Sperm cryopreservation of the Indian major carp, Labeo calbasu: Effects of cryoprotectants, cooling rates and thawing rates on egg fertilization

M Nahiduzzaman, Md Hassan, Pankoz Roy, Mohammed Akhter Hossain, Terrence Tiersch

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    9 Citations (Scopus)

    Abstract

    A sperm cryopreservation protocol for the Indian major carp, Labeo calbasu, was developed for long-term preservation and artificial fertilization. Milt collected from mature male fish were placed in Alsever's solution (296mOsmolkg-1) to immobilize the sperm. Cryoprotectant toxicity was evaluated by motility assessment with dimethyl sulfoxide (DMSO) and methanol at 5, 10 and 15% concentrations. DMSO was more toxic at higher concentrations than methanol, and consequently 15% DMSO was excluded from further study. A one-step cooling protocol (from 5 to 80°C) with two cooling rates (5 and 10°C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL® CL-3300; Australia). Based on post-thaw motility, the 10°C/min cooling rate with either 10% DMSO or 10% methanol yielded significantly higher (P=0.011) post-thaw motility than the other rate and cryoprotectant concentrations. Sperm thawed at 40°C for 15s and fresh sperm were used to fertilize freshly collected L. calbasu eggs and significant differences were observed (P=0.001) in percent fertilization between cryopreserved and fresh sperm as well as among different sperm-to-egg ratios (P=0.001). The highest fertilization and hatching rates were observed for thawed sperm at a sperm-to-egg ratio of 4.1×105:1. The cryopreservation protocol developed can facilitate hatchery operations and long-term conservation of genetic resources of L. calbasu.

    Original languageEnglish
    Pages (from-to)133-138
    Number of pages6
    JournalANIMAL REPRODUCTION SCIENCE
    Volume136
    Issue number1-2
    DOIs
    Publication statusPublished - Dec 2012

    Keywords

    • Endangered species
    • Fertilization
    • Hatching
    • L. calbasu
    • Sperm cryopreservation
    • Sperm-to-egg ratio

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