Rozanska-Wrobel, J.; Przesmycka, K.; Wasilewska, J.; Grzybek, M.; Notarnicola, R. F.; Bajer, A.; Dwuznik-Szarek, D.; Alsarraf, M.; Behnke-Borowczyk, J.; Behnke, J. M.; Radwan, J.

Background: Lyme borreliosis is a common tick-borne disease in Europe caused by spirochetes of the Borrelia burgdorferi sensu lato complex, including Borrelia afzelii, which is maintained in nature through interactions with rodent reservoir hosts. These spirochetes have evolved several surface proteins to manipulate rodent host immunity, some of which remain polymorphic in Borrelia populations. Among these proteins, OspE, which binds the host complement-regulating factor CFH to evade destruction by complement, is one of the most variable. Yet, what evolutionary forces maintain this polymorphism is not well understood. Motivated by a recent discovery of CFH polymorphism in the bank vole (Clethrionomys glareolus), the main reservoir host of B. afzelii, we hypothesized that the polymorphism is maintained by host-parasite coevolution involving specific associations between host and parasite genetic variants. Methods: We analyzed associations between bank vole CFH alleles and B. afzelii OspE variants across three datasets sampled in Poland. Selection acting on OspE was evaluated using omegaMap. Host-pathogen genotype associations were tested using partial redundancy analysis (RDA), and co-structure was assessed using co-correspondence analysis (CoCA). Results: We found that OspE evolves under positive selection, however, we found no evidence for an association between OspE and host CFH variants at the individual level based on RDA or at the population level based on CoCA. Conclusions: Despite evidence of positive selection acting on OspE, we found no support for specific genetic matching between B. afzelii and its bank vole host at the CFH-OspE interface. These results suggest that the evolution of CFH and OspE may be shaped by broader selective pressures, potentially including interactions with multiple host species.