奶牛乳腺炎源金黄色葡萄球菌噬菌体P82的分离及特性研究

doi:10.16431/j.cnki.1671-7236.2022.06.032

Abstract

Abstract: 【Objective】 The study was aimed to provide biological materials for developing phage "cocktail" preparations for the prevention and treatment of dairy mastitis.【Method】 Staphylococcus aureus 82 isolated from cow mastitis was used as the host bacteria, the phages were isolated and purified from the sewage, abandoned milk and feces of dairy farms by drip method and double-layer plate method, the morphological characteristics of the phage were observed by plaque and transmission electron microscope.The nucleic acid type of the phage was analyzed by nuclease treatment.The pyrolysis spectrum, MOI, one-step growth curve, thermal stability and pH stability were studied and analyzed.【Result】 The lytic phage P82 was isolated and screened.Under electron microscope, the head of the lytic phage P82 was icosahedral with a size of about 120 nm×83 nm, and the length of the lytic phage P82 was about 126 nm, it belonged to the order of Endurophage, and the family of Myoendurophage.The nucleic acid type was identified as dsDNA by agarose gel electrophoresis.The biological characteristics of the phage were determined:The best MOI of P82 was 0.001, and the cleavage rate of Staphylococcus aureus was 36.51%(23/63).The host spectrum was wide.The incubation period of growth was about 25 min, the outbreak period was about 45 min, and the cleavage volume was about 74 PFU/cell.P82 was stable at pH 4.0-12.0, and had high thermal stability.It completely lost activity at pH 2.0 and 13.0.It still had cracking ability at 70 ℃ for 60 min, but lost cracking ability at 80 ℃ for 40 min.【Conclusion】 The phage P82 isolated in this study had high titer, strong cleavage ability, fast proliferation speed, wide phage spectrum and good tolerance to different temperatures and pH.It could be used as a phage candidate strain for specific cleavage of Staphylococcus aureus from cow mastitis.It could be mixed with other phages to make a phage "cocktail" for the prevention and treatment of cow mastitis and provide materials for phage therapy of cow mastitis.

Key words: Staphylococcus aureus; cow mastitis; phage

CLC Number:

S852.65

DANG Ruiying, CHANG Junshuai, LIANG Yan, QU Yonggang, LI Yanfang, CHENG Lulu, LIU Ruotong, YANG Shengyuan, WANG Ziyang, ZHANG Xiaoyu, ZHAO Yu, ZHOU Haiqin, XIE Fangde. Isolation and Characterization of Staphylococcus aureus Phage P82 from Dairy Mastitis[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2318-2325.

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Isolation and Characterization of Staphylococcus aureus Phage P82 from Dairy Mastitis

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