台湾泥鳅精子超低温冷冻保存技术研究

doi:10.16378/j.cnki.1003-1111.23052

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Abstract To establish cryopreservation method of sperm for Taiwan loach Paramisgurnus dabryanus ssp. taiwan, effects of four extenders (HBSS, D-15, D-17, and Ringer), five cryoprotectants [methanol (MeOH), ethylene glycol (EG), propylene glycol (PG), dimethyl sulfoxide (DMSO), and N, N-dimethylformamide (DMF)] at four volume concentrations (2.5%, 5.0%, 7.5%, and 10.0%), different equilibrium time (0, 10, 20, 30, 40 and 50 min), fumigation height and time (5, 7, 9, 11 cm and 5, 10, 15 min), and thawing temperature (17, 27 and 37 ℃) and time (5, 10 and 15 s) on the sperm viability of Taiwan loach was observed by microscopy and the plasma membrane integrity was measured by flow cytometry (FCM). The results revealed that the sperm with HBSS had the maximal motility (45.28±5.75)%, and the sperm with 5% methanol had the maximal motility of (81.70±2.35)% and the plasma membrane integrity of (51.25±5.03)%, with the best equilibrium time of 30 min, and the best fumigation height of 9 cm for 10 min. The better thawing effect was observed by thawing in 37 ℃ water bath for 10 s, with the fertilization rate of (68.94±6.22)% at artificial insemination with thawed Taiwan loach sperm. In summary, the fresh sperm of Taiwan loach was diluted with HBSS and mixed with MeOH to a final concentration of 5% at 4 ℃ for 30 min, then put it into straws, fumigated at 9 cm above the liquid nitrogen surface for 10 min, and then was stored in liquid nitrogen. When thawing, good results were observed when the fresh sperm of Taiwan loach was thawed in a 37 ℃ water bath for 10 s. The findings provided data foundation with artificial breeding and germplasm resource protection of Taiwan loach.

Key words： Taiwan loach sperm cryopreservation flow cytometry propidium iodide

Received: 03 July 2023

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	WANG Yinjie
	WANG Pin
	CAI Kejun

Cite this article:

WANG Yinjie,WANG Pin,CAI Kejun. Cryopreservation Technology of Taiwan Loach Sperm[J]. Fisheries Science, 2024, 43(6): 915-924.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.23052 OR https://www.shchkx.com/EN/Y2024/V43/I6/915

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