title: Characterization and complete genome sequence analysis of a novel virulent Siphoviridae phage against Staphylococcus aureus isolated from bovine mastitis in Xinjiang, China.
abstract: Bovine mastitis is one of the most costly diseases in dairy cows worldwide. It can be caused by over 150 different microorganisms, where Staphylococcus aureus is the most frequently isolated and a major pathogen responsible for heavy economic losses in dairy industry. Although antibiotic therapy is most widely used, alternative treatments are necessary due to the increasing antibiotic resistance. Using phage for pathogen control is a promising tool in the fight against antibiotic resistance. Mainly using high-throughput sequencing, bioinformatics and our proposed phage termini identification method, we have isolated and characterized a novel virulent phage, designated as vB_SauS_IMEP5, from manure collected from dairy farms in Shihezi, Xinjiang, China, for use as a biocontrol agent against Staphylococcus aureus infections. Its latent period was about 30 min and its burst size was approximately 272PFU/cell. Phage vB_SauS_IMEP5 survives in a wide pH range between 3 and 12. A treatment at 70 °C for 20 min can inactive the phage. Morphological analysis of vB_SauS_IMEP5 revealed that phage vB_SauS_IMEP5 morphologically resembles phages in the family Siphoviridae. Among our tested multiplicity of infections (MOIs), the optimal multiplicity of infection (MOI) of this phage was determined to be 0.001, suggesting that phage vB_SauS_IMEP5 has high bacteriolytic potential and good efficiency for reducing bacterial growth. The complete genome of IME-P5 is a 44,677-bp, linear, double-stranded DNA, with a G+C content of 34.26%, containing 69 putative ORFs. The termini of genome were determined with next-generation sequencing data using our previously proposed termini identification method, which suggests that this phage has non-redundant termini with 9nt 3' protruding cohesive ends. The genomic and proteomic characteristics of IMEP5 demonstrate that this phage does not belong to any of the previously recognized Siphoviridae Staphylococcus phage groups, suggesting the creation of a new lineage, thus adding to the knowledge on the diversity of Staphylococcus phages. An N-acetylmuramoyl-L-alanine amidase gene and several conserved genes were predicted, while no virulence or antibiotic resistance genes were identified. This study isolated and characterized a novel S. aureus phage vB_SauS_IMEP5, and our findings suggest that this phage may be potentially utilized as a therapeutic or prophylactic candidate against S.aureus infections.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, 832003, Xinjiang, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Shaozhen
authors.initials: S
authors.lastname: Xing
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Guangqian
authors.initials: G
authors.lastname: Pei
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Shi
authors.initials: S
authors.lastname: Cheng
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Yannan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: An
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China. zhangxianglilan@gmail.com.
authors.forename: Xianglilan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, 832003, Xinjiang, China. quyonggang@shzu.edu.cn.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Qu
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China. tong62035@163.com.
authors.forename: Yigang
authors.initials: Y
authors.lastname: Tong
mesh_terms: D000818:Animals; D001433:Bacteriolysis; D001482:Base Composition; D061046:Biological Control Agents; D002417:Cattle; D002681:China; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D016679:Genome, Viral; D059014:High-Throughput Nucleotide Sequencing; D008414:Mastitis, Bovine / Q000382:microbiology*; D000071059:Phage Therapy / Q000379:methods; D017422:Sequence Analysis, DNA; D017906:Siphoviridae / Q000235:genetics* / Q000302:isolation & purification*; D013203:Staphylococcal Infections / Q000382:microbiology / Q000662:veterinary*; D013204:Staphylococcus Phages / Q000235:genetics*; D013211:Staphylococcus aureus / Q000254:growth & development / Q000302:isolation & purification / Q000821:virology*; D014774:Virulence; D000073336:Whole Genome Sequencing
chemical_list: D061046:Biological Control Agents; D004279:DNA, Viral
keywords: Bovine mastitis; Complete genome; Genome termini; Phage; Staphylococcus aureus