Detection and Characterization of Alongshan Virus in Ticks and Tick Saliva from Lower Saxony, Germany with Serological Evidence for Viral Transmission to Game and Domestic Animals.

The newly discovered group of Jingmenviruses has been shown to infect a wide range of hosts and has been associated with febrile illness in humans. During a survey for Jingmenviruses in ticks from Lower Saxony, Germany, Alongshan virus (ALSV) was identified in Ixodes spp. ticks. Additional virus screenings revealed the presence of ALSV in the bodies and saliva of ticks collected at several locations in Lower Saxony. Vector competence studies that included Ixodes ricinus and Dermacentor reticulatus validated the replication of ALSV within those tick species. In vitro feeding experiments with ALSV-injected Ixodes ricinus demonstrated effective viral transmission during blood feeding. To evaluate the potential viral transmission during a natural blood meal, sera from wild game and domestic animals were investigated. One serum sample from a red deer was found to be positive for ALSV RNA, while serological screenings in game and domestic animals revealed the presence of ALSV-specific antibodies at different locations in Lower Saxony. Overall, those results demonstrate the broad distribution of ALSV in ticks in Lower Saxony and hypothesize frequent exposure to animals based on serological investigations. Hence, its potential risk to human and animal health requires further investigation.

Regional, seasonal, biennial and landscape-associated distribution of Anaplasma phagocytophilum and Rickettsia spp. infections in Ixodes ticks in northern Germany and implications for risk assessment at larger spatial scales.

Tick-associated Rickettsiales are important pathogens with relevance for public and animal health; therefore, knowledge regarding their distribution is essential for risk assessment and disease prevention. To investigate the prevalence of Anaplasma phagocytophilum and Rickettsia spp. in northern Germany, Ixodes ticks were flagged monthly from April to October in 2018 and 2019 at three collection sites each in the regions of Bremen, Emsland, Hanover, Kassel and Uelzen. A total of 3150 ticks (1052 females, 1048 males and 1050 nymphs) were individually examined for rickettsial infections using probe-based quantitative real-time PCR. Overall prevalence of A. phagocytophilum was 6.4 % (202/3150; 6.7 % [71/1052] in females, 7.5 % [79/1048] in males and 5.0 % [52/1050] in nymphs). For Rickettsia spp., the overall prevalence was 29.6 % (931/3150; 33.4 % [351/1052] in females, 28.3 % [297/1048] in males and 27.0 % [283/1050] in nymphs). Rickettsia species identification by real-time pyrosequencing on a subset of 409 positive samples was successful in 407 cases (99.5 %). Rickettsia helvetica was the predominant species with a detection rate of 99.8 % (406/407). Additionally, Rickettsia monacensis was detected in one tick (0.2 %). Generalized linear mixed models showed significant regional as well as monthly differences regarding the prevalence of both pathogens. In addition, the prevalence of both pathogens was significantly higher in 2018 (A. phagocytophilum: 8.0 % [126/1575], Rickettsia spp.: 35.4 % [558/1575]) than in 2019 (A. phagocytophilum: 4.8 % [76/1575], Rickettsia spp.: 23.9 % [373/1575]). In contrast, no effect of landscape type on pathogen prevalence was found. As Rickettsia spp.-detection was based on the single-copy gene gltA, it was possible to calculate the individual pathogen load per tick, which was significantly higher in female ticks than in nymphs (mean values: 8.19 × 104 vs. 9.58 × 103). Regional, seasonal and biennial prevalence differences of tick-transmitted Rickettsiales show the necessity to investigate ticks from multiple locations, over several months and in more than one year to reliably assess the infection risk on a larger geographical scale.

Ixodes frontalis: a neglected but ubiquitous tick species in Germany.

The ornithophilic tick species Ixodes frontalis (Panzer) is spatially distributed in Europe, Asia and northern Africa. It can serve as a carrier of different bacteria and viruses, but little is known of its actual vector competence. In addition, adult females of this species are associated with the avian "tick-related syndrome" (TRS). Like most ornithophilic tick species, I. frontalis is usually collected from bird nests or directly from their hosts. Reports of I. frontalis being collected with the flagging method are scarce. In Germany, the species is considered as very rare. In the few reports that are available, the majority of ticks has been discovered on migrating birds. Therefore, knowledge of the actual distribution of this species in Germany was strictly limited. In this study, we report the finding of 1084 individuals of I. frontalis collected with the flagging method in urban areas and gardens in several regions of Germany. Furthermore, the species was discovered on 35 dead Eurasian blackbirds (Turdus merula) from all over Germany. Finally, we report of five new possible cases of TRS, three in free-ranging birds and two in captive birds, including a Harris's hawk (Parabuteo unicinctus) kept for falconry.