Mycobacterium bovis Persistence in two Different Captive Wild Animal Populations in Germany: A Longitudinal Molecular Epidemiological Study Revealing Pathogen Transmission by Whole Genome Sequencing

Bovine tuberculosis (bTB) caused by Mycobacterium bovis is a transmissible disease notifiable to the World Organization for Animal Health and to European Union, with ongoing efforts of surveillance and eradication in every EU member state. In Germany, a country which has been declared officially free from bovine tuberculosis since 1997 by the EU, M. bovis infections still occur sporadically in cattle and other mammals, including humans. Here, the transmission routes of a bovine TB outbreak in a wildlife park in Germany affecting different cervid species, bison, lynx, and pot-bellied pig were followed employing whole genome sequencing (WGS) combined with spoligotyping and MIRU-VNTR-typing. One single M. bovis strain persisted from 2002 – 2015, and transmission between the park and a distantly located captive cervid farm was verified. The spoligotyping pattern remained identical while MIRU-VNTR-typing of 24 loci of the standardized panel and locus 2163a as additional locus revealed one change at locus 2165 in a strain from a fallow deer, and one at locus 2461 in isolates from red deer over the whole time period. WGS analysis confirmed close relatedness of the isolates, with a maximum of 12 SNPs detected between any two sequenced isolates. In conclusion, our data confirm a longitudinal outbreak of M. bovis in a German wildlife park and provide first insights into the dynamics of different genotyping markers in M. bovis.