Adapting field-mosquito collection techniques in a perspective of near-infrared spectroscopy implementation

Near-infrared spectroscopy (NIRS) has the potential to be a useful tool for assessing key entomological parameters of malaria-transmitting mosquitoes, including age, infectious status and species identity. However, before NIRS can be reliably used in the field at scale, methods for killing mosquitoes and conserving samples prior to NIRS scanning need to be further optimized. Historically, mosquitoes used in studies have been killed with chloroform, although this approach is not without health hazards and should not be used in human dwellings. For the application of NIRS scanning it is also unclear which mosquito preservation method to use. The aim of the study reported here was to investigate the use of pyrethrum spray, a commercially available insecticide spray in Burkina Faso, for killing mosquitoes METHODS: Laboratory-reared Anopheles gambiae and Anopheles coluzzii were killed using either a pyrethrum insecticide spray routinely used in studies involving indoor mosquito collections (Kaltox Paalga®; Saphyto, Bobo-Dioulasso, Burkina Faso) or chloroform ("gold standard"). Preservative methods were also investigated to determine their impact on NIRS accuracy in predicting the species of laboratory-reared Anopheles and wild-caught mosquito species. After analysis of fresh samples, mosquitoes were stored in 80% ethanol or in silica gel for 2 weeks and re-analyzed by NIRS. In addition, experimentally infected An. coluzzii and wild-caught An. gambiae sensu lato (s.l.) were scanned as fresh samples to determine whether they contained sporozoites, then stored in the preservatives mentioned above for 2 weeks before being re-analyzed. The difference in the accuracy of NIRS to differentiate between laboratory-reared An. gambiae mosquitoes and An. coluzzii mosquitoes killed with either insecticide (90%) or chloroform (92%) was not substantial. NIRS had an accuracy of 90% in determining mosquito species for mosquitoes killed with chloroform and preserved in ethanol or silica gel. The accuracy was the same when the pyrethrum spray was used to kill mosquitoes followed by preservation in silica gel, but was lower when ethanol was used as a preservative (80%). Regarding infection status, NIRS was able to differentiate between infected and uninfected mosquitoes, with a slightly lower accuracy for both laboratory and wild-caught mosquitoes preserved in silica gel or ethanol. The results show that NIRS can be used to classify An. gambiae s.l. species killed by pyrethrum spra