Next-generation sequencing analysis of bacterial flora in bovine Prototheca mastitic milk

Rui Kano, Yuki Kobayashi, Akitoyo Nishikawa, Ryo Murata, Takuya Itou, Takaaki Ito, Kazuyuki Suzuki, Hiroshi Kamata

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Prototheca zopfii is an achlorophyllic algae that causes bovine mastitis, resulting in a reduction in milk production and the secretion of thin, watery milk with white flakes. The aim of the present study was to evaluate the bacterial flora in the udder environment in protothecal mastitis. We used metagenomic next-generation sequencing (NGS) analysis to identify 16S rRNA genes from bacterial flora present in milk samples from protothecal mastitic dairy cows. Seven clinical strains of P. zopfii genotype 2 were isolated from 7 milk samples from 7 cases (Holstein cow) of protothecal mastitis; another 9 milk samples were obtained from 9 normal Holstein cows. The samples were collected in 2017 from cows in one dairy located in the Kushiro region in Hokkaido, Japan, which had a history of protothecal mastitis infection. The NGS produced 10,000 to 15,000 sequences in each DNA sample. To facilitate comparison, we grouped the sequencing results according to the culture-based protothecal mastitis diagnosis; sequences derived from the milk samples obtained from healthy cows were grouped separately. Sequences classified as Streptococcus spp., Pseudomonas spp. and Sphingomonas spp., Caulobacter segnis, Macrococcus caseolyticus, Methylobacterium tarhaniae, andSphingomonas leidyi were the main sequences detected in the groups of samples from cows characterized by culture as having protothecal mastitis. Notably, Calothrix desertica (a cyanobacterium) sequences showed higher prevalence in these samples. To our knowledge, this is the first study to report that C. desertica sequences, effectively absent in the samples derived from healthy cows, are detected at high prevalence in samples from protothecal mastitic animals.

Original languageEnglish
Pages (from-to)E41-E46
JournalMedical Mycology Journal
Volume59
Issue number3
DOIs
StatePublished - 2018
Externally publishedYes

Keywords

  • Bacterial flora
  • Bovine prototheca mastitis
  • Next-generation sequencing
  • Prototheca zopfii

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