Modification of contact avoidance behaviour associated with pyrethroid resistance in Anopheles sinensis (Diptera: Culicidae)

BACKGROUND: Anopheles sinensis is the primary vector of vivax malaria in China and its control is under great threat as the development of insecticide resistance. In contrast to physiological resistance, there is no report of behavioural modifications of resistant An. sinensis after long-term insecticide use, despite their huge potential impact on malaria transmission. METHODS: Larvae or pupae of An. sinensis were collected from Yuanyang, Bishan, and Wuhe counties from southwestern to eastern China. Resistance to deltamethrin was assayed using the standard World Health Organization (WHO) susceptibility test. The frequency distribution of the kdr allele of the para-type sodium channel gene was determined by polymerase chain reaction (PCR) amplification and DNA sequencing. Contact repellency to deltamethrin-impregnated bed nets was evaluated using a modified WHO cone bioassay. RESULTS: All contemporary field populations for all three geographic locations were resistant to deltamethrin, with mortality ranging from 6.00 to 26.79%. Three kdr genotypes with either an L1014F or L1014C substitution with frequencies of 76.10–100% were identified in the Bishan and Wuhe populations, but no kdr mutations were detected in the Yuanyang samples despite high phenotypic resistance. The susceptible mosquitoes exhibited significantly longer flying time and more takeoffs on deltamethrin-treated bed nets (DTN) than on untreated bed nets (UTN), suggestive of robust avoidance behaviour. However, no significant increases in the frequency of takeoffs or flying time were observed in deltamethrin-resistant An. sinensis populations when exposed on DTNs, regardless of the presence of a kdr mutation. Moreover, the first takeoff from DTNs by resistant mosquitoes significantly lagged behind compared to susceptible mosquitoes. CONCLUSION: The An. sinensis populations were highly resistant to deltamethrin and exhibited decreased avoidance behaviour. Behavioural modification significantly associated with deltamethrin resistance, but not directly related to the presence of kdr mutations, indicating that there are additional factors contributing to the changes.