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Continuous subcutaneous infusion of follicle stimulating hormone as a method of superovulating dairy cows
THERIOGENOLOGY
CONTINUOUS
SUBCUTANEOUS AS A METHOD
A. WubishetI , C.N.
INFUSION OF FOLLICLE STIMULATING OF SUPEROVULATING DAIRY cows Graves,
S.L. Spahr
and D.J.
HORMONE
Kesler
Department of Animal Sciences University of Illinois Urbana, IL 61801 Received
for publication: Accepted:
Se@ember
6, 1985
April.16, 1986
ABSTRACT Twenty-eight Holstein cows were randomly assigned to two treatGroup I received 50 mg of follicle stimulating hormone-pituitary ments. (FSH-P) intramuscularly (i.m.) in declining doses in 5 d. Group II received 49 mg of FSH-P in 7 d via constant-infusion, subcutaneously Cows received 25 mg prostaglandin F2a (s.c.) implanted osmotic pumps. 48 h after initiation of treatment and were inseminated at estrus. Intramuscular injections resulted in higher numbers of corpora lutea These differences were largely (CL) and embryos than the S.C. infusion. attributed to differences in the doses of FSH-P administered in the two It was concluded that continuously infusing FSH-P S.C. via groups. osmotic pumps can be used as a method of superovulating dairy cows. Key words:
superovulation,
embryo,
infusion,
bovine
INTRODUCTION The most commonly used superovulatory agent in the cow is FSH. Several investigators (l-3) have indicated that the use of FSH results in high numbers of CL and recovered embryos. However, due to the short half-life of this hormone (4), multiple injections are necessary over 4 or 5 d, requiring handling of cows several times during each superovulation. Though other investiqators (5. 6) had ureviouslv reoorted observing an increased number of large‘follicles and a higher number of CL when FSH was continuously infused directly into the jugular veins, information on the superovulatory effects ofcontinuous S.C. FSH infusion is lacking. This project was therefore undertaken to investigate whether continuous FSH infusion via S.C. implanted osmotic pumps can result in satisfactory superovulation of dairy cows and thereby reduce the labor required in administering FSH.
1
Present
JUNE
1986
address:
Alcorn
VOL.25N0.6
State
University,
Lorman,
MS
39096.
809
THERIOGENOLOGY
MATERIALS
AND METHODS
Twenty-eight multiparous Holstein cows were randomly assiqned to Some cows were synchronized with two treatment groups of 14 cows each. prostaglandin (Lutalyse, Upjohn, Kalamazoo, MI), while others showed Supernatural estrus before they were admitted into the experiment. ovulation of cows in both treatment groups was started on Day 9 of the Cows varied from two to eight years of age and were on estrous cycle. the average 71 d postpartum when treatments were initiated. Cows in Group I (control) were injected with a total of 50 mg of FSH (FSH-P, Burns-Biotec, Omaha, NB) i.m. twice daily at 12-h intervals in declining doses of 7, 6, 5, 4, and 3 mg over 5 d. Those in Group II received FSH in Model 2MLl osmotic pumps (ALZA Corporation, Palo Alto, CA) implanted in the area anterio-dorsal to the scapula. The osmotic pumps were filled with 2 ml of saline solution containing 50 mg of FSH 1 d The osmotic pumps were taken out on before they were to be implanted. Day 8. These pumps delivered FSH at the rate of 7 mg/d; therefore, this group received 35 mg FSH in 5 d and a total of only 49 mg in 7 d. All cows were injected i.m. with 25 mg Lutalyse 48 h after initiaAll cows were inseminated with frozen semen from tion of FSH treatment. the same bull at 12 and 24 h after the start of heat., Cows that did not exhibit estrus by 2000 h on Day 5 of superovulation were inseminated once at this time and again 24 h later. Efficiency of superovulation was determined by estimatinq the number of embryos recovered following nonsurgical embryo recovery 8 d after insemination. The number of CL palpated rectally and embryos recovered were then compared by t-test (7). Fertilization rates were compared by the Chi-square test (8). RESULTS
AND DISCUSSION
Twenty-four (86%) of the 28 cows (I3 from Group I and 11 from Group II) showed estrus. A total of 225 CL was estimated by rectal palpation, and 174 developing embryos and unfertilized ova were recovered nonsurgitally from cows in both treatments for a recovery rate of 78%. Out of these, 135 (78%) were developing embryos, while 39 (22%) were unfertilized Of the developing embryos 71% were blastocyst and 29% were morulae. ova. Three or more embryos were recovered from a higher (P < 0.01) proportion (79%) of cows in Group I than from those in Group II (57%). The results of rectal palpation and nonsurgical embryo recovery are shown (Table 1). The number of CL palpated varied from 0 to 22 in Group I and from 0 to 19 in Group II. The mean number of CL (10.4tl.7) palpated in Group I was higher (P < 0.01) than that (5.721.4) in Likewise, the mean numbers of developing embryos and unferGroup II. tilized ova recovered from Group I (8.221.5) were higher (P < 0.01) than those (4.2t1.3) recovered from Group II. However, there was no difference (P > 0.05) in the fertilization rate between the two treatments (76% vs 81% for Groups I and II respectively).
810
JUNE 1986 VOL. 25 NO. 6
THERIOGENOLOGY Table
1.
Superovulatory results from Holstein cows to which administered in two different methods
Group I Multiple Injection No. of cows Total no. of CL Mean no. of CL (?SE) Total no. of ova and embryos Mean no. of ova and embryos (?SE) Total no. of unfertilized ova Total no. of developing embryos Fertilization rate (%) Total no. of morulae Total no. of blastocysts ayb Figures in the same row with different (P < 0.01).
Group
II
It&scion 14 80 5.7bf1.4 58 4.zb+1.3 11 48 81 10 38
2:: 10.4f1.7 115 8.Za+l.5 :: 76 28 59 superscripts
FSH was
are different
The lower superovulatory responses [numbers of CL palpated and embryos recovered) in Group II may be largely attributed to the lower doses of FSH Cows in Group I received all of their 50 mg this group of cows received. superovulation, while those in Group of FSH during the first 5 d of II received only 35 mg during the same period. A similar decrease in CL number (6 vs 10.4) has been reported (9) when 32 and 50 mg FSH were compared. The results of this study indicate that continuous infusion of FSH via s.c.-implanted osmotic pumps can be used as a method of superovulation. The method required less labor than the twice-daily injection technique, since each cow was handled only three times (one time each to implant and remove the osmotic pump and once for the PGF2a) as contrasted to 11 times in the twice-daily injection method. However, to prove which of the two methods is more efficient, further investigation with equal doses of the hormone over the same length of time must be conducted. REFERENCES 1.
Critser, J.K., Rowe, R.F., Del Campo, M.R., and Ginther, O.J. Embryo transfer in cattle: factors affecting superovulatory response, number of transferable embryos and length of post-treatment estrous cycles. Theriogenology -13:397-406 (1980).
2.
Elsden, R.P., with follicle gonadotropin.
3.
Monniaux, D., Chupin, D. and Saumande, J. Superovulatory Theriogenology of cattle. -19:55-&l (1983).
4.
Laster. D.B. ewe a&l cow.
5.
Laster, D.B. Follicular development in heifers infused with follicle stimulating hormone. J. Reprod. Fertil. (1972). -28:285-289
Nelson, L.D. and Seidel, G.E. Superovulating cows stimulating hormone and pregnant mare's serum Theriogenology 2:17-26 (1978). responses
Disaooearance and uptake of I25I FSH in rat, rabbit, J. Reprod. Fertil. (1972). -30:407-415
JUNE 1986 VOL. 25 NO. 6
811
THEFUOGENOLOGY
6.
Bercovitz, A-6. and Lambeth, V.A. Continuous infusion Godke, LA., methods for superovulating beef and dairy cattle. The Louisiana Cattleman -13:4-6 (1980).
7.
Principles and Procedures of Steel, R.G.D. and Torrie, J.H. McGraw-Hill Book Company, Inc., New York, 1960, Statistics. pp. 67-81.
8.
Huntsberger, D.V. and BillingsTey, Inference. Allyn and Bacon, Inc.,
9.
812
P.
Elements of Statistical 1970, pp. 200-211.
Boston,
A., Kazmer, G.W., Wade, R.J. Barnes, M.A., Martinez-Castellana, and Halman, R.D. Effect of exogenous FSH on estrus, ovulation and Theriogenology endogenous hormone release in dairy cows. -18:311-322 (1982).
JUNE 1986VOL. 25N0.6
CONTINUOUS
SUBCUTANEOUS AS A METHOD
A. WubishetI , C.N.
INFUSION OF FOLLICLE STIMULATING OF SUPEROVULATING DAIRY cows Graves,
S.L. Spahr
and D.J.
HORMONE
Kesler
Department of Animal Sciences University of Illinois Urbana, IL 61801 Received
for publication: Accepted:
Se@ember
6, 1985
April.16, 1986
ABSTRACT Twenty-eight Holstein cows were randomly assigned to two treatGroup I received 50 mg of follicle stimulating hormone-pituitary ments. (FSH-P) intramuscularly (i.m.) in declining doses in 5 d. Group II received 49 mg of FSH-P in 7 d via constant-infusion, subcutaneously Cows received 25 mg prostaglandin F2a (s.c.) implanted osmotic pumps. 48 h after initiation of treatment and were inseminated at estrus. Intramuscular injections resulted in higher numbers of corpora lutea These differences were largely (CL) and embryos than the S.C. infusion. attributed to differences in the doses of FSH-P administered in the two It was concluded that continuously infusing FSH-P S.C. via groups. osmotic pumps can be used as a method of superovulating dairy cows. Key words:
superovulation,
embryo,
infusion,
bovine
INTRODUCTION The most commonly used superovulatory agent in the cow is FSH. Several investigators (l-3) have indicated that the use of FSH results in high numbers of CL and recovered embryos. However, due to the short half-life of this hormone (4), multiple injections are necessary over 4 or 5 d, requiring handling of cows several times during each superovulation. Though other investiqators (5. 6) had ureviouslv reoorted observing an increased number of large‘follicles and a higher number of CL when FSH was continuously infused directly into the jugular veins, information on the superovulatory effects ofcontinuous S.C. FSH infusion is lacking. This project was therefore undertaken to investigate whether continuous FSH infusion via S.C. implanted osmotic pumps can result in satisfactory superovulation of dairy cows and thereby reduce the labor required in administering FSH.
1
Present
JUNE
1986
address:
Alcorn
VOL.25N0.6
State
University,
Lorman,
MS
39096.
809
THERIOGENOLOGY
MATERIALS
AND METHODS
Twenty-eight multiparous Holstein cows were randomly assiqned to Some cows were synchronized with two treatment groups of 14 cows each. prostaglandin (Lutalyse, Upjohn, Kalamazoo, MI), while others showed Supernatural estrus before they were admitted into the experiment. ovulation of cows in both treatment groups was started on Day 9 of the Cows varied from two to eight years of age and were on estrous cycle. the average 71 d postpartum when treatments were initiated. Cows in Group I (control) were injected with a total of 50 mg of FSH (FSH-P, Burns-Biotec, Omaha, NB) i.m. twice daily at 12-h intervals in declining doses of 7, 6, 5, 4, and 3 mg over 5 d. Those in Group II received FSH in Model 2MLl osmotic pumps (ALZA Corporation, Palo Alto, CA) implanted in the area anterio-dorsal to the scapula. The osmotic pumps were filled with 2 ml of saline solution containing 50 mg of FSH 1 d The osmotic pumps were taken out on before they were to be implanted. Day 8. These pumps delivered FSH at the rate of 7 mg/d; therefore, this group received 35 mg FSH in 5 d and a total of only 49 mg in 7 d. All cows were injected i.m. with 25 mg Lutalyse 48 h after initiaAll cows were inseminated with frozen semen from tion of FSH treatment. the same bull at 12 and 24 h after the start of heat., Cows that did not exhibit estrus by 2000 h on Day 5 of superovulation were inseminated once at this time and again 24 h later. Efficiency of superovulation was determined by estimatinq the number of embryos recovered following nonsurgical embryo recovery 8 d after insemination. The number of CL palpated rectally and embryos recovered were then compared by t-test (7). Fertilization rates were compared by the Chi-square test (8). RESULTS
AND DISCUSSION
Twenty-four (86%) of the 28 cows (I3 from Group I and 11 from Group II) showed estrus. A total of 225 CL was estimated by rectal palpation, and 174 developing embryos and unfertilized ova were recovered nonsurgitally from cows in both treatments for a recovery rate of 78%. Out of these, 135 (78%) were developing embryos, while 39 (22%) were unfertilized Of the developing embryos 71% were blastocyst and 29% were morulae. ova. Three or more embryos were recovered from a higher (P < 0.01) proportion (79%) of cows in Group I than from those in Group II (57%). The results of rectal palpation and nonsurgical embryo recovery are shown (Table 1). The number of CL palpated varied from 0 to 22 in Group I and from 0 to 19 in Group II. The mean number of CL (10.4tl.7) palpated in Group I was higher (P < 0.01) than that (5.721.4) in Likewise, the mean numbers of developing embryos and unferGroup II. tilized ova recovered from Group I (8.221.5) were higher (P < 0.01) than those (4.2t1.3) recovered from Group II. However, there was no difference (P > 0.05) in the fertilization rate between the two treatments (76% vs 81% for Groups I and II respectively).
810
JUNE 1986 VOL. 25 NO. 6
THERIOGENOLOGY Table
1.
Superovulatory results from Holstein cows to which administered in two different methods
Group I Multiple Injection No. of cows Total no. of CL Mean no. of CL (?SE) Total no. of ova and embryos Mean no. of ova and embryos (?SE) Total no. of unfertilized ova Total no. of developing embryos Fertilization rate (%) Total no. of morulae Total no. of blastocysts ayb Figures in the same row with different (P < 0.01).
Group
II
It&scion 14 80 5.7bf1.4 58 4.zb+1.3 11 48 81 10 38
2:: 10.4f1.7 115 8.Za+l.5 :: 76 28 59 superscripts
FSH was
are different
The lower superovulatory responses [numbers of CL palpated and embryos recovered) in Group II may be largely attributed to the lower doses of FSH Cows in Group I received all of their 50 mg this group of cows received. superovulation, while those in Group of FSH during the first 5 d of II received only 35 mg during the same period. A similar decrease in CL number (6 vs 10.4) has been reported (9) when 32 and 50 mg FSH were compared. The results of this study indicate that continuous infusion of FSH via s.c.-implanted osmotic pumps can be used as a method of superovulation. The method required less labor than the twice-daily injection technique, since each cow was handled only three times (one time each to implant and remove the osmotic pump and once for the PGF2a) as contrasted to 11 times in the twice-daily injection method. However, to prove which of the two methods is more efficient, further investigation with equal doses of the hormone over the same length of time must be conducted. REFERENCES 1.
Critser, J.K., Rowe, R.F., Del Campo, M.R., and Ginther, O.J. Embryo transfer in cattle: factors affecting superovulatory response, number of transferable embryos and length of post-treatment estrous cycles. Theriogenology -13:397-406 (1980).
2.
Elsden, R.P., with follicle gonadotropin.
3.
Monniaux, D., Chupin, D. and Saumande, J. Superovulatory Theriogenology of cattle. -19:55-&l (1983).
4.
Laster. D.B. ewe a&l cow.
5.
Laster, D.B. Follicular development in heifers infused with follicle stimulating hormone. J. Reprod. Fertil. (1972). -28:285-289
Nelson, L.D. and Seidel, G.E. Superovulating cows stimulating hormone and pregnant mare's serum Theriogenology 2:17-26 (1978). responses
Disaooearance and uptake of I25I FSH in rat, rabbit, J. Reprod. Fertil. (1972). -30:407-415
JUNE 1986 VOL. 25 NO. 6
811
THEFUOGENOLOGY
6.
Bercovitz, A-6. and Lambeth, V.A. Continuous infusion Godke, LA., methods for superovulating beef and dairy cattle. The Louisiana Cattleman -13:4-6 (1980).
7.
Principles and Procedures of Steel, R.G.D. and Torrie, J.H. McGraw-Hill Book Company, Inc., New York, 1960, Statistics. pp. 67-81.
8.
Huntsberger, D.V. and BillingsTey, Inference. Allyn and Bacon, Inc.,
9.
812
P.
Elements of Statistical 1970, pp. 200-211.
Boston,
A., Kazmer, G.W., Wade, R.J. Barnes, M.A., Martinez-Castellana, and Halman, R.D. Effect of exogenous FSH on estrus, ovulation and Theriogenology endogenous hormone release in dairy cows. -18:311-322 (1982).
JUNE 1986VOL. 25N0.6