某鸡场大肠杆菌噬菌体的分离鉴定及其生物学特性研究

doi:10.16431/j.cnki.1671-7236.2021.06.034

Abstract

Abstract: In order to explore the morphological and biological characteristics of coliphages,55 strains of avian pathogenic Escherichia coli stored in the laboratory were used as the host bacteria in this study.A coliphage with a wide lysis spectrum was obtained from the sewage and sludge of a chicken farm by the double-layer agar plate method,named Bp16,after negative staining with 2% phosphotungstic acid,the morphology and structure were observed by transmission electron microscope.The temperature and pH stability,optimal multiplicity of infection (MOI),and one-step growth curve were measured by the double-layer agar plate method.Transmission electron microscopy showed that the isolated broad lysis spectrum phage Bp16 had an icosahedral structure in the head,with tails and filaments,and the body length was about 220 nm.According to the classification characteristics of Virus Classification—The 9th Report of the International Committee on Classification of Viruses,the bacteriophage was a member of the Caudovirales and the Mycobacteriophages family.Using 55 avian pathogenic Escherichia coli stored in the laboratory as indicator bacteria,its cracking efficiency was only 12.73%.The results of temperature stability showed that the titer of the phage remained basically unchanged at 37-60 ℃ for 30 min or 1 h,and the titer could still reach more than 1×10⁴ PFU/mL after treatment at 80 ℃ for 1 h,indicating that the phage was not sensitive to temperature changes and had heat resistance.The results of pH stability showed that the titer remained basically unchanged between pH 5.0 to 10.0,and it was relatively tolerant to acid and alkali.The results of the best MOI showed that the best MOI was 1.One-step growth curve according to the best MOI was drew,and it was measured that the incubation period was 20 min,the outbreak period was 50 min,and the lysis amount was 73 PFU/cell.In summary,the bacteriophage Bp16 had thermal stability,acid-base stability and strong lysis ability,which laid the foundation for the development of coliphage preparations for subsequent experiments.

Key words: phage; isolation and identification; biological characteristics; Escherichia coli

CLC Number:

S852.61

ZHAO Zunfu, YANG Tingting, ZHANG Huanrong. Isolation,Identification and Biological Characteristics of Coliphage from a Chicken Farm[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2204-2211.

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Isolation,Identification and Biological Characteristics of Coliphage from a Chicken Farm

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