Effect of Silencing subolesin and enolase impairs gene expression, engorgement and reproduction in Haemaphysalis longicornis (Acari: Ixodidae) ticks

Importance: Haemaphysalis longicornis is an obligate blood-sucking ectoparasite that has gained attention due its role of transmitting medically and veterinary significant pathogens and it is the most common tick species in Republic of Korea.The preferred strategy for controlling ticks is a multi-antigenic vaccination.Testing the efficiency of a combination antigen is a promising method for creating a tick vaccine.Objective: The aim of the current research was to analyze the role of subolesin and enolase in feeding and reproduction of H. longicornis by gene silencing.Methods: In this study, we used RNA interference to silence salivary enolase and subolesin in H. longicornis.Unfed female ticks injected with double-stranded RNA targeting subolesin and enolase were attached and fed normally on the rabbit's ear.Real-time polymerase chain reaction was used to confirm the extent of knockdown.Results: Ticks in the subolesin or enolase dsRNA groups showed knockdown rates of 80% and 60% respectively.Ticks in the combination dsRNA (subolesin and enolase) group showed an 80% knockdown.Knockdown of subolesin and enolase resulted in significant depletion in feeding, blood engorgement weight, attachment rate, and egg laying.Silencing of both resulted in a significant (p < 0.05) reduction in tick engorgement, egg laying, egg hatching (15%), and reproduction.Conclusions and Relevance: Our results suggest that subolesin and enolase are an exciting target for future tick control strategies.