›› 2018, Vol. 45 ›› Issue (11): 3211-3220.doi: 10.16431/j.cnki.1671-7236.2018.11.028

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Isolation, Identification and Biological Characteristic of Donkey Mastitis Escherichia coli

GAO Jingwen, ZENG Zongbo, ZHANG Li, QI Yayin   

  1. College of Animal Science & Technology, Shihezi University, Shihezi 832000, China
  • Received:2018-03-01 Online:2018-11-20 Published:2018-11-20

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Abstract:

In order to investigate the main pathogenic bacteria of mastitis and biological characteristics from a large-scale donkey farm in Xinjiang,routine microbiological methods were used to isolate pathogens,biochemical identification and 16S rRNA PCR amplification methods were used to identify pathogens,K-B susceptibility paper method was used to determine their drug resistance,D600 nm value was measured,bacterial growth curves were drawn,the homology was analyzed,and the phylogenetic tree was constructed.The results showed that the isolates were short gram-negative bacilli,could grow pink and metallic luster colonies on MacConkey culture medium and Eosin-Meilan medium,respectively.The biochemical identification of isolates of phosphohydrazine,lysine,ornithine,raffinose,sorbitol,side ginsenoside and xyloglucan tested positively,while the phenylalanine and hydrogen sulfide tests were negative,and after the PCR amplification,the 205 bp No.1 strain and the 203 bp No.2 strain were obtained,their characteristics accorded with the characteristics of Escherichia coli(E.coli).Antibiotic sensitive test showed that the isolates were highly sensitive to streptomycin and chloramphenicol and resistant to penicillin,amoxicillin and gentamicin.The growth curve showed that the isolates were highly prolificated at 2 to 16 h,stabilized at 16 to 32 h,and entered the recession after 32 h.The homology of No.1 and No.2 strains was as high as 99%.The homology with E.coli 11J and RCB800 was the highest (100%);The No.2 strain had the highest homology with E.coli 675SK2 and DTU-1 (100%).From the phylogenetic tree,No.1 strain was in the same small branch with E.coli 11J and RCB800,and No.2 strain was in the same small branch with E.coli 675SK2 and DTU-1.Therefore,the pathogen of the donkey mastitis was E.coli,and its biological characteristics laid a solid theoretical basis for the further systematic study of the pathogenic mechanism of donkey mastitis E.coli.

Key words: donkey mastitis; Escherichia coli; isdation and identification; biological characteristics

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