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Open Access Research article

Phylogeny of Mycoplasma bovis isolates from Hungary based on multi locus sequence typing and multiple-locus variable-number tandem repeat analysis

Kinga M Sulyok1, Zsuzsa Kreizinger1, Lilla Fekete1, Szilárd Jánosi2, Nóra Schweitzer2, Ibolya Turcsányi2, László Makrai3, Károly Erdélyi2 and Miklós Gyuranecz1*

Author Affiliations

1 Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Hungária körút 21, Budapest 1143, Hungary

2 Veterinary Diagnostic Directorate, National Food Chain Safety Office, Tábornok utca 2, Budapest 1143, Hungary

3 Faculty of Veterinary Science, Szent István University, Hungária körút 23-25, Budapest 1143, Hungary

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BMC Veterinary Research 2014, 10:108  doi:10.1186/1746-6148-10-108

Published: 7 May 2014

Abstract

Background

Mycoplasma bovis is an important pathogen causing pneumonia, mastitis and arthritis in cattle worldwide. As this agent is primarily transmitted by direct contact and spread through animal movements, efficient genotyping systems are essential for the monitoring of the disease and for epidemiological investigations. The aim of this study was to compare and evaluate the multi locus sequence typing (MLST) and the multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) through the genetic characterization of M. bovis isolates from Hungary.

Results

Thirty one Hungarian M. bovis isolates grouped into two clades by MLST. Two strains had the same sequence type (ST) as reference strain PG45, while the other twenty nine Hungarian isolates formed a novel clade comprising five subclades. Isolates originating from the same herds had the same STs except for one case. The same isolates formed two main clades and several subclades and branches by MLVA. One clade contained the reference strain PG45 and three isolates, while the other main clade comprised the rest of the strains. Within-herd strain divergence was also detected by MLVA. Little congruence was found between the results of the two typing systems.

Conclusions

MLST is generally considered an intermediate scale typing method and it was found to be discriminatory among the Hungarian M. bovis isolates. MLVA proved to be an appropriate fine scale typing tool for M. bovis as this method was able to distinguish closely related strains isolated from the same farm. We recommend the combined use of the two methods for the genotyping of M. bovis isolates. Strains have to be characterized first by MLST followed by the fine scale typing of identical STs with MLVA.

Keywords:
Cattle; Genotyping; MLST; MLVA; Mycoplasma bovis; Phylogeny; VNTR