Pituitary hormone and insulin responses to infusion of amino acids and N-methyl-D,L-aspartate in horses

Thirty-nine adult light horse mares, geldings, and stallions were used in two experiments to assess the pituitary hormone and insulin responses to infusions of arginine, aspartic acid, lysine, glutamic acid, and N-methyl-D,L-aspartate (NMA). In Exp. 1, 27 horses were assigned to one of three infusion treatments: 1) physiological saline (1 L); 2) 2.855 mmol of arginine/kg BW in 1 L of water; or 3) 2.855 mmol of aspartic acid/kg BW in 1 L of water. In Exp. 2, 12 horses were assigned, in a multiple-square 4 x 4 Latin square design, to one of four infusion treatments: 1) 2 mL of saline/kg BW; 2) 2.855 mmol of lysine/kg BW in water; 3) 2.855 mmol of glutamic acid/kg BW in water; or 4) 1 mg of NMA/kg BW in water. In Exp. 1, an acute (within 20 min) release of growth hormone (GH) was induced (P = 0.002) by aspartic acid. In contrast, acute release of prolactin (P = 0.001) and insulin (P = 0.002) was induced only by arginine; moreover, the arginine effect on insulin was present only in mares (P = 0.011). In Exp. 2, an acute release of GH was induced (P = 0.001) by glutamic acid and NMA. In males, the glutamic acid-induced GH release was greater than that of NMA; in mares, the NMA-induced GH release was greater than that of glutamic acid (P = 0.069). Both lysine and glutamic acid induced (P = 0.001) acute release of prolactin, whereas an acute release of insulin was elicited (P = 0.002) only by lysine. The NMA-induced LH response was due almost entirely to the response in mares and stallions (P = 0.016), and the NMA-induced FSH release was due almost entirely to the response in mares (reproductive status effect; P = 0.004). In the horse, aspartic acid, glutamic acid, and NMA seem to stimulate GH release; arginine and lysine seem to stimulate prolactin and insulin release; and NMA seems to stimulate LH and FSH release. It seems that N-methyl-D-aspartate glutamate receptors are involved in controlling GH, LH, and FSH secretion, whereas other mechanisms are involved with prolactin secretion. These results also indicate that gonadal steroids interact with amino acid-induced pituitary hormone release in adult horses.     

Effects of lactation on metabolic and reproductive hormones in Lipizzaner mares

In this study, growth hormone (GH), insulin-like growth factor 1 (IGF-1), leptin, luteinising hormone (LH) and prolactin were analyzed in mares from late pregnancy throughout lactation (group 1, n=46) and in non-lactating mares (group 2, n=11). Plasma GH concentrations in group 1 mares during gestation and lactation were lower than in mares of group 2 (P<0.05). Highest IGF-1 levels were found in lactating mares in the week of foaling. IGF-1 concentrations decreased continuously thereafter. Plasma leptin concentrations decreased after foaling and, for 4 weeks, were lower in lactating than in non-lactating mares (P<0.05). Reduced leptin concentrations may promote feed intake and allow lactating mares to avoid an energy deficit. In group 1 mares, prolactin concentrations reached a maximum in the week of foaling and decreased rapidly thereafter. Plasma LH concentrations in group 1 mares before foaling were lower than at corresponding times in group 2 (P<0.05). LH concentrations then increased and did no longer differ from group 2 until week 2 postpartum. This increase may contribute to the resumption of cyclic ovarian activity in postpartum mares. Subsequently, LH levels in lactating mares decreased again (P<0.05). Increased IGF-1 concentrations early postpartum might contribute to ovarian stimulation while reduced IGF-1 and GH concentrations later in lactation might cause reduced stimulation. The changes in somatotrophic hormones could thus explain, at least in part, a more pronounced stimulation of ovarian function early postpartum than during the following months of lactation.     

Glucose clearance in grazing mares is affected by diet,pregnancy, and lactation

The glucose tolerance test in the horse may be used to determine metabolic responses to diet, disease, or physiologic state. The objective of this study was to determine the effect of reproductive stage (gestation and lactation) and supplemental dietary energy source (sugar and starch [SS] or fiber and fat [FF]) on glucose metabolism in grazing mares using an oral glucose tolerance test. Twelve mares, six on each supplement, were examined on three occasions: one in the third trimester of pregnancy, the second in early lactation, and the third in late lactation. During each test, venous samples were taken at 30 and 1 min before, and 30, 60, 90, 120, 150, 180, 240, and 300 min after a nasogastric dose of glucose at 0.2 g/kg of BW. Plasma was assayed for glucose, insulin, and cortisol. Statistical analysis was a mixed model with repeated measures with horse, diet, and reproductive stage as fixed effects. The incremental glucose area under the curve (AUC) in response to oral glucose was lower in SS than in FF mares (P = 0.022). Mares tended to have a lower incremental glucose AUC in early lactation than in late gestation (P = 0.057), and insulin AUC was lower in early lactation than in late gestation (P = 0.002) and late lactation (P = 0.013). Glucose clearance was more rapid (P = 0.007) in SS than in FF mares. The glycemic response to the oral glucose tolerance test was consistent with adaptation to dietary sugar and starch as well as metabolic changes associated with pregnancy and lactation. Feeding twice-daily grain meals rich in SS influenced glucose metabolism in horses to an extent that the natural adaptation of glucose metabolism to pregnancy was moderated. Feeding a diet rich in FF more closely mimics the natural grazing state of pasture and allows for adaptation of glucose metabolism to pregnancy and lactation.     

Effects of Exogenous Insulin on Luteolysis and Reproductive Cyclicity in the Mare

Insulin is a pancreatic hormone that classically regulates carbohydrate and fat metabolism, but also appears to play a role in various reproductive processes. A preliminary study suggested insulin production by day 10 to 18 equine conceptuses. The aim of the present study was to examine the hypothesis that insulin is the conceptus signal responsible for maternal recognition of pregnancy (MRP) in the mare, or otherwise influences reproductive cyclicity during the MRP period. Six Warmblood mares were treated daily during days 7 to 17 after ovulation of two successive oestrous cycles with either (short and intermediate acting) insulin or control saline. Mares were assigned randomly to treatment, and crossed over during the subsequent cycle. Time of ovulation and corpus luteum surface area were determined by serial transrectal ultrasonographic examination of the mares' ovaries, and daily jugular vein blood samples were analysed for progesterone and luteinizing hormone (LH) concentrations. On day 14 of dioestrus, the luteolytic drive was examined by measuring systemic 15-ketodihydroprostaglandin F_{2 α} (PG-metabolite) release in response to oxytocin challenge. In addition, yolk sac fluid recovered from 32 day 10 to 14 equine conceptuses was analysed for insulin concentrations. Insulin administration did not affect luteal size, dioestrus length, the interovulatory interval, or circulating LH concentrations. Insulin administration also failed to suppress oxytocin-induced PGF_{2 α} release, and tended to depress systemic progesterone concentrations. Finally, insulin could not be detected in the yolk sac fluid of day 10 to 14 equine conceptuses by radio-immunoassay. It is concluded that insulin administered daily during days 7 to 17 of dioestrus has little or no effect on reproductive cyclicity in the mare, and is unlikely to be the MRP signal.     

The relationship between body condition,leptin, and reproductive and hormonal characteristics of mares during the seasonal anovulatory period

An experiment was conducted to determine the effects of high vs low body condition scores (BCS) produced by restricted feeding on reproductive characteristics, hormonal secretion, and leptin concentrations in mares during the autumnal transition and winter anovulatory period. Mares with BCS of 6.5 to 8.0 were maintained on pasture and/or grass hay, and starting in September, were full fed or restricted to produce BCS of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) by December. All but one mare with high BCS continued to ovulate or have follicular activity during the winter, whereas mares with low BCS went reproductively quiescent. Plasma leptin concentrations varied widely before the onset of restriction, even though all mares were in good body condition. During the experiment, leptin concentrations gradually decreased (P < 0.0001) over time in both groups, but were higher (P < 0.009) in mares with high vs low BCS after 6 wk of restriction, regardless of initial concentration. No differences (P > 0.1) between groups were detected for plasma concentrations of LH, FSH, TSH, GH, glucose, or insulin in samples collected weekly; in contrast, plasma prolactin concentrations were higher (P < 0.02) in mares with high BCS, but also decreased over time (P < 0.008). Plasma IGF-I concentrations tended (P = 0.1) to be greater in mares with high vs low BCS. The prolactin response to sulpiride injection on January 7 did not differ (P > 0.1) between groups. During 12 h of frequent blood sampling on January 12, LH concentrations were higher (P < 0.0001), whereas GH concentrations (P < 0.0001) and response to secretagogue (EP51389; P < 0.03) were lower in mares with high BCS. On January 19, the LH response to GnRH was higher (P < 0.02) in mares with high BCS; the prolactin response to TRH also was higher (P < 0.01) in mares with high BCS. In conclusion, nutrient restriction resulting in low BCS in mares resulted in a profound seasonal anovulatory period that was accompanied by lower leptin, IGF-I, and prolactin concentrations. All but one mare with high BCS continued to cycle throughout the winter or had significant follicular activity on the ovaries. Although leptin concentrations on average are very low in mares with low BCS and higher in well-fed mares, there is a wide variation in concentrations among well-fed mares, indicating that some other factor(s) may determine leptin concentrations under conditions of high BCS.     

The effects of equine somatotropin (eST) on follicular development and circulating plasma hormone profiles in cyclic mares treated during different stages of the estrous cycle

The effects of exogenous equine somatotropin (eST) administration on ovarian activity and plasma hormone levels were evaluated on horse and pony mares. The objectives of this study were to determine the effects of eST on follicular development and circulating concentrations of leutinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor I (IGF-I) in cyclic horse and pony mares. Sixteen mares received daily injections (i.m.) of eST at a concentration of 25 micrograms/kg body weight on either Days 6 through 12 (Treatment A) or 13 through 19 (Treatment B) postovulation. In addition, contemporary mares were similarly given the carrier vehicle and served as controls (Treatments C and D). Blood samples were collected at 24-hr intervals and ultrasonographic evaluations were performed on the ovaries of each mare at 48-hr intervals beginning on the first day of treatment and ending either on the day of ovulation or 5 d postovulation. Circulating levels of insulin-like growth factor-I (IGF-I) were increased in treated mares by Day 3 post-treatment (P < 0.05). Also, mares in Treatment B exhibited a decrease in plasma estradiol concentrations (P < 0.05) when compared with control mares on Days 1 through 5 postovulation of the post-treated estrous cycle. In addition, circulating leutinizing hormone levels were different for mares in Treatment A compared with controls on Days--8 through--1 pre-ovulation (P < 0.05). All follicles present on the ovaries of each mare were measured and placed into one of five categories based on their diameter. Neither the mean number of follicles per size category > or = 8 mm in diameter nor the mean follicular diameter within each size category differed among treatment and control mares. However, eST treatment significantly increased the number of follicles < or = 7 mm on the ovaries of mares treated early in the estrous cycle when compared with control mares on Days 3 and 7 post-treatment and at the onset of standing estrus.     

Effects of Body Condition and Protein Supplementation on LH Secretion and Luteal Function in Sheep

In ruminants, nutrition is one of the exogenous inputs affecting reproductive function at different levels of the hypothalamic-hypophyseal-gonadal axis. However, the exact mechanisms or even the identification of the signalling metabolic compounds by which nutrition affects reproductive function still need further clarification. The role of static body condition (BC) and its interaction with a short-term protein supplementation (PL), on secretion of metabolic hormones [growth hormone (GH), insulin and insulin-like growth factor-1 (IGF-1)], as well as on secretion of LH and progesterone (P4) was evaluated in sheep. Twenty-four Rambouillet ewes divided into two groups, with lower (LBC) and higher body condition (HBC), were randomly assigned within BC to one of two PL levels: low (LPL, 24% of crude protein; 14 g/animal/day), and high (HPL, 44% of crude protein; 30 g/animal/day). The secretion of GH, insulin, IGF-1 and LH was evaluated on day 10 of the oestrous cycle; appearance and timing of oestrous behaviour were previously detected using rams. Progesterone secretion was evaluated on day 13 of the same cycle. No differences were found (p > 0.05) between PL groups on serum GH concentrations during the sampling period (overall mean of 4.0 +/- 0.3 ng/ml), but a trend for lower values in HBC sheep was found (3.6 +/- 0.4 vs 4.4 +/- 0.4 ng/ml, p = 0.06). A BC effect was observed (p < 0.05) on serum IGF-1 level, with higher values in HBC sheep (p < 0.05). Neither BC nor PL affected (p > 0.05) secretion of LH and the number of corpora lutea, nor serum P4 and insulin concentrations. Results indicate a predominance of the static component of nutrition on sheep metabolic hormone responses, GH and IGF-1, with no effect of short-term PL on secretion of pituitary and ovarian hormones as well as luteal number and activity.     

Concentrations of prolactin, luteinizing hormone and follicle stimulating hormone in pituitary and serum of horses: effect of sex, season and reproductive state

Pituitary and serum from 86 male or female horses of various reproductive states were collected in the normal breeding season (summer) and in the nonbreeding season (winter) at a commercial slaughterhouse. Concentrations of prolactin (PRL), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay. Concentrations of pregnant mare serum gonadotropin and reproductive steroids in serum and gross appearance of the reproductive tract and gonads were used to catagorize reproductive state. Concentrations of PRL were higher (P less than .01) in summer than in winter in pituitary and serum of mares, stallions and geldings. In summer, mares had higher (P less than .01) concentrations of PRL in serum than stallions. In mares, concentrations of LH in pituitary were higher (P less than .05) in summer than in winter. Concentrations of LH in serum were higher (P less than .01) in summer than in winter in mares and geldings, higher (P less than .01) in mares than in stallions in summer, higher (P less than .01) in geldings than in stallions in summer and higher (P less than .01) in mares with low serum progesterone (P) concentrations than in mares with high P concentrations in summer. Concentrations of FSH in pituitary and serum did not differ between summer and winter for any type of horse.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Pattern of Embryonic Fixation and its Relationship to Pregnancy Loss in Thoroughbred Mares

Ultrasonographic pregnancy records of 195 mares from six Thoroughbred stud farms, over a period of 7 years were retrospectively analysed to assess the effect of various factors on embryonic vesicle (EV) fixation pattern and pregnancy outcome. Of the total of 746 pregnancies analysed, significantly (p < 0.01) more EV fixations were evident in the right uterine horn than in the left (53.35% vs 46.65% respectively). There was no significant effect of either, the side of ovulation, or age of the mare, on the side of EV fixation. However, EV fixation, was significantly (p < 0.001) more likely to occur in the right uterine horn in maiden and barren mares (65.75% vs 57.45% respectively). The age and reproductive status of the mare as well as foal heat breeding failed to demonstrate a consistent effect on pregnancy loss relative to the side of EV fixation. In lactating and foal heat bred mares, EVs were significantly (p < 0.0001) more frequently established in the contralateral horn to the one from which the mare delivered her most recent foal. In lactating mares, significantly (p < 0.05) higher embryonic and pregnancy losses were observed in the ipsilateral horn. In conclusion, (a) side of EV fixation was (i) independent of the side of ovulation and mare age (ii) significantly (p < 0.001) affected by reproductive status, (b) neither age of mare nor reproductive status had any effect on pregnancy loss rates relative to the side of EV fixation and (c) in lactating mares the EV had a greater chance of fixation and survival in the horn contralateral to the one from which the mare delivered her most recent foal.     

Differences in diagnostic test results and hematologic data between aged and young horses

Hematologic data and results of diagnostic tests were compared between aged (greater than or equal to 20 years old) and young (less than or equal to 5 years old) horses to identify hematologic and metabolic changes associated with aging. Initial data were obtained from 8 aged and 6 young mares (group 1). Similar data were collected from a second group of aged (3 mares and 3 geldings) and young (1 mare and 5 geldings) horses (group 2). Dexamethasone suppression tests (DST) and necropsies were performed on 6 additional mares and mare 8 from group 1 (group 3). Complete blood counts and serum biochemical profiles were compared between young and aged horses of groups 1 and 2. Mean corpuscular volume was higher (P less than 0.05) in aged horses. Oral glucose tolerance and insulin response to orally administered glucose were measured in 13 aged horses (groups 1 and 2) and 6 young mares of group 1. In group 1, plasma ascorbic acid values were lower (P less than 0.05) in aged horses than in young horses maintained under the same conditions and feeding regimens. An apparent age-related hyperinsulinemic response to orally administered glucose identified in group-1 mares was probably a result of a high occurrence of subclinical hypophyseal and/or thyroid adenomas. Of 13 aged horses necropsied (groups 2 and 3), 10 had hypophyseal and/or thyroid adenomas that, in group 2, were consistently associated (P less than 0.05) with hyperinsulinemic responses to orally administered glucose. All horses in groups 2 and 3 were given a 24-hour DST.(ABSTRACT TRUNCATED AT 250 WORDS)     

Determinants of attenuated LH-release associated with the first ovulation of the equine breeding season

An attenuated ovulatory rise in circulating concentrations of LH is characteristically associated with the first seasonal reproductive cycle of horse mares. Unlike ovulations (OV) of subsequent estrous cycles, the first OV of the breeding season (OV1) is not preceded by elevated concentrations of progesterone (PROG). Hence, the ability of pretreatment with PROG to abolish attenuation of LH-secretion associated with OV1 was investigated. Ten nonpregnant anestrous mares were randomly divided into 2 groups; control (C) and treated (T). Per individual, when diameter of the largest follicle was consistently greater than or equal to 35 mm, C mares received 3 times daily injections of cottonseed oil (IM), for 15 d or until OV, while T mares received exogenous PROG (IM) for 15 d, in a manner designed to mimic a diestrous pattern of release. Jugular blood samples were collected daily from onset of treatment through 10 d following the third OV (OV3). Repeated measures analyses of area under the ovulatory LH-rise (AUC) and the maximum concentration of LH associated with OV (MAX) revealed a significant main effect of OV (P less than .005) but no main effect of group or OV by group interaction (P greater than or equal to .5). When groups were combined, a significant increase in mean AUC and MAX from OV1 to OV3 was observed (P less than .01). To evaluate the influence of hypothalamic-hypophyseal recrudescence on ovulatory LH-release at OV1, 16 mares were bred to foal, and subsequently initiate reproductive activity, early (E; mid April; n = 8) or late (L; mid July; n = 8) in the year. A significant OV (1, 2 or 3) by group (E or L) interaction was observed for AUC (P less than .06) and MAX (P less than .04). Mean AUC and MAX increased progressively from OV1 to OV3 in E mares (P less than .05). In L mares, neither AUC or MAX changed from OV1 to OV3 (P greater than .4). Based on these data, we suggest that attenuation of the LH-rise at OV1 in the mare, is a consequence of incomplete recrudescence of the hypothalamic-pituitary axis.     

Hyperleptinemia in Mares: Prevalence in Lactating Mares and Effect on Rebreeding Success

Previous research from our laboratory showed that approximately one third of obese, nonfoaling mares displayed a condition of hyperleptinemia coupled with hyperinsulinemia that resembled type 2 diabetes in humans. The current study was performed to evaluate the prevalence of the hyperleptinemic syndrome in lactating mares and its possible impact on their rebreeding success. Additionally, we investigated possible relationships between leptin levels in lactating versus nonlactating mares. In experiment 1, jugular blood samples were collected from 198 lactating mares on two occasions approximately 2 weeks apart. The mares resided on eight farms in Louisiana; breeds included Thoroughbred (n = 86), Quarter Horse (n = 71), Warmblood (n = 24), and draft-type (n = 17). Body condition scores (BCS) were measured at the time of blood sampling; plasma samples were assessed for leptin and progesterone concentrations. Reproductive and medical histories, as well as feeding regimens, were compiled on each mare. Based on our previous reports and examination of the current data, a mare was considered hyperleptinemic if her plasma samples contained greater than10 ng/mL leptin; normal was considered 6.0 ng/mL or less; mares with levels above 6.0 and 10 ng/mL or greater were classified as intermediate. Overall mean leptin concentration was 4.7 ng/mL, and average BCS was 5.5. After analysis, 24 mares were classified as hyperleptinemic (12%), 138 were classified as normal (70%), and 36 were classified as intermediate (18%). Leptin concentrations were affected by BCS (P = .08), with higher concentrations in mares with higher body condition; however, there were hyperleptinemic mares with BCS of 4 to 5.5. Feeding regimen affected leptin concentrations (P < .01), with mares on pasture full-time having the highest concentrations. There was no effect of breed, mare age, number of years the mare had been bred, number of live foals, progesterone concentrations, or last foaling date on leptin concentrations. Rebreeding success averaged 81% overall and was not affected by leptin classification. In experiment 2, nonfoaling mares kept on pasture had mean leptin concentrations of 7.0 ng/mL; 8 of 31 mares (26%) displayed hyperleptinemia. Mean leptin concentration was correlated with BCS (R² = 0.65; P < .02) but was not affected by age of the mare. It was concluded that the hyperleptinemic condition occurs in lactating broodmares, even at BCS as low as 4. The overall incidence appears to be lower in broodmares than in nonfoaling mares, likely because of their lower BCS in general and the energy demands of lactation. Hyperleptinemia did not affect rebreeding success at the end of the breeding season.     

Oral and intravenous carbohydrate challenges decrease active ghrelin concentrations and alter hormones related to control of energy metabolism in horses

This study tested the hypothesis that grain and intravenous dextrose challenges would alter plasma concentrations of active ghrelin, adiponectin, leptin, glucose, insulin, and cortisol in Standardbred mares. To deliver 0.5 g of glucose (dextrose solution for the intravenous test)/kg of BW, mares received intravenous dextrose (50% solution) or oral grain administration in 2 trials. In response to the oral grain challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 56 and 802%, respectively. Plasma ghrelin concentration initially decreased (P < 0.001) by 40%, then subsequently increased (P < 0.05) from its nadir by 259%. Plasma leptin concentration decreased (P = 0.002) 17% compared with baseline. There was no change (P = 0.34) in plasma adiponectin concentration in response to oral grain challenge; however, plasma cortisol concentrations decreased (P < 0.001) by 24%. In response to the intravenous dextrose challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 432 and 395%, respectively. Plasma active ghrelin concentration initially decreased (P < 0.001) by 56%, then subsequently increased (P < 0.001) from its nadir by 314%. Plasma leptin concentration also increased (P < 0.001) by 33% compared with baseline. There was no change (P = 0.18) in plasma adiponectin concentration throughout the dextrose challenge. Plasma cortisol concentration increased (P = 0.027) by 20%. Hence, oral grain and intravenous nutrient challenges have the ability to alter variables potentially related to energy metabolism in mares, with acute changes in glucose and insulin possibly modulating changes in ghrelin and leptin.     

Testosterone effects on gonadotropin response to GNRH: cows and pony mares

Effects of testosterone propionate (TP) treatment on plasma concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) before and after an injection of gonadotropin releasing hormone (GnRH) were studied using ovariectomized cows and pony mares. An initial injection of GnRH (1 microgram/kg of body weight) was followed by either TP treatment or control injections for 10 (cows) or 11 (ponies) d. A second GnRH injection was administered 1 d after the last TP or oil injection. Concentrations of LH and FSH were determined in samples of plasma taken before and after each GnRH injection. Control injections did not alter the response to GnRH (area under curve) nor the pre-GnRH concentrations of LH and FSH in ovariectomized cows or ponies. Testosterone treatment increased (P less than .01) the FSH release in response to GnRH in ovariectomized mares by 4.9-fold; there was no effect in cows, even though average daily testosterone concentrations were 59% higher than in pony mares. Testosterone treatment reduced the LH release in response to GnRH by 26% in ovariectomized mares (P less than .05) and by 17% in ovariectomized cows (P approximately equal to .051). These results are consistent with a model that involves ovarian androgens in the regulation of FSH secretion in the estrous cycle of the mare, but do not support such a model in the cow.     

Hormonal patterns in normal and hyperleptinemic mares in response to three common feeding-housing regimens

We previously reported that a rise in plasma leptin concentrations followed the rise in insulin and glucose in meal-fed horses, whereas horses maintained on pasture had little fluctuations in hormonal patterns. We have also described a hyperleptinemic-hyperinsulinemic condition that occurs in about 30% of our light horse mares of high body condition maintained on pasture. The present experiment was designed to 1) study the effect of 3 common feeding-housing regimens on leptin and other metabolic hormones in mares and 2) determine whether the hyperleptinemic condition interacted with these regimens. Six light horse mares with high body condition (average score = 7) were assigned to 2 simultaneous 3 x 3 Latin squares, 1 with normal mares (leptin = 0.1 to 6 ng/mL) and 1 with mares displaying hyperleptinemia (>10 ng/mL). Three feeding-housing regimens were compared: ad libitum pasture, ad libitum native grass hay in an outdoor paddock, and single morning feedings of a pelleted concentrate and hay at 0700 in a barn. Five days of acclimation to the feeding regimens were followed by a 36-h period of hourly blood collection to characterize the hormonal characteristics. Leptin concentrations were elevated (P < 0.001) in mares predetermined to be hyperleptinemic compared with normal mares, regardless of the feeding regimen. Leptin was greatest (P < 0.01) in mares on pasture and least in mares fed hay. Variations over time (P < 0.01) were present for all hormones and metabolites studied. Glucose and insulin concentrations were greatest (P < 0.01) in mares on pasture, with meal-fed mares exhibiting an immediate rise in plasma concentrations of both after feeding. Mares on hay had low and constant concentrations of glucose, insulin, and leptin, with no apparent fluctuations. Cortisol, prolactin, and IGF-I did not differ with leptin status, whereas GH differed due to feeding-housing regimen (P < 0.02); there was also an interaction of leptin status and feeding-housing regimen for GH concentrations (P = 0.094). It was concluded that 1) estimates of hormonal secretion in horses based on frequent sampling, depending upon the hormone in question, can be profoundly affected by the feeding-housing regimens, and 2) the hyperleptinemic condition persists under differing conditions of feeding-housing.     

Diabetes mellitus in a domesticated Spanish mustang

An 18-year-old Spanish Mustang mare was referred for evaluation of progressive weight loss and persistent hyperglycemia. Clinicopathologic abnormalities included marked hyperglycemia and glycosuria. Serum cortisol concentration was appropriately decreased following administration of dexamethasone, indicating that the horse did not have pituitary pars intermedia dysfunction. Serum insulin and plasma C-peptide concentrations were low, suggesting that hyperglycemia was a result of decreased secretion of insulin by pancreatic beta cells. In addition, glucose concentration did not return to the baseline concentration until 5 hours after i.v. administration of a glucose bolus, suggesting that insulin secretion, insulin effect, or both were reduced. However, i.v. administration of insulin caused only a slight decrease in the plasma glucose concentration, giving the impression that the action of insulin was impaired. Within 5 hours after administration of a combination of glyburide and metformin, which is used to treat diabetes mellitus in humans, the glucose concentration was within reference limits. The horse was euthanized, and a postmortem examination was done. Immunohistochemical staining of sections of the pancreas revealed attenuation of the pancreatic islet beta-cell population, with beta cells that remained generally limited to the periphery of the islets. These findings indicate that, albeit rare, pancreatic beta-cell failure may contribute to the development of diabetes mellitus in horses.     

马卵母细胞体外成熟的研究进展

随着全球马产业的发展,马发挥的经济价值越来越大。辅助生殖技术有利于发挥优良马匹的潜在价值。马卵母细胞体外成熟(IVM)是辅助生殖技术重要的组成部分,卵母细胞的获取是体外成熟的前提,切刮法能从离体卵巢中获得较多的马卵母细胞,而活体采卵技术(OPU)则能持续地获得卵母细胞,并能较好的保存马卵母细胞的发育能力。扩张型卵母细胞的成熟率高于紧密型卵母细胞,母马的年龄会影响到其卵母细胞的质量。马卵母细胞体外存放较长时间不会影响其发育能力,现在已有较为成熟的体系能使马卵母细胞在体外保存24 h以上而不影响其成熟率。在马卵母细胞成熟体系中常用的基础培养液是M199,添加胎牛血清(FBS)、促卵泡素(FSH)、促黄体生成素(LH)、胰岛素样生长因子-1(IGF-1)等物质能显著提高成熟率,常用培养环境为38~39℃,5%CO2饱和湿度下培养,培养时间30 h。成熟的卵母细胞有扩张的卵丘细胞和极体,且成熟的卵母细胞的细胞骨架及微管结构也会发生变化。本文针对马卵母细胞的采集和体外成熟培养的相关研究进行总结,重点阐述了不同采集技术的回收率以及影响马卵母细胞体外成熟率的关键因素,以期对今后马卵母细胞体外成熟的进一步研究及后期体外受精技术的发展提供借鉴与参考。     

Effects of a Streptococcus equi infection--mediated nutritional insult during mid-gestation in primiparous Thoroughbred fillies. Part 1: placental and fetal development

REASONS FOR PERFORMING STUDY: There is a paucity of information on factors that influence placental development in the horse. Hypothesis: Changes in nutrition, particularly around the time of proliferative placental growth, can affect development of the placenta and fetal growth. OBJECTIVE: To investigate the effects of 2 planes of nutrition and an unforeseen infection-mediated nutritional insult on placental and fetal development in the mare. METHODS: Twenty maiden Thoroughbred fillies, age 3 or 4 years, mated to one Thoroughbred stallion, were maintained on either High or Moderate food intake throughout pregnancy. In mid-gestation all the mares unexpectedly became infected with Streptococcus equi and suffered varying degrees of weight loss as a consequence. Gross and stereological measurements of the placenta and measurements of foal birthweight, crown-rump length, ponderal index and plasma IGF-1 concentration were made at term. RESULTS: Gains in bodyweight during gestation were significantly higher in the High vs. the Moderate nutrition groups. Placental and fetal growth parameters were not influenced by the plane of nutrition. However, transient weight loss in mid-gestation from acutely reduced food intake resulted in morphological changes to the allantochorion and decreased foal birthweight. CONCLUSIONS: Excessive weight gain during gestation in maiden Thoroughbred fillies does not result in nutrient partitioning to the dam at the expense of the fetus. However, sudden weight loss in mid-gestation has detrimental effects on placental development which results in reduced fetal growth. POTENTIAL RELEVANCE: Clinicians should appreciate the likely effects of maternal infection and resulting weight loss on pregnancy outcome.     

Passive immunization of cyclic mares against androgen: gonadotropin and progesterone concentrations and estrous characteristics

Antiserum generated in a horse against testosterone conjugated to bovine serum albumin (BSA) was administered to six lighthorse mares (androgen-immunized mares) 1 to 3 d before a prostaglandin-induced estrus and twice again at 2-d intervals. Six control mares were administered antiserum generated against BSA on the same schedule. Relative to testosterone, cross-reactivities of other steroids with the testosterone antiserum were (%): dihydrotestosterone, 52; 5 alpha-androstane-3 alpha,17 beta-diol, 8.6; androst-4-ene-3,17-dione, 1.2; and all others tested less than .1. Tritiated testosterone binding in plasma increased (P less than .01) in androgen-immunized mares within 1 h and remained elevated (P less than .01) relative to controls for greater than 21 d. There was no effect (P greater than .10) of passive immunization against androgen on interval to estrus after prostaglandin injection, duration of estrus, ovarian volume, number of palpable follicles or follicular volume during estrus. In contrast, concentrations of luteinizing hormone (LH) were higher (P less than .05) in androgen-immunized mares than in control mares during estrus and early diestrus. Concentrations of follicle stimulating hormone (FSH) and progesterone at those times were not affected (P greater than .10). From these data, we conclude that androgens in the mare during estrus may be involved with the regulation of LH secretion. In contrast, no involvement with FSH secretion was apparent under these short-term conditions.     

Response to metabolic challenges in early lactation dairy cows during treatment with bovine somatotropin

Milk production is increased in lactating cows treated with bovine somatotropin (bST) because a greater portion of absorbed nutrients are partitioned for milk synthesis. This homeorhetic action may be caused by alterations in response of key tissues to homeostatic signals. To examine this theory, acute metabolic challenges were administered to 8 multiparous Holstein cows (61 +/- 2 days postpartum) receiving daily subcutaneous injections of pituitary-derived bST (26.3 mg) or excipient during two 14-day treatment periods (crossover experimental design). Treatment with bST increased milk yield 12%. Feed intake did not change so that net energy balance decreased (+ .5 vs. -4.3 Mcal/day). Plasma concentrations of nonesterified fatty acids (NEFA) were chronically elevated in bST-treated cows, consistent with energy balance differences. However, baseline concentrations of glucose, insulin, and glucagon in plasma did not differ. On the last 3 days of treatment, individual metabolic challenges were administered via jugular cannulas: epinephrine (700 ng/kg BW), glucose (250 mg/kg BW), insulin (1.0 micrograms/kg BW), and glucagon (175 ng/kg BW). Plasma glucose was reduced after the insulin challenge to a lesser extent during bST treatment. In bST-treated cows, the increase in plasma NEFA in response to epinephrine was greater, and NEFA concentrations were lowered to a greater extent after insulin and glucose challenges. Glucose, insulin, and glucagon removal rates were not altered, nor was plasma glucose response to epinephrine or glucagon challenges. Treatment of lactating cows with bST primarily altered the response of adipose tissue to homeostatic signals which affect lipid metabolism.