Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

Abstract

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.

Assessment of viability of sperm cells of Litopenaeus vannamei on cryopreservation

“…In the present work, the mortality of sperm cells changed as the cryoprotectant concentrations and exposition times increased and the optimal concentration and equilibration time were 10% and 10 minutes, respectively. Barth et al (2006) while working with the giant tiger prawn, P. monodon, used DMSO at 5 and 10% as cryoprotectant. Results showed that DMSO at 5% offered better results when sperm survival reached 79.7%.…”

“…The development of methods of semen preservation and the implementation of effective artificial reproduction protocols require a good knowledge of the reproductive biology and seminal characteristics of the targeted species (Ninhaus-Silveira et al 2006). For marine shrimps, cryoprotectants dimethyl sulfoxide (DMSO) and ethylene glycol (EG) are widely used (Anchordoguy et al 1988;Barth et al 2006;Vuthiphandchai et al 2007). DMSO, when compared with glycerol, proline and trehalose, presents low toxicity to the spermatic cells of Sicyonia ingentis (Anchordoguy et al 1988).…”

Effect of Different Cryoprotectants and Sperm Densities of Orange Mud Crab, Scylla olivacea (Herbst, 1796) for Long-Term Storage of Spermatozoa

“…The Ca-Fsaline (21.63 g NaCl, 1.12 g KCl, 0.53 g H 3 BO 3 , 0.19 g NaOH and 4.93 g MgSO 4 .7H 2 O in 1 L sterile distilled water (adjusted to pH 7.4 with 1 NHCl)) was prepared as the extender medium (Bart et al 2006;Memon et al 2012d). The extender was prepared one day before the collection of samples and stored in the refrigerator.…”

“…The samples were kept in LN for 30 days (Memon et al 2012b). On day 30, thawing was done by immersing the cryovial in a water bath at 30ºC for 2 min (Bart et al 2006;Memon et al 2012b). After thawing, the samples were examined under the electron microscope.…”

Induction of acrosome reaction of spermatozoa in Eriocheir sinensis by low temperature

“…Temperature, which is one of the most important factors that affect the physiology of cell (Boon et al 2008;Fan et al 2008;Jong et al 2009;Oguchi et al 2006), is one of them (Martin et al 2001;Nagao and Munehara 2007). Low temperature has been widely used for in vitro spermatozoa storage and artificial fertilization of aquatic crustaceans (Bart et al 2006;Magda et al 2004;Nimrat et al 2006;Thomas et al 1988;Vuthiphandchai et al 2007). However, the information on the effects of temperature on AR and capacitation in crabs are sparse (Bhavanishankar and Subramoniam1997;Kang et al 2009) although aquaculture of crabs has become very important.…”