Cryopreservation of Anopheles gambiae Larvae.

Mosquitoes remain a significant global health concern due to their role in transmitting deadly diseases including malaria, dengue, and yellow fever. Recent advances in genetic modification technologies have enabled researchers to better study disease transmission and is an emerging approach for vector control. However, a major bottleneck is the need for continuous rearing of each new strain, an intensive process requiring substantial manual effort and space with inherent risks of cross-contamination, genetic drift, and catastrophic loss. Cryopreservation offers a potential solution, where strains could be stored indefinitely and revived on an as-needed basis. Here, we report a cryopreservation protocol that results in the recovery of viable Anopheles gambiae larvae. After thawing, 82% of larvae showed signs of viability, including wriggling and mouth brush movements. Approximately 15% further exhibited coordinated swimming behavior. Although post-thaw survival was limited to 24 h, this study provides the first evidence that mosquito larvae can survive cryopreservation, representing a key milestone toward long-term storage of mosquito lines.