Repeatability of Prolactin Responses to a Small Dose of Sulpiride in Estrogen-Primed Geldings in Spring and in Mares During the Estrous Cycle in Summer

Two experiments were conducted to assess the repeatability of prolactin responses to a small dose of sulpiride in estrogen-primed geldings in spring and in mares during the estrous cycle in summer. Six long-term geldings each received a single intramuscular injection of 100 mg of estradiol cypionate on March 31, 2011, and were then challenged with an intravenous injection of dl-sulpiride (5 μg/kg of body weight of the racemic mixture) every other day for a total of 8 days. Jugular blood was collected at 0, 10, 20, 40, and 60 minutes after the injection of sulpiride for prolactin measurement. The experiment was repeated with six mares during the summer (July), except that the number of challenges was extended to 15 over 30 days so that any effect of estrous cycle stage could be assessed. Prolactin responses in geldings during April were robust and were varied in a quadratic manner (P < .003) over the eight sulpiride injections, increasing linearly to a plateau by the fourth injection. Mares also displayed robust prolactin responses to sulpiride injections in July, and there was no effect (P > .1) of day of injection and no effect of stage of estrous cycle (follicular phase, early diestrus, or late diestrus). We concluded that prolactin responses to this dose of sulpiride were sufficiently robust and repeatable for use as a paradigm for studies of the relative competitive efficacy and duration of action of various dopaminergic compounds and their vehicular formulations.     

Inhibitory Effects of Pergolide and Cabergoline Formulations on Daily Plasma Prolactin Concentrations in Geldings and on the Daily Prolactin Responses to a Small Dose of Sulpiride in Mares

Two experiments were conducted to assess the efficacy and duration of action of two dopaminergic compounds, pergolide and cabergoline, on daily prolactin secretion in geldings and on prolactin responses to a small dose of sulpiride over 10 days. In the first experiment, oral administration of 2 mg of pergolide was compared to a single injection of 2 mg of pergolide in a slow-release vehicle and a single injection of 5 mg of cabergoline in slow-release vehicle. Controls received vehicle only. All drug treatments reduced (P < .05) prolactin concentrations relative to that in controls but differed substantially in duration of action (oral pergolide approximately 6 hours or less, injected pergolide 6 to 24 hours, and injected cabergoline at least 6 days). In the second experiment, repeated small doses of sulpiride (2 μg/kg of body weight intravenously) were used to stimulate prolactin release in mares, and the ability of seven daily injections of pergolide (2 mg each) and a single injection of cabergoline (5 mg) in slow-release vehicle to suppress this release were compared. Control mares receiving vehicle injections had robust prolactin responses to the sulpiride injections on all days of injection (days 1, 0, 1, 2, 3, 4, 6, 8, and 10 relative to treatment). Prolactin responses were muted (P < .05) by pergolide and cabergoline treatments on the first day of injection (day 0, 30 min after treatment) and were basically absent on days 1 to 8. The single injection of cabergoline continued to be suppressive through day 10, whereas mares previously treated with pergolide (through day 6) had begun to recover a prolactin response by day 10. We conclude that either daily 2-mg pergolide injections in slow-release vehicle or a single injection of 5 mg of cabergoline in slow-release vehicle is an effective way to apply dopaminergic activity to horses for approximately 7 to 10 days and may have application in the treatment of pituitary pars intermedia dysfunction in affected horses.     

Pharmacokinetics of Sulpiride After Intravenous,Intramuscular, and Oral Single-Dose Administration in Nurse Mares

Sulpiride (SLP) is an antipsychotic drug used in humans. Although no pharmacokinetic data are available for horses, it is commonly used to encourage ovulation in noncycling mares and to stimulate lactation in adoptive mares. The aim of this study is to assess the pharmacokinetics profile of SLP after intravenous (IV), intramuscular (IM), and oral (PO) administrations in normal horses. Animals (n = 6) were treated with 1 mg/kg SLP, administered by IV, IM, and PO routes according to a randomized crossover design (3 × 3 Latin square). Blood samples (5 mL) were collected at a programmed time and analyzed using a validated with fluorescence detection method. SLP was present at a detectable concentration up to 24 hours postadministration for all routes, except for one animal in the PO group. IV and IM administrations gave similar curves, with an IM average bioavailability of 118.0%. These high values were mainly the result of the profile generated by two horses, in which a secondary concentration peak occurred in the terminal phase of the curve. After PO administration, AUC_0-∞, and consequently bioavailability (20.4%), was low. This finding could be owing to the physicochemical features of the drug. Indeed, considering that SLP is a weak base, existing in the ionized form at gastric and physiological pH, it is unsurprising that it is poorly absorbable, especially in horses with a particularly acidic gastric pH. In conclusion, injective routes are definitely preferable to PO dosing because of the low bioavailability using this route.     

Dose-Response of Prolactin to Increasing Doses of the Dopamine Receptor Antagonist,Sulpiride, in Horses: Effect of Season in Mares and Stallions of Estradiol Pretreatment in Geldings

Two experiments were performed to determine whether dopaminergic input to the adenohypophysis (1) differs across seasons in mares and stallions proportionally with changes in prolactin secretion and (2) is altered by estradiol administration in geldings. In experiment 1, prolactin responses to increasing doses of l-sulpiride in eight mares and eight stallions in March, June, September, and December were used to estimate the theoretical dose equivalent to 50% of maximal response. Prolactin areas increased (P < .001) with increasing doses of sulpiride and were greatest (P < .05) in March for stallions, but in June for mares. Mean half-maximal dose, which was assumed to be proportional to the dopaminergic input to the pituitary, was lowest (P < .05) in June and greatest in September. Experiment 2 used the same approach to determine whether the stimulatory effect of estradiol pretreatment on prolactin secretion was associated with an alteration of the half-maximal response. Geldings (n = 6/group) were administered 100 mg of estradiol cypionate in oil, or oil alone, on day 0 (October 3) and increasing doses of l-sulpiride starting on day 6. Estradiol treatment increased (P < .08) the prolactin response to l-sulpiride at 0.41 μg/kg body weight and all higher doses (P < .05); mean half-maximal dose did not differ (P > .1) between groups. We conclude that dopaminergic input to the adenohypophysis of mares and stallions varies with season and that the stimulatory effect of estradiol on prolactin secretion is not associated with a decrease in dopaminergic input to the adenohypophysis.     

Prolactin and Gonadotropin Responses in Geldings to Injections of Estradiol Benzoate in Oil, Estradiol Benzoate in Biodegradable Microspheres, and Estradiol Cypionate

We previously reported success in inducing early ovulation in seasonally anovulatory mares with a combination of estradiol pretreatment followed by daily administration of a dopamine antagonist (sulpiride). Although every-other-day injections of estradiol benzoate (EB) were effective in that experiment, practical application of this technology would require simplification of the treatment regimen. The current experiment was designed to compare, in a gelding model, the biologic responses of two alternative, one-injection regimens for estradiol delivery to the established EB treatment used previously. Fifteen long-term geldings were sampled via jugular venipuncture from November 5 to 7, 2006, and were then administered intramuscular injections of vegetable oil (n = 4); EB, 11 mg in oil (n = 4; controls); EB in biodegradable microspheres (300 mg; n = 3); or estradiol cypionate, 100 mg in oil (n = 4). Injections of EB in oil were repeated every other day for a total of 10 injections, as was done in our previous experiment. Jugular blood samples were drawn from all geldings at 3, 6, 12, 24, 36, and 48 hours relative to injections, and then on the mornings of days 3, 4, 6, 8, 10 to 18, 22, 26, and 30. On days 10 through 13, all geldings received subcutaneous injections of 125 mg sulpiride, a dopamine receptor antagonist, to stimulate prolactin secretion. On day 12, each gelding received an intravenous injection of 30 μg gonadotropin-releasing hormone (GnRH) analog and 3 mg thyrotropin-releasing hormone (TRH); frequent blood samples were drawn to characterize the luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin responses. Relative to geldings receiving oil, all geldings receiving estradiol injections had a rise (P < .05) in estradiol concentrations lasting at least 12 days. Daily LH concentrations increased (P < 0.01) in all treated groups, but the response was delayed approximately 14 days in the geldings receiving EB in microspheres. Daily FSH concentrations decreased (P < .01) in all treated groups, with the greatest response in the geldings receiving EB in microspheres. Prolactin in daily samples increased (P < .01) similarly in all estradiol-treated groups after injection of sulpiride. The LH response to GnRH analog was greatest (P < .05) in geldings receiving EB in oil and estradiol cypionate; the FSH response was not altered by treatment. The prolactin response to TRH was greater (P < .01) in estradiol-treated geldings relative to controls, but did not differ among groups. Compared with the responses to every-other-day EB injections in oil, as we used previously, a single injection of 100 mg estradiol cypionate gave the most similar and consistent responses. Because of these similar responses in this gelding model, it is likely that a single injection of 100 mg estradiol cypionate can be used in lieu of every-other-day injections of EB in oil in the treatment regimen we reported previously for stimulating ovarian activity in seasonally anovulatory mares.     

Insulin Sensitivity in Thoroughbred Geldings: Effect of Weight Gain,Diet, and Exercise on Insulin Sensitivity in Thoroughbred Geldings

The effects of dietary energy source, controlled weight gain, and exercise restriction on insulin sensitivity (SI) were studied in mature Thoroughbred geldings with body condition scores (BCS) of 4.3 ± 0.1. Two dietary energy sources were used, one high in starch and sugar (HS; n = 9) and one high in fat and fiber (HF; n = 7), and horses were fed 20 Mcal digestible energy (DE)/day above maintenance requirements to encourage weight gain. Using the minimal model of glucose and insulin dynamics, no differences in SI between groups were noted before initiation of treatment concentrate feeding. After dietary acclimation, SI was decreased in HS (P < 0.01) as compared with HF. After 32 weeks of controlled weight gain (90.8 kg; final BCS, 7.0 ± 0.1), SI remained lower in HS (P = 0.07) but did not change from the preweight gain value. SI in HF did not change between the start and end of weight gain. After completion of weight gain, exercise was restricted for 2 weeks, resulting in a reduction in SI in HF (P = 0.03) but no change in HS. It was concluded that dietary energy source may be more influential than weight gain on SI in the mature Thoroughbred gelding between BCS 4 and 7. The higher SI found in horses consuming the HF diet appeared to be partially dependent on some level of physical activity.     

Physical, Hematological, and Biochemical Responses to Acute Intense Exercise in Polo Horses

The aim of this study was to evaluate the response of physical, hematological, and biochemical parameters after acute intense exercise in polo horses playing in an outdoor international competition. The game consisted of four periods (chukkas) and each period consisted a playing time of 7 minutes. Two matches were played everyday for a week. A total of 12 horses were examined. Each “high-goal” polo horse played one chukka a day for 4 days. Horses were clinically examined the day before the games started and then daily during the 4 days of their participation in the games. During these days, physical examination was performed and blood sample was collected at rest (T0), immediately (T1) after exercise, and after 30 minutes of exercise (T2). Blood samples were analyzed for total cell counts and for determination of creatine kinase, lactate dehydrogenase (LDH), aspartate aminotransferase, lactate, total proteins, calcium, magnesium, phosphorus , and cortisol. Data were evaluated using two-way analysis of variance. Exercise caused significant dehydration (P < .01), mucous membranes congestion, increased heart rate (P < .001), and capillary refill time (P < .001). It also caused increased value of the following parameters: hematocrit (P < .001), red blood cells (P < .001), hemoglobin (P < .001), white blood cells (P < .05), lymphocyte (P < .001), total proteins (P < .001), creatine kinase (P < .05), LDH (P < .01), lactate (P < .001), and cortisol (P < .01), and a decrease in the platelet count (P < .001), calcium (P < .01), phosphorus (P < .001), and magnesium (P < .001). All parameters returned within or near the reference range by 30 minutes postexercise. On the basis of these observations, data were considered indicative of a good response to an acute intense exercise. Moreover statistical results obtained were typical of a mixed aerobic/anaerobic metabolic pathway that is prevailing in this sport.     

Description of the Responses of Some Blood Constituents to Rodeo Exercise in Chilean Creole Horses

The aim of this study was to assess the effect of the Chilean rodeo exercise on some blood constituents, classically used to assess welfare, during rest; training with and without steer; and before and after official competitions. During the training season, 13 horses were assessed at their farms of origins, and samples were taken at rest and after training with and without a steer; then during the competition season, 16 horses were assessed at four different times, one before and three after the competition. The blood constituents assessed were packed cell volume (PCV), total proteins, fibrinogen, cortisol, neutrophil:lymphocyte ratio, creatine phosphokinase, aspartate transaminase, glutathione peroxidase, and serum amyloid A. Analysis of variance for repeated measures was applied using the Statistix 8.0 software, and a significance level of P < .05 was applied. For horses assessed during exercise with and without a steer, significant differences were observed for PCV, which were significantly lower at rest than after both exercises, and total proteins, which showed a significant increase only after exercise with steer. In the case of official competitions, PCV was significantly higher immediately after rodeo, and the lowest value for cortisol was observed the evening after competition (P < .05); among the enzymes assessed, creatine phosphokinase plasma concentrations immediately after rodeo and the evening after competition were significantly higher than concentrations the next morning. The use of physiological indicators did not seem to be sufficient to assess the overall effect of rodeo on the welfare of these horses, and other types of indicators should be considered in future studies.     

早期断奶应激对仔猪某些血清激素水平及细胞免疫功能的影响

2窝(每窝8头)21日龄健康仔猪16头，随机分成吮乳组和断奶组，每组8头(每窝各4头)。断奶组仔猪21日龄断奶后转栏，吮乳组仔猪留原栏继续哺乳。在断奶时(21日龄)和断奶后1、3d(22、24日龄)，仰卧前腔静脉采血，进行血清皮质醇、生长激素、胰岛素、T3、T4、FT3、FT4水平及全血PHAT-淋巴细胞转化率和白细胞吞噬率检测。结果表明，与断奶前相比，断奶组仔猪断奶后1、3d血清皮质醇水平极显著升高(P<0.01)；断奶后3d血清T3、FT3、生长激素、胰岛素水平显著降低(P<0.05)；断奶后PHAT-淋巴细胞转化率和白细胞吞噬率显著或极显著降低(P<0.05或P<0.01)。与21日龄测定值相比，吮乳组仔猪在22、24日龄时的上述血清激素水平及细胞免疫功能无显著变化(P>0.05)。     

Leptin and Ghrelin and the Indices of Lipid Metabolism as Related to Sex Steroid Hormones in Trotters

This study examined the influence of sex steroid hormones on lipid metabolism in horses. The group of 34 clinically healthy Standardbred trotters aged 2 to 4 years was studied during an exercise test. The horses were divided into groups according to their sex. These groups were: 11 stallions, 16 mares, and seven geldings. Concentrations of testosterone, 17-β-estradiol, leptin, ghrelin, glycerol, free fatty acids (FFA), and triacylglycerols (TG) were measured in plasma obtained from blood samples taken at rest and after the end of the exercise. At rest, plasma ghrelin concentration was significantly higher in geldings than in stallions and mares (1,541 ± 206 vs 1,280 ± 288 and 1,310 ± 267 pg/mL, respectively; P = .012). Leptin was lower in geldings than in mares (2.65 ± 0.93 vs 4.70 ± 2.31 ng/mL; P = .036). The post-exercise rise in plasma ghrelin and TG concentrations was significantly higher in mares than in geldings (+220 ± 330 vs -25 ± 206 pg/mL; P = .049 and 0.31 ± 0.14 vs 0.13 ± 0.15 mmol/L; P = .016, respectively). The increase in plasma FFA level was higher in geldings than in stallions (535 ± 178 vs 334 ± 191 μmol/L, P = .046). In conclusion, lipolysis rate in geldings is higher than in noncastrated trotters.     

Relationship between Testosterone Responses to Administration of GnRH and Breeding Performance in Boars

The aim of the present study was to determine the relationship between peripheral testosterone responses to the administration of GnRH and breeding performance in boars. Twelve, sexually mature Large White boars were used. The first blood samples were collected at 9.00. At that time all boars received 100 μg GnRH (Ovurelin inj. A.U.V. Reanal). The second blood samples were taken 2.5 hours after the injection of GnRH. Serum was analyzed for testosterone by radioimmunoassay. Breeding performance was evaluated over a 150-day breeding season. The basal and the stimulated testosterone levels were related to the breeding performance. The correlation coefficient between breeding performance and post-GnRH peripheral testosterone concentration was significant (r = 0.61; P ≤ 0.05). This data suggests that the increase of the peripheral testosterone concentration may be one of the indicators for breeding performance in boars.     

Long-term Treatment of Insulin-insensitive Mares with Cabergoline: Effects on Prolactin and Melanocyte Stimulating Hormone Responses to Sulpiride and on Indices of Insulin Sensitivity

The main experiment assessed whether the inhibitory effects of the dopamine agonist, cabergoline, on prolactin and α-melanocyte stimulating hormone (MSH) concentrations would persist throughout a longer-term administration (65 days). The possible effect of cabergoline on insulin sensitivity was also studied. Ten mares known to be insulin insensitive were allotted to two groups (treated vs. control). An insulin challenge, a glucose tolerance test, and a sulpiride challenge were administered before treatment. On day 0, treated mares (n = 5) received an injection of 5 mg cabergoline in slow-release vehicle; control mares (n = 5) received an equivalent vehicle injection. Injections were repeated every 10 days for a total of seven injections. Sulpiride challenges were done 1 day before each cabergoline treatment to assess possible refractoriness to the treatment. Behavior and hair coat density were also monitored. Plasma prolactin was suppressed (P < .01) to undetectable levels in mares receiving cabergoline; control mares had robust prolactin responses to each sulpiride injection. There was no indication of refractoriness to cabergoline over time. Plasma MSH concentrations after sulpiride were also suppressed (P < .05) by cabergoline. After treatment, neither the glucose response to insulin nor the insulin response to glucose differed (P > .1) between groups. No behavioral changes were noted because of treatment. Weight of hair samples indicated that cabergoline perturbed (P < .05) winter coat growth. It is concluded that 5 mg of cabergoline in slow-release vehicle administered every 10 days is an effective way of delivering dopaminergic activity to mares that results in no noticeable detrimental effects and no refractoriness to the drug.     

粗酶制剂在大麦日粮中对产蛋鸡的影响

就粗酶制剂在大麦日粮中对产蛋鸡产蛋性能的影响进行了两次试验。按０．１％比例的酶制剂添加于含大麦２８．５％和５０％的日粮中,分别饲喂３３０和２４２日龄的产蛋鸡。结果添加酶制剂对鸡产蛋率无明显提高,而对每个蛋重和鸡蛋质量有所改善,同时使产蛋鸡外周血液中促甲状腺激素和生长激素显著提高,而使甲状腺素显著降低。     

通过式固相萃取-高效液相色谱-串联质谱法测定牛乳和乳粉中4种类固醇激素

建立通过式固相萃取-高效液相色谱-串联质谱法同时测定牛乳和乳粉样品中4 种类固醇激素的方法,采用内标法定量,样品用乙腈提取,经PRiME HLB固相萃取小柱净化,采用Acquity BEH C18色谱柱（2.1 mm×100 mm,2.5 μm）分离,乙腈和水作为流动相,使用电喷雾离子源负离子模式检测。结果表明：4 种类固醇激素在质量浓度0.2～100.0 ng/mL范围内线性关系良好,检出限为0.4～2.0 μg/kg,定量限为0.8～4.0 μg/kg;在牛乳基质中加标回收率为87.7%～110.8%,相对标准偏差为0.85%～4.56%,在乳粉基质中加标回收率为89.2%～107.4%,相对标准偏差为1.22%～4.22%;该方法分析速度快、灵敏度高,可用于牛乳和乳粉中4 种类固醇激素的定性及定量分析。     

Ovarian and Hormonal Responses to Follicular Phase Administration of Investigational Metastin/Kisspeptin Analog,TAK‐683, in Goats

This study evaluated the effects of follicular phase administration of TAK‐683, an investigational metastin/kisspeptin analog, on follicular growth, ovulation, luteal function and reproductive hormones in goats. After confirmation of ovulation by transrectal ultrasonography (Day 0), PGF_2α (2 mg/head of dinoprost) was administered intramuscularly on Day 10 to induce luteal regression. At 12 h after PGF_2α administration, intravenous administration of vehicle or 35 nmol (50 μg)/head of TAK‐683 was performed in control (n = 4) and treatment (n = 4) groups, respectively. Blood samples were collected at 6‐h intervals for 96 h and then daily until the detection of subsequent ovulation (second ovulation). After the second ovulation, ultrasound examinations and blood sampling were performed every other day or daily until the subsequent ovulation (third ovulation). Mean concentrations of LH and FSH in the treatment group were significantly higher 6 h after TAK‐683 treatment than those in the control group (12.0 ± 10.7 vs 1.0 ± 0.7 ng/ml for LH, 47.5 ± 28.2 vs 15.1 ± 3.4 ng/ml for FSH, p < 0.05), whereas mean concentrations of oestradiol in the treatment group decreased immediately after treatment (p < 0.05) as compared with the control group. Ovulation tended to be delayed (n = 2) or occurred early (n = 1) in the treatment group as compared with the control group. For the second ovulation, ovulatory follicles in the treatment group were significantly smaller in maximal diameter than in the control group (3.8 ± 0.5 vs 5.4 ± 0.2 mm, p < 0.05, n = 3). Administration of TAK‐683 in the follicular phase stimulates gonadotropin secretion and may have resulted in ovulation of premature follicles in goats.     

豆禾混播牧草碳氮磷钾生态化学计量特征对施肥与混播比例的响应

本研究以紫花苜蓿（Medicago sativa）与无芒雀麦（Bromus inermis）混播草地为对象,研究了施用不同比例氮（Nitrogen,N）磷（Phosphorus,P）钾（Potassium,K）肥与不同混播比例对牧草营养元素含量及生态化学计量特征的影响。结果表明：施肥对1~3茬无芒雀麦P,K含量、C （碳,Carbon）：P和K：P以及1茬N含量、2茬N：P、3茬C含量和C：N,N：P影响极显著（P<0.01）,对1茬和2茬苜蓿C,P,K含量和C：P,N：P以及3茬N,P,K含量和C：P,C：N,N：P,K：P影响极显著（P<0.01）;混播比例对1茬无芒雀麦P含量和C：P,2茬C含量和3茬C：N,N：P影响极显著（P<0.01）;施肥与混播比例互作对1~3茬无芒雀麦C含量、苜蓿P含量和C：P,1茬与2茬无芒雀麦P含量和C：P以及苜蓿N：P影响极显著（P<0.01）。禾草N：P在3.53~7.60之间,苜蓿在8.37~12.32之间,表明无芒雀麦生长主要受N素影响,苜蓿生长同时受N素和P素的影响。     

河西内陆盐渍区碱茅属植物建植及草地对生长条件的反应

在河西走廊地区建植碱茅（Ｐｕｃｃｉｎｅｌｌｉａｔｅｎｕｉｆｌｏｒａ－Ｐ．ｃｈｉｎｕｍｐｏｅｎｓｉｓ）人工草地，做收种和刈割兼用。撒播方式下，播量达２２．５ｋｇ／ｈｍ２时，种子落种数为２０００／ｍ２，田间出苗数７９４／ｍ２，出苗率为１３％，定苗数受地表积盐程度的影响（Ｐ＜０．０１），地表盐斑为５０％，９０％的幼苗数为９８８株／ｍ２和３０１株／ｍ２；碱茅草地盖度，种子产量和牧草产量均与土壤全盐含量呈负线性关系（ｒ－－０．８０５１，ｒ＝－０．８１５８，ｒ＝－０．９２９２），但种子发芽率和千粒重似乎不受立地土壤盐分的影响。由于受田间管理措施和碱茅自身发育节律的影响，生长第３年的草地盖度、种子产量和牧草产量均比第２年有不同幅度的下降，据此，适时灌水，加强施肥等田间管理措施对维持草地基况良好，获得高产有重要作用。     

The Effect of Endotoxin on the Liver Lipid Peroxide Level and on the Liver and Plasma Zinc Concentrations in Rats As Related to Time After Administration

The lipid peroxide level of rat liver increased due to i.p. endotoxin injection, reaching its maximum, value 24 h after administration. After that the level declined, reaching approximately the initial value at 48 h. The liver Zn concentration increased and the plasma Zn concentration rapidly decreased after the injection. The liver Zn concentration then gradually declined and the plasma Zn respectively increased near the initial level 48 h after the injection.The highest lipid peroxide level occurred at 24 h when the mean Zn concentration of the plasma and liver were already relatively near the initial level, suggesting a regulative role of Zn in lipid peroxidation.