Table 1.
Descriptive statistics of major computer-assisted semen analysis (CASA) measurements of the main trial (mean values ± standard deviation) at each observation time (0–4 h). Number of replicates = 10 in each test.
| Treatments # | 0 h | 1 h | 2 h | 3 h | 4 h | p * (DMSO vs. Mycotoxin(s) Treatment) |
|---|---|---|---|---|---|---|
| Immotile Spermatozoa (%) | ||||||
| DMSO | 2.76 ± 1.72 | 4.01 ± 3.11 | 5.48 ± 3.08 | 8.67 ± 4.18) | 10.70 ± 5.34 | |
| DON | 3.95 ± 2.43 | 6.11 ± 2.94 | 8.32 ± 3.84 | 12.09 ± 5.28 | 17.63 ± 10.90 | 0.001 |
| ZEN | 13.87 ± 10.98 | 21.95 ± 16.75 | 31.11 ± 19.02 | 32.37 ± 20.55 | 38.83 ± 18.69 | <0.001 |
| DON + ZEN | 19.22 ± 9.55 | 19.17 ± 11.82 | 25.77 ± 8.16 | 34.13 ± 14.27 | 35.98 ± 15.51 | <0.001 |
| Non-progressive Motile Spermatozoa (%) | ||||||
| DMSO | 19.35 ± 3.82 | 22.37 ± 6.03 | 21.67 ± 4.25 | 20.60 ± 3.52 | 21.19 ± 4.55 | |
| DON | 19.88 ± 2.18 | 24.18 ± 6.13 | 24.79 ± 2.83 | 24.76 ± 4.10 | 25.33 ± 8.43 | 0.001 |
| ZEN | 34.02 ± 5.94 | 34.20 ± 11.17 | 33.81 ± 9.21 | 36.40 ± 11.52 | 33.21 ± 11.33 | <0.001 |
| DON + ZEN | 41.31 ± 9.91 | 39.64 ± 12.04 | 44.07 ± 8.32 | 39.87 ± 10.90 | 38.50 ± 8.87 | <0.001 |
| Progressive Motile Spermatozoa (%) | ||||||
| DMSO | 77.88 ± 4.68 | 73.62 ± 7.13 | 72.85 ± 6.53 | 70.73 ± 6.12 | 68.11 ± 6.64 | |
| DON | 76.17 ± 3.53 | 69.71 ± 8.28 | 66.89 ± 5.70 | 63.14 ± 7.65 | 57.04 ± 16.61 | 0.056 |
| ZEN | 52.11 ± 14.24 | 43.85 ± 21.09 | 35.08 ± 18.95 | 31.24 ± 19.52 | 27.95 ± 17.81 | <0.001 |
| DON + ZEN | 39.48 ± 18.70 | 41.19 ± 20.20 | 30.16 ± 11.62 | 26.00 ± 14.57 | 25.52 ± 16.43 | <0.001 |
| Rapid (%) | ||||||
| DMSO | 73.75 a ± 15.03 | 61.51 a ± 18.80 | 53.72 a ± 17.03 | 47.18 a ± 15.10 | 43.65 a ± 12.82 | |
| DON | 74.02 a ± 15.48 | 53.86 a ± 18.39 | 44.95 a ± 16.06 | 39.33 a ± 12.86 | 33.47 a ± 13.16 | 4 h: 0.063 ** |
| ZEN | 42.00 b ± 13.66 | 31.43 b ± 20.68 | 22.99 b ± 15.56 | 20.84 b ± 16.26 | 19.80 b ± 14.31 | Each time point: <0.001 ** |
| DON + ZEN | 28.24 c ± 16.05 | 31.40 b ± 21.67 | 16.93 b ± 10.90 | 15.65 b ± 11.15 | 15.01 b ± 10.61 | Each time point: <0.001 ** |
| Medium (%) | ||||||
| DMSO | 17.36 a ± 9.5 | 24.35 a ± 9.86 | 27.91 ac ± 7.59 | 31.87 a ± 6.6 | 31.44 a ± 8.14 | |
| DON | 15.79 a ± 8.07 | 26.68 a ± 9.47 | 31.26 a ± 7.88 | 31.78 a ± 5.01 | 32.31 a ± 8.29 | |
| ZEN | 23.17 a ± 5.97 | 22.34 a ± 6.51 | 18.96 c ± 7.92 | 20.00 b ± 7.76 | 18.14 b ± 7.53 | 0 h: 0.08, 2 h: 0.008, 3 h, 4 h: <0.001 ** |
| DON + ZEN | 22.17 a ± 6.96 | 21.02 a ± 4.85 | 21.89 bc ± 6.24 | 19.11 b ± 6.87 | 21.37 b ± 8.13 | 2 h: 0.07, 3 h & 4 h: <0.001 ** |
| Slow (%) | ||||||
| DMSO | 6.12 a ± 4.89 | 10.14 a ± 7.40 | 12.88 a ± 7.43 | 12.27 a ± 5.19 | 12.21 a ± 3.07 | |
| DON | 6.24 a ± 5.91 | 13.34 a ± 8.03 | 15.47 a ± 6.50 | 16.79 a ± 5.82 | 16.59 ab ± 4.12 | 1 h: 0.062 ** |
| ZEN | 20.95 b ± 7.15 | 24.28 b ± 9.38 | 26.94 b ± 9.64 | 26.79 b ± 10.37 | 23.22 bc ± 8.20 | 0 h, 1 h, 2 h, 3 h: <0.001 ** 4 h: 0.006 |
| DON + ZEN | 30.37 b ± 11.70 | 28.41 b ± 11.62 | 35.40 b ± 7.18 | 31.11 b ± 8.79 | 27.63 c ± 6.77 | Each time point: <0.001 ** |
| VCL (Curvilinear Velocity; μm/s) | ||||||
| DMSO | 74.36 a ± 19.10 | 61.90 a ± 15.31 | 59.21 a ± 21.78 | 51.08 a ± 9.86 | 48.12 a ± 5.88 | |
| DON | 74.01 a ± 16.57 | 57.27 a ± 13.69 | 51.44 a ± 10.90 | 45.22 a ± 8.12 | 41.88 ab ± 7.99 | 2 h: 0.098 ** |
| ZEN | 50.09 b ± 10.69 | 43.02 b ± 13.77 | 37.74 b ± 10.65 | 35.72 b ± 12.42 | 35.55 bc ± 9.96 | 0 h, 1 h, 2 h: <0.001, 3 h: 0.001, 4 h: 0.008 ** |
| DON + ZEN | 39.96 c ± 11.87 | 44.38 b ± 18.49 | 33.92 b ± 9.59 | 32.63 b ± 9.19 | 32.43 c ± 7.46 | 0 h, 1 h, 2 h, 3 h: <0.001 ** 4 h: 0.001 |
| VSL (Straight-line Velocity; μm/s) | ||||||
| DMSO | 32.46 ± 3.55 | 32.46 ± 2.60 | 33.58 ± 5.71 | 30.83 ± 3.81 | 28.81 ± 5.08 | |
| DON | 32.71 ± 2.16 | 30.57 ± 3.31 | 29.35 ± 2.18 | 27.18 ± 5.29 | 24.58 ± 7.46 | |
| ZEN | 18.57 ± 4.82 | 19.05 ± 8.91 | 15.74 ± 7.74 | 14.46 ± 10.03 | 12.33 ± 8.90 | <0.001 |
| DON + ZEN | 14.43 ± 7.52 | 19.12 ± 12.25 | 13.60 ± 6.22 | 12.74 ± 9.04 | 10.47 ± 6.45 | <0.001 |
| VAP (Average Path Velocity; μm/s) | ||||||
| DMSO | 45.19 ± 5.96 | 42.18 ± 4.42 | 42.59 ± 9.02 | 38.36 ± 4.73 | 36.21 ± 5.14 | |
| DON | 45.78 ± 4.66 | 38.91 ± 4.03 | 37.43 ± 3.34 | 33.81 ± 6.08 | 30.72 ± 7.81 | 0.089 |
| ZEN | 28.19 ± 6.32 | 26.33 ± 10.39 | 22.06 ± 8.19 | 20.60 ± 10.84 | 18.20 ± 9.10 | <0.001 |
| DON + ZEN | 22.07 ± 9.57 | 26.90 ± 14.05 | 19.70 ± 6.74 | 18.53 ± 9.47 | 16.31 ± 6.85 | <0.001 |
| LIN (Linearity; %) | ||||||
| DMSO | 45.62 a ± 9.8 | 54.94 a ± 12.1 | 60.16 a ± 12.73 | 62.08 a ± 12.03 | 60.21 a ± 10.48 | |
| DON | 46.15 a ± 10.14 | 56.25 a ± 14.69 | 59.59 a ± 14.06 | 61.21 a ± 12.35 | 59.49 a ± 17.55 | |
| ZEN | 37.39 b ± 7.47 | 43.35 b ± 14.24 | 41.06 b ± 15.28 | 38.43 b ± 20.01 | 33.68 b ± 18.74 | 0 h: 0.028, 1 h: 0.002, 2 h,3 h, 4 h: <0.001 ** |
| DON + ZEN | 34.16 b ± 10.36 | 41.30 b ± 14.47 | 40.52 b ± 17.56 | 37.21 b ± 19.45 | 31.98 b ± 17.6 | 0 h: 0.002, 1 h,2 h,3 h, 4 h: <0.001 ** |
| STR (Straightness; %) | ||||||
| DMSO | 72.08 a ± 4.07 | 77.20 a ± 3.64 | 79.37 a ± 4.25 | 80.45 a ± 3.62 | 79.16 a ± 3.79 | |
| DON | 71.76 a ± 4.06 | 78.64 a ± 4.72 | 78.66 a ± 5.06 | 80.29 a ± 3.92 | 78.85 a ± 7.96 | |
| ZEN | 65.50 b ± 4.16 | 70.09 b ± 8.27 | 68.12 b ± 12.13 | 63.74 b ± 16.32 | 62.87 b ± 12.78 | 0 h: 0.033, 1 h: 0.022, 3 h, 4 h: <0.001 ** |
| DON + ZEN | 62.61 b ± 7.70 | 67.64 b ± 10.08 | 66.23 b ± 12.06 | 63.12 b ± 14.46 | 59.75 b ± 14.63 | 0 h & 1 h: 0.002, 2 h,3 h, 4 h: <0.001 ** |
| Wobble (WOB; %) | ||||||
| DMSO | 62.92 a ± 10.79 | 70.66 a ± 12.26 | 75.26 a ± 12.23 | 76.72 a ± 11.6 | 75.68 a ± 10.18 | |
| DON | 63.89 a ± 10.78 | 70.79 a ± 14.43 | 75.06 a ± 13.11 | 75.75 a ± 12.13 | 74.03 a ± 16.52 | |
| ZEN | 56.70 a ± 7.84 | 60.55 b ± 13.47 | 58.34 b ± 13.92 | 56.44 b ± 18.55 | 50.45 b ± 17.86 | 0 h: 0.057, 1 h: 0.002, 2 h,3 h, 4 h: <0.001 ** |
| DON + ZEN | 53.49 b ± 10.45 | 59.39 b ± 13.52 | 58.69 b ± 16.25 | 55.43 b ± 17.88 | 50.09 b ± 16.84 | 0 h: 0.004, 1 h: 0.001, 2 h,3 h, 4 h: <0.001 ** |
| ALH (Amplitude of Lateral Head Displacement; μm) | ||||||
| DMSO | 1.89 a ± 0.26 | 1.64 a ± 0.2 | 1.60 a ± 0.4 | 1.41 a ± 0.14 | 1.39 a ± 0.07 | |
| DON | 1.90 a ± 0.24 | 1.58 a ± 0.2 | 1.48 a ± 0.17 | 1.32 a ± 0.17 | 1.26 a ± 0.18 | |
| ZEN | 1.74 a ± 0.12 | 1.51 a ± 0.19 | 1.51 a ± 0.14 | 1.40 a ± 0.09 | 1.43 a ± 0.19 | 0 h: 0.074 ** |
| DON + ZEN | 1.59 b ± 0.24 | 1.57 a ± 0.18 | 1.42 b ± 0.17 | 1.41 a ± 0.13 | 1.36 a ± 0.17 | 0 h: <0.001, 2 h: 0.033 ** |
| BCF (Beat/Cross Frequency; Hz) | ||||||
| DMSO | 13.13 ± 2.81 | 11.80 ± 2.41 | 10.60 ± 2.25 | 10.50 ± 1.79 | 10.32 ± 1.83 | |
| DON | 12.70 ± 2.03 | 11.14 ± 2.37 | 10.05 ± 1.87 | 9.73 ± 1.47 | 9.36 ± 1.72 | |
| ZEN | 9.72 ± 1.58 | 9.04 ± 2.60 | 9.17 ± 2.63 | 7.95 ± 3.51 | 7.64 ± 2.19 | 0.001 |
| DON + ZEN | 7.93 ± 2.08 | 8.3 ± 11.97 | 8.00 ± 2.97 | 7.13 ± 2.39 | 6.05 ± 2.53 | <0.001 |
| Hyperactive (%) | ||||||
| DMSO | 0.031 a ± 0.02 | 0.022 a ± 0.017 | 0.022 a ± 0.033 | 0.007 a ± 0.008 | 0.005 a ± 0.005 | |
| DON | 0.031 a ± 0.019 | 0.018 ab ± 0.014 | 0.010 b ± 0.010 | 0.006 a ± 0.006 | 0.004 a ± 0.004 | 2 h: 0.024 ** |
| ZEN | 0.010 b ± 0.005 | 0.011 b ± 0.013 | 0.005 b ± 0.007 | 0.003 a ± 0.004 | 0.001 a ± 0.001 | 0 h: 0.000, 1 h: 0.036, 2 h: 0.001 ** |
| DON + ZEN | 0.009 b ± 0.011 | 0.016 ab ± 0.02 | 0.004 b ± 0.007 | 0.002 a± 0.003 | 0.001 a ± 0.001 | 0 h: 0.000, 2 h: 0.001 ** |
a,b,c Mean values with different superscripts in the same column differ significantly (p < 0.05). * When p-values are reported without superscripts in relevant parameter mean values, they refer to statistically significant main effect (p < 0.05) or trend towards statistical significance (0.05 < p < 0.1) of treatment (mycotoxin) on the response variable, without significant interaction term treatment × time, thus differences refer to the total observation period. ** When p-values are reported and superscripts (a,b,c) are placed in relevant parameter mean values, they refer to statistically significant main effect (p < 0.05) or trend towards statistical significance (0.05 < p < 0.1) of treatment (mycotoxin) on the response variable, with a statistically significant interaction term treatment × time on the response variable present, thus differences refer to specific time points (0–4 h of investigation). # Treatments: dimethyl sulfoxide (DMSO) = 0.7% (v/v); deoxynivalenol (DON) = 50.6 μM; zearalenone (ZEN) = 62.8 μM; DON + ZEN = 50.6 μM DON + 62.8 μM ZEN.