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Effects of the slow cooling during cryopreservation on the survival and morphology of Taiwan shoveljaw carp (Varicorhinus barbatulus) spermatozoa

Published online by Cambridge University Press:  23 December 2009

Sujune Tsai ,
Emma Spikings ,
Ching-Chiang Hwang  and
Chiashin Lin
Sujune Tsai
Affiliation:
Department of biotechnology, Mingdao University, 369 Wenhua Road, Peetow, Chang Hua 52345, Taiwan
Emma Spikings
Affiliation:
LIRANS Institute of research in the applied natural sciences, University of Bedfordshire, 250 Butterfield, Great Marlings, Luton, Bedfordshire LU2 8DL, UK
Ching-Chiang Hwang
Affiliation:
LIRANS Institute of research in the applied natural sciences, University of Bedfordshire, 250 Butterfield, Great Marlings, Luton, Bedfordshire LU2 8DL, UK
Chiashin Lin
Affiliation:
National museum of marine biology & Aquarium, 2 Houwan Road, Checheng, Pingtung 944, Taiwan
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Abstract

Over the past decades, pollution, overfishing, and habitat degradation have driven the population size of Taiwan shoveljaw carp down markedly in Taiwan. Cryopreservation is a useful tool which could be used to maintain genetic resources to protect and preserve this endemic species. Four cryoprotectants [dimethyl sulphoxide (DMSO), dimethylacetamide (DMA), glycerol and methanol] and six freezing rates (0.5, 1, 2, 4, 8, 16 °C min-1) were tested in order to develop an optimal controlled slow-freezing protocol for Taiwan shoveljaw carp spermatozoa. Samples were subsequently examined under the scanning electron microscope to reveal whether cryopreservation had affected their ultrastructural morphology. The highest survival rate (50.1 ± 2.0%) was observed with a freezing rate of 8 °C min-1 in 1M DMSO, using SYBR-14 + PI staining. Fertility and hatching rate results using frozen-thawed spermatozoa (90.2 ± 2.2% and 22.3 ± 2.5%, respectively) were not significantly different from results with fresh spermatozoa. After cryopreservation, 21.0 ± 1.6% of frozen-thawed spermatozoa had mid-piece swelling and rupture of the head. Cryopreservation might, therefore, slightly affect Taiwan shoveljaw carp spermatozoa in terms of morphological change. However, these alterations could be compensated by using large enough numbers of normally functioning frozen-thawed spermatozoa to achieve a standard equal to fresh spermatozoa. This is the first report of successful cryopreservation of Taiwan shoveljaw carp spermatozoa using a controlled slow-cooling method.

Keywords

CryopreservationScanning electron microscopeFreezing rateFertilisationTaiwan shoveljaw carpCyprinidae
Type
Note
Information
Aquatic Living Resources , Volume 23 , Issue 1 , January 2010 , pp. 119 - 124
Copyright
© EDP Sciences, IFREMER, IRD, 2009

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