Assessment of viability of sperm cells of Litopenaeus vannamei on cryopreservation

Abstract

Aiming at assessing the cryopreservation potential of

Testes de toxicidade, congelamento e refrigeração em sêmen de espécie nativa

“…Cryopreservation in liquid nitrogen (-196°C) with 20% ethylene glycol also led to a decrease in survival over time, and the authors found a sperm survival rate of 75% at 200 th day of storage; however, no embryo was produced with the cryopreserved sperm during this period. Decreased survival was also reported by NIMRAT et al (2006) and UBERTI et al (2013) for Litopenaeus vannamei (Boone 1931), and by BAMBOZZI et al (2014), and CASTELO BRANCO et al (2014) for L. schmitti (Burkenroad 1936).…”

“…However, these authors noted a decrease in sperm quality when the spermatophores were cryopreserved with 20% ethylene glycol at -196°C for a long period. UBERTI et al (2013) used ethylene glycol and 10% DMSO with an equilibrium time of 10 minutes for the cryopreservation of L. vannamei semen and did not report a significant difference between these two cryoprotectants in terms of sperm survival.…”

“…The protocol adopted in this study (30°C for 4 min) is similar to the protocol used by AKARASANON et al (2004) for M. rosenbergii (30°C for 5 min) and VUTHIPHANDCHAI et al (2007) for P. monodon (30°C for 2 min); however, the results were not positive, which is inconsistent with those studies. UBERTI et al (2013) and CASTELO BRANCO et al (2014) used protocols of 20-25°C for 40 seconds and 20°C for 10 seconds, and reported excellent sperm survival rates for L. vannamei and L. schimitti, respectively. BHAVANISHANKAR and SUBRAMONIAM (1997) thawed semen of S. serrata at 55°C for 10-15 seconds and obtained positive sperm survival results.…”

Recent developments in male fertility evaluation, sperm cryopreservation and artificial fertilisation, and their potential application to decapod crustacean aquaculture

“…By contrast, PI binds to nucleic acids but can only penetrate cells that have ruptured membranes, causing the nucleus of dead spermatozoa to stain fluorescent red 91–93 . In crustaceans, fluorescent dyes coupled with flow cytometry showed 33%–89% post‐thaw viability in whiteleg shrimp spermatozoa after exposure to different cryopreservation protocols 24,54,94 . In freshwater crabs, PI staining and flow cytometry showed that sperm plasma membranes were significantly damaged (17%–20% dead spermatozoa) when exposed to high concentrations of lead 95 …”

“…[91][92][93] In crustaceans, fluorescent dyes coupled with flow cytometry showed 33%-89% post-thaw viability in whiteleg shrimp spermatozoa after exposure to different cryopreservation protocols. 24,54,94 In freshwater crabs, PI staining and flow cytometry showed that sperm plasma membranes were significantly damaged (17%-20% dead spermatozoa) when exposed to high concentrations of lead. 95 These results show that fluorescent stains coupled with flow cytometry represent a sensitive tool for assessing sperm cell viability in decapod crustaceans.…”