title: Molecular analysis of Coxiella burnetii in Germany reveals evolution of unique clonal clusters.
abstract: The causative agent of Q fever, Coxiella burnetii, is a query agent occurring naturally all over the world. We studied 104 German Coxiella burnetii strains/DNA samples obtained between 1969 and 2011 using a 14 microsatellite marker Multiple-locus variable-number of tandem repeat (VNTR) analysis (MLVA) technique. We were able to divide our collection into 32 different genotypes clustered into four major groups (A-D). Two of these (A and C) formed predominant clonal complexes that covered 97% of all studied samples. Group C consisted exclusively of cattle-associated isolates/DNA specimens, while group A comprised all other affected species including all sheep-derived strains/DNA samples. Within this second cluster, two major genotypes (A1, A2) were identified. Genotype A2 occurred in strains isolated from ewes in northern and central Germany, whereas genotype A1 was found in most areas of Germany. MLVA analysis of C. burnetii strains from neighbouring countries revealed a close relationship to German strains. We thus hypothesize that there is a western and central European cluster of C. burnetii. We identified predominant genotypes related to relevant host species and geographic regions which is in line with findings of the Dutch Q fever outbreak (2007-2010). Furthermore three of our analyzed German strains are closely related to the Dutch outbreak clone. These findings support the theory of predominant genotypes in the context of regional outbreaks. Our results show that a combination of 8 MLVA markers provides the highest discriminatory power for attributing C. burnetii isolates to genotypes. For future epidemiological studies we propose the use of three MLVA markers for easy and rapid classification of C. burnetii into 4 main clusters.
authors.affiliation: Bundeswehr Institute of Microbiology, Munich, Germany. Electronic address: DimitriosFrangoulidis@Bundeswehr.org.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Frangoulidis
authors.affiliation: Helmholtz Center for Health and Environment, Neuherberg, Germany; Department of Genome-Oriented Bioinformatics, Technische Universität München, Freising, Germany.
authors.forename: Mathias C
authors.initials: MC
authors.lastname: Walter
authors.affiliation: Bundeswehr Institute of Microbiology, Munich, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Antwerpen
authors.affiliation: Bavarian Health and Food Safety Authority, Oberschleissheim, Germany.
authors.forename: Pia
authors.initials: P
authors.lastname: Zimmermann
authors.affiliation: Bavarian Animal Health Service, Poing, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Janowetz
authors.affiliation: Bavarian Animal Health Service, Poing, Germany.
authors.forename: Michaela
authors.initials: M
authors.lastname: Alex
authors.affiliation: Bavarian Animal Health Service, Poing, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Böttcher
authors.affiliation: Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, Wusterhausen, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Henning
authors.affiliation: Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, Wusterhausen, Germany.
authors.forename: Angela
authors.initials: A
authors.lastname: Hilbert
authors.affiliation: Clinic for Swine and Small Ruminants, University of Veterinary Medicine Hannover Foundation, Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Ganter
authors.affiliation: Lower Saxony State Office for Consumer Protection and Food Safety (LAVES), Food and Veterinary Institute, Braunschweig/Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Runge
authors.affiliation: Helmholtz Center for Health and Environment, Neuherberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Münsterkötter
authors.affiliation: Bundeswehr Institute of Microbiology, Munich, Germany.
authors.forename: Wolf D
authors.initials: WD
authors.lastname: Splettstoesser
authors.affiliation: Bundeswehr Institute of Microbiology, Munich, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Hanczaruk
mesh_terms: D000818:Animals; D002417:Cattle; D016997:Coxiella burnetii / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D014644:Genetic Variation; D005838:Genotype; D005858:Germany; D006801:Humans; D018598:Minisatellite Repeats; D058889:Molecular Typing; D058974:Phylogeography; D011778:Q Fever / Q000382:microbiology* / Q000662:veterinary*; D012756:Sheep
keywords: Germany; MLVA; Molecular epidemiology; Q fever; Typing; VNTR