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Physiological responses to cryoprotectant treatment in an early larval stage of the malaria mosquito, Anopheles gambiae
- Source :
- Cryobiology. 99:114-121
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- The development of cryopreservation protocols for Anopheles gambiae could significantly improve research and control efforts. Cryopreservation of any An. gambiae life stage has yet to be successful. The unique properties of embryos have proven to be resistant to any practical cryoprotectant loading. Therefore, we have chosen to investigate early non-feeding first instar larvae as a potential life stage for cryopreservation. In order to determine an appropriate cryoprotective compound, larvae were treated with progressively better glass-forming cryoprotective mixtures. Toxicity evaluation in combination with calorimetry-based water content and supercooling point depression assessments were used to determine the cryoprotectants that could be used for cryostorage of viable larvae. Approximately 35–75% of the larvae were viable after reasonably high osmotic and biochemical challenge. This study provides ample evidence for an active osmoregulatory response in the Anopheles larvae to counter the permeation of cryoprotectants from the surrounding medium. The data show a strong correlation between the larval mortality and water content, indicating an osmoregulatory crisis in the larva due to certain cryoprotectants such as the higher concentrations of ethane diol (ED). The observations also indicate that the ability of the larvae to regulate permeation and water balance ceases at or within 20 min of cryoprotectant exposure, but this is strongly influenced by the treatment temperature. Among the compound cryoprotectants tested, 25% ED + 10% dimethyl sulfoxide (DMSO) and 40% ED + 0.5 M trehalose seem to present a compromise between viability, larval water content, supercooling point depression, and glass forming abilities.
- Subjects :
- Cryoprotectant
Anopheles gambiae
Biology
General Biochemistry, Genetics and Molecular Biology
Cryopreservation
Andrology
03 medical and health sciences
chemistry.chemical_compound
Cryoprotective Agents
0302 clinical medicine
Anopheles
parasitic diseases
Animals
Larva
030219 obstetrics & reproductive medicine
Dimethyl sulfoxide
fungi
0402 animal and dairy science
04 agricultural and veterinary sciences
General Medicine
biology.organism_classification
040201 dairy & animal science
Trehalose
chemistry
Instar
General Agricultural and Biological Sciences
Subjects
Details
- ISSN :
- 00112240
- Volume :
- 99
- Database :
- OpenAIRE
- Journal :
- Cryobiology
- Accession number :
- edsair.doi.dedup.....b7e6beef56fec85834775cdbceef4a5a