@article{MAKHILLJAVA201211103445,
    title = {Optimization of Spermatophores Cryopreservation Protocol of Banana Shrimp (<I>Penaeus merguiensis</I>) (De Man, 1888)},
    journal = {Journal of Animal and Veterinary Advances},
    volume = {11},
    number = {10},
    pages = {1688-1704},
    year = {2012},
    issn = {1680-5593},
    doi = {javaa.2012.1688.1704},
    url = {https://makhillpublications.co/view-article.php?issn=1680-5593&doi=javaa.2012.1688.1704},
    author = {A.J.,A.D.,M.I.,M.O.,J.,A.B. and},
    keywords = {Banana shrimp,Penaeus merguiensis,spermatophore,cryopreservation,fertilization,hatching},
    abstract = {The objectives of this study were to determine the effects of different cryoprotectants on sperm viability and optimization of spermatophore cryopreservation protocol for durable storage of Banana shrimp (<I>Penaeus merguiensis</I>). Spermatophore suspended for 15 min in Calcium-Free saline (Ca-F saline), used CPA MgCl<SUB>2</SUB> and with concentration (15%), thawing temperature was 27&deg;C. Use 15 min equilibration in room temperature (25&deg;C) overall. Exposure and cooling rate selected as 25, 20, 16, 4, 2, -4, -20, -80, -150&deg;C/10 min. Examination of sperm viability used a modified eosin-nigrosin staining technique. The smallest reductions in apparent sperm viability occurred with MgCl<SUB>2</SUB>, however freezing protocol was developed using Ca-F saline containing 15% MgCl<SUB>2</SUB>. Spermatophores were cryopreserved using above exposure/cooling rate and -196&deg;C in liquid nitrogen up to 180 days. Mean sperm viability for fresh (93.8&plusmn;1.3%) and cryopreserved spermatophore held for 24 h and 60 days was 83.5&plusmn;0.6 and 61&plusmn;1.2 did not differ (p&gt;0.05), however that for spermatophore stored in liquid nitrogen between 90 and 180 days were lower (p&lt;0.05) and varied from 55.4&plusmn;0.3-16.4&plusmn;1.2. Spermatophores earlier held in liquid nitrogen for 60 and 90 days. However, storage beyond 90 days caused a significant decline (p&lt;0.05) in sperm viability. Spermatophores kept for 120 and 150 days had viabilities of 48.9&plusmn;0.9 and 32.4&plusmn;0.9%, respectively. Cryopreserved spermatophore stored in liquid nitrogen from 150-180 days had low viabilities (&lt;35%). Mean fertilization rate of <I>P. merguiensis</I> females artificially inseminated with cryopreserved spermatophore that had been stored in liquid nitrogen for 7-30 days and for 60-90 days were 73.9&plusmn;1.5-66.7&plusmn;3.1 and 67.3&plusmn;3-64.1&plusmn;2.1%, respectively whereas that of fresh spermatophore was 88.2&plusmn;1.5%. Hatching rates of eggs fertilized with cryopreserved spermatophore kept for 7-30 days and for 60-90 days were 77.6&plusmn;2.5-72.7&plusmn;3.5 and 81.5&plusmn;12.1-62.5&plusmn;1.5 which were not different (p&gt;0.05) from those of the control group 76.2&plusmn;13.5%, respectively. In conclusion, Cryopreserved spermatophore held in liquid nitrogen l&lt;90 days revealed high sperm viability although, for longer periods, sperm viability declined at 180 days.}
    }