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Applied and Environmental Microbiology, February 2006, p. 1420-1428, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1420-1428.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

First Insights into the Evolution of Streptococcus uberis: a Multilocus Sequence Typing Scheme That Enables Investigation of Its Population Biology

Tracey J. Coffey,1 Gillian D. Pullinger,1 Rachel Urwin,2 Keith A. Jolley,3 Stephen M. Wilson,1 Martin C. Maiden,3 and James A. Leigh1*

Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, United Kingdom,1 Pennsylvania State University, 0208 Mueller Lab, University Park, Pennsylvania 16802,2 The Peter Medawar Building for Pathogen Research, University of Oxford, South Parks Road, Oxford OX1 3SY, United Kingdom3

Received 30 September 2005/ Accepted 29 November 2005

Intramammary infection with Streptococcus uberis is a common cause of bovine mastitis throughout the world. Several procedures to differentiate S. uberis isolates have been proposed. However, all are prone to interlaboratory variation, and none is suitable for the description of the population structure. We describe here the development of a multilocus sequence typing (MLST) scheme for S. uberis to help address these issues. The sequences of seven housekeeping gene fragments from each of 160 United Kingdom milk isolates of S. uberis were determined. Between 5 and 17 alleles were obtained per locus, giving the potential to discriminate between 1.3 x 107 sequence types. In this study, 57 sequence types (STs) were identified. Statistical comparisons between the maximum-likelihood trees constructed by using the seven housekeeping gene fragments showed that the congruence was no better than that between each tree and trees of random topology, indicating there had been significant recombination within these loci. The population contained one major lineage (designated the ST-5 complex). This dominated the population, containing 24 STs and representing 112 isolates. The other 33 STs were not assigned to any clonal complex. All of the isolates in the ST-5 lineage carried hasA, a gene that is essential for capsule production. There was no clear association between ST or clonal complex and disease. The S. uberis MLST system offers researchers a valuable tool that allows further investigation of the population biology of this organism and insights into the epidemiology of this disease on a global scale.

* Corresponding author. Present address: Nuffield Department of Clinical Laboratory Sciences, John Radcliffe Hospital, University of Oxford, Oxford OX3 9DU, United Kingdom. Phone: 44 1865 221226. Fax: 44 1865 764192. E-mail: james.leigh{at}ndcls.ox.ac.uk.

Applied and Environmental Microbiology, February 2006, p. 1420-1428, Vol. 72, No. 2
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.2.1420-1428.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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