Evaluation of sperm cryopreservation effectiveness for largemouth bass(Micropterus salmoides) based on computer-assisted sperm analysis

doi:10.3969/j.issn.1004-1524.20240769

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (7): 1417-1423.DOI: 10.3969/j.issn.1004-1524.20240769

• Animal Science • Previous Articles Next Articles

Evaluation of sperm cryopreservation effectiveness for largemouth bass(Micropterus salmoides) based on computer-assisted sperm analysis

LI Fan¹(), ZHANG Xiaofeng², LI Jianying³, CHEN Xiaoming², LIU Wei², ZHOU Fan², MENG Qinghui²^,^*()

1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
2. Zhejiang Fisheries Technical Extension Center, Hangzhou 311122, China
3. College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 311121, China

Received:2024-08-30 Online:2025-07-25 Published:2025-08-20

Abstract

Abstract:

To evaluate and screen out suitable sperm preservation solutions to extend the storage time of largemouth bass (Micropterus salmoides) sperm, the effects of four preservation solutions, i.e. Cortland saline, Ringer’s saline, Zebrafish E3 culture solution, and PBS solution, on sperm motility were compared during the storage at 4 ℃. The changes on key motility parameters, including the percentage of motile (MOT), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP) and beatingcross frequency (BCF) of the origin sperm and the sperm stored with preservation solution were assessed using a computer-assisted sperm analysis (CASA) system. It was shown that there was a gradual decline in MOT with the extended storage time. The cryopreservation effectiveness of preservation solutions varied. Among the preservation solutions, Ringer’s saline and PBS solution demonstrated superior performance, with higher MOT and other motion parameters. When the sperm was stored with Ringer’s saline, the MOT could maintain over 95% after 72 h. These findings suggest that Ringer’s saline could be used for the short-term refrigeration of largemouth bass sperm.

Key words: largemouth bass (Micropterus salmoides), computer-assisted sperm analysis, sperm, refrigerated preservation

CLC Number:

S961.2

LI Fan, ZHANG Xiaofeng, LI Jianying, CHEN Xiaoming, LIU Wei, ZHOU Fan, MENG Qinghui. Evaluation of sperm cryopreservation effectiveness for largemouth bass(Micropterus salmoides) based on computer-assisted sperm analysis[J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1417-1423.

Figures/Tables 4

References 31

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保存液 Preservation solution	NaCl	KCl	MgSO₄·7H₂O	NaHCO₃	NaH₂PO₄·2H₂O	CaCl₂	KH₂PO₄	葡萄糖 Glucose
Cortland	7.250	0.380	0.230	1.000	0.410	0.162	—	1.000
Ringer	6.500	0.140	—	0.200	0.010	0.120	—	—
E3	0.344	0.015	0.098	—	—	0.058	—	—
PBS	8.000	0.200	—	—	1.440	—	0.240	—

保存液 Preservation solution	NaCl	KCl	MgSO₄·7H₂O	NaHCO₃	NaH₂PO₄·2H₂O	CaCl₂	KH₂PO₄	葡萄糖 Glucose
Cortland	7.250	0.380	0.230	1.000	0.410	0.162	—	1.000
Ringer	6.500	0.140	—	0.200	0.010	0.120	—	—
E3	0.344	0.015	0.098	—	—	0.058	—	—
PBS	8.000	0.200	—	—	1.440	—	0.240	—

t/h	MOT/%	STR/%	VCL/(μm·s^-1)	VSL/(μm·s^-1)	VAP/(μm·s^-1)	BCF/Hz	MAD/(°)	DHS/μm
2	99.98±5.24	59.93±2.24	202.06±22.08	46.53±15.64	77.64±18.57	38.82±5.66	20.97±5.76	60.17±15.71
6	96.26±5.61	77.94±6.87	110.29±17.81	45.67±16.32	58.60±16.89	29.30±6.91	19.87±5.23	44.88±11.35
24	80.00±4.77	70.39±8.53	47.00±11.66	15.14±9.47	21.51±8.83	10.76±1.24	11.47±3.43	24.51±8.52
48	63.58±3.26	55.95±3.49	28.54±3.97	5.78±1.87	10.33±2.16	5.16±0.36	7.40±2.17	16.10±3.18
72	54.13±6.31	76.16±9.82	25.26±3.75	9.84±3.24	12.92±2.99	6.46±1.08	6.58±1.11	19.23±2.60

t/h	MOT/%	STR/%	VCL/(μm·s^-1)	VSL/(μm·s^-1)	VAP/(μm·s^-1)	BCF/Hz	MAD/(°)	DHS/μm
2	99.98±5.24	59.93±2.24	202.06±22.08	46.53±15.64	77.64±18.57	38.82±5.66	20.97±5.76	60.17±15.71
6	96.26±5.61	77.94±6.87	110.29±17.81	45.67±16.32	58.60±16.89	29.30±6.91	19.87±5.23	44.88±11.35
24	80.00±4.77	70.39±8.53	47.00±11.66	15.14±9.47	21.51±8.83	10.76±1.24	11.47±3.43	24.51±8.52
48	63.58±3.26	55.95±3.49	28.54±3.97	5.78±1.87	10.33±2.16	5.16±0.36	7.40±2.17	16.10±3.18
72	54.13±6.31	76.16±9.82	25.26±3.75	9.84±3.24	12.92±2.99	6.46±1.08	6.58±1.11	19.23±2.60

保存液	t/h	VCL/	VSL/	VAP/	DHS/μm	MAD/(°)	BCF/Hz	STR/%
Preservation solution		(μm·s^-1)	(μm·s^-1)	(μm·s^-1)
Ringer	2	74.49±14.32	30.43±11.84	39.24±12.33	31.82±18.57	15.90±3.66	19.62±6.16	75.90±5.27
	72	70.78±15.84	26.18±12.38	35.10±13.01	26.46±2.43	15.78±4.08	17.55±6.51	72.08±7.26
PBS	2	78.31±15.61	30.47±15.34	40.04±15.12	29.81±13.07	16.43±2.39	20.02±7.56	73.01±8.89
	72	75.76±13.63	29.52±15.90	38.77±15.40	37.80±12.67	13.77±3.42	19.39±7.70	72.34±9.63
E3	2	27.42±1.79	6.16±0.27	10.41±0.53	40.67±4.67	6.64±0.59	5.20±0.26	59.19±1.37
	72	10.21±0.95^**	1.43±0.27^**	3.18±0.16^**	24.25±4.54^**	2.86±0.47^**	1.59±0.08^**	44.74±7.41^*
Cortland	2	24.81±3.82	5.59±0.53	9.44±1.15	38.01±7.47	5.31±0.51	4.72±0.57	59.42±2.54
	72	11.30±0.36^**	1.29±0.13^**	3.29±0.05^**	19.38±2.95^**	3.47±0.33^**	1.65±0.02^**	39.12±3.53^**

Evaluation of sperm cryopreservation effectiveness for largemouth bass(Micropterus salmoides) based on computer-assisted sperm analysis

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