Male Reproductive Transcript Silencing Reduces Fitness in Anopheles funestus

Male mosquito biology remains underrepresented in studies of host traits influencing vector populations and parasite transmission. Anopheles funestus is a major African malaria vector, yet the molecular basis of male reproductive function in this species is poorly understood. Transcript EZ976679 is strongly male-biased and shares high sequence identity with a juvenile hormone esterase-like gene, suggesting a potential role in reproductive physiology.

In this study, EZ976679 expression was profiled across developmental stages, sexes, and dissected tissues using quantitative PCR. Functional characterisation was performed using dsRNA-mediated knockdown in adult males, with transcript abundance assessed at multiple time points post-injection. Reproductive and longevity assays were conducted to evaluate the effects of EZ976679 knockdown on mating success, fecundity, fertility and adult male survival.

EZ976679 expression was age-dependent and highly enriched in the testes and accessory glands of 3–5-day-old males, with minimal expression in females and immature stages. RNA interference knockdown reduced transcript levels by more than 80% for up to five days, followed by partial recovery at later time points. Knockdown males exhibited significantly reduced mating success, decreased female fecundity and reduced adult male survival compared with controls, while fertility remained unaffected.

These findings identify EZ976679 as a male-specific host factor required for optimal reproductive performance in An. funestus. By modulating male survival, mating success and female reproductive output, this transcript may influence vector population dynamics. Targeting male reproductive biology represents a complementary and underexplored avenue for innovative malaria vector control strategies.