Collection of immature Dermacentor reticulatus (Fabricius, 1794) ticks from vegetation and detection of Rickettsia raoultii in them

It is commonly assumed that Dermacentor reticulatus immature life stages are nidicolous and therefore cannot be collected from vegetation. However, in June and July of 2018 and 2019, a total of 47 questing D. reticulatus larvae and two nymphs were collected by the flagging method in two different sites close to the city of Leipzig, Germany. To confirm their role in the transmission of tick-borne pathogens, 45 larvae (pooled by 2 in 21 pools and 1 pool with three individuals) and one nymph were tested either by conventional or real-time PCR for the presence of Bartonella spp., Neoehrlichia mikurensis, Rickettsia spp., and Babesia spp. All samples tested negative for Bartonella spp., N. mikurensis, and Babesia spp.; while the minimal infection rate of larvae for Rickettsia spp. was 42%, and the one tested nymph was also positive. Sequencing partial ompB genes revealed the presence of Rickettsia raoultii in larvae and nymph. Further research needs to be done to determine under which circumstances immature D. reticulatus ticks are found outside the burrows of their hosts and can be collected from vegetation.