Abstract

Mycoplasma hyorhinis is one of the causative agents of polyserositis and arthritis in post-weaning pigs. Here we describe the development of a multi-locus sequence typing (MLST) protocol for the characterization of M. hyorhinis field isolates. A total of 104 field isolates from different geographical locations, swine production systems, and clinical backgrounds, were analyzed. Twenty-seven genes, including housekeeping and those encoding surface proteins, were evaluated to index diversity. Genes encoding surface proteins were included to increase the discriminatory power of the MLST. Four target gene fragments were selected to be included in the final MLST-s (surface) protocol: pdhB, p95, mtlD and ung. Within each locus the nucleotide variation ranged from 1.4% to 20%. The 104 field isolates were classified into 39 distinct sequence types (STs). Multiple STs were found within the same production system and within the same pig. The majority of STs grouped strains from the same production system; however, cases existed where multiple systems shared a ST, indicating potential relationships between pig flows. The majority of the nucleotide changes observed in these genes generated synonymous changes, while non-synonymous changes were exclusively in the mtlD gene fragment, suggesting that this protein is undergoing selection. Molecular typing of M. hyorhinis will primarily aid swine practitioners with pig flow management and identifying sources of infection during outbreaks.

Highlights

  • In recent years, Mycoplasma hyorhinis has been recognized as an important cause of mortality in nursery pigs [1,2]

  • We describe the development of a novel multi-locus sequence typing (MLST) assay based on housekeeping and surface-encoding proteins to differentiate M. hyorhinis strains, termed MLST-s to reference the addition of surface proteins

  • While variation was observed within the nrdf and lspA surface protein genes when comparing the 4 annotated genomes, testing of the initial 6 M. hyorhinis isolates revealed no nucleotide sequence variation and were not included for further testing

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Summary

Introduction

Mycoplasma hyorhinis has been recognized as an important cause of mortality in nursery pigs [1,2]. This pathogen colonizes the nasal and oropharyngeal epithelial surface of pigs. Piglets presumably become colonized through contact with sows, and the bacterium is transmitted via nose-to-nose contact among pigs following colonization and infection [3]. Molecular epidemiology of M. hyorhinis field isolates

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