Antioxidant effect of Cymbopogon citratus and Hypericum perforatum extracts in boar semen like a new alternative for conservation

Figures & data

Table 1. Concentration series for calibration curve of Trolox.

Concentration (µM)	Trolox (µl)	80% Methanol (µl)
0	0	80
80	4	76
160	8	72
320	16	64
480	24	56
640	32	48
960	48	32
1280	64	16

Figure 1. Average of temporal evolution of the antioxidant activity of natural extracts and BTS. Blue corresponds to Cymbopogon citratus, black for Hypericum perforatum and red corresponds to BTS.

Table 2. Total phenolic contents of Cymbopogon citratus and Hypericum perforatum extracts.

Cymbopogon citratus	160	±	1.6	mg GAE/100g
Hypericum perforatum	264	±	0.9	mg GAE/100g

Gallic acid equivalents (GAE), Media ± standar deviation, mg = milligrams, g = grams.

Figure 2. A peak corresponding to fructose in Hypericum perforatum can be observed under the limit quantification 4 min after release.

Figure 3. A peak corresponding to glucose in Cymbopogon citratus can be observed under the limit quantification 3.7 min after release.

Table 3. Seminal motility averages for all times and extender types.

	Time of motility (h)†
Seminal extender (µl)	24 (%) SD±	48 (%) SD±	72 (%) SD±
T1	55.0^a ± 4.5	43.3^a ± 8.2	33.3^a ± 5.2
T2	52.5^a ± 4.2	16.7^b c ± 4.1	9.2^b ± 3.8
T3	36.7^b c ± 5.2	20.0^b c ± 6.3	5.8^b c ± 2.0
T4	30.8^c d ± 5.9	21.7^b ± 2.6	2.5^c d ± 0.5
T5	25.0^d ± 5.5	13.3^c d ± 4.1	2.5^c d ± 0.8
T6	24.2^d ± 5.9	6.7^d ± 2.6	0.8^d ± 0.8
T7	51.7^a ± 2.6	18.3^b c ± 2.6	5.8^b c  ±  2.0
T8	42.5^b ± 2.7	12.5^c d ± 2.7	1.7^c d  ±  0.5
T9	31.7^c d ± 2.6	7.5^d ± 2.7	0.7^d ± 1.0
P value	0.000	0.000	0.000

†: Averages values (%): Media ± standard deviation. BTS: Beltsville Thawing Solution.

a,b,c,d: Values with different literals are different between the same column (P < 0.05).

T1 = BTS 0.250 µl, T2 = BTS 0.125 µl, T3 = BTS 0.0625 µl, T4 = Hp 0.250 µl, T5 = Hp 0.125 µl, T6 = Hp 0.0625 µl, T7 = Cc 0.250 µl, T8 = Cc 0.125 µl, T9 = Cc 0.0625 µl.

Table 4. Seminal viability averages for all times and extender types.

	Time of viability (h)†
Seminal extender (µl)	24 (%) SD ±	48 (%) SD ±	72 (%) SD ±
T1	83.3^a ± 4.3	84.5^a b ± 5.8	83.5^a ± 5.7
T2	83.1^a ± 5.5	85.3^a ± 2.4	85.1^a ± 5.5
T3	76.6^a ± 4.4	76.8^a b c ± 6.1	73.1^a ± 14.7
T4	76.8^a ± 4.2	81.5^a b c ± 3.1	83^a ± 4.7
T5	82.1^a ± 3.9	80.7^a b c ± 5.1	85.3^a ± 2.7
T6	83^a ± 6.0	80.8^a b c ± 6.1	85.1^a ± 3.3
T7	76.8^a ± 6.7	72.1^c ± 8.8	69.3^a ± 16.3
T8	80.1^a ± 7.6	74.8^b c ± 3.9	74.5^a ± 12.4
T9	79^a ± 6.8	73.1^c ± 5.0	76.1^a ± 13.9
P value	0.136	0.000	0.042

†: Averages values (%): Media ± standar deviation. BTS: Beltsville Thawing Solution.

a,b,c,d: Values with different literals are different between the same column (P < 0.05).

T1 = BTS 0.250 µl, T2 = BTS 0.125 µl, T3 = BTS 0.0625 µl, T4 = Hp 0.250 µl, T5 = Hp 0.125 µl, T6 = Hp 0.0625 µl, T7 = Cc 0.250 µl, T8 = Cc 0.125 µl, T9 = Cc 0.0625 µl.

Table 5. Average of primary morphologies of seminal doses preserved with different extenders and their specific concentrations at 24 and 72 h.

	N†		DA†		HD†
Seminal extender (µl)	24(%)	72(%)	24(%)	72(%)	24(%)	72(%)
T1	93.1^a ± 1.2	72.9^d ± 5.7	2.5 ^a ± 2.1	18.2^a ± 7.7	0.1^a ± 0.4	0.5^c ± 0.6
T2	91.7^a ± 1.5	81^a b c ± 1.3	2.9 ^a ± 1.9	12.8^a b c ± 0.7	0.3^a ± 0.5	0.6^c ± 0.5
T3	93.3^a ± 3.4	79.5^b c d ± 6.0	2 ^a ± 2.6	13.1^a b c ± 7.1	0.4^a ± 0.6	0.3^c ± 0.5
T4	93.5^a ± 1.4	74.3^c d ± 7.2	2.1 ^a ± 0.9	15.9^a b ± 4.9	0.1^a ± 0.4	0.8^c ± 0.8
T5	94.2^a ± 2.5	80.8^a b c d ± 2.3	2.0 ^a ± 2.5	6.7^a b c ± 2.1	0.5^a ± 0.6	3.3^a b ± 1.9
T6	93.4^a ± 2.2	87.6^a ± 2.4	3.7 ^a ± 2.1	6.5 ^c ± 2.3	0.2^a ± 0.4	1.4^b c ± 1.5
T7	95^a ± 1.1	86.1^a b ± 2.6	1.3^a ± 0.9	7.8^b c ± 2.9	0.5^a ± 0.4	1^b c ± 0.6
T8	92.9^a ± 1.7	83.3^a b ± 4.1	1.7^a ± 1.7	8.3^b c ± 4.6	0.9^a ± 0.8	0.5^c ± 0.5
T9	94.1^a ± 2.6	82.3^a b c ± 1.7	2^a ± 1.9	5.8^c ± 1	1^a ± 0.6	4.5^a ± 2.7
P Value	0.327	0.000	0.617	0.000	0.116	0.000

†: Averages values (%): Media ± standar deviation. BTS: Beltsville Thawing Solution.

a,b,c,d: Values with different literals are different between the same column (P < 0.05).

T1 = BTS 0.250 µl, T2 = BTS 0.125 µl, T3 = BTS 0.0625 µl, T4 = Hp 0.250 µl, T5 = Hp 0.125 µl, T6 = Hp 0.0625 µl, T7 = Cc 0.250 µl, T8 = Cc 0.125 µl, T9 = Cc 0.0625 µl.

N = Normal, DA = Damaged acrosomes, HD = Heads detached.

Data availability statement (DAS)

The data that support the findings of this study are available in Figshare at https://doi.org/10.6084/m9.figshare.13363559.