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Characterization and complete genome sequence analysis of a novel virulent Siphoviridae phage against Staphylococcus aureus isolated from bovine mastitis in Xinjiang, China

Virus Genes volume 53, pages 464–476 (2017)Cite this article

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.

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Abbreviations

S. aureus:

Staphylococcus aureus

S. pneumoniae :

Streptococcus pneumoniae

MOI:

Optimal multiplicity of infection

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Authors’ contributions

Yigang Tong and Yonggang Qu conceived and designed the experiments and critically evaluated the manuscript. Xianglilan Zhang carried out the data analysis and wrote the manuscript. Qian Zhang isolated and identified the phage and carried out the experiments. Shaozhen Xing extracted the phage nucleotide. Qiang Sun analyzed the phage sequences. Guangqian Pei, Shi Cheng, Yannan Liu, and Xiaoping An conducted the sequencing experiments. All authors read and approved the final manuscript.

Funding

This research was supported by grants from The National Key Research and Development Program of China (2015AA020108, 2016YFC1202705, AWS16J020, and AWS15J006), the National Natural Science Foundation of China (81572045, 81672001, and 81621005), and the State Key Laboratory of Pathogen and biosecurity (SKLPBS1518).

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Affiliations

  1. College of Animal Science and Technology, Shihezi University, Shihezi, 832003, Xinjiang, China

    Qian Zhang & Yonggang Qu

  2. State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China

    Shaozhen Xing, Qiang Sun, Guangqian Pei, Shi Cheng, Yannan Liu, Xiaoping An, Xianglilan Zhang & Yigang Tong

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  1. Qian Zhang
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Correspondence to Xianglilan Zhang, Yonggang Qu or Yigang Tong.

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Additional information

Genome accession number

The GenBank accession number for its complete nucleotide sequence is [GenBank: KX156762].

Edited by Joachim Jakob Bugert.

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Zhang, Q., Xing, S., Sun, Q. et al. Characterization and complete genome sequence analysis of a novel virulent Siphoviridae phage against Staphylococcus aureus isolated from bovine mastitis in Xinjiang, China. Virus Genes 53, 464–476 (2017). https://doi.org/10.1007/s11262-017-1445-z

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  • DOI: https://doi.org/10.1007/s11262-017-1445-z

Keywords

  • Bovine mastitis
  • Staphylococcus aureus
  • Phage
  • Genome termini
  • Complete genome