Borrelia prevalence and species distribution in ticks removed from humans in Germany, 2013-2017

Lyme borreliosis caused by spirochaetes of the Borrelia burgdorferi sensu lato (s.l.) complex is the most common tick-borne disease in Europe. In addition, the relapsing-fever spirochaete Borrelia miyamotoi, which has been associated with febrile illness and meningoencephalitis in immunocompromised persons, is present in Europe. This study investigated Borrelia prevalence and species distribution in ticks removed from humans and sent as diagnostic material to the Institute for Parasitology, University of Veterinary Medicine Hannover, in 2013-2017. A probe-based real-time PCR was carried out and Borrelia-positive samples were subjected to species determination by reverse line blot (RLB), including a B. miyamotoi-specific probe. The overall Borrelia-infection rate as determined by real-time PCR was 20.02 % (510/2547, 95 % CI: 18.48-21.63 %), with annual prevalences ranging from 17.17 % (90/524, 95 % CI: 14.04-20.68 %) in 2014 to 24.12 % (96/398, 95 % CI: 19.99-28.63 %) in 2015. In total, 271/475 (57.1 %) positive samples available for RLB were successfully differentiated. Borrelia afzelii was detected in 30.53 % of cases (145/475, 95 % CI: 26.41-34.89), followed by B. garinii/B. bavariensis (13.26 % [63/475], 95 % CI: 10.34-16.65). Borrelia valaisiana occurred in 5.89 % (28/475, 95 % CI: 3.95-8.41), B. spielmanii in 4.63 % (22/475, 95 % CI: 2.93-6.93), B. burgdorferi sensu stricto (s.s.)/B. carolinensis in 2.32 % (11/475, 95 % CI: 1.16-4.11), B. lusitaniae in 0.63 % (3/475, 95 % CI: 0.13-1.83) and B. bisettiae in 0.42 % (2/475, 95 % CI: 0.05-1.51) of positive ticks. Borrelia kurtenbachii was not detected, while B. miyamotoi was identified in 7.37 % (35/475, 95 % CI: 5.19-10.10) of real-time PCR-positive samples. Sanger sequencing of B. garinii/B. bavariensis-positive ticks revealed that the majority were B. garinii-infections (50/52 successfully amplified samples), while only 2 ticks were infected with B. bavariensis. Furthermore, 6/12 B. burgdorferi s.s./B. carolinensis-positive samples could be differentiated; all of them were identified as B. burgdorferi sensu stricto. Thirty-nine ticks (8.21 %, 95 % CI: 5.90-11.05) were coinfected with two different species. Comparison of the species distribution between ticks removed from humans in 2015 and questing ticks collected in the same year and the same area revealed a significantly higher B. afzelii-prevalence in diagnostic tick samples than in questing ticks, confirming previous observations. The obtained data indicate that Borrelia prevalence fluctuated in the same range as observed in a previous study, analysing the period from 2006 to 2012. Detection of B. miyamotoi in 7.37 % of Borrelia-positive samples points to the fact that clinicians should be aware of this pathogen as a differential diagnosis in cases of febrile illness.