Evaluation of progesterone deficiency as a cause of fetal death in mares with experimentally induced endotoxemia

The role of decreased luteal activity in embryonic loss after induced endotoxemia was studied in mares 21 to 35 days pregnant. Fourteen pregnant mares were treated daily with 44 mg of altrenogest to compensate for the loss of endogenous progesterone secretion caused by prostaglandin F2 alpha (PGF2 alpha) synthesis and release following intravenous administration of Salmonella typhimurium endotoxin. Altrenogest was administered daily from the day of endotoxin injection until day 40 of gestation (group 1; n = 7), until day 70 (group 2; n = 5), or until day 50 (group 3; n = 2). In all mares, secretion of PGF2 alpha, as determined by the plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations, followed a biphasic pattern, with an initial peak at 30 minutes followed by a second, larger peak at 105 minutes after endotoxin injection. Plasma progesterone concentrations decreased in all mares to values less than 1 ng/ml within 24 hours after endotoxin injection. In group 1, progesterone concentrations for all mares were less than 1 ng/ml until the final day of altrenogest treatment. In 6 of 7 mares in group 1, the fetuses died within 4 days after the end of treatment, with progesterone concentrations less than 1 ng/ml at that time. In the mare that remained pregnant after the end of treatment, plasma progesterone concentration was 1.6 ng/ml on day 41 and increased to 4.4 ng/ml on day 44. In group 2, all mares remained pregnant, even though plasma progesterone concentrations were less than 1 ng/ml in 4 of 5 mares from the day after endotoxin injection until after the end of altrenogest treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in plasma progesterone concentrations from days 17 to 42 of gestation in mares maintaining or losing pregnancy

Plasma progesterone concentrations were measured in 179 mares bled on alternate days commencing with a positive pregnancy diagnosis on Days 17 to 18 after ovulation and concluding on Days 42 to 45. During this period 17 mares (10 per cent) lost their pregnancies, 11 before Day 25. In 15 mares the timing of the pregnancy loss could be determined with adequate accuracy; in only one did a decline in progesterone precede the loss. Thus pregnancy loss between Days 17 and 42 was rarely caused by a fall in plasma progesterone.     

Plasma prolactin concentrations in mares and their neonates after oxytocin induction of parturition

Studies were undertaken to investigate the effects of oxytocin induction on prolactin release in term (Group II) and preterm (Group III) mares and to compare these effects to spontaneously foaling mares (Group I). Since physiological concentrations of prolactin in blood have not been measured in the neonatal foal, experiments were designed to monitor prolactin in the cord artery and jugular blood of the foals from all groups of mares. Although prolactin levels varied in term mares (Group I and II) during the last 11 days of pregnancy, an increase was observed between Day -6 and Day 0 (2.7 and 11.9 ng/ml respectively; P less than 0.1). The average concentration of prolactin over the last 4 days (Days -3 to 0) had increased by 40% when compared to the average concentration on Days -6, -5, and -4. These findings indicate a rising trend which appears to occur concomitantly with changes in concentrations of 2 mammary components tested, sodium and potassium. Prolactin concentrations did not significantly increase in term mares after oxytocin treatment or in spontaneously foaling mares. However, the preterm induced mares had higher prolactin concentrations during the first stage of labor (19.3 +/- 7.2 ng/ml) than prior to treatment with oxytocin (4.7 +/- 2.0 ng/ml; P less than 0.01). Levels of prolactin in all groups significantly declined by 20-min post-placental expulsion. For the first 30 min after birth, prolactin concentrations in foals from oxytocin-induced mares appeared to be 2-fold higher than those from spontaneously foaling mares. Thereafter, prolactin values declined to baseline values by 48 hrs. When comparing cord arterial plasma with cord venous plasma in each group, prolactin concentrations were similar. However, the average prolactin levels in both the cord artery and vein appeared higher (ave: 1.1 ng/ml) in Group II and III than in Group I (less than 0.5 ng/ml). From these results, the authors suggest that 1) prolactin may have a role in regulating mammary secretory products in mares just prior to parturition; 2) oxytocin may increase prolactin secretion in preterm induced mares; 3) oxytocin induction may have a short term effect to increase circulatory prolactin concentrations in neonates in utero regardless whether their dams were treated preterm or term.     

Effects of flunixin meglumine on endotoxin-induced prostaglandin F2 alpha secretion during early pregnancy in mares

The role of prostaglandin F2 alpha (PGF2 alpha) in embryonic loss following induced endotoxemia was studied in mares that were 21 to 44 days pregnant. Thirteen pregnant mares were treated with a nonsteroidal anti-inflammatory drug, flunixin meglumine, to inhibit the synthesis of PGF2 alpha caused by Salmonella typhimurium endotoxin given IV. Flunixin meglumine was administered either before injection of the endotoxin (group 1, -10 min; n = 7), or after endotoxin injection into the mares (group 2, 1 hour, n = 3; group 3, 2 hours, n = 3); 12 pregnant mares (group 4) were given only S typhimurium endotoxin. In group 4, the secretion of PGF2 alpha, as determined by plasma 15-keto-13,14-dihydro-PGF2 alpha concentrations, was biphasic, initially peaking at 30 minutes followed by a second, larger peak approximately 105 minutes after the endotoxin was given IV. When flunixin meglumine was administered at -10 minutes, synthesis of PGF2 alpha was inhibited for several hours, after administration of flunixin meglumine at 1 hour, the second secretory surge of PGF2 alpha was blocked, and administration of the drug at 2 hours did not substantially modify the secretion of PGF2 alpha. Plasma progesterone concentrations were unchanged after endotoxin injections were given in group 1. In group 2, progesterone values decreased less than 2 ng/ml and remained low for several days. In group 3 and group 4, progesterone concentrations decreased to values less than 0.5 ng/ml by 48 hours after endotoxin injections were given.(ABSTRACT TRUNCATED AT 250 WORDS)     

Enzymeimmunoassay of oestrone sulphate concentrations in faeces for non-invasive pregnancy determination in mares

AIM: To determine the suitability of measuring faecal oestrone sulphate (OS) by enzymeimmunoassay as a means of determining pregnancy status in mares bred under New Zealand conditions. METHODS: An antibody-coated microtitre plate-based enzymeimmunoassay was used to determine the concentration of OS in faecal and plasma samples obtained from pregnant and non-pregnant mares. RESULTS: In non-pregnant mares, the mean faecal OS concentration was 34 ng/g, and the value three standard deviations above this was 80 ng/g. None of 427 faecal samples collected from 116 non-pregnant mares over a l-year period had an OS concentration >80 ng/g. Only five samples from three mares had an OS concentration >65 ng/g, the value two standard deviations above the mean non-pregnant value. Analysis of faecal OS concentrations in 532 faecal samples collected from 39 pregnant mares showed that as pregnancy progressed, an increasing proportion of faecal samples had OS concentrations >80 ng/g. None of the mares 150 days or more pregnant had faecal OS concentrations <50 ng/g, and 204/220 samples obtained from these mares had faecal OS concentrations >80 ng/g. Following foaling or foetal death, elevated faecal OS concentrations returned quickly to non-pregnant levels. The mean +/- s.e.m. plasma level of OS in five mares bled daily throughout one oestrous cycle was 1.7 +/- 0.2 ng/ml. Sixty-eight blood samples from pregnant mares bled up to five times between 92 days after mating and foaling all had plasma OS concentrations >30 ng/ml, with 64/68 being >50 ng/ml. CONCLUSIONS: This study shows that measuring faecal OS concentrations by enzymeimmunoassay offers a convenient, accurate, non-invasive means of determining pregnancy status in mares from 150 days after mating onwards. Mares with faecal OS concentrations <50 ng/g can be considered not pregnant, while mares with faecal OS concentrations >80 ng/g can be considered pregnant. Those few mares returning a faecal OS concentration between 50 and 80 ng/g should be retested to obtain a conclusive result. Measuring plasma OS concentrations allows pregnancy status to be determined earlier (from 100 days after mating). Moreover, the discrimination between non-pregnant and pregnant levels is greater for OS in plasma than in faeces. CLINICAL RELEVANCE: Measurement of OS concentrations in faeces provides an alternative and non-invasive means of determining pregnancy status in mares from 150 days after mating.     

Lactation and reproductive performance of mares fed added dietary fat during late gestation and early lactation

Eighteen pregnant mares were randomly allotted to one of two treatment groups. The control group was fed a conventional concentrate and the fat group was fed a concentrate containing 5% feed-grade rendered fat. Both concentrates had the same nutrient:calorie ratio and were fed in amounts required to maintain zero change in percent body fat of the mares. During the study, which began 60 d prior to expected foaling date and ended 60 d postpartum, mares were monitored for feed intake, body weight, rump fat thickness, ration digestibility, plasma glucose and lipid concentrations, milk composition and reproductive efficiency. Birth weight, growth rate, and plasma glucose and lipid concentrations were also measured in foals. Mares fed fat consumed less concentrate (P<.09 during the last 60 d of gestation but consumed similar amounts of concentrate over 60 d of lactation. Protein and ether extract digestion was higher (P<.05 and P<.09, respectively) in the mares fed fat during the postpartum period. Dietary treatment had no influence on plasma glucose or lipid concentrations of the mares or plasma glucose concentrations of their foals, but foals whose dams were fed fat had higher plasma lipid concentrations at birth (P<.01), d 10 and d 30 (P<.05). Percent fat was higher in milk samples from mares fed fat at d 10 (P<.09, 1.23 vs .99%) and d 60 (P<.01, .72 vs .43%) of lactation. Reproductive performance was not significantly different between treatment groups, however there was a trend for a shorter postpartum interval and fewer cycles per pregnancy in the mares fed added fat. Foals from both groups were of similar size and weight at birth and had similar weight gains over 60 d, however, foals nursing mares fed fat tended to gain more weight in the first week of life (1.85 vs 1.56 kg/d) and have more rump fat at d 60 (.53 vs .44 cm) than foals nursing control mares.     

Effects of short-term early gestational exposure to endophyte-infected tall fescue diets on plasma 3,4-dihydroxyphenyl acetic acid and fetal development in mares

Consumption of wild-type (toxic) endophyte-infected tall fescue (E+) by horses during late gestation is known to adversely affect pregnancy outcome; however, little is known of the potential disruptive consequences of E+ consumption by mares during the critical phases of placentation and fetal development in early pregnancy. The objective of this study was to evaluate the detrimental effects of feeding E+ to mares during early gestation. Mares (n = 12) paired by stage of gestation (d 65 to 100) were assigned to diets (six per diet) consisting of endophyte-free (E-) or E+ tall fescue seed (50% E- or E+ tall fescue seed, 45% sweet feed, and 10% molasses fed at 1.0% of BW/d). Mares also had ad libitum access to E+ or E- annual ryegrass hay, and were fed diets for 10 d. Following removal from the tall fescue diet on d 11, mares were placed on common bermudagrass pasture and monitored until d 21. Morning and evening rectal temperatures were recorded and daily blood samples were collected for progesterone and prolactin (PRL) analyses, whereas samples for 3,4-dihydroxyphenyl acetic acid (a catecholamine metabolite) analysis were collected on alternate days. For clinical chemistry analysis, blood samples were collected on d 0, 5, 10 and 21. Daily urine samples were collected for ergot alkaloid analysis, and ultrasonography was performed for presence of echogenic material in fetal fluids. Rectal temperatures (E+ 37.76+/-0.03; E- 37.84+/-0.03 degrees C) and serum PRL concentrations (E+ 14.06< or =0.76; E- 12.11+/-0.76 ng/mL) did not differ (P = 0.96) between treatments. Measuring the change in basal serum concentration from d 0 over time, progesterone concentrations did not differ (-0.64 +/-1.49 and -0.55+/-1.47 ng/mL for E+ and E- mares, respectively). There was no negative pregnancy outcome, and ultrasonography indicated no increase in echogenic material in fetal fluids. Plasma 3,4-dihydroxyphenyl acetic acid concentrations decreased (P < 0.05) in E+ compared with E- mares (2.1+/-0.14 and 4.4+/0.43 ng/mL, respectively). Urinary ergot alkaloid concentration was greater (P < 0.01) in mares consuming E+ compared with E- (532.12+/- 52.51 and 13.36+/-2.67 ng/mg of creatinine, respectively). Although no fetal loss was observed during the current study, elevated concentrations of urinary ergot alkaloid were consistent with depressed endogenous catecholamine activity, suggestive of an endocrine disruptive effect of hypothalamic origin.     

Measurement of equine prolactin with an equine-canine radioimmunoassay: seasonal effects on the prolactin response to thyrotropin releasing hormone

A radioimmunoassay (RIA) based on anti-equine prolactin antiserum and radioiodinated canine prolactin was used to assess the dose response of plasma prolactin to thyrotropin releasing hormone (TRH) in mares in the nonbreeding season (winter) and in mares in estrus in the breeding season (summer). Mares were administered TRH intravenously and blood samples were collected via jugular catheters at −15, 0, 15, 30, 45, 60, 90, 120, 180 and 240 min relative to injection. Doses of TRH were 0, .08, .40, 2.0 and 10.0 mg per mare (n = 3 per dose within each season). The prolactin response was assessed by absolute hormonal concentrations before and after TRH injection and by net area under the curve. Prolactin concentrations in plasma before injection of TRH were higher (P < .01) in estrous mares in summer than in anestrous mares in winter (4.8 vs 1.3 ng/ml). Moreover, there was a greater (P < .01) response to TRH injection in estrous mares than in anestrous mares. Based on areas under the curve, there was an effect of season (P < .01) and of TRH dose (P < .01) as well as a season-dose interaction (P < .01). In general, there was little or no prolactin response to any dose of TRH in anestrous mares in winter when pre-TRH concentrations were low. In contrast, there was an increase in the prolactin response with increasing doses of TRH up to 2.0 mg in estrous mares in summer; 2.0 and 10.0 mg of TRH resulted in similar prolactin secretion. We conclude 1) that prolactin secretion in the horse is stimulated by TRH as has been reported for other species and 2) that prolactin concentrations and the TRH-induced secretion of prolactin are greater in estrous mares in summer than in anestrous mares in winter.     

A preliminary study of the efficacy of fluphenazine as a treatment for fescue toxicosis in gravid pony mares

This study was designed to determine the efficacy of a single injection of the long-acting D₂-dopamine receptor antagonist, fluphenazine deconoate, on the clinical symptoms and plasma prolactin concentrations of mares grazing endophyte-infected tall fescue. Twelve mares were maintained on an 80% endophyte-infected tall fescue pasture. Group T (n=6) received 25 mg of fluphenazine deconoate i.m. on day 320 of gestation, while group C (n=6) served as untreated controls. Daily plasma samples were obtained and analyzed for prolactin using a homologous equine radioimmunassay. The prolactin data were then grouped according to week after treatment (day 320 to 327=Week 1; day 328 to 335=Week 2) and to week prior to parturition (day 0 to −7=Week −1; day −8 to −14=Week −2). A single injection of fluphenazine had no effect on experiment-wide plasma prolactin concentrations or on prolactin concentrations relative to week of treatment or parturition. However, a treatment by time period interaction was observed. Plasma prolactin concentrations were higher (p<.05) in treated mares (35.6±29.1 ng/ml) on week 2 compared to control mares (14.9±14.2 ng/ml). In addition, there was a trend (p=.09) for fluphenazine treated mares (68.1±64.2 ng/ml) to exhibit higher prolactin concentrations one week prior to parturition when compared to control animals (27.8±27.4 ng/ml). Peak prolactin was higher (p<.05) in fluphenazine treated mares and in mares which foaled normally. Gestation lengths were shorter (p<.05) in group T (330±2.9 days) compared to group C (341±3.2 days). It appeared that treated mares exhibited fewer clinical signs of fescue toxicosis than mares in group C. Four out of six of the untreated mares exhibited at least one of the clinical signs of fescue toxicosis (agalactia, stillbirth, thickened placenta, retained placenta) while only one treated mare showed obvious clinical symptoms. These data indicate that a single injection of a long-acting dopamine receptor antagonist may be beneficial in reducing the effects of fescue toxicosis in pregnant mares grazing endophyte-infected tall fescue pastures. However, additional research is needed to determine the most effective dosage and administration times.     

Kontrolle der luteolytischen Wirkung eines neuen Prostaglandin F2α-Analogs bei der Behandlung von Stuten mit Anostrie and Dyszyklie unter fortlaufender Bestimmung des Progesteronspiegels im Plasma

Inhalt 15 Stuten mit Anöstrie bzw. Dyszyklie wurden unter Kontrolle ihres Plasmaprogesteronspiegels mit einem Prostaglandinanalog (Hoe 837 V; 0,1 mg/ml) behandelt. Die Progesteronwerte lagen vor der Behandlung zwischen 1,6 ng/ml and 13,6 ng/ml. Eine ausreichende Luteolyse lieβ sich bei 14 Stu ten mit einer einzigen intramuskulären Injektion von 3,3 ml induzieren. Die Marke von 1 ng/ml war bei 9 Stuten schon am ersten and bei weiteren 5 Stuten am zweiten Tag nach der PG-Injektion erreicht. Die während der Rosse gemessenen Progesteronwerte entsprachen weitgehend den von anderen Autoren bestimmten. Dauer and Verlauf der Rosse waren wie bei nicht behandelten Stuten. 2–7 Tage nach Rossebeginn bzw. zwischen dem 5. and 12. Tag nach der PG-Behandlung stiegen die Progesteronwerte wieder über 1 ng/ml. In der Lutealphase erreichten sie bei den tragend gewordenen Stuten schnell einen Spitzenwert um 8 ng/ml (x?). Die Verlaufskurven zeigten groβe individuelle Unterschiede hinsichtlich des Progesteronniveaus. Bei Stuten mit späterer Fruchtresorption wurde zwischen dem 5./6. bis 13./14. Tag ein deutliches Progesterondefizit erkennbar. Von 2 Stuten, die in der induzierten Rosse nicht konzipierten, hatte die eine einen verkürzten, die andere einen spontan verlängerten Diöstrus. Die Frühkonzeptionsrate betrug 84,6 %. Nachkontrollen ergaben, daβ 61,5 % der behandelten Stuten tragend wurden. Pro Trächtigkeit waren durchschnittlich 1,6 Sprünge erforderlich. Contents Control of the luteolytic effect of a new prostaglandin F₂α-analogue during the treatment of anestrous and dyscyclic mares under continual measurement of the progesterone level in the blood plasma. 15 mares in clinical anestrus and dyscyclia were treated with the prostaglandin F₂α-analogue (Hoe 837 V; 0,1 mg/ml) under control of the plasma progesterone level. Before treatment the plasma progesterone values ranged between 1.6 ng/ml and 13.6 ng/ml. A satisfactory luteoly tic response was obtained with a single intramusculary dose of 3.3 ml in 14 mares. 24–48 h after the PG-injection progesterone declined to baseline levels of 1 ng/ml. The mares came to estrus 1–2 days after falling of progesterone < 1 ng/ml or 1–6 days after the PG-injection. Progesterone values measured in the estrus-period were like those determined from other authors. Duration and performance of estrus were as in untreated mares. 2–7 days from the onset of estrus and between five to twelve days after treatment, the progesterone levels again arose >1 ng/ml. In the lutealphase in pregnant mares they reached a peak of 8 ng/ml (x?) quickly. The progesterone courses showed great individual difference in their basal levels. In mares undergoing embryonic resorption a progesterone deficiency is seen in the early lutealphase between the 5.16. day and 13.114. day. From the two mares which did not conceive in the induced estrus one had a shortened and another a prolonged diestrus. The conception rate for early pregnancy was 84.6 %, and still 61.5 % for late pregnancy. In the average for each pregnancy 1.6 copulations were needed.     

Plasma progesterone in mares showing oestrus during early pregnancy (author's transl)]

Sixtyfour mares were examined 3 and 6 weeks after mating. Progesterone was measured in 22 mares 3 weeks after mating in order to see if this could be of any help in the oestrous diagnosis. None of the pregnant mares had plasma progesterone below 2 ng/ml. Pregnant mares that did not show oestrus had higher levels of plasma progesterone than pregnant mares showing signs of oestrus 3 weeks after mating. Clinical findings in pregnant and nonpregnant mares 3 weeks after mating is compared, and oestrus in pregnant mares is discussed.     

Progesterone and equine chorionic gonadotropin concentrations around the time of pregnancy loss in mares impregnated by donkeys or stallions

The objective of this study was to compare the incidence of pregnancy loss of mares carrying a mule embryo with that of mares carrying a horse embryo. The possible causes of such mortality were evaluated through serial ultrasonographic evaluations and hormonal monitoring, paying special attention to the role of premature regression of the endometrial cups and its relation to inadequate luteal function.

Twenty-eight mares impregnated by stallions and 19 mares impregnated with donkey semen were evaluated ultrasonographically every week from day 20 to day 150 of pregnancy. The viability of the product was assessed each time, and the diameter of the embryonic vesicle was measured from day 25 to day 60. Blood samples for progesterone and equine chorionic gonadotropin (eCG) determination were taken every week.

Both progesterone and eCG concentrations during normal pregnancies wegre significantly lower in the mares inseminated with donkey semen than in the mares impregnated by stallions (P<.05). In 7 of the mares carrying mule conceptuses to term, the concentrations of eCG remained basal throughout the study. In the other animals from this group, the levels of this hormone did increase but returned to baseline much earlier (on day 77 of pregnancy) than in the mares served by stallions (on day 126 day of pregnancy). There was no significant difference between the growth rate of embryonic vesicles of mares carrying mule embryos and that of mares carrying horse embryos (P>.05).

The incidence of pregnancy loss was significantly higher (P<.05) in mares carrying a mule embryo (36.8 %) than in mares carrying a horse embryo (21.4%); it occurred on average on day 93 of pregnancy in mares carrying mule embryos and on day 43 on mares carrying horse embryos. There was only 1 case in which pregnancy loss was associated with concentrations of both eCG and progesterone that were much lower than the average for the normal pregnancies of the same group, and this was in a mare carrying a horse embryo. The most frequent cause of pregnancy loss was premature luteal regression due to primary luteolysis, as evaluated via peripheral progesterone concentrations. This occurred in 2 mares carrying horse embryos and in 4 mares carrying mule embryos. Three mares carrying mule embryos and 1 carrying a horse embryo had abortions that were not preceded or accompanied by any alteration in progesterone or eCG levels and were thus classified as fetal deaths of non-endocrine origin.

It is concluded that the incidence of pregnancy loss is higher in mares carrying a mule embryo than in mares carrying a horse embryo. However, this is not due to the low progesterone concentrations associated with the premature regression of the endometrial cups that occurs in mares with interspecific pregnancy.     

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.     

Estradiol and progesterone concentrations resulting from the administration of an exogenous hormone regimen in diestrous embryo transfer recipients

Estradiol and progesterone concentrations were evaluated from diestrous embryo transfer recipient mares (5 to 14 days post-ovulation) which were treated with an exogenous hormone regimen. Upon detection of the donor mare's ovulation (0 hours), 10 mg PGF_2α was given to the recipient mare; at 12, 24 and 36 hours 20 mg estradiol cypionate; at 48 hours, 500 mg progesterone in oil and then 22 mg altrenogest at 60, 72 and 96 hours. Altrenogest (22 mg/day) was continued until end of the trial (detection of a fetal heart beat). Embryos were transferred non-surgically 6 or 7 days after the start of treatment.Plasma samples were evaluated over three periods; period 1-between recipient mare ovulation and prior to PGF_2α period 2-between PGF and embryo transfer and period 3-post-transfer. During periods 2 and 3, estradiol was higher (P<.05) for mares which were 10 to 14 days post-ovulation (late diestrous) as compared to mares which were 5 to 9 days post ovulation (mid-diestrous) when treatment began. Progesterone concentrations were higher (P<.05) for the mid-diestrous mares in the same periods. The pregnancy rate was higher for the late diestrous mares than the mid-diestrous mares (58% (7/12) vs 10% (1/10)). However, no difference (P>.05) was detected in estradiol or progesterone in the late diestrous mares which were pregnant or open. During period 2, estradiol was higher (P<.05) in the pregnant than open mares. Whereas, during period 3, progesterone was higher (P<.05) in the open mares.These data suggest that estradiol is important for the establishment of pregnancy in the mare. Furthermore, hormone treatment developed in this study appears to have some potential in synchronization of diestrus mares to be used as embryo recipients.     

Induced abortion with two Prostaglandin F2α analogues in mares: Plasma progesterone changes

Three experiments were conducted to test the abortifacient effects of PGF2α analogues on mares during midgestation (average gestation length 141.5 days). The progesterone concentration was measured by radioimmunoassay. In experiment I, five mares recieved an injection of PGF2α analogue (fluprostenol: 500 μg intramuscularly) and a second injection either at 24, 48, of 72 h. Although the progesterone concentration decreased (P < 0.05) an average of 44 per cent in 24 h, none of the pregnancies were terminated. In experiment 2, beginning at least 10 days after experiment I, the same five mares were given PGF2α analogue as follows: 250 μg intravaginally and 500 μg intramuscularly. The treatment was repeated 48 h later. Progesterone concentrations had not increased since experiment 1 and dit not decrease during the 48 h following either injection. In experiment 3, six mares (average gestation length 162 days) were treated every 6 or 12 h with PGF2α analogue (cloprostenol: 375 μg) until expulsion of the fetus occurred at 47 ± 25 h after the initial injection; the mares received an average of 5 treatments. The progesterone concentration averaged 22 ± 7 ng/ml before the initial PGF2α treatment, decreased (P<0.05) to 8.4 ±2.7 ng/ml by 12 h before expulsion and 1–8 ±0.4 ng/ml 12 h after fetal expulsion. The progesterone concentration remained below 1.0 ng/ml for the next 4 days. However, only one of six mares exhibited estrual behavior after induced abortion.     

Use of a Compounded Long-Acting Progesterone Formulation for Equine Pregnancy Maintenance

The objective of this study was to test the efficacy of a compounded long-acting progesterone formulation (BioRelease P4 LA 150; BETPHARM, Lexington, KY) containing 150 mg progesterone/ml for pregnancy maintenance in mares after prostaglandin (PG) F_2α-induced luteolysis. On day 18 of gestation, mares were randomly assigned to one of four groups (n = 7/group): (1) saline-treated control (Saline); (2) PGF_2α-treated control (PGF); (3) PGF_2α- and Regu-Mate-treated (Regu-Mate); and (4) PGF_2α- and BioRelease P4 LA 150-treated (BioRelease). On day 18, Saline mares received 1 ml sterile saline IM, whereas PGF, Regu-Mate, and BioRelease mares received 250 μg cloprostenol IM. Beginning on day 18, Regu-Mate mares received 10 ml Regu-Mate orally once daily and BioRelease mares received 10 ml BioRelease P4 LA 150 containing 150 mg/ml progesterone IM once every 7 days; treatments were continued until day 45 or until pregnancy loss occurred. Pregnancy diagnosis was performed every 3 days between days 18 and 45 (or until pregnancy loss). Pregnancy loss was defined as complete absence of a discernible embryonic vesicle as determined with transrectal ultrasonography. Pregnancy loss rates between days 18 and 45 were: Saline, 1/7; PGF, 7/7; Regu-Mate, 1/7; and BioRelease, 0/7. The pregnancy loss rate was higher (P < .01) in PGF_2α-treated control mares compared with the other groups. There were no differences (P > .1) in pregnancy loss rates among the saline-treated control, Regu-Mate-treated, and BioRelease P4 LA 150-treated mares. These results indicate that intramuscular administration of BioRelease P4 LA 150 containing a total of 1.5 g progesterone every 7 days provided a sufficient level of progesterone to maintain pregnancy between days 18 and 45 of gestation in mares that lacked an endogenous source of progesterone; therefore, this long-acting formulation of progesterone appears to be an efficacious and suitable alternative to currently available progesterone formulations that require daily administration.     

Daily rhythm of prolactin and corticosterone in unrestrained, incubating turkey hens

Plasma concentrations of prolactin and corticosterone were determined in hourly samples collected over a 25-hr period from unrestrained turkey hens exhibiting incubation (broody) behavior. Hens were maintained in cages on a 14 hr light: 10 hr dark photoperiod. Mean plasma prolactin concentration increased significantly late in the dark period to reach maximum daily levels of 496 ng/ml at the start of the photophase. However, a well-defined daily rhythm in prolactin secretion was not evident. Plasma corticosterone concentration showed a significant daily rhythm characterized by a major secretory peak (7.14 ng/ml) in the middle of the light period and a smaller, less well-defined peak (4.11 ng/ml) during the dark period. Both prolactin and corticosterone secretion ranged widely throughout the day in individual hens in a manner suggestive of pulsatile hormone secretion. This study demonstrates that the extremely high prolactin concentrations characteristic of the incubating turkey hen reflect large, dynamic changes in plasma prolactin concentrations occurring throughout the photoperiod, rather than a constantly high level of prolactin secretion.     

Factors affecting pregnancy rate to estrous synchronization and fixed-time artificial insemination in beef cattle

Application of AI in extensive beef cattle production would be facilitated by protocols that effectively synchronize ovarian follicular development and ovulation to enable fixed-time AI (TAI). The objectives were to determine whether use of a controlled internal drug release (CIDR) device to administer progesterone in a GnRH-based estrous synchronization protocol would optimize blood progesterone concentrations, improve synchronization of follicular development and estrus, and increase pregnancy rates to TAI in beef cows. Beef cows (n = 1,240) in 6 locations within the US Meat Animal Research Center received 1 of 2 treatments: 1)?an injection of GnRH [100 μg intramuscularly (i.m.)] followed by PGF(2α) (PGF; 25 mg i.m.) 7 d later (CO-Synch), or 2) CO-Synch plus a CIDR during the 7 d between GnRH and PGF injections (CO-Synch + CIDR). Cows received TAI and GnRH (100 μg i.m.) at 60 h after PGF. Progesterone was measured by RIA in blood samples collected 2 wk before and at initiation of treatment (d 0) and at PGF injection (d 7). Estrous behavior was monitored by Estrotect Heat Detectors. Pregnancy was diagnosed by ultrasonography 72 to 77 d after TAI. Plasma progesterone concentrations did not differ (P > 0.10) between synchronization protocols at first GnRH injection (d 0), but progesterone was greater (P < 0.01) at PGF injection (d 7) in cows receiving CO-Synch + CIDR vs. CO-Synch as a result of fewer CIDR-treated cows having progesterone ≤1 ng/mL at PGF (10.7 vs. 29.6%, respectively). A greater (P < 0.01) proportion of CO-Synch + CIDR vs. CO-Synch cows were detected in estrus within 60 h after PGF (66.7 vs. 57.8 ± 2.6%, respectively) and a greater (P < 0.01) proportion were pregnant to TAI (54.6 vs. 44.3 ± 2.6%, respectively). For both synchronization protocols, cows expressing estrus within 60 h before TAI had a greater pregnancy rate than cows without estrus. For cows with plasma progesterone ≤1 ng/mL at PGF injection, CO-Synch + CIDR increased pregnancy rate (65.2 ± 5.9 vs. 30.8 ± 3.4% with vs. without CIDR), whereas pregnancy rates did not differ (P > 0.10) between protocols (52.1 ± 2.1 vs. 50.0 ± 2.4%, respectively) when progesterone was >1 ng/mL (treatment × progesterone; P < 0.01). Inclusion of a CIDR in the synchronization protocol increased plasma progesterone concentration, proportion of cows detected in estrus, and pregnancy rate; however, the increase in pregnancy rate from inclusion of the CIDR was primarily in cows with decreasing or low endogenous progesterone secretion during treatment.     

Plasma progesterone, oestradiol-17β and total oestrogen profiles in relation to oestrous behaviour during induced ovulation in Murrah buffalo heifers

The objectives of this study were to establish the characteristics of oestrous behaviour in Ovsynch (induction of ovulation through administration of GnRH-PGF2-GnRH in a systemic manner on 0, seventh and ninth day respectively) and Ovsynch plus Norprolac (Quinagolide hydrochloride – an inhibitor of prolactin secretion) treated Murrah buffalo heifers and to determine the relationships between this behaviour and the plasma concentrations of oestradiol-17β (E2), total oestrogen, and progesterone. Oestrus was detected by visual observations of oestrus signs, per rectal examination of genitalia and bull parading thrice a day during treatment period. Among all the symptoms, it was observed that bull mounting of heifers in oestrus was highest. Examination of genital tracts per rectum revealed that the cervix was relaxed, uterus was turgid and ovaries had palpable follicle in animals with oestrus. The peak concentrations of E2 (10.81 ± 0.62 pg/ml) and total oestrogen (17.11 ± 1.21 pg/ml) occurred at 9.45 ± 0.85 and 9.64 ± 0.93 h after second GnRH administration, respectively, in Ovsynch treated animals. However, the peak levels of E2 (20.02 ± 2.87 pg/ml) and total oestrogen (32.71 ± 3.15 pg/ml) occurred at 10.18 ± 0.50 and 10.36 ± 0.75 h after second GnRH administration, respectively, in Ovsynch plus Norprolac treated animals. Plasma progesterone concentration was basal (0.20 ± 0.001 ng/ml) during the peri-oestrus period. The plasma progesterone concentration was the lowest on the day of oestrus and increased to register a peak on day 13 ± 2 of the cycle. Oestrous behaviour was positively correlated with the peak concentration of E2 (p < 0.001) and total oestrogen (p < 0.001) during the peri-oestrus period. Inhibition of prolactin by Norprolac administration significantly increased the concentration of E2 and total oestrogen during oestrus in buffaloes in comparison to those recorded in animals subjected to Ovsynch protocol alone. In conclusion, our results suggest that the peak concentrations of E2 and total oestrogen and mean level of E2 and total oestrogen during the peri-oestrus period are the important factors contributing the behavioural manifestation of oestrus in buffalo cows.     

Responses of seasonally anovulatory mares to daily administration of thyrotropin-releasing hormone and(or) gonadotropin-releasing hormone analog

Seventeen seasonally anovulatory light horse mares were treated daily, starting January 5 (d 1), for 28 d with GnRH analog (GnRH-A; 50 ng/kg BW) and(or) thyrotropin-releasing hormone (TRH; 5 microg/kg BW) in a 2 x 2 factorial arrangement of treatments to test the hypothesis that combined treatment may stimulate follicular growth and development. Ovaries were examined via ultrasonography and jugular blood samples were collected every 3 d. Frequent blood samples were collected after treatment injections on d 1, 2, 4, 7, 11, 16, and 22; on d 29, all mares received an i.v. mixture of GnRH, TRH, sulpiride, and EP51389 (a growth hormone secretagogue) to assess pituitary responsiveness. No consistent effects (P > 0.1) of treatment were observed for plasma LH, FSH, prolactin, or thyroxine concentrations in samples collected every 3 d. The only effect on ovarian follicle numbers was a reduction in number of follicles 11 to 19 mm in diameter due to TRH treatment (P = 0.029). No mare ovulated during treatment. On the days of frequent sampling, mean LH (P = 0.0001) and FSH (P = 0.001) concentrations were higher in mares receiving GnRH-A and tended to increase from d 1 through 7. In contrast, mean prolactin (P = 0.001) and thyroid-stimulating hormone (P = 0.0001) concentrations were high in mares receiving TRH on d 1 but rapidly decreased thereafter. When mares were administered the secretagogue mixture on d 29, the LH response was greater (P = 0.0002) in mares that had previously received GnRH-A but the FSH response was not affected (P > 0.1); the prolactin response was greater (P = 0.014) and the TSH response was smaller (P = 0.0005) in mares that had previously received TRH. Surprisingly, an immediate growth hormone response to EP51389 was absent in all mares. In conclusion, daily GnRH-A treatment stimulated plasma LH and FSH concentrations immediately after injection; although no long-term elevation in preinjection concentrations was achieved, the responses gradually increased over time, indicating a stimulation of gonadotropin production and storage. Daily treatment with TRH stimulated plasma TSH and prolactin concentrations, but the response diminished rapidly and was minimal within a few days, indicating a depletion of pituitary stores and little or no stimulation of production. There was no beneficial effect of adding TRH treatment to the daily GnRH-A regimen.