eaeA^a, exhA^a and stx1 genes,but no cnf1, aer, estA and stx2 genes were detected.【Conclusion】 The main pathogens causing mastitis in large-scale sheep farms were Staphylococcus aureus and Escherichia coli,which carried rich virulence genes.It was recommended to prioritize the use of ciprofloxacin and levofloxacin for treatment.This article provided scientific basis for clinical treatment of this disease.

Key words: Hu sheep; mastitis; pathogenic bacteria; drug resistance

CLC Number:

S852.61

WANG Yu, SHI Song, JIN Panhao, LI Jinxin, ZHANG Mingxi, LI Tuo, LI Longping, QU Lei, DONG Shuwei. Isolation,Identification and Detection of Resistance and Virulence Genes of Pathogenic Bacteria Causing Mastitis in Some Areas of Yulin[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1746-1756.

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Isolation,Identification and Detection of Resistance and Virulence Genes of Pathogenic Bacteria Causing Mastitis in Some Areas of Yulin

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