Table 3.
Steroid concentrations determined by GC/MS/MS in the preovulatory follicular fluid of ewes according to their metabolic status and dietary exposure to BPS.
| Steroids | POOL SAMPLES | ||||||
|---|---|---|---|---|---|---|---|
| Mean concentration (ng/mL) ± SEM | p-value | ||||||
| R0 | R50 | WF0 | WF50 | Diet effect | BPS effect | Diet x BPS effect | |
| Precursors | |||||||
| PREG | 194.800 ± 61.869 | 103.200 ± 28.385 | 124.467 ± 24.461 | 153.460 ± 7.076 | 0.76 | 0.37 | 0.072 |
| 20α-DHPREG | < 0.050 | < 0.050 | < 0.050 | < 0.050 | NA | NA | NA |
| 17α-OH PREG | 38.417 ± 9.359 a | 20.597 ± 4.952 ab | 11.145 ± 5.073 b | 23.330 ± 5.057 ab | 0.07 | 0.65 | 0.028 |
| Progestogens (C21) | |||||||
| Progesterone | 13.550 ± 3.580 ab | 8.517 ± 1.763 a | 20.013 ± 6.793 ab | 14.398 ± 1.632 b | 0.25 | 0.34 | 0.99 |
| 5α-DHP | 7.950 ± 1.748 | 8.590 ± 2.758 | 12.310 ± 4.412 | 11.302 ± 2.827 | 0.35 | 0.94 | 0.81 |
| 5β-DHP | 0.177 ± 0.125 | 0.178 ± 0.132 | 0.113 ± 0.084 | 0.171 ± 0.088 | 0.75 | 0.64 | 0.73 |
| 20α-DHP | 2.677 ± 0.737 | 1.315 ± 0.397 | 2.977 ± 0.794 | 2.526 ± 0.245 | 0.28 | 0.17 | 0.51 |
| 3α5α-THP | 6.902 ± 1.000 | 5.052 ± 2.015 | 5.263 ± 1.096 | 3.732 ± 1.770 | 0.33 | 0.27 | 0.96 |
| 3α5β-THP | 1.222 ± 0.738 | 1.105 ± 0.389 | 1.568 ± 0.575 | 1.046 ± 0.444 | 0.80 | 0.59 | 0.75 |
| 3β5α-THP | 0.294 ± 0.098 | 0.574 ± 0.410 | 0.747 ± 0.448 | 1.053 ± 0.875 | 0.48 | 0.64 | 0.99 |
| 3α5α20α-HHP | < 0.010 | < 0.010 | < 0.010 | < 0.010 | NA | NA | NA |
| 3α5β20α-HHP | 0.131 ± 0.043 a | 0.111 ± 0.051 ab | 0.086 ± 0.008 ab | 0.048 ± 0.024 b | 0.11 | 0.38 | 0.75 |
| 3α5β20β-HHP | < 0.200 | < 0.200 | < 0.200 | < 0.200 | NA | NA | NA |
| 3β5α20β-HHP | 0.684 ± 0.307 | 1.723 ± 1.247 | 1.181 ± 0.838 | 2.867 ± 2.744 | 0.77 | 0.52 | 0.88 |
| 3β5α20α-HHP | 0.165 ± 0.078 | 0.609 ± 0.377 | 0.531 ± 0.340 | 0.466 ± 0.336 | 0.76 | 0.63 | 0.43 |
| 17α-OHP | 37.185 ± 7.076 a | 22.625 ± 4.275 b | 27.062 ± 6.900 ab | 26.688 ± 2.748 ab | 0.62 | 0.22 | 0.24 |
| 16α-OHP | 11.582 ± 3.468 ab | 11.390 ± 1.994 a | 6.346 ± 1.204 b | 7.668 ± 1.463 ab | 0.040 | 0.80 | 0.71 |
| Androgens (C19) | |||||||
| Testosterone | 5.800 ± 1.783 a | 5.897 ± 1.628 a | 1.690 ± 0.689 b | 3.658 ± 1.010 ab | 0.060 | 0.52 | 0.54 |
| 5α-DHT | 0.200 ± 0.056 a | 0.053 ± 0.016 b | 0.076 ± 0.016 b | 0.084 ± 0.008 b | 0.08 | 0.004 | 0.001 |
| 5β-DHT | 0.239 ± 0.114 a | 1.150 ± 0.298 b | 0.855 ± 0.308 ab | 0.707 ± 0.164 b | 0.76 | 0.16 | 0.057 |
| DHEA | 3.760 ± 1.157 | 3.265 ± 0.261 | 1.247 ± 0.802 | 2.422 ± 0.959 | 0.08 | 0.72 | 0.37 |
| ADIOL | 2.335 ± 0.436 a | 2.070 ± 0.426 a | 0.465 ± 0.265 b | 1.838 ± 0.826 ab | 0.07 | 0.33 | 0.15 |
| ADIONE | 4.027 ± 1.068 | 3.787 ± 0.989 | 2.323 ± 1.049 | 3.202 ± 1.505 | 0.37 | 0.77 | 0.64 |
| 5α-DHADIONE | 0.154 ± 0.107 | 0.106 ± 0.026 | 0.083 ± 0.023 | 0.101 ± 0.013 | 0.55 | 0.88 | 0.65 |
| Estrogens (C18) | |||||||
| Estradiol | 92.270 ± 7.836 a | 76.205 ± 12.855 a | 36.427 ± 11.181 b | 100.340 ± 24.113 a | 0.34 | 0.16 | 0.020 |
| E1 | 40.110 ± 5.168 a | 46.685 ± 14.805 a | 18.360 ± 6.138 b | 44.520 ± 11.877 ab | 0.21 | 0.10 | 0.39 |
| E3 | 0.410 ± 0.077 ab | 0.445 ± 0.032 a | 0.300 ± 0.061 b | 0.356 ± 0.090 ab | 0.19 | 0.55 | 0.88 |
| 2-ME2 | 0.095 ± 0.033 | 0.171 ± 0.039 | 0.097 ± 0.048 | 0.088 ± 0.043 | 0.36 | 0.46 | 0.37 |
| 2-OH-E2 | 9.242 ± 2.515 | 10.730 ± 0.782 | 4.630 ± 2.183 | 6.924 ± 3.109 | 0.10 | 0.46 | 0.88 |
| Corticoids (C21) | |||||||
| Glucocorticoids | |||||||
| Cortisol (F) | < 2.000 | < 2.000 | < 2.000 | < 2.000 | NA | NA | NA |
| Cortisone | < 2.000 | < 2.000 | < 2.000 | < 2.000 | NA | NA | NA |
| 11-deoxyF | < 2.000 | < 2.000 | < 2.000 | < 2.000 | NA | NA | NA |
| 3α5β-THF | < 2.000 | < 2.000 | < 2.000 | < 2.000 | NA | NA | NA |
| Mineralocorticoids | |||||||
| Corticosterone (B) | 0.842 ± 0.200 | 0.805 ± 0.158 | 0.662 ± 0.136 | 0.650 ± 0.348 | 0.5 | 0.92 | 0.99 |
| 11-dehydroB | 1.050 ± 0.313 a | 0.250 ± 0.161 b | 0.822 ± 0.383 ab | 0.247 ± 0.124 b | 0.76 | 0.027 | 0.69 |
| 3α5α-THDOC | 0.036 ± 0.016 ab | 0.022 ± 0.014 a | 0.070 ± 0.018 ab | 0.088 ± 0.016 b | 0.013 | 0.88 | 0.36 |
Ewes were either restricted (R groups) or well-fed (WF groups) and received BPS 0 or 50 µg/kg/day, for at least three months before the start of plasma/fluid collection. Thus, four groups were formed: R0 (n = 4), R50 (n = 4), WF0 (n = 6) and WF50 (n = 5). Results are presented as mean (ng/mL) ± SEM. Tukey post-tests are indicated by superscript letters, different letters indicating significantly different conditions (p < 0.05). Two-way ANOVA p-values are presented for the effects of diet, dietary exposure to BPS and the interaction of these effects. Bold text indicates significant differences (p < 0.05). See Table 1 for the steroid abbreviations. NA, not available.