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Open Access Highly Accessed Research article

Molecular typing of Coxiella burnetii from animal and environmental matrices during Q fever epidemics in the Netherlands

Arnout de Bruin1*, Pleunie TW van Alphen2, Rozemarijn QJ van der Plaats1, Lianne ND de Heer1, Chantal BEM Reusken1, Bart J van Rotterdam1 and Ingmar Janse1

Author Affiliations

1 National Institute for Public Health and the Environment (RIVM), Centre for infectious Disease Control (Cib), Laboratory for Zoonoses and Environmental Microbiology (LZO), PO Box 1, Bilthoven, 3720, BA, the Netherlands

2 Laboratories for Pathology and Medical Microbiology (PAMM Foundation), PO Box 2, Veldhoven, 5500, AA, the Netherlands

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BMC Veterinary Research 2012, 8:165  doi:10.1186/1746-6148-8-165

Published: 18 September 2012

Abstract

Background

The bacterium Coxiella burnetii has caused unprecedented outbreaks of Q fever in the Netherlands between 2007 and 2010. Since 2007, over 4000 human cases have been reported, with 2354 cases in 2009 alone. Dairy goat farms were identified as most probable sources for emerging clusters of human Q fever cases in their vicinity. However, identifying individual farms as primary source for specific clusters of human cases remains a challenge, partly due to limited knowledge of the different C. burnetii strains circulating in livestock, the environment and humans.

Results

We used a multiplex multi-locus variable number of tandem repeats analysis (MLVA) assay to investigate the genotypic diversity of C. burnetii in different types of samples that were collected nationwide during the Dutch Q fever outbreaks between 2007 and 2010. Typing was performed on C. burnetii positive samples obtained from several independent studies investigating C. burnetii presence in animals and the environment. Six different genotypes were identified on 45 farm locations, based on sequence-confirmed estimates of repeat numbers of six MLVA markers. MLVA genotype A was observed on 38 of the 45 selected farm locations in animals and in environmental samples.

Conclusions

Sequence confirmation of the numbers of tandem repeats within each locus and consensus about repeat identification is essential for accurate MLVA typing of C. burnetii. MLVA genotype A is the most common genotype in animal samples obtained from goat, sheep, and rats, as well as in environmental samples such as (aerosolized) dust, which is considered to be the major transmission route from animals via the environment to humans. The finding of a single dominant MLVA genotype in patients, the environment, and livestock complicates accurate source-finding. Pinpointing individual sources in the Netherlands requires discrimination of genotypes at a higher resolution than attained by using MLVA, as it is likely that the dominant C. burnetii MLVA type will be detected on several farms and in different patients in a particular area of interest.

Keywords:
Coxiella burnetii; Q fever; Molecular typing; MLVA; Environment; Goat; Sheep