China Animal Husbandry and Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (3): 1352-1359.doi: 10.16431/j.cnki.1671-7236.2025.03.036

• Basic Veterinary Medicine • Previous Articles    

Isolation,Identification and Drug Resistance Analysis of a Sheep-derived Strain of Clostridium perfringens Type D

LI Na1, LIU Chongyang2, ZHANG Jingjing2, MALIYA Qiqige2, ZHU Na2, LU Bin3, HAI Ying2   

  1. 1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
    2. Wushen County Animal Disease Prevention and Control Center, Ordos 017300, China;
    3. Wushen County Rural Revitalization Comprehensive Development Center, Ordos 017300, China
  • Received:2024-06-29 Published:2025-02-22

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Abstract: 【Objective】 Clostridium perfringens type D was a Gram-positive anaerobic bacterium,which could cause enterotoxaemia in animals such as goats,sheep and cattle,which had a great impact on the health of ruminants.The aim of this study was to understand the drug resistance of Clostridium perfringens type D in Wushen county, Inner Mongolia, and provide a scientific basis for the treatment of diseases caused by Clostridium perfringens type D in this region.【Method】 The isolated strain was initially identified by culture characteristics and Gram staining microscopy.The suspected strain was further identified by 16S rRNA sequence comparison and PCR amplification of toxin gene.And the drug sensitivity test was carried out by K-B paper slide method,and PCR was used to detect the drug resistance gene of the isolated strain.【Result】 The isolated strain grew black round colonies in TSC medium,and the Gram staining microscopy showed that the bacterium was thick and short,and became a single or double arrangement of Gram-positive rectobacteria,and the morphological and microscopic results were consistent with the characteristics of Clostridium perfringens.The results of 16S rRNA sequence alignment showed that the similarity of the isolated strain and 16S rRNA gene sequences of Clostridium perfringens published in NCBI database was more than 99%,and the strain was identified as Clostridium perfringens.The PCR detection results of toxin genes showed that cpa and etx genes were amplified,indicating that the strain was Clostridium perfringens type D.The results of drug sensitivity tests showed that the isolated strain was resistant to 16 antimicrobials including ceftriaxone,cefuroxime and gentamicin.The results of resistance gene detection showed that 4 resistance genes,blaCTX-M,blaSHV,qnrA and aac(6')-Ⅰb-cr,were detected,and 2 resistance genes,blaTEM and qnrS,were not detected.【Conclusion】 In this experiment,a strain of Clostridium perfringens type D,which was multi-drug resistant,was successfully isolated from sheep.The results of this study provided a reference basis for the diagnosis,epidemiological investigation,and treatment of Clostridium perfringens type D infected sheep.

Key words: sheep; Clostridium perfringens type D; isolation and identification; drug resistance

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