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Effect of injection time of anti-PMSG antiserum on ovulation rate and quality of embryos in superovulated cows
THERIOGENOLOGY EFFECT OF INJECTION TIME OF ANTI-PMSG ANTISERUM ON OVULATION RATE AND QUALITY OF EMBRYOS IN SUPEROVULATED COWS
J. Saumande, R. Procureur
and D. Chupin
I.N.R.A. Station de Physiologie de la Reproduction 37380 Nouzilly, France
Received
for publication: Accepted:
September 12, 1983 March 15, 1984
ABSTRACT Eighteen cows were superovulated by injecting 3000 IU of PMSG during the luteal phase, followed 48h later with an injection of Estrumate. They were then placed in a control group or were given anti-PMSG antiserum at either 12h or 24h after the onset of oestrus. Sixteen of these animals were used for the same experiment five months later. The results were pooled because they were not significantly different between the two treatment periods The timing of the injection ot anti-PMSG antiserum, either 12h (II cows) or 24h (12 cows) after the onset of oestrus,did not significantly affect the ovulation rate, the number of embryos collected or the number of good embryos. The antiserum significantly increased the number of good embryos but did not affect the ovulation rate or embryo recovery. It is concluded that even with a moderate dose of PMSG, the use of anti-PMSG at l2h or 24h after the beginning of oestrus improves the quality of embryos. The mean number of embryos to be transferred (5.5) is in the range of those obtained after the FSH treatments,but the procedure required only three injections compared with nine for the FSH treatment.
INTRODUCTION When superovulation is induced in cattle by injecting PMSG, large follicles are often observed on the ovary at the time of embryo recovery (1)2). Such follicles are observed more frequently at the high doses of PMSG (3), and their presence is associated with a lower quality of recovered embryos (4). Under these conditions, the number of embryos to be transferred cannot be improved by increasing ovulation rate with a larger dose of PMSG. The injection of anti-PMSG in order to control the duration of follicular growth stimulation has been shown to improve the quality of embryos (5) as previously hypothesized (6). In superovulated animals, ovulation is spread over 72 hours (7). Anti-PMSG could interfere with ovulation since such antisera are known to crossreact with hypophyseal gonadotrophins (8). The purpose of this experiment was to test the effect of time of anti-PMSG injection on the number of good embryos recovered and to assess the effect of such treatment when moderate doses of PMSG are used.
MAY 1984VOL. 21 NO. 5
727
THERIOGENOLOGY MATERIALS AND METHODS Eighteen lactating Friesian cows were injected with PMSG (3000 IU) during the luteal phase (between day 8 and day 14) and 4Sh later with Img of Estrumate. Oestrus was detected twice a day with a vasectomized bull; the day of oestrus was day 0. Cows were inseminated at 12h and 24h after the beginning of oestrus. Six cows were used as controls and received no antiserum. The remaining 12 cows were injected intravenously with 5ml anti-PMSG*. Six cows received the antiserum 12h and six 24h after the beginning of oestrus. The same treatment was applied five months later to 16 Of these animals(fivecontrol, five injected with anti-PMSG at 12h and six injected at 24h after the beginning of oestrus).Care was taken to allot the cows so that each group included animals of the other groups of the first trial. After the first superovulation, embryos were collected nonsurgically on day 7 or day 8 and ovarian responses were measured by rectal palpation; after the second, superovulation, collection and ovarian examination were done after slaughter for six animals and by nonsurgical recovery for the other ten. The quality of embryos was assessed through morphological criteria (2) using a stereo-microscope (10 to 40 magnification). Results were compared with the Mann-Whitney U test (for two samples) and the Kruskal and Wallis one-way analysis of variance.
RESULTS The results obtained five months apart were not different for any of the parameters studied (P (O.OOl), so the data were pooled and are presented in Table 1. Whether anti-PMSG was injected 12h or 24h after the beginning of oestrus did not affect either the ovulation rate, the number of embryos collected or the percentage of good embryos (P< 0.001). Comparison of the results obtained after injection of anti-PMSG with those of the control group showed that the injection of antiserum did not affect ovulation rate (P(O.05) or the number of embryos collected (P(O.02); however, the percentage of good embryos obtained after injection of anti-PMSG was increased (P
DISCUSSION The ovulation rate was not affected by the injection of anti-PMSG 12h or 24h after the beginning of oestrus. In both cases we can assume that the antiserum was injected after the preovulatory peak of gonado-
x This antiserum was a gift of Prof. R. Bouters, Gand, Belgium.
728
MAY 1984VOL. 21 NO. 5
THERIOGENOLOGY
MAY 1984VOL. 21 NO. 5
729
THERIOGENOLOGY
trophins, which occurs at the beginning of oestrus (9, IO, 11). On the other hand, when antiserum was injected at the beginning of oestrus in four cows that had received 3000 IU of PMSG, only one follicle was ovulated in each animal (unpublished work). These results all agree with the current hypothesis on the ovulation process: once the LH-FSH discharge has occurred, the events which lead to ovulation are no longer gonadotrophin-dependent (12). However, this still leaves unexplained the fact that ovulation can be spread over a long time (7) . Our data also show that for a moderate dose of PMSG, the injection of anti-PMSG increased the percentage of good embryos. With these conditions, even though neither the ovulation rate nor the recovery rate was changed, the improvement in the quality of embryos led to an increase in the number of embryos to be transferred (5.5 vs 3.5). When lactating Friesian cows are treated with the most efficent of the FSH treatments, a mean of 5.3 good embryos is obtained (13). Therefore,the same number of good embryos can be obtained with both treatments. However, from a practical point of view, only three injections are needed when anti-PMSG is used (PMSG, prostaglandin and antiserum) compared with nine for an FSH treatment (height injection of gonadotrophin and prostaglandin).
In conclusion, PMSG is an efficient hormone for the production of embryos provided the duration of its stimulatory effect is controlled with an antiserum. This treatment is more practical than those with FSH because the number of injections is reduced and the time of antiPMSG administration after oestrus is not critical.
REFERENCES 1.
Booth, W.D., Newcomb, R., Strangle, H., Rowson, L.E.A. and Sacher, H.B Plasma oestrogen and progesterone in relation to superovulation and egg recovery in the cow. Vet. Rec. -97: 366-369 (1975).
2.
Renard, J.P., Menezo, Y., Saumande, 3. and Heyman, Y. Attempts to check the viability of embryos produced by superovulated heifers. In Control of Reproduction in the Cow, Ed. J.M. Sreenan. Nijhoff M., xe Hague. 1978, pp.398-417.
3.
Mauldon, P., Mariana, J.C., Benoit, M., Solari, A. and Chupin,D. Action upon the ovulation number and yield of French Friesian cows of different PMSG and HCG doses injected during the follicular phase of the oestrous cycle. Ann. Biol. Anim. Bioch. Biophys. lo, Suppl. I: 31-46 (1980).
4.
normality of Boland, M.P., Crosby, T.F. and Gordon, I. Morphological cattle embryos following superovulation using PMSG. Theriogenology (1978). -IO: 175-180
5.
Kummer, V., Zraly, Z., Holcak, V., Veznik, Z., Schlegelova, J. and Hruska, K. Superovulation in cattle: Effect of goat anti-PMSG serum. Theriogenology (1980). -14: 383-390
730
MAY 1984VOL. 21 NO. 5
THERIOGENOLOGY
6.
Saumande, J. Relationships between ovarian stimulation by PMSG and steroid secretion. In Control of Reproduction in the Cow, Ed. J.M. Sreenan. MFNijhoff, The Hague. 1978, pp.l69-194.
7.
Moncada Angel, A.H. Endoscopic examination of cattle with particular reference to superovulation. Anim. Breed. Abstr. -48: 698 (Abstr.) (1980).
8.
Dhondt, D., Bouters, R., Spincemaille, J., Coryn, M., and Vandeplassche, M. The control of superovulation in the bovine with a PMSG-antiserum. Theriogenology 2: 529-534 (1978).
9.
Swanson, L.V. and Hafs, H.D. LH and prolactin in blood serum from oestrus to ovulation in Holstein heifers. J. Anim. Sci. -33: 1078-1041 (1971).
10.
Lemon, M., Pelletier, J., Saumande, J. and Signoret,J.P. Peripheral plasma concentrations of progesterone, oestradiol -178 and luteinizing hormone around oestrus in the cow. J. Reprod. Fert. (1975). -42: 137-140
11.
Dobson, H. Plasma gonadotrophins and oestradiol the cow. J. Reprod. Fert. -52: 51-53 (1978).
12.
Parr, E.L. Rupture of ovarian follicles at ovulation. 3. Reprod. Fert. Suppl. 22: 1-22 (1975).
13.
Chupin, D. and Procureur, R. Efficiency of pituitary extracts for induction of superovulation in cattle. Anim. Reprod. Sci. 6: II-23 (1983).
MAY 1984VOL. 21 NO. 5
during
oestrus
in
(FSH)
731
J. Saumande, R. Procureur
and D. Chupin
I.N.R.A. Station de Physiologie de la Reproduction 37380 Nouzilly, France
Received
for publication: Accepted:
September 12, 1983 March 15, 1984
ABSTRACT Eighteen cows were superovulated by injecting 3000 IU of PMSG during the luteal phase, followed 48h later with an injection of Estrumate. They were then placed in a control group or were given anti-PMSG antiserum at either 12h or 24h after the onset of oestrus. Sixteen of these animals were used for the same experiment five months later. The results were pooled because they were not significantly different between the two treatment periods The timing of the injection ot anti-PMSG antiserum, either 12h (II cows) or 24h (12 cows) after the onset of oestrus,did not significantly affect the ovulation rate, the number of embryos collected or the number of good embryos. The antiserum significantly increased the number of good embryos but did not affect the ovulation rate or embryo recovery. It is concluded that even with a moderate dose of PMSG, the use of anti-PMSG at l2h or 24h after the beginning of oestrus improves the quality of embryos. The mean number of embryos to be transferred (5.5) is in the range of those obtained after the FSH treatments,but the procedure required only three injections compared with nine for the FSH treatment.
INTRODUCTION When superovulation is induced in cattle by injecting PMSG, large follicles are often observed on the ovary at the time of embryo recovery (1)2). Such follicles are observed more frequently at the high doses of PMSG (3), and their presence is associated with a lower quality of recovered embryos (4). Under these conditions, the number of embryos to be transferred cannot be improved by increasing ovulation rate with a larger dose of PMSG. The injection of anti-PMSG in order to control the duration of follicular growth stimulation has been shown to improve the quality of embryos (5) as previously hypothesized (6). In superovulated animals, ovulation is spread over 72 hours (7). Anti-PMSG could interfere with ovulation since such antisera are known to crossreact with hypophyseal gonadotrophins (8). The purpose of this experiment was to test the effect of time of anti-PMSG injection on the number of good embryos recovered and to assess the effect of such treatment when moderate doses of PMSG are used.
MAY 1984VOL. 21 NO. 5
727
THERIOGENOLOGY MATERIALS AND METHODS Eighteen lactating Friesian cows were injected with PMSG (3000 IU) during the luteal phase (between day 8 and day 14) and 4Sh later with Img of Estrumate. Oestrus was detected twice a day with a vasectomized bull; the day of oestrus was day 0. Cows were inseminated at 12h and 24h after the beginning of oestrus. Six cows were used as controls and received no antiserum. The remaining 12 cows were injected intravenously with 5ml anti-PMSG*. Six cows received the antiserum 12h and six 24h after the beginning of oestrus. The same treatment was applied five months later to 16 Of these animals(fivecontrol, five injected with anti-PMSG at 12h and six injected at 24h after the beginning of oestrus).Care was taken to allot the cows so that each group included animals of the other groups of the first trial. After the first superovulation, embryos were collected nonsurgically on day 7 or day 8 and ovarian responses were measured by rectal palpation; after the second, superovulation, collection and ovarian examination were done after slaughter for six animals and by nonsurgical recovery for the other ten. The quality of embryos was assessed through morphological criteria (2) using a stereo-microscope (10 to 40 magnification). Results were compared with the Mann-Whitney U test (for two samples) and the Kruskal and Wallis one-way analysis of variance.
RESULTS The results obtained five months apart were not different for any of the parameters studied (P (O.OOl), so the data were pooled and are presented in Table 1. Whether anti-PMSG was injected 12h or 24h after the beginning of oestrus did not affect either the ovulation rate, the number of embryos collected or the percentage of good embryos (P< 0.001). Comparison of the results obtained after injection of anti-PMSG with those of the control group showed that the injection of antiserum did not affect ovulation rate (P(O.05) or the number of embryos collected (P(O.02); however, the percentage of good embryos obtained after injection of anti-PMSG was increased (P
DISCUSSION The ovulation rate was not affected by the injection of anti-PMSG 12h or 24h after the beginning of oestrus. In both cases we can assume that the antiserum was injected after the preovulatory peak of gonado-
x This antiserum was a gift of Prof. R. Bouters, Gand, Belgium.
728
MAY 1984VOL. 21 NO. 5
THERIOGENOLOGY
MAY 1984VOL. 21 NO. 5
729
THERIOGENOLOGY
trophins, which occurs at the beginning of oestrus (9, IO, 11). On the other hand, when antiserum was injected at the beginning of oestrus in four cows that had received 3000 IU of PMSG, only one follicle was ovulated in each animal (unpublished work). These results all agree with the current hypothesis on the ovulation process: once the LH-FSH discharge has occurred, the events which lead to ovulation are no longer gonadotrophin-dependent (12). However, this still leaves unexplained the fact that ovulation can be spread over a long time (7) . Our data also show that for a moderate dose of PMSG, the injection of anti-PMSG increased the percentage of good embryos. With these conditions, even though neither the ovulation rate nor the recovery rate was changed, the improvement in the quality of embryos led to an increase in the number of embryos to be transferred (5.5 vs 3.5). When lactating Friesian cows are treated with the most efficent of the FSH treatments, a mean of 5.3 good embryos is obtained (13). Therefore,the same number of good embryos can be obtained with both treatments. However, from a practical point of view, only three injections are needed when anti-PMSG is used (PMSG, prostaglandin and antiserum) compared with nine for an FSH treatment (height injection of gonadotrophin and prostaglandin).
In conclusion, PMSG is an efficient hormone for the production of embryos provided the duration of its stimulatory effect is controlled with an antiserum. This treatment is more practical than those with FSH because the number of injections is reduced and the time of antiPMSG administration after oestrus is not critical.
REFERENCES 1.
Booth, W.D., Newcomb, R., Strangle, H., Rowson, L.E.A. and Sacher, H.B Plasma oestrogen and progesterone in relation to superovulation and egg recovery in the cow. Vet. Rec. -97: 366-369 (1975).
2.
Renard, J.P., Menezo, Y., Saumande, 3. and Heyman, Y. Attempts to check the viability of embryos produced by superovulated heifers. In Control of Reproduction in the Cow, Ed. J.M. Sreenan. Nijhoff M., xe Hague. 1978, pp.398-417.
3.
Mauldon, P., Mariana, J.C., Benoit, M., Solari, A. and Chupin,D. Action upon the ovulation number and yield of French Friesian cows of different PMSG and HCG doses injected during the follicular phase of the oestrous cycle. Ann. Biol. Anim. Bioch. Biophys. lo, Suppl. I: 31-46 (1980).
4.
normality of Boland, M.P., Crosby, T.F. and Gordon, I. Morphological cattle embryos following superovulation using PMSG. Theriogenology (1978). -IO: 175-180
5.
Kummer, V., Zraly, Z., Holcak, V., Veznik, Z., Schlegelova, J. and Hruska, K. Superovulation in cattle: Effect of goat anti-PMSG serum. Theriogenology (1980). -14: 383-390
730
MAY 1984VOL. 21 NO. 5
THERIOGENOLOGY
6.
Saumande, J. Relationships between ovarian stimulation by PMSG and steroid secretion. In Control of Reproduction in the Cow, Ed. J.M. Sreenan. MFNijhoff, The Hague. 1978, pp.l69-194.
7.
Moncada Angel, A.H. Endoscopic examination of cattle with particular reference to superovulation. Anim. Breed. Abstr. -48: 698 (Abstr.) (1980).
8.
Dhondt, D., Bouters, R., Spincemaille, J., Coryn, M., and Vandeplassche, M. The control of superovulation in the bovine with a PMSG-antiserum. Theriogenology 2: 529-534 (1978).
9.
Swanson, L.V. and Hafs, H.D. LH and prolactin in blood serum from oestrus to ovulation in Holstein heifers. J. Anim. Sci. -33: 1078-1041 (1971).
10.
Lemon, M., Pelletier, J., Saumande, J. and Signoret,J.P. Peripheral plasma concentrations of progesterone, oestradiol -178 and luteinizing hormone around oestrus in the cow. J. Reprod. Fert. (1975). -42: 137-140
11.
Dobson, H. Plasma gonadotrophins and oestradiol the cow. J. Reprod. Fert. -52: 51-53 (1978).
12.
Parr, E.L. Rupture of ovarian follicles at ovulation. 3. Reprod. Fert. Suppl. 22: 1-22 (1975).
13.
Chupin, D. and Procureur, R. Efficiency of pituitary extracts for induction of superovulation in cattle. Anim. Reprod. Sci. 6: II-23 (1983).
MAY 1984VOL. 21 NO. 5
during
oestrus
in
(FSH)
731