Background: Ixodes ricinus is the predominant tick species in Europe and the primary pathogen vector for both humans and animals. These ticks are frequently involved in the transmission of Borrelia burgdorferi (sensu lato), the causative agents of Lyme borreliosis. While much more is known about I. ricinus tick-borne pathogen composition, information about temporal tick-borne pathogen patterns remain scarce. These data are crucial for predicting seasonal/annual patterns which could improve understanding and prevent tick-borne diseases. Methods: We examined tick-borne pathogen (TBP) dynamics in I. ricinus collected monthly in a peri-urban forest over three consecutive years. In total, 998 nymphs were screened for 31 pathogenic species using high-throughput microfluidic real-time PCR. Results: We detected DNA from Anaplasma phagocytophilum (5.3%), Rickettsia helvetica (4.5%), Borrelia burgdorferi (s.l.) (3.7%), Borrelia miyamotoi (1.2%), Babesia venatorum (1.5%) and Rickettsia felis (0.1%). Among all analysed ticks, 15.9% were infected by at least one of these microorganisms, and 1.3% were co-infected. Co-infections with B. afzeli/B. garinii and B. garinii/B. spielmanii were significantly over-represented. Moreover, significant variations in seasonal and/or inter-annual prevalence were observed for several pathogens (R. helvetica, B. burgdorferi (s.l.), B. miyamotoi and A. phagocytophilum). Conclusions: Analysing TBP prevalence in monthly sampled tick over three years allowed us to assess seasonal and inter-annual fluctuations of the prevalence of TBPs known to circulate in the sampled area, but also to detect less common species. All these data emphasize that sporadic tick samplings are not sufficient to determine TBP prevalence and that regular monitoring is necessary.

Birds play a role in maintaining tick-borne diseases by contributing to the multiplication of ticks and pathogens on a local scale during the breeding season. In the present study, we describe the diversity of tick and pathogen species of medical and veterinary importance in Europe hosted by 1040 captured birds (56 species) during their breeding season in France. Of the 3114 ticks collected, Ixodes ricinus was the most prevalent species (89.5%), followed by I. frontalis (0.8%), I. arboricola (0.7%), Haemaphysalis concinna (0.5%), H. punctata (0.5%), Hyalomma spp. (0.2%), and Rhipicephalus spp. (0.06%). Because they may be representative of the bird infection status for some pathogen species, 1106 engorged tick larvae were screened for pathogens. Borrelia burgdorferi sensu lato was the most prevalent pathogen genus in bird-feeding larvae (11.7%), followed by Rickettsia spp. (7.4%), Anaplasma spp. (5.7%), Babesia spp. (2.3%), Ehrlichia spp. (1.4%), and B. miyamotoi (1%). Turdidae birds (Turdus merula and T. philomelos), Troglodytes troglodytes, and Anthus trivialis had a significantly higher prevalence of B. burgdorferi s.l.-infected larvae than other pathogen genera. This suggests that these bird species could act as reservoir hosts for B. burgdorferi s.l. during their breeding season, and thus play an important role in acarological risk.

The geographical distribution and impact on animal and human health of both West Nile and Usutu viruses, two flaviviruses of the Japanese encephalitis complex, have been increasing during the past two decades. Both viruses circulate in Europe and Africa within a natural cycle between wild birds and mosquitoes, mainly from the Culex genus. We retrospectively analyzed sera from domestic and wild birds sampled in 2008 in two wetlands, namely the Inner Niger Delta, Mali, and the Lake Alaotra area, Madagascar. Sera were first tested using a commercial ID Screen West Nile Competition Multi-species ELISA kit. Then, positive sera and sera with insufficient volume for testing with ELISA were tested with a Microneutralization Test. In Mali, the observed seroprevalence in domestic birds was 28.5% [24.5; 32.8] _95%CI, 3.1 % [1.8; 5.2] _95%CI, 6.2% [3.4; 10.2] _95%CI and 9.8 % [7.3; 12.8] _95%CI, for West Nile virus (WNV), Usutu virus (USUV), undetermined flavivirus, and WNV/USUV respectively. Regarding domestic birds of Madagascar, the observed seroprevalence was 4.4 % [2.1; 7.9]_95%CI for WNV, 0.9% [0.1; 3.1] _95%CI for USUV, 1.3% [0.5; 2.8] _95%CI for undetermined flavivirus, and null for WNV/USUV. Among the 150 wild birds sampled in Madagascar, two fulvous whistling-ducks (Dendrocygna bicolor) were positive for WNV and two for an undetermined flavivirus. One white-faced whistling-duck (Dendrocygna viduata) and one Hottentot teal (Spatula hottentota) were tested positive for USUV. African and European wetlands are linked by wild bird migrations. This first detection of USUV—as well as the confirmed circulation of WNV in domestic birds of two wetlands of Mali and Madagascar—emphasizes the need to improve the surveillance, knowledge of epidemiological patterns, and phylogenetic characteristics of flavivirus in Africa, particularly in areas prone to sustained, intense flavivirus transmission such as wetlands.

Ticks are the most important vectors of pathogens affecting both domestic and wild animals worldwide. Hard tick feeding is a slow process—taking up to several days—and necessitates extended control over the host response. The success of the feeding process depends upon injection of tick saliva, which not only controls host hemostasis and wound healing, but also subverts the host immune response to avoid tick rejection that creates a favorable niche for the survival and propagation of diverse tick-borne pathogens. Here, we report on the molecular and biochemical features and functions of an Ixodes ricinus serine protease inhibitor (IrSPI). We characterize IrSPI as a Kunitz elastase inhibitor that is overexpressed in several tick organs—especially salivary glands—during blood-feeding. We also demonstrated that when IrSPI is injected into the host through saliva, it had no impact on tissue factor pathway-induced coagulation, fibrinolysis, endothelial cell angiogenesis or apoptosis, but the protein exhibits immunomodulatory activity. In particular, IrSPI represses proliferation of CD4⁺ T lymphocytes and proinflammatory cytokine secretion from both splenocytes and macrophages. Our study contributes valuable knowledge to tick-host interactions and provides insights that could be further exploited to design anti-tick vaccines targeting this immunomodulator implicated in I. ricinus feeding.