羊呼吸道溶血性曼氏杆菌的分离鉴定及其致病性和耐药性研究

doi:10.16431/j.cnki.1671-7236.2025.05.044

Abstract

Abstract: 【Objective】 The purpose of this experiment was to understand the main pathogens causing respiratory tract diseases in sheep in Inner Mongolia,and explore the pathogenicity and drug resistance,so as to provide reference for the control drugs selection of related disease.【Method】 The pathogenic bacteria were isolated,cultured,biochemical and molecular identified from the lung tissue of dead sheep,and the pathogenicity test was carried out in mice.The virulence genes of the isolates were detected by PCR.The drug susceptibility of isolates was detected by K-B method,and the drug resistance genes of the isolates was detected by PCR method.The in vitro inhibitory effects of berberine,citraldehyde and thymol on the isolated strains were determined by Oxford cup method.【Result】 The isolated strains formed β-hemolysis grey white colonies on the blood agar plate medium,and two suspected strains were isolated，and named Mh1-1 and Mh1-2,respectively.Biochemical identification showed that maltose,mannose,sucrose,xylose and sorbitol of the isolates were positive.PCR amplified the specific gene of Mannheimia haemolytica with the size of 453 bp and the capsular serotype A1 band with the size of 306 bp.The similarity comparison results showed that the similarity between the isolate Mh1-1 and Mannheimia haemolytica GDZJcattle2014 (accession No.:KM576848.1) was 98.49%,and the similarity between the isolate Mh1-2 and Mannheimia haemolytica 38599 (accession No.:CP017491.1) was 98.14%.The two isolates were identified as serotype A1 Mannheimia haemolytica.The results of pathogenicity test in mice showed that the LD₅₀ of Mh1-1 was 2.2×10⁷ CFU/mL,and the LD₅₀ of Mh1-2 was 1.0×10⁷ CFU/mL.Virulence genes gs60,lktC,lktA,tbpB,adh and nmaA were detected in both isolates.The results of drug sensitivity test showed that Mh1-1 was resistant to tetracycline and lincomycin,and intermediated to streptomycin.Mh1-2 showed resistance to streptomycin and lincomycin,and intermediated to tetracycline.The two isolates were sensitive to ofloxacin,ciprofloxacin,enrofloxacin,cotrimoxazole,cefotaxime and flufenicol.The results of drug resistance gene detection showed that the two isolates carried strA,strB and tetA genes.The results of in vitro bacteriostatic test showed that berberine and citraldehyde had good inhibitory effects on the isolates.【Conclusion】 In this study,two strains of serotype A1 Mannheimia haemolytica were isolated from the lung tissues of dead sheep with respiratory diseases.Two isolates were pathogenic and carried a variety of virulence genes，and resistant to lincomycin,and carried resistance genes such as strA.The results provided reference for rational selection of antibiotics in clinical,and provided ideas for the development and research of subsequent therapeutic drugs.

Key words: Mannheimia haemolytica; isolation and identification; pathogenicity; drug resistance

CLC Number:

S855.1

WU Jiaxin, SUN Yue, MAO Wei, LIU Shuying, YIN Kaiwen, ZHANG Zhidan, HAN Kaifan, ZHAO Hongxia. Isolation and Identification of Mannheimia haemolytica from Sheep Respiratory Tract and Its Pathogenicity and Drug Resistance[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(5): 2421-2431.

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Isolation and Identification of Mannheimia haemolytica from Sheep Respiratory Tract and Its Pathogenicity and Drug Resistance

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