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新分离的感染金黄色葡萄球菌的烈性噬菌体KMSP1的特性及其在乳制品中的应用

Characterization of KMSP1, a newly isolated virulent bacteriophage infecting Staphylococcus aureus, and its application to dairy products.

作者信息

Kwak Hyerim, Kim Jinshil, Ryu Sangryeol, Bai Jaewoo

机构信息

Division of Applied Food System, Major in Food Science & Technology, Seoul Women's University, Seoul 01797, Republic of Korea.

Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea; Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea.

出版信息

Int J Food Microbiol. 2023 Apr 2;390:110119. doi: 10.1016/j.ijfoodmicro.2023.110119. Epub 2023 Feb 3.

Abstract

Staphylococcus aureus is one of the major pathogens causing foodborne outbreaks and severe infections worldwide. Generally, various physical and chemical treatments have been applied to control S. aureus in the food industry. However, conventional treatments usually affected food quality and often produced toxic compounds. Therefore, bacteriophage (phage), a natural antimicrobial agent, has been suggested as an alternative strategy to control foodborne pathogens including S. aureus. In this study, KMSP1, a bacteriophage infecting S. aureus was isolated from a raw milk sample and characterized. Transmission electron microscopy (TEM) analysis revealed that phage KMSP1 belongs to the Myoviridae family. Phage KMSP1 efficiently inhibited bacterial growth for >28 h post-infection. In addition, phage KMSP1 could infect a broad spectrum of S. aureus strains, including methicillin-resistant S. aureus (MRSA) strains. Whole-genome sequence analysis showed that KMSP1 is a lytic phage with the absence of genes related to lysogen formation, toxin production, and antibiotics resistance, respectively. In the genome of KMSP1, the presence of putative tail lysin containing a cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) domain could be one of the reasons for the effective antimicrobial activity of KMSP1. Furthermore, high stability of phage KMSP1 at temperature ranging from 4 to 55 °C and pH ranging from 5 to 11, suggested its potential use in various food systems. Receptor analysis revealed that KMSP1 utilized cell wall teichoic acid (WTA), one of the major virulence factors of S. aureus, as a host receptor. Application of phage KMSP1 at an MOI of 10 achieved a significant reduction of log 8.8 CFU/mL of viable cell number in pasteurized milk and log 4.3 CFU/cm in sliced cheddar cheese after 24 h. Taken together, the strong antimicrobial activity of phage KMSP1 suggested that it could be developed as a biocontrol agent in dairy products to control S. aureus contamination.

摘要

金黄色葡萄球菌是全球范围内引起食源性疾病暴发和严重感染的主要病原体之一。一般来说,食品工业中已采用各种物理和化学处理方法来控制金黄色葡萄球菌。然而,传统处理方法通常会影响食品质量,且常常产生有毒化合物。因此,噬菌体作为一种天然抗菌剂,已被提议作为控制包括金黄色葡萄球菌在内的食源性病原体的替代策略。在本研究中,从一份生乳样品中分离并鉴定了一株感染金黄色葡萄球菌的噬菌体KMSP1。透射电子显微镜(TEM)分析表明,噬菌体KMSP1属于肌尾噬菌体科。噬菌体KMSP1在感染后>28小时能有效抑制细菌生长。此外,噬菌体KMSP1可感染多种金黄色葡萄球菌菌株,包括耐甲氧西林金黄色葡萄球菌(MRSA)菌株。全基因组序列分析表明,KMSP1是一种裂解性噬菌体,分别不存在与溶原形成、毒素产生和抗生素抗性相关的基因。在KMSP1的基因组中,存在含有半胱氨酸/组氨酸依赖性酰胺水解酶/肽酶(CHAP)结构域的推定尾溶素,这可能是KMSP1具有有效抗菌活性的原因之一。此外,噬菌体KMSP1在4至55°C的温度范围和5至11的pH范围内具有高稳定性,表明其在各种食品体系中的潜在应用价值。受体分析表明,KMSP1利用细胞壁磷壁酸(WTA)作为宿主受体,WTA是金黄色葡萄球菌的主要毒力因子之一。以10的感染复数(MOI)应用噬菌体KMSP1后,经过24小时,巴氏杀菌乳中的活菌数显著降低了log 8.8 CFU/mL,切达干酪切片中的活菌数显著降低了log 4.3 CFU/cm。综上所述,噬菌体KMSP1强大的抗菌活性表明它可被开发为乳制品中的生物防治剂,以控制金黄色葡萄球菌污染。

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