What is it about?
In this research work, we have successfully cryopreserved the spermatozoa of two endangered fish species, Tor khudree and Tor putitora as an ex situ conservation biotechnology tool
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Why is it important?
In this research work, for the first time we have 1) proved the efficacy of using combination of DMSO and Glycerol which has got better cryoprotective role due to the synergistic effect 2) Also, we have successfully used Nitro Blue Tetrazolium (NBT) Assay for the first time for fish spermatozoa for quantitative assessment of sperm mitochondrial activity to determine extent of damage due to cryopreservation-thawing cycle
Perspectives
In this research work, we have successfully 1) used a combination of DMSO and Glycerol for the first time with better post-thaw hatching rates proving the synergistic effect of the cryoprotectants 2) used NBT test for the first time for fish spermatozoa for quantitative assessment of mitochondrial enzyme complex
Dr RAVINDRAGOUDA PATIL
FRIC, KVAFSU, Hesaraghatta
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This page is a summary of: Effect of cryoprotectants, equilibration periods and freezing rates on cryopreservation of spermatozoa of mahseer, Tor khudree (Sykes) and T. putitora (Hamilton), Aquaculture Research, November 2005, Wiley,
DOI: 10.1111/j.1365-2109.2005.01331.x.
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Cryopreservation of fish spermatozoa
Mahseers, a group of important food and game ¢shes of India, formed a major ¢shery of the rivers and streams of the Shivalik Himalayas and the lower reaches of uplands of the deccan plateau (Mahanta, Kapoor, Pandey, Srivastava, Dayal, Patiyal, Joshi, Singh & Paul 1998). They are currently enlisted as a group of threatened ¢shes (Ogale1994; Basavaraja & Keshavanath 2000). Cryopreservation of ¢sh spermatozoa has many advantages over other forms of live gene banking and has gained much acceptance in the past few years (Lakra1993; Rana1995). However, detailed studies with respect to various parameters involved in cryopreservation of mahseer spermatozoa are lacking (Ponniah, Lakra & Ogale 1999a; Ponniah, Sahoo, Dayal & Barat1999b; Basavaraja, Hegde, Akash & Udupa 2002). Even though few studies have been conducted on these lines (Basavaraja et al. 2002), no e¡ort has been made so far to evaluate the e¡ect of cryoprotectants in di¡erent combinations. Hence, two species of mahseer, viz., deccan mahseer, Tor khudree (Sykes), and golden mahseer, T. putitora (Hamilton), were selected for the standardization of cryopreservation protocol for the spermatozoa with respect to the e¡ect of cryoprotectants, di¡erent equilibration times and freezing rates. ravi.patil30atgmail.com
Cryopreservation of fish spermatozoa
Abstract A study was conducted to standardize a protocol for cryopreservation of spermatozoa of the endangered mahseer, Tor khudree (Sykes) and T. putitora (Hamilton). The suitability of the cryoprotectants, dimethyl sulphoxide (DMSO) and glycerol, and the combination of the twowere tested. Four equilibration periods and four freezing rates were also tested for their standardization. A combination of 9% DMSO and 11% glycerol gave signi¢cantly higher mean percentage of hatching in both T. khudree (45.59 1.86%) (control 71.08 0.59%) and T. putitora (45.00 1.25%) (control 73.48 1.19%) among the eight di¡erent treatments. Among the four di¡erent equilibration periods tested, the equilibration period of 30min1 yielded the highest mean hatching percentage in T. khudree (39.46 1.94%) (control 71.70 0.61%) and T. putitora (38.28 1.06%) (control 73.11 0.82%). Freezing straws at a height of 8 cm above LN2 surface for 10min1 gave higher hatching percentages for both T. khudree (41.75 1.72%) (control 73.99 1.17%) and T. putitora (41.34 2.04%) (control 72.48 1.51%). The study reports the superior performance of the combination of DMSO and glycerol for the ¢rst time. Keywords: cryopreservation, mahseer, spermatozoa, cryoprotectants, equilibration time, freezing rate
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