- Email: [email protected]
Efficacy of short-term administration of altrenogest to postpone ovulation in mares
I:qulno lQuttltlon and Physiology (~oeloty Annual (~gmpor May 2 ~ ' - g l , 1 9 9 7 . i:ort Worth, Taxam.
EFFICACY OF SHORT-TERM ADMINISTRATION OF ALTRENOGESTTO POSTPONE OVULATION IN MARES A.N. James, M.M. Vogelsang, D.W. Forrest, G.G. Stott
SUMMARY Estrogen from a growing follicle stimulates the preovulatory surge of luteinizing hormone (LH) while progesterone (P) is known to suppress LH. The possibility exists that administration of P, in the presence of an ovulatory follicle, would sufficiently suppress LH and, therefore, delay ovulation. The objective of this research was to elucidate the potential for oral administration of altrenogest (17-Allyl- 1713-hydroxyestra-4,9,11-trien-3-one) to postpone ovulation of a preovulatory follicle (35 mm) for approximately two days. Fourteen light-horse mares, ranging in age from two to 19 years, were randomly assigned to one of three treatments (A - .044 mg/kg BW altrenogest for two days; B - .088 mg/kg BW altrenogest for two days; and C - no altrenogest). Mares began treatment when a 35-mm or greater follicle was observed via real-time transrectal ultrasonography. Both number of days until ovulation and follicular maintenance differed between treated and control mares. Number of days until ovulation was increased (P<.05) for mares in treatment A when compared with the control mares. Follicular diameter maintenance, a measurement of follicular diameter throughout treatment, also increased (P<.05) for mares in treatment A when compared with the control mares. Mean LH concentration was not different between mares treated with altrenogest at either treatment dose when compared with the control mares. Pregnancy rates and embryonic vesicle size change were also measured to determine potential effects of altrenogest administration. No differences (P>.05) were found in either characteristic. Short-term administration of altrenogest increased the number of days to ovulation. Further study is warranted to prove conclusively that altrenogest increases follicular maintenance, alters the preovulatory LH surge, and has no detrimental effects upon reproductive efficiency.
Authors' Addresses: TexasA&M University.CollegeStation.TX 778432471 Volume 18, Number 5, 1998
INTRODUCTION In fulfillment of the requirements of certain breed registries, a stallion must live-cover the mares which are in his book. Frequently, there are more mares that meet criteria for breeding than a stallion can successfully cover in one day. Similarly, some stallions on a rigorous show schedule often have more mares to breed when they arrive at the breeding farm than is possible from one day's spermatozoal output even with the use of artificial insemination. Further, the use of transported cooled semen somewhat limits availability of semen since larger insemination doses are utilized. For these reasons, it would be beneficial to be able to control the time of ovulation in a mare in order to optimize use of the stallion or to minimize the number of cycles per conception. Methods currently exist to induce ovulation in a mare within a given time frame (using human chorionic gonadotropin, gonadotropin releasing hormone, or prostaglandin), but no protocols exist to briefly prolong the interval from the beginning of standing estrus to ovulation. Development of such a tool would be beneficial for breeders who have large numbers of mares that could ovulate in a short time frame since it would extend the window of time in which a mare could be bred or inseminated.
MATERIALS AND METHODS Animals. Fourteen mares of light-horse type, ranging in age from two to 19 years, were randomly assigned to one of three treatment groups as follows, A (n=5), .044 mg/kg BW altrenogest for two days, B (n=5), .088 mg/kg B W altrenogest for two days, and C (n=4), no altrenogest. All mares were fed to meet maintenance requirements as recommended. 1 Treatment. Mares received the altrenogest when a
329
follicle of _> 35 mm (Day 1) was observed via real-time transrectal ultrasonography a performed every other day. Mares in group C were inseminated when a 35-mm follicle was first detected and every other day until ovulation. Mares in groups A and B received altrenogest b (17-Allyl17B-hydroxyestra-4,9,11-trien-3-one) orally for two days and were then inseminated every other day starting the third day following altrenogest withdrawal until ovulation. All mares were bred to the same stallion of proven fertility. Semen was processed the same for each insemination to minimize variation in conception rates due to semen quality. Each mare was inseminated with a minimum dose of 500 million motile sperm extended with an equal volume of skim milk extender.
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Data Collection. Mares were examined by ultrasonography on Day 12 post-ovulation for detection of pregnancy and on Days 14 and 16 to assess embryonic vesicle growth. Plasma samples were collected via jugular venipuncture into heparinized evacuated tubes every other day until altrenogest administration. Samples were collected daily from Day 1 until two days post-ovulation for treated and control mares. Plasma was pipetted into 6-ml tubes for storage at -20~ Samples were assayed for LH using a double-antibody RIA2 validated in this laboratory. 3 Data were subjected to analysis of variance using procedures appropriate for an incomplete randomized design to account for the uneven numbers. All data were also subjected to T tests (LSD) to determine which treatments were different.
RESULTS AND DISCUSSION
Mean days until ovulation, from Day 1 of treatment, for treatments A, B, C were 6.60, 8.20, and 5.00, respectively (Table 1). The number of days from observation of a 35 mm follicle (Day 1) until ovulation was larger (P < .05) for both altrenogest treated groups when compared to the untreated controls. Number of days until ovulation, ascertained from the day a _>35 mm follicle was first observed, was affected by aAIoka| 210, Corometrics Medical Systems, Inc., Carrollton, TX. bRegumate| Hoechst Roussel, Somerville, NJ.
Table 1. Mean number of days until ovulation, and follicular diameter at time of ovulation in mares treated with altrenogest versus controls (_+ SEM). Follicular Number of Days Diameter Treatment Number of Until Ovulation at Time of Groups Mares from D1 Ovulation (mm) A (.044 mg/kg) 5 6.60-2:0.40a 41.60 + 3.28 a B (.088 mg/kg) 5 8.20 + 1.20a 38.80 + 4.24a Control 4 5.00 • 0.00b 40.25 +_2.75a a,bMeanslackinga commonsuperscriptwithin columnsdiffer (P<.05).
330
Figure 1 . Luteinizing hormone concentrations of mares fed altrenogest (A = .044 mg/kg BW; B = .088 mglkg BW) for two days after attainment of a 35-mm follicle (Day 1) versus control mares. Mean days of ovulation were 6.6, 8.2, and 5.0 for groups A,B, and C, respectively.
administration of altrenogest. The most consistent response was observed when mares were treated with .044 mg/kg BW of altrenogest, which increased number of days until ovulation over the control group by approximately two days. Although mares on treatment B (.088 mg/kg BW) did demonstrate an increased number of days until ovulation, the response times were more variable than for mares on treatment A. Small treatment numbers are the most likely cause of this chance variability. The altrenogest dose used in treatment B increases the number of days until ovulation but the length of delay exhibited could increase the possibilities of decreased ova fertility. Mean follicular diameters at ovulation for treatments A, B, and C were 41.60, 38.80, and 40.25 mm, respectively (Table 1). Mares showed no difference (P = .65) between the treated and control groups with respect to size of the follicle at ovulation. It appears, therefore, that treatment with altrenogest during the latter stage of estrus does not alter the ultimate size a follicle will reach prior to ovulation, but will slow the rate at which it attains ovulatory size. Luteinizing hormone concentrations were measured every day during the treatment period and Day 1 through Day 13 values were analyzed (Figure 1). Day 13 was two days after the last mare ovulated in treatment B. Measurements for LH were not taken for the rest of the mares this far from the start of treatment because they ovulated at earlier times. Due to the variation in the number of days to ovulation, there were different numbers of samples to be analyzed for LH concentration. The LH curve for the control mares resembled a normal LH profile whereas the curve for both altrenogest treated groups was lengthened and did not demonstrate the normal peak. The treated mares peaked later and at a lesser magnitude than did the control mares. This agrees with Wiepz et al., 4 who found JOURNAL OF EQUINE VETERINARY SCIENCE
that altrenogest can suppress or lower LH concentration in transitional mares. Pregnancy rates were evaluated to assess the effect of ovulation delay using altrenogest on reproductive efficiency. No difference in pregnancy rates at Day 14 were detected, with 2/5 (40%), 3/5 (60%), and 3/4 (75%) pregnancies achieved in groups A, B, and C, respectively. This is in agreement with previous research by Allen et al. 5 and Squires et al. 6'7 who reported no detrimental effects on conception rates for mares which had been treated with altrenogest previous to the estrous cycle in which they had been inseminated. Ultrasonographic evaluation of embryonic vesicles was conducted on Days 12, 14, and 16 to determine whether administration of altrenogest had any detrimental effects on embryonic growth. Embryonic vesicle growth was monitored between Days 12 to 16 for treatment A, treatment B, and control mares and changes in embryonic vesicle size were 7.17, 9.00, and 8.00 mm, respectively. There was no difference (P > .05) in size of the embryonic vesicle among treated and control mares. Administration of altrenogest appeared to have no detrimental effects of pregnancy rates nor on early embryonic vesicle growth. Some general observations were made during this study which did not lend themselves to statistical analysis. These observations included estrous behavior during treatment, reproductive tract changes during treatment, and criteria that may have affected the response of certain mares to the treatment protocol. Before administration of altrenogest, mares exhibited normal signs of estrus at each mare's individual intensity. While mares were undergoing treatment, they demonstrated a decrease in intensity of behavioral estrus similar to that observed in diestrus or pregnant mares. When administration of altrenogest was discontinued, the mares continued to exhibit diestrus behavior for an average of two days following cessation of treatment. After withdrawal of treatment, mares eventually returned to behavioral estrus but with a decreased intensity. During estrus, mares' cervixes and uteri exhibited a characteristic flaccid tone. These structures became turgid, typical ofdiestrus, following altrenogest treatment. The mares retumed to estrous uterine tone after altrenogest was discontinued. In addition, mares which demonstrated a normal estrus of six to eight days had less variable responses to altrenogest treatment than mares that demonstrated a normal estrus of nine to ten days.
sw, Hunt MDN, Digby NJW: Preliminary studies on the use of an oral progestagen to induce oestrus and ovulation in seasonally anoestrous thoroughbred mares. Equine Vet J 1980;12:141-145. 6. Squires EL, Stevens WB, McGIothin DE, Pickett BW: Effect of an oral progestin on the estrous cycle and fertility of mares. JAnim Sci 1979;49:729735. 7. Squires EL, Hesseman CP, Webel SK, Voss JL: Relationship of altrenogest to ovarian activity hormone concentrations and fertility of mares. J Anim Sci 1983;56:901-910. Dry coat, non-sweater, anhidrosis? Many horses are afflicted with the genetic condition known as anhidrosis, which is a lack of or reduced ability to sweat. This can seriously reduce performance during hot humid weather. Now a nutriceutical is available to reduce the affects of this condition. MPCO, LLC 13150 N. 2 2 n d Ave. Phoenix, AZ 8 5 0 2 9 - 1 5 0 7
CONCLUSIONS
Results indicate that short-term administration of altrenogest does delay ovulation. Although trends are suggested by the present study, further studies with larger populations are needed to prove conclusively that administration of altrenogest increases follicular maintenance, decreases the preovulatory LH surge, and does not affect fertility.
REFERENCES 1. NRC:Nutrient Requirements ofHorses(5th Ed). Washington, DC: NationalAcademy Press, 1989. 2. RoserJF, Hughes JP: Prolonged pulsatile administration of gonadotropin-releasing hormone (GnRH) to fertile stallions during winter and summer. J Reprod Fertil, Suppl 1991;44:155-168. 3. Lang AL, Vogelsang MM, Potter GD: Circadian patterns of plasma LH and testosterone concentrations in stallions in summer and winter. Proc 14th ENPS 1995;284289. 4. Wiepz GJ, Squires EL, Chapman PL: Effects of norgestomet, altrenogest, and/or estradiol on follicular and hormonal characteristicsof late transitional mares. Theriogenology 1988;30:181-193. 5. Allen WR, Urwin V, Simson DJ, Greenwood RES, Crowhurst RC, Ellis DR, Ricketts
Volume 18, Number 5, 1998
PERFORMANCE & THERAPEUTIC HORSESHOE SHOCK ABSORBING TREAD ULTRA WIDE WEB BAR SHOE EFFECTIVE & ECONOMICAL FOR MORE INFO CONTACT
EQUITHOTICS INC. 7580 A C A C I A LN. V A C A V I L L E CA. 95688 PH. 707-449-3570 FAX 707-452-8394 [email protected]
331
EFFICACY OF SHORT-TERM ADMINISTRATION OF ALTRENOGESTTO POSTPONE OVULATION IN MARES A.N. James, M.M. Vogelsang, D.W. Forrest, G.G. Stott
SUMMARY Estrogen from a growing follicle stimulates the preovulatory surge of luteinizing hormone (LH) while progesterone (P) is known to suppress LH. The possibility exists that administration of P, in the presence of an ovulatory follicle, would sufficiently suppress LH and, therefore, delay ovulation. The objective of this research was to elucidate the potential for oral administration of altrenogest (17-Allyl- 1713-hydroxyestra-4,9,11-trien-3-one) to postpone ovulation of a preovulatory follicle (35 mm) for approximately two days. Fourteen light-horse mares, ranging in age from two to 19 years, were randomly assigned to one of three treatments (A - .044 mg/kg BW altrenogest for two days; B - .088 mg/kg BW altrenogest for two days; and C - no altrenogest). Mares began treatment when a 35-mm or greater follicle was observed via real-time transrectal ultrasonography. Both number of days until ovulation and follicular maintenance differed between treated and control mares. Number of days until ovulation was increased (P<.05) for mares in treatment A when compared with the control mares. Follicular diameter maintenance, a measurement of follicular diameter throughout treatment, also increased (P<.05) for mares in treatment A when compared with the control mares. Mean LH concentration was not different between mares treated with altrenogest at either treatment dose when compared with the control mares. Pregnancy rates and embryonic vesicle size change were also measured to determine potential effects of altrenogest administration. No differences (P>.05) were found in either characteristic. Short-term administration of altrenogest increased the number of days to ovulation. Further study is warranted to prove conclusively that altrenogest increases follicular maintenance, alters the preovulatory LH surge, and has no detrimental effects upon reproductive efficiency.
Authors' Addresses: TexasA&M University.CollegeStation.TX 778432471 Volume 18, Number 5, 1998
INTRODUCTION In fulfillment of the requirements of certain breed registries, a stallion must live-cover the mares which are in his book. Frequently, there are more mares that meet criteria for breeding than a stallion can successfully cover in one day. Similarly, some stallions on a rigorous show schedule often have more mares to breed when they arrive at the breeding farm than is possible from one day's spermatozoal output even with the use of artificial insemination. Further, the use of transported cooled semen somewhat limits availability of semen since larger insemination doses are utilized. For these reasons, it would be beneficial to be able to control the time of ovulation in a mare in order to optimize use of the stallion or to minimize the number of cycles per conception. Methods currently exist to induce ovulation in a mare within a given time frame (using human chorionic gonadotropin, gonadotropin releasing hormone, or prostaglandin), but no protocols exist to briefly prolong the interval from the beginning of standing estrus to ovulation. Development of such a tool would be beneficial for breeders who have large numbers of mares that could ovulate in a short time frame since it would extend the window of time in which a mare could be bred or inseminated.
MATERIALS AND METHODS Animals. Fourteen mares of light-horse type, ranging in age from two to 19 years, were randomly assigned to one of three treatment groups as follows, A (n=5), .044 mg/kg BW altrenogest for two days, B (n=5), .088 mg/kg B W altrenogest for two days, and C (n=4), no altrenogest. All mares were fed to meet maintenance requirements as recommended. 1 Treatment. Mares received the altrenogest when a
329
follicle of _> 35 mm (Day 1) was observed via real-time transrectal ultrasonography a performed every other day. Mares in group C were inseminated when a 35-mm follicle was first detected and every other day until ovulation. Mares in groups A and B received altrenogest b (17-Allyl17B-hydroxyestra-4,9,11-trien-3-one) orally for two days and were then inseminated every other day starting the third day following altrenogest withdrawal until ovulation. All mares were bred to the same stallion of proven fertility. Semen was processed the same for each insemination to minimize variation in conception rates due to semen quality. Each mare was inseminated with a minimum dose of 500 million motile sperm extended with an equal volume of skim milk extender.
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.
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.
6
.
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.
8
.
:
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10
11
12
: 13
Days
Data Collection. Mares were examined by ultrasonography on Day 12 post-ovulation for detection of pregnancy and on Days 14 and 16 to assess embryonic vesicle growth. Plasma samples were collected via jugular venipuncture into heparinized evacuated tubes every other day until altrenogest administration. Samples were collected daily from Day 1 until two days post-ovulation for treated and control mares. Plasma was pipetted into 6-ml tubes for storage at -20~ Samples were assayed for LH using a double-antibody RIA2 validated in this laboratory. 3 Data were subjected to analysis of variance using procedures appropriate for an incomplete randomized design to account for the uneven numbers. All data were also subjected to T tests (LSD) to determine which treatments were different.
RESULTS AND DISCUSSION
Mean days until ovulation, from Day 1 of treatment, for treatments A, B, C were 6.60, 8.20, and 5.00, respectively (Table 1). The number of days from observation of a 35 mm follicle (Day 1) until ovulation was larger (P < .05) for both altrenogest treated groups when compared to the untreated controls. Number of days until ovulation, ascertained from the day a _>35 mm follicle was first observed, was affected by aAIoka| 210, Corometrics Medical Systems, Inc., Carrollton, TX. bRegumate| Hoechst Roussel, Somerville, NJ.
Table 1. Mean number of days until ovulation, and follicular diameter at time of ovulation in mares treated with altrenogest versus controls (_+ SEM). Follicular Number of Days Diameter Treatment Number of Until Ovulation at Time of Groups Mares from D1 Ovulation (mm) A (.044 mg/kg) 5 6.60-2:0.40a 41.60 + 3.28 a B (.088 mg/kg) 5 8.20 + 1.20a 38.80 + 4.24a Control 4 5.00 • 0.00b 40.25 +_2.75a a,bMeanslackinga commonsuperscriptwithin columnsdiffer (P<.05).
330
Figure 1 . Luteinizing hormone concentrations of mares fed altrenogest (A = .044 mg/kg BW; B = .088 mglkg BW) for two days after attainment of a 35-mm follicle (Day 1) versus control mares. Mean days of ovulation were 6.6, 8.2, and 5.0 for groups A,B, and C, respectively.
administration of altrenogest. The most consistent response was observed when mares were treated with .044 mg/kg BW of altrenogest, which increased number of days until ovulation over the control group by approximately two days. Although mares on treatment B (.088 mg/kg BW) did demonstrate an increased number of days until ovulation, the response times were more variable than for mares on treatment A. Small treatment numbers are the most likely cause of this chance variability. The altrenogest dose used in treatment B increases the number of days until ovulation but the length of delay exhibited could increase the possibilities of decreased ova fertility. Mean follicular diameters at ovulation for treatments A, B, and C were 41.60, 38.80, and 40.25 mm, respectively (Table 1). Mares showed no difference (P = .65) between the treated and control groups with respect to size of the follicle at ovulation. It appears, therefore, that treatment with altrenogest during the latter stage of estrus does not alter the ultimate size a follicle will reach prior to ovulation, but will slow the rate at which it attains ovulatory size. Luteinizing hormone concentrations were measured every day during the treatment period and Day 1 through Day 13 values were analyzed (Figure 1). Day 13 was two days after the last mare ovulated in treatment B. Measurements for LH were not taken for the rest of the mares this far from the start of treatment because they ovulated at earlier times. Due to the variation in the number of days to ovulation, there were different numbers of samples to be analyzed for LH concentration. The LH curve for the control mares resembled a normal LH profile whereas the curve for both altrenogest treated groups was lengthened and did not demonstrate the normal peak. The treated mares peaked later and at a lesser magnitude than did the control mares. This agrees with Wiepz et al., 4 who found JOURNAL OF EQUINE VETERINARY SCIENCE
that altrenogest can suppress or lower LH concentration in transitional mares. Pregnancy rates were evaluated to assess the effect of ovulation delay using altrenogest on reproductive efficiency. No difference in pregnancy rates at Day 14 were detected, with 2/5 (40%), 3/5 (60%), and 3/4 (75%) pregnancies achieved in groups A, B, and C, respectively. This is in agreement with previous research by Allen et al. 5 and Squires et al. 6'7 who reported no detrimental effects on conception rates for mares which had been treated with altrenogest previous to the estrous cycle in which they had been inseminated. Ultrasonographic evaluation of embryonic vesicles was conducted on Days 12, 14, and 16 to determine whether administration of altrenogest had any detrimental effects on embryonic growth. Embryonic vesicle growth was monitored between Days 12 to 16 for treatment A, treatment B, and control mares and changes in embryonic vesicle size were 7.17, 9.00, and 8.00 mm, respectively. There was no difference (P > .05) in size of the embryonic vesicle among treated and control mares. Administration of altrenogest appeared to have no detrimental effects of pregnancy rates nor on early embryonic vesicle growth. Some general observations were made during this study which did not lend themselves to statistical analysis. These observations included estrous behavior during treatment, reproductive tract changes during treatment, and criteria that may have affected the response of certain mares to the treatment protocol. Before administration of altrenogest, mares exhibited normal signs of estrus at each mare's individual intensity. While mares were undergoing treatment, they demonstrated a decrease in intensity of behavioral estrus similar to that observed in diestrus or pregnant mares. When administration of altrenogest was discontinued, the mares continued to exhibit diestrus behavior for an average of two days following cessation of treatment. After withdrawal of treatment, mares eventually returned to behavioral estrus but with a decreased intensity. During estrus, mares' cervixes and uteri exhibited a characteristic flaccid tone. These structures became turgid, typical ofdiestrus, following altrenogest treatment. The mares retumed to estrous uterine tone after altrenogest was discontinued. In addition, mares which demonstrated a normal estrus of six to eight days had less variable responses to altrenogest treatment than mares that demonstrated a normal estrus of nine to ten days.
sw, Hunt MDN, Digby NJW: Preliminary studies on the use of an oral progestagen to induce oestrus and ovulation in seasonally anoestrous thoroughbred mares. Equine Vet J 1980;12:141-145. 6. Squires EL, Stevens WB, McGIothin DE, Pickett BW: Effect of an oral progestin on the estrous cycle and fertility of mares. JAnim Sci 1979;49:729735. 7. Squires EL, Hesseman CP, Webel SK, Voss JL: Relationship of altrenogest to ovarian activity hormone concentrations and fertility of mares. J Anim Sci 1983;56:901-910. Dry coat, non-sweater, anhidrosis? Many horses are afflicted with the genetic condition known as anhidrosis, which is a lack of or reduced ability to sweat. This can seriously reduce performance during hot humid weather. Now a nutriceutical is available to reduce the affects of this condition. MPCO, LLC 13150 N. 2 2 n d Ave. Phoenix, AZ 8 5 0 2 9 - 1 5 0 7
CONCLUSIONS
Results indicate that short-term administration of altrenogest does delay ovulation. Although trends are suggested by the present study, further studies with larger populations are needed to prove conclusively that administration of altrenogest increases follicular maintenance, decreases the preovulatory LH surge, and does not affect fertility.
REFERENCES 1. NRC:Nutrient Requirements ofHorses(5th Ed). Washington, DC: NationalAcademy Press, 1989. 2. RoserJF, Hughes JP: Prolonged pulsatile administration of gonadotropin-releasing hormone (GnRH) to fertile stallions during winter and summer. J Reprod Fertil, Suppl 1991;44:155-168. 3. Lang AL, Vogelsang MM, Potter GD: Circadian patterns of plasma LH and testosterone concentrations in stallions in summer and winter. Proc 14th ENPS 1995;284289. 4. Wiepz GJ, Squires EL, Chapman PL: Effects of norgestomet, altrenogest, and/or estradiol on follicular and hormonal characteristicsof late transitional mares. Theriogenology 1988;30:181-193. 5. Allen WR, Urwin V, Simson DJ, Greenwood RES, Crowhurst RC, Ellis DR, Ricketts
Volume 18, Number 5, 1998
PERFORMANCE & THERAPEUTIC HORSESHOE SHOCK ABSORBING TREAD ULTRA WIDE WEB BAR SHOE EFFECTIVE & ECONOMICAL FOR MORE INFO CONTACT
EQUITHOTICS INC. 7580 A C A C I A LN. V A C A V I L L E CA. 95688 PH. 707-449-3570 FAX 707-452-8394 [email protected]
331