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多价溶菌噬菌体 GSP044 的宿主受体鉴定,及其在清除沙门氏菌中的疗效初步评估。

Host receptor identification of a polyvalent lytic phage GSP044, and preliminary assessment of its efficacy in the clearance of Salmonella.

机构信息

National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan, Hubei 430070, People's Republic of China.

National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, People's Republic of China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan, Hubei 430070, People's Republic of China.

出版信息

Microbiol Res. 2023 Aug;273:127412. doi: 10.1016/j.micres.2023.127412. Epub 2023 May 17.

Abstract

Salmonella and pathogenic Escherichia coli are important foodborne pathogens. Phages are being recognized as potential antibacterial agents to control foodborne pathogens. In the current study, a polyvalent broad-spectrum phage, GSP044, was isolated from pig farm sewage. It can simultaneously lyse many different serotypes of Salmonella and E. coli, exhibiting a broad host range. Using S. Enteritidis SE006 as the host bacterium, phage GSP044 was further characterized. GSP044 has a short latent period (10 min), high stability at different temperatures and pH, and good tolerance to chloroform. Genome sequencing analysis revealed that GSP044 has a double-stranded DNA (dsDNA) genome consisting of 110,563 bp with G + C content of 39%, and phylogenetic analysis of the terminase large subunit confirmed that GSP044 belonged to the Demerecviridae family, Epseptimavirus genus. In addition, the genomic sequence did not contain any lysogenicity-related, virulence-related, or antibiotic resistance-related genes. Analysis of phage-targeted host receptors revealed that the outer membrane protein (OMP) BtuB was identified as a required receptor for phage infection of host bacteria. The initial application capability of phage GSP044 was assessed using S. Enteritidis SE006. Phage GSP044 could effectively reduce biofilm formation and degrade the mature biofilm in vitro. Moreover, GSP044 significantly decreased the viable counts of artificially contaminated S. Enteritidis in chicken feed and drinking water. In vivo tests, a mouse model of intestinal infection demonstrated that phage GSP044 was able to reduce the number of colonized S. Enteritidis in the intestine. These results suggest that phage GSP044 may be a promising candidate biologic agent for controlling Salmonella infections.

摘要

肠炎沙门氏菌和致病性大肠杆菌是重要的食源性病原体。噬菌体被认为是控制食源性病原体的潜在抗菌剂。在本研究中,从猪场污水中分离到一种多价广谱噬菌体 GSP044,它可以同时裂解多种不同血清型的沙门氏菌和大肠杆菌,表现出广泛的宿主范围。使用肠炎沙门氏菌 SE006 作为宿主菌,进一步对噬菌体 GSP044 进行了表征。GSP044 的潜伏期短(10 分钟),在不同温度和 pH 值下稳定,对氯仿有良好的耐受性。基因组测序分析表明,GSP044 具有一个由 110,563 bp 组成的双链 DNA (dsDNA)基因组,G + C 含量为 39%,末端酶大亚基的系统发育分析证实 GSP044 属于 Demerecviridae 科,Epseptimavirus 属。此外,基因组序列不包含任何溶原性、毒性或抗生素耐药性相关基因。噬菌体靶向宿主受体的分析表明,外膜蛋白 (OMP) BtuB 被鉴定为噬菌体感染宿主细菌所必需的受体。使用肠炎沙门氏菌 SE006 评估了噬菌体 GSP044 的初步应用能力。噬菌体 GSP044 可以有效地减少体外生物膜的形成并降解成熟的生物膜。此外,GSP044 显著降低了鸡饲料和饮用水中人工污染肠炎沙门氏菌的活菌数。体内试验中,肠道感染的小鼠模型表明,噬菌体 GSP044 能够减少肠道中定植的肠炎沙门氏菌数量。这些结果表明,噬菌体 GSP044 可能是控制沙门氏菌感染的一种有前途的生物制剂候选物。

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