Ramirez, Gabriela, authorDobos, Karen, advisorEbel, Greg, committee memberBarfield, Jennifer, committee memberBroeckling, Corey, committee member2024-09-092025-08-162024https://hdl.handle.net/10217/239291The lipid profile and cryopreservation methods of Anopheles stephensi mosquitoes have significant implications for mosquito management and research. The intricate life cycle of mosquitoes is governed by lipid metabolism, involving lipogenesis, lipolysis, and fatty acid metabolism, which are critical for transitioning between life stages. Despite the importance of these processes, mosquito cryopreservation has faced challenges, mainly due to the impenetrable nature of mosquito eggs to traditional cryoprotective agents. While other insects like honeybees and fruit flies have seen some success in cryopreservation, mosquitoes have posed unique difficulties. Initial attempts to cryopreserve mosquito eggs were unsuccessful, and there remains a need for effective cryopreservation protocols that maintain the viability and normal development of mosquitoes post-thaw. This study investigates lipid metabolism across mosquito life stages and advances cryopreservation techniques. The lipid profile analysis focused on major lipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelins (SM), and triacylglycerides (TGs), as well as non-bilayer lipids like diacylglycerol (DG) and lysophospholipids (LysoPC, LysoPE). For cryopreservation, a new protocol using methylformamide (MF) with trehalose was developed for first-instar larvae (L1s) older than 1.5 hours post-hatching. The hypotheses were that 1) Lipid metabolism plays a crucial role in the successful development and reproduction of mosquitoes, and 2) Inducing diapause and using suitable cryoprotectants can improve the recovery rates of cryopreserved mosquitoes. The lipid profile analysis revealed that PE is vital for protein anchoring required for embryogenesis and immune responses and that steroids like 20-hydroxyecdysone (20E) are crucial for molting and development. The cryopreservation study showed that supercooling larvae for up to 60 minutes resulted in normal development to adulthood, although longer durations inhibited adult emergence. Supercooled larvae and their offspring exhibited typical sex ratios and developmental patterns, indicating genetic and phenotypic stability. This research underscores the well-orchestrated metabolic strategies in mosquitoes. It suggests that understanding these biochemical processes is essential for effective cryopreservation, potentially paralleling natural cold survival strategies seen in other insects.born digitaldoctoral dissertationsengCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.TextEmbargo expires: 08/16/2025.