Affiliations 

  • 1 School of Fisheries and Aquaculture Sciences, Universiti Malaysia Terengganu, 21030, Kuala Nerus, Terengganu, Malaysia
  • 2 Institute of Tropical Aquaculture (AKUATROP), Universiti Malaysia Terengganu, 21030, Kuala Nerus, Terengganu, Malaysia
  • 3 School of Fundamental Science, Universiti Malaysia Terengganu, 21030, Kuala Nerus, Terengganu, Malaysia
  • 4 Fisheries Research Institute (FRI) Tanjung Demong, 22200, Besut, Terengganu, Malaysia
  • 5 School of Fisheries and Aquaculture Sciences, Universiti Malaysia Terengganu, 21030, Kuala Nerus, Terengganu, Malaysia; Institute of Tropical Aquaculture (AKUATROP), Universiti Malaysia Terengganu, 21030, Kuala Nerus, Terengganu, Malaysia. Electronic address: ivankcc@umt.edu.my
Cryobiology, 2018 04;81:168-173.
PMID: 29355519 DOI: 10.1016/j.cryobiol.2018.01.005

Abstract

This study developed the cryopreservation of brown-marbled grouper spermatozoa for practical application. We examined 32 cryodiluents, developed from four types of cryoprotectants [propylene glycol (PG), dimethyl-sulphoxide (Me2SO), dimethyl-acetamide (DMA) and ethylene glycol (EG)] at four concentrations of 5, 10, 15 and 20% in combination with two extenders [Fetal bovine serum (FBS) and artificial seminal plasma (ASP). Cooling rates were examined by adjusting the height of straws (2.5-12.5 cm) from the liquid nitrogen (LN) vapor and cooled for 5 min before immersion into LN. DNA laddering was used to detect DNA damage in cryopreserved sperm. In fertilization trials, 0.5 g of eggs was mixed with cryopreserved sperm stored for 30 days in LN. The best motility of post-thaw sperm was achieved using 15% PG + 85% FBS (76.7 ± 8.8%); 10% PG + 90% FBS was also effective as cryodiluent. Generally, FBS gave better post-thaw motility compared to ASP. The optimum cooling rate was at 17.6 °C min-1 obtained by freezing at the height of 7.5 cm surface of LN. The results obtained showed that cryopreserved sperm of brown-marbled grouper suffered slight DNA fragmentation, which resulted in significantly lower motility. However, the fertilization (90.9 ± 0.5%), hatching (64.5 ± 4.1%) and deformity rates (3.8 ± 0.2%) obtained from cryopreserved sperm showed no significant difference with fresh sperm. These findings show that the developed protocol for cryopreservation of brown-marbled grouper sperm was viable and will be useful for successful breeding and seed production of brown-marbled grouper.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.