Sperm cryopreservation of the critically endangered olive barb (Sarpunti) Puntius sarana (Hamilton, 1822)

Document Type

Article

Publication Date

2-1-2011

Abstract

The present study focused on development of a sperm cryopreservation protocol for the critically endangered olive barb Puntius sarana (Hamilton, 1822) collected from two stocks within Bangladesh and reared in the Fisheries Field Laboratory, Bangladesh Agricultural University (BAU). The sperm were collected in Alsever's solution prepared at 296mOsmolkg-1. Sperm were activated with distilled water (24mOsmolkg-1) to characterize motility. Maximum motility (90%) was observed within 15s after activation, and sperm remained motile for 35s. Sperm activation was evaluated in different osmolalities and motility was completely inhibited when osmolality of the extender was 287mOsmolkg-1. To evaluate cryoprotectant toxicity, sperm were equilibrated with 5%, 10% and 15% each of dimethyl sulfoxide (DMSO) and methanol. Sperm motility was noticeably reduced within 10min, when sperm were equilibrated with 15% DMSO, indicating acute toxicity to spermatozoa and therefore this concentration was excluded in further trials. Sperm were cryopreserved using DMSO at concentrations of 5% and 10% and methanol at 5%, 10% and 15%. The one-step freezing protocol (from 5°C to -80°C at 10°C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL® CL-3300; Australia) and 0.25-ml straws containing spermatozoa were stored in liquid nitrogen for 7-15days at -196°C. The highest motility in thawed sperm 61±8% (mean±SD) was obtained with 10% DMSO. The fertilization and hatching rates were 70% and 37% for cryopreserved sperm, and 72% and 62% for fresh sperm. The protocol reported here can be useful for hatchery-scale production of olive barb. The use of cryopreserved sperm can facilitate hatchery operations, and can provide for long-term conservation of genetic resources to contribute in the recovery of critically endangered fish such as the olive barb. © 2010 Elsevier Inc.

Publication Source (Journal or Book title)

Cryobiology

First Page

62

Last Page

67

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