Cooling of porcine semen in an extender supplemented with isoespintanol

Betancur, Giovanni Restrepo; Zapata, Kelly Vanessa; Vidal, Paola Andrea Colorado; Sánchez, Yudith; Rojano, Benjamín Alberto

doi:10.1590/0103-8478cr20220508

ABSTRACT:

Spermatozoa experience oxidative, osmotic, chemical, and thermal stresses when cooled, which degrade the quality and fertilizing capacity of the cells. Adding antioxidants to the sperm extender mitigates these alterations. This study evaluated the effect of isoespintanol (ISO) on boar semen subjected to cooling. Fifteen ejaculates from five boars (Susscrofadomestica) were extended in Beltsville thawing solution (BTS) supplemented with 0 µM (control), 5 µM (ISO5), 10 µM (ISO10), 15 µM (ISO15), 20 µM (ISO20), 25 µM (ISO25), and 30 µM (ISO30) of ISO, which were then cooled for five days at 16 °C. Sperm kinetics, total motility (TM), and progressive motility (PM) were evaluated every 24 h using an IVOS computer-assisted sperm analysis (CASA) system. On day 1 and day 5 of cooling, a hypoosmotic test, spectrofluorometry, and flow cytometry were performed to evaluate the following: membrane functionality, measured as a function of hypoosmotic swelling (HOS); total antioxidant capacity (TAC); reactive oxygen species (ROS); and mitochondrial membrane potential (Δ¥M). Regression analysis and comparison of means using the Duncan test were performed. The ISO added had a slight impact on sperm motility, as evidenced by a reduction in TM at 24 h of cooling (but not prior) with the addition of 20 µM of ISO. Similarly, no effect of the ISO on the kinetics and functional integrity of the sperm membrane was observed at 96 h of cooling; however, the regression coefficients indicated that the ISO lowered the rate of decrease in sperm motility and the proportion of rapid spermatozoa relative to the concentration of ISO used. The ISO did not affect the TAC of the cooled semen; however, different concentrations of ISO lowered ROS production in the semen after 96 h of cooling. ISO also impacted the Δ¥M of the spermatozoa at 0 h of cooling, increasing the proportion of low Δ¥M cells and decreasing the proportion of high Δ¥M cells. In conclusion, ISO can reduce the loss of quality and oxidative stress occurring in boar semen during cooling and can modulate the mitochondrial activity of sperm.

Key words:
antioxidant; boar; preservation; semen quality; spermatozoa

[1] E-mail: grestre0@unal.edu.com. *Corresponding author.

	VAP	VSL	VCL	ALH	BCF	STR	LIN	RAP
---------------------------------------------------------------------------------0 h-------------------------------------------------------------------------------
CONTROL	50.7 ± 3.2	42.9 ± 2.3	96.7 ± 4.0^ab	5.0 ± 0.2	42.4 ± 0.7^a	78.2 ± 1.7^a	46.0 ± 2.1^a	52.4 ± 4.6
ISO5	49.9 ± 3.9	39.7 ± 2.4	100.4 ± 5.0^ab	5.3 ± 0.2	42.3 ± 0.6^ab	75.4 ± 1.4^ab	40.6 ± 1.0^b	51.7 ± 6.8
ISO10	51.5 ± 2.6	38.4 ± 2.1	95.8 ± 4.3^ab	5.3 ± 0.2	40.2 ± 0.7^b	75.6 ± 1.5^ab	43.0 ± 1.4^ab	43.0 ± 4.7
ISO15	51.8 ± 2.3	39.7 ± 1.6	94.1 ± 3.8^ab	5.2 ± 0.2	40.3 ± 0.8^ab	77.2 ± 1.1^ab	44.5 ± 1.3^ab	46.1 ± 4.9
ISO20	51.4 ± 3.4	66.0 ± 27.0	95.0 ± 5.1^ab	5.3 ± 0.2	41.0 ± 0.8^ab	76.4 ± 1.3^ab	43.0 ± 1.5^ab	43.9 ± 6.7
ISO25	49.9 ± 3.6	38.4 ± 2.8	92.1 ± 5.5^b	5.1 ± 0.2	41.0 ± 0.5^ab	77.6 ± 1.4^a	43.4 ± 1.5^ab	40.5 ± 6.2
ISO30	53.6 ± 2.1	39.3 ± 1.4	103.2 ± 4.5^a	5.5 ± 0.2	41.4 ± 1.0^ab	74.0 ± 1.3^b	40.2 ± 1.3^b	48.8 ± 3.8
-------------------------------------------------------------------------------24 h-------------------------------------------------------------------------------
CONTROL	50.8 ± 2.8^a	36.4 ± 1.9^a	97.2 ± 5.6	5.7 ± 0.2	38.1 ± 1.1	73.0 ± 1.8	40.7 ± 1.8	39.0 ± 5.2^a
ISO5	52.8 ± 2.0^a	38.9 ± 1.3^a	100.3 ± 4.8	5.5 ± 0.3	39.4 ± 1.2	74.6 ± 1.8	42.1 ± 1.9	42.4 ± 4.2^a
ISO10	45.7 ± 2.5^b	33.9 ± 2.0^b	88.4 ± 3.7	5.5 ± 0.2	39.0 ± 1.3	74.8 ± 1.4	40.8 ± 1.6	31.0 ± 4.9^b
ISO15	46.1 ± 2.3^b	31.8 ± 1.6^b	91.8 ± 4.0	6.1 ± 0.2	37.9 ± 0.7	72.7 ± 1.6	39.0 ± 1.7	33.2 ± 4.8^b
ISO20	46.8 ± 2.5^b	33.5 ± 1.5^b	94.2 ± 4.5	6.0 ± 0.2	65.5 ± 26.9	73.1 ± 1.6	38.8 ± 1.6	34.4 ± 4.8^a
ISO25	49.2 ± 2.1^a	35.9 ± 1.6^a	96.7 ± 3.9	5.5 ± 0.3	63.4 ± 23.1	74.2 ± 2.0	40.4 ± 2.0	39.7 ± 5.2^a
ISO30	47.5 ± 2.2^b	33.7 ± 1.5^b	165.4 ± 2.4	5.8 ± 0.2	63.7 ± 24.3	72.6 ± 2.1	39.4 ± 2.4	33.0 ± 4.0^b
-------------------------------------------------------------------------------48 h-------------------------------------------------------------------------------
CONTROL	45.1 ± 3.2	33.3 ± 2.5	91.5 ± 5.6	5.7 ± 0.3	38.7 ± 1.2^b	74.0 ± 1.7	38.8 ± 1.7	29.4 ± 5.1
ISO5	47.1 ± 2.8	34.5 ± 1.9	91.7 ± 5.2	5.7 ± 0.3	82.5 ± 29.7^a	74.2 ± 1.5	67.2 ± 27.1	31.4 ± 4.4
ISO10	48.4 ± 2.7	31.7 ± 1.8	98.3 ± 4.5	6.2 ± 0.2	40.1 ± 1.2^b	72.4 ± 1.2	37.9 ± 1.3	32.9 ± 4.8
ISO15	43.4 ± 2.8	31.2 ± 1.8	89.4 ± 4.9	5.8 ± 0.2	40.3 ± 1.0^b	73.5 ± 1.5	37.8 ± 1.3	26.5 ± 4.7
ISO20	43.7 ± 3.0	32.0 ± 2.3	88.4 ± 5.7	5.4 ± 0.2	40.1 ± 1.0^b	74.8 ± 2.0	40.3 ± 2.5	29.9 ± 5.7
ISO25	46.9 ± 2.9	33.0 ± 1.8	94.7 ± 5.4	5.9 ± 0.2	40.1 ± 1.0^b	72.6 ± 2.2	38.2 ± 2.1	32.4 ± 4.4
ISO30	45.9 ± 2.8	32.4 ± 1.7	88.8 ± 5.8	5.4 ± 0.4	38.9 ± 1.1^b	74.5 ± 2.4	41.5 ± 2.4	32.1 ± 4.8
-------------------------------------------------------------------------------72 h-------------------------------------------------------------------------------
CONTROL	46.7 ± 3.4	33.4 ± 1.9	91.2 ± 7.7	6.2 ± 0.3^a	36.0 ± 2.0	73.7 ± 2.8^a	44.0 ± 3.9	27.2 ± 5.7
ISO5	51.1 ± 3.7	36.3 ± 2.4	101.4 ± 7.4	6.0 ± 0.2^a	38.8 ± 0.9	73.0 ± 2.0^a	40.2 ± 2.7	30.7 ± 5.2
ISO10	49.3 ± 3.9	37.9 ± 3.3	91.3 ± 6.6	5.3 ± 0.3^a	38.5 ± 1.1	77.6 ± 1.9 ^a	45.3 ± 2.6	32.0 ± 6.8
ISO15	47.8 ± 3.7	34.3 ± 2.6	95.0 ± 7.0	6.0 ± 0.3^a	38.3 ± 0.9	73.7 ± 2.0^a	41.0 ± 2.5	29.2 ± 4.7
ISO20	51.6 ± 3.6	35.4 ± 2.4	100.0 ± 7.6	5.1 ± 0.5^b	40.1 ± 1.6	70.4 ± 3.6^a	39.8 ± 3.8	32.4 ± 5.1
ISO25	49.7 ± 3.5	36.2 ± 2.7	102.7 ± 8.3	5.9 ± 0.3^a	38.8 ± 1.1	73.5 ± 1.8^a	38.2 ± 2.0	33.0 ± 6.2
ISO30	44.4 ± 4.7	31.5 ± 3.4	89.0 ± 9.2	4.9 ± 0.6^b	36.2 ± 2.9	68.0 ± 5.7^b	37.3 ± 4.6	24.8 ± 4.4
-------------------------------------------------------------------------------96 h-------------------------------------------------------------------------------
CONTROL	45.5 ± 3.3	34.0 ± 2.9	89.6 ± 4.6	5.6 ± 0.3^a	39.1 ± 1.1	76.1 ± 1.8	40.5 ± 1.7	24.0 ± 6.5
ISO5	49.3 ± 3.7	34.6 ± 2.6	95.8 ± 6.6	6.1 ± 0.3^a	36.9 ± 1.2	73.3 ± 1.8	40.6 ± 2.0	28.3 ± 5.8
ISO10	47.1 ± 3.3	33.3 ± 2.5	91.5 ± 6.1	5.2 ± 0.6^a	35.0 ± 2.2	74.8 ± 2.7	41.7 ± 2.7	25.7 ± 5.2
ISO15	46.5 ± 3.1	33.4 ± 2.4	94.3 ± 5.9	5.9 ± 0.3^a	39.6 ± 1.2	74.3 ± 2.3	38.9 ± 1.6	23.0 ± 2.9
ISO20	48.6 ± 3.7	33.5 ± 3.1	95.8 ± 6.6	5.3 ± 0.4^a	39.6 ± 1.8	70.7 ± 3.6	37.6 ± 2.5	25.8 ± 4.4
ISO25	50.0 ± 4.7	36.2 ± 3.5	98.0 ± 8.9	5.3 ± 0.5^a	37.7 ± 2.3	74.7 ± 2.92	40.3 ± 2.5	30.5 ± 5.5
ISO30	44.7 ± 5.2	32.1 ± 3.2	88.6 ± 9.4	4.5 ± 0.7^b	39.9 ± 1.8	75.8 ± 2.9	40.1 ± 3.0	26.7 ± 6.9

COOLING TIME (h)	-------------------TREATMENT-----------------------	---------------------------HOS---------------------------
0	CONTROL	49.80 ± 2.06 ^a
	ISO5	48.30 ± 2.08 ^ab
	ISO10	43.90 ± 2.14 ^b
	ISO15	49.70 ± 2.77 ^a
	ISO20	50.10 ± 2.91 ^a
	ISO25	45.30 ± 2.45 ^ab
	ISO30	44.90 ± 1.97 ^ab
96	CONTROL	42.77 ± 2.91
	ISO5	41.33 ± 3.71
	ISO10	36.44 ± 3.73
	ISO15	42.44 ± 2.45
	ISO20	42.33 ± 4.12
	ISO25	39.78 ± 2.92
	ISO30	39.22 ± 3.88

VARIABLE	CONTROL	ISO5	ISO10	ISO15	ISO20	ISO25	ISO30
TM	-0.372^*	-0.175	-0.185^*	-0.212^*	-0.227^*	-0.172^*	-0.247^*
PM	-0.314^*	-0.266^*	-0.144	-0.222^*	-0.209	-0.082	-0.219^*
VAP	-0.048	-0.007	-0.008	-0.044	-0.036	0.033	-0.057
VSL	-0.101	-0.055	-0.057	-0.068	-0.076	-0.008	-0.052
VCL	-0.115	-0.098	0.063	0.058	0.023	0.131	-0.113
ALH	0.001	0.004	0.006	0.006	0.001	0.006	-0.008
BCF	-0.046^*	-0.056^*	-0.036	-0.005	-0.014	-0.045	-0.024
STR	-0.009	-0.016	-0.063	-0.048	-0.091	-0.049	0.002
LIN	-0.043	0.018	-0.062	-0.090^*	-0.083	-0.041	0.008
RAP	-0.328^*	-0.266^*	-0.109	-0.213^*	-0.174	-0.047	-0.213^*
HOS	-0.073	-0.076	-0.078	-0.076	-0.081	-0.058	-0.059

Brasil

Brasil

Cooling of porcine semen in an extender supplemented with isoespintanol

Resfriamento de sêmen suíno em um diluente suplementado com isoespintanol

ABSTRACT: