China Animal Husbandry and Veterinary Medicine ›› 2020, Vol. 47 ›› Issue (1): 258-266.doi: 10.16431/j.cnki.1671-7236.2020.01.030

• Basic Veterinary Medicine • Previous Articles     Next Articles

Isolation,Identification and Probiotic Characteristics of a Clostridium butyricum

WANG Qian1, LI Haihua2, DOU Caixia1, LIU Lili1, QIAO Jiayun1   

  1. 1. College of Life Sciences, Tianjin Normal University, Tianjin 300387, China;
    2. College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China
  • Received:2019-08-13 Online:2020-01-20 Published:2020-01-17

Abstract: This study explored the probiotic characteristics of Clostridium butyricum isolates and aimed to provide a theoretical basis for their application in piglet diets.Clostridium butyricum was isolated by routine methods and cultured in pure culture.The isolates were identified by biochemical detection and 16S rRNA gene sequencing;The viable cell count method and the Oxford cup method were used to study the inhibitory effect of the culture supernatant of the isolate on the three pathogenic bacteria,the adhesion of the isolated pig intestinal epithelial cells (IPEC-J2) and its inhibitory effect on the adherent cells of pathogenic bacteria;The effect of the isolate on the growth of IPEC-J2 was studied by cell counting method;The cytokine level in the culture supernatant was determined by ELISA after treatment of the isolate.The results showed that a strain of Clostridium butyricum was successfully isolated in this study.Compared with the control group,the antibacterial effect of Clostridium butyricum culture supernatant on the three pathogenic bacteria was significant (P<0.05).Clostridium butyricum could adhere to IPEC-J2,and the optimal multiplicity of infection(MOI) was 50,and the optimal treatment time was 3 h;Clostridium butyricum had inhibitory effect on three pathogenic adhesion IPEC-J2 cells (P<0.05).When the MOI of Clostridium butyricum was 1,10 and 50,the cell growth was normal and the morphology was intact.When MOI was 100,the growth of IPEC-J2 was significantly inhibited,and some cell death occurred at the same time.There was no difference in cytokine production levels when MOI was 1,and cytokine production levels were significantly increased at MOI of 10,50,and 100 (P<0.05).In conclusion,a Clostridium butyricum strain was successfully isolated in this study,which has good probiotic characteristics.The results provided a theoretical basis for the application of the isolated strain in production.

Key words: Clostridium butyricum; isolation and identification; pig intestinal epithelial cells; probiotic characteristics

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