PGFM response to exogenous oxytocin and determination of the half-life of oxytocin in nonpregnant mares.

We investigated the half-life of oxytocin in reproductively normal mares and the prostaglandin response after oxytocin administrations. Mares were given oxytocin, 10 or 25 iu, i.v., on the day of, or 2 days after, ovulation, and frequent jugular blood samples were collected for analysis of oxytocin and Prostaglandin F metabolite (PGFM) by RIA. Neither dose of oxytocin nor day of treatment affected the half-life of the exogenous oxytocin, which was determined to be 6.8 min. A significant increase in PGFM was observed within 6 min of oxytocin administration and peak values were observed within 10 min. PGFM response after oxytocin administration on the day of ovulation appeared elevated compared to the response 2 days after ovulation.     

Effect of oxytocin on suppression of oestrus in mares exhibiting normal oestrous cycles

Abstract

AIM: To compare the efficacy of oxytocin given once daily, either I/V or I/M, on Days 7–14 post-ovulation, on the expression of oestrus in mares through to 65 days post-ovulation.

METHODS: Eighteen mares of various breeds that were displaying normal oestrous cycles were randomly assigned to one of three treatment groups on the day of ovulation (Day 0), detected using transrectal ultrasonography. Mares in the control group (n = 6) were given 1 mL saline I/V; mares in the I/V and I/M groups (n = 6 per group) were injected with 10 IU oxytocin I/V and I/M, respectively. All treatments were given once daily on Days 7–14. Mares were teased by a stallion three times per week, up to 65 days post-ovulation, to detect oestrous or dioestrous behaviour. Ovarian follicular and luteal activity were monitored using transrectal ultrasonography three times weekly, and daily when a follicle >30 mm diameter was present until ovulation. Blood samples were collected weekly for analysis of concentrations of progesterone in serum. Prolonged dioestrus was defined as a period of >30 days of dioestrous behaviour after Day 0, confirmed by detection of corpora lutea and concentrations of progesterone in serum >4 nmol/L.

RESULTS: Overall, 8/18 (44%) mares showed prolonged dioestrus. These included 2/6 (33%) mares in the control group, compared with 5/6 (83%) and 1/6 (16%) mares in the I/V and I/M groups, respectively (p = 0.11). The median duration of the first dioestrus was longer for the I/V group (64 (min 16, max 66) days) compared with the control group (18 (min 12, max 64) days) (p = 0.05), but was not different between the control group and the I/M group (16 (min 13, max 65) days) (p = 0.57). For all mares there was strong agreement between teasing behaviours, ultrasonographic assessment of ovarian activity, and concentration of progesterone in serum.

CONCLUSIONS AND CLINICAL RELEVANCE: This study found that low doses of oxytocin did not increase the proportion of mares with prolonged dioestrus, compared with controls, although I/V oxytocin did increase the median duration of dioestrus. The results must be interpreted with some caution as group numbers were small, and a variety of breeds were used. Further investigation of oxytocin given I/V may be warranted as a potential method of oestrus suppression in mares exhibiting oestrous cycles that is low cost, safe and well-tolerated, and potentially reversible with prostaglandin.     

Epidemiological studies on abortions in Indian thoroughbred mares

Young mares up to 8 years of age accounted for 41.55 percent of abortions. Forty and one quarter (40.25) percent of abortions were recorded in the 8th month of gestation. Twin abortions were 25.98 per cent and correlated directly to overall abortions.     

Nicotinic acid supplementation at a supraphysiological dose increases the bioavailability of NAD+ precursors in mares

NAD⁺ deficiency has recently been linked with increased occurrences of congenital abnormalities and embryonic death in human and animal subjects. Early embryonic death is a major component of pregnancy loss in mares and very little is known regarding the requirement for NAD⁺ in horses. The aim of this study was to quantify NAD⁺ and its metabolites in the plasma and urine of mares after orally administering an acute dose of nicotinic acid and determine the absorption, metabolism and excretion of this essential precursor for NAD⁺ biosynthesis. Nicotinic acid (5 g per os) was administered to four mares via a dosing syringe. Blood samples were collected at 0, 0.25, 0.5, 1, 2, 4, 6 and 22 h, and urine samples were collected at 0, 3, 6 and 22 h. The samples were processed and analysed by mass spectrometry. A general additive model was applied to all metabolite concentration values followed by a post-hoc multiple comparisons test. Nicotinic acid was rapidly absorbed into peripheral blood within 15 min of administration and the concentrations of nicotinic acid, nicotinamide (NAM), nicotinuric acid, nicotinic acid mononucleotide and nicotinic acid adenine dinucleotide (NaAD) increased significantly in plasma at 30 min. The concentrations of NAM, nicotinic acid riboside and NaAD increased significantly in urine at 3 h. The levels of NAM and NaAD remained significantly elevated in plasma at 22 h, sixfold and ninefold greater, respectively, than the basal levels at 0 h. While the extracellular levels of NAD⁺ in the samples remained undetected, the large, sustained elevation of NaAD levels in plasma indicates that the NAD⁺ levels were boosted within the cellular compartments. The results show that nicotinic acid supplementation increases the bioavailability of NAD⁺ precursors in mares, which is proposed to be beneficial during periods of peak NAD⁺ demand, such as during early embryo development.     

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.     

Effect of repeated administration of oxytocin during diestrus on duration of function of corpora lutea in mares

OBJECTIVE: To determine whether IM administration of exogenous oxytocin twice daily on days 7 to 14 after ovulation blocks luteolysis and causes prolonged function of corpora lutea (CL) in mares. DESIGN: Prospective study. ANIMALS: 12 mares. PROCEDURES: Beginning on the day of ovulation (day 0), jugular blood samples were collected every other day until day 40 for determination of progesterone concentration. On day 7, mares (n = 6/group) were treated with saline (0.9% NaCl) solution (control group) or oxytocin. Beginning on day 7, control mares received 3 mL of sterile saline solution every 12 hours, IM, and oxytocin-treated mares received 60 units of oxytocin every 12 hours, IM, through day 14. Mares were considered to have prolonged CL function if progesterone concentration remained > 1.0 ng/mL continuously through day 30. RESULTS: The proportion of mares with prolonged CL function was significantly higher in the oxytocin-treated group (6/6), compared with the control group (0/6). All control mares underwent luteolysis by day 16, at which time their progesterone concentrations were < 1.0 ng/mL. In contrast, all 6 oxytocin-treated mares maintained progesterone concentrations > 1.0 ng/mL continuously through day 30. CONCLUSIONS AND CLINICAL RELEVANCE: IM administration of 60 units of oxytocin twice daily on days 7 to 14 after ovulation was an efficacious method of inhibiting luteolysis and extending CL function in mares. Disrupting luteolysis by administering exogenous oxytocin during diestrus appears to be a plausible and practical method of long-term suppression of estrus in mares.     

Twinning in mares: A review of recent studies

Recent studies on twinning are reviewed. Multiple ovulations were more frequent in thoroughbreds (19%) than in quarter horses (9%) and Appaloosas (8%). The multiple ovulation rate was reduced approximately 50% in foaling mares compared to barren and maiden mares. There was a high degree of repeatability of double ovulations and twin pregnancies within mares and within family lines.Only one embryo was found in each of 23 pituitary extract-treated mares with multiple, synchronous ovulations (<2 days apart) and in each of 39 brood-farm mares with double, synchronous ovulations. Pregnancy rates (number of mares pregnant, regardless of number of embryos/mare) were significantly higher for double, synchronous ovulations than for single ovulations in artificially stimulated mares (58% vs 38%) and in brood-farm mares (83% vs 54%). The results indicated that ova produced by synchronous, double ovulations are viable and fertilizable (indicated by the higher pregnancy rates), but that one of the resulting embryos is eliminated (indicated by the absence of twins).Synchronous, double ovulations were not recorded in association with any of 107 sets of natrually occurring twins. Most (76%) of the twin sets were associated with one detected ovulation. The remaining twins were associated either with one ovulation, but a large unovulated follicle was present at the time of the last examination (10%), or with asynchronous, double estrous ovulations (14%). Twins originated more frequently (P < .05) from asynchronous, double estrous ovulations (9/57) than from synchronous, double ovulations (0/39).Approximately 50% of the mares in which twin embryos were diagnosed rectally before day 31, had 1 foal. However, mares in which twins were recorded as present at day 32–36 and day 40–42 had a single foal in only 17% and 6% of the mares, respectively. The methods used for intervention when twins were diagnosed were unsatisfactory. Complete termination of pregnancy with a prostaglandin or intrauterine flushing resulted in failure to establish a singleton pregancy during the remaining breeding season in 10/11 mares. Attempts to eliminate one embryo resulted in loss of both embryos in 6/7 mares.     

Morphological studies on the fetal membranes of the normal singleton foal at term.

Of 211 consecutive thoroughbred foalings, 145 satisfied a set of criteria for normal parturition and foal viability. The fetal membranes from these and from 10 pony foalings have been systematically examined morphologically and quantitatively and the findings compared to those of other authors. Five sites on the allantochorion were consistently devoid of villi. Expulsion usually occurred with the non-villous side outermost. In 24 per cent of allantochorions the non-pregant horn was of equal length or longer than the pregnant horn. These placentae tended to be shed with the villous side outermost. All membranes contained a hippomane in the allantoic cavity. Remnants of the yolk sac placenta and extra-embryonic coelom were always present. Large vestigeal yolk sacs have previously been confused with anomalous twins. In most cases the endometrial cup sites were site recognisable, usually arranged around the umbilical cord attachment. This attachment was found on the dorsal aspect of the uterine cavity either at the junction of the horns or within the pregnant hor; rarely, it was in the non-pregnant horn. A degree of twistng of the umbilical cord is normal. In 6.2 per cent of cords small urachal dilatations were noted, resulting from flight stenosis of the urachal luman at the site of a twist. Ninety-five per cent of cords measured between 36 and 83 cm.     

Pharmacokinetics and physiologic effects of alprazolam after a single oral dose in healthy mares

The objective of this study was to evaluate the pharmacokinetic properties and physiologic effects of a single oral dose of alprazolam in horses. Seven adult female horses received an oral administration of alprazolam at a dosage of 0.04 mg/kg body weight. Blood samples were collected at various time points and assayed for alprazolam and its metabolite, α‐hydroxyalprazolam, using liquid chromatography/mass spectrometry. Pharmacokinetic disposition of alprazolam was analyzed by a one‐compartmental approach. Mean plasma pharmacokinetic parameters (±SD) following single‐dose administration of alprazolam were as follows: C_max 14.76 ± 3.72 ng/mL and area under the curve (AUC_0–∞) 358.77 ± 76.26 ng·h/mL. Median (range) T_max was 3 h (1–12 h). Alpha‐hydroxyalprazolam concentrations were detected in each horse, although concentrations were low (C_max 1.36 ± 0.28 ng/mL). Repeat physical examinations and assessment of the degree of sedation and ataxia were performed every 12 h to evaluate for adverse effects. Oral alprazolam tablets were absorbed in adult horses and no clinically relevant adverse events were observed. Further evaluation of repeated dosing and safety of administration of alprazolam to horses is warranted.     

Fertility at the first post partum estrous compared with fertility at the following estrous cycles in foaling mares and with fertility in nonfoaling mares

A retrospective study on the reproductive performance of 401 artificially inseminated trotter mares during six breeding seasons is presented. Mares, 279 post partum (PP) and 122 maiden and barren, or nonlactating (NL), were inseminated with fresh semen obtained from four fertile stallions of the same breed. Pregnancy rate (PR) of mares inseminated at the foal heat (182/253, 71.9%) was lower, but not significantly different, than the PR (22/26, 84.6%) of mares inseminated for the first time at the second post partum cycle and similar to the PR at the first and second cycle of NL mares (95/112, 77.8% and 25/33, 75.7%, respectively). PR of mares inseminated at the foal heat was higher, but nonsignificantly different, from PR of the post partum mares not pregnant after artificial insemination (AI) at foal heat and inseminated again at the following estrous cycle. The PR after AI at the foal heat was significantly higher than the PR when the AI was performed at the third or later cycle in NL mares (71.9% vs 22.2%, P<0.01). The estrus cycle/pregnancy ratio for the PP mares inseminated, for the first time at the foal heat or at the second heat and for NL mares was, respectively, 1.4, 1.2, 1.3 at first cycle and 1.4, 1.3 and 1.4 at the end of the season or when mares left the stud. The proportion of open mares at the end of the season or when leaving the stud was 7.1% (18/253), 3.8% (1/26) and 4.1% (5/122) for PP mares first inseminated at the first or second post partum cycle and for NL mares, respectively; the total rate of open mares was 6% (24/401). The foaling rate (FR) following conception at the foal heat (72.1%) was not statistically different from the FR following conception at any other cycle (50–100%). Based on the absence of significant differences in fertility at the first post partum estrus cycle versus any other estrus cycle, we conclude that breeding at the foal heat should be advisable.     

Retrospective study of dystocia in mares at a referral hospital

REASONS FOR PERFORMING STUDY: The period between the onset of dystocia and its resolution has an important bearing on fetal outcome. There are few published data on which to base decisions regarding optimum management of cases in practice. OBJECTIVES: To evaluate and compare the effects of a coordinated dystocia management protocol (CDMP) with that of a previous protocol of random management on time to resolution and outcome in both an emergency dystocia referral population of mares (referred emergency cases: EM) and in a population of mares residing in hospital due to high risk pregnancy (HRP) concerns that then experience dystocia at parturition. METHODS: Retrospective study performed at a university hospital referral centre of cases presenting from 1991-2004 divided into Group 1 (pre-CDMP) and Group 2 (CDMP). RESULTS: Medical records of 71 cases with dystocia were retrieved and data recorded. For referred emergency cases (EM), time from hospital presentation to resolution decreased significantly by 32 min (P = 0.03) after institution of CDMP. Survival rate of mares at discharge was 86%. Survival of EM foals was low, with 10% in Group 1 and 13% in Group 2, surviving to discharge. For EM foals delivered alive, survival to discharge was 30% and 43% in Groups 1 and 2, respectively. Median Stage II was significantly (P < 0.001) different at 71 and 282 min for EM foals delivered alive vs. those not alive at delivery, respectively. Median duration of Stage II was also significantly (P < 0.001) different between EM foals surviving and not surviving to discharge, at 44 and 249 min, respectively. Survival of HRP dystocia foals to discharge was 79%. CONCLUSIONS: Although CDMP reduced the time from presentation at the hospital to resolution significantly for EM, total duration of Stage II for EM was unchanged, as was foal outcome. POTENTIAL RELEVANCE: Very early referral of mares with dystocia to referral centres with dystocia management protocols may improve fetal outcome as increased duration of Stage II in the horse affects fetal outcome negatively.     

Long term consequences of low birthweight on postnatal growth, adiposity and brain weight at maturity in sheep

Low birth weight (LBW) as a result of restricted fetal growth increases the risk for later metabolic diseases and adiposity. However the relationship between LBW and postnatal growth and adult body composition has not been fully investigated. We have used sheep to determine the effects of LBW on postnatal growth and body composition at maturity. LBW was induced by twinning and placental embolization. At birth, LBW lambs were 38% lighter than controls (2.8 +/- 0.2 vs 4.4 +/- 0.3 kg, P<0.05), but had caught up in bodyweight by 8 weeks after birth. At approximately 2.3 years, bodyweights were not different between groups, but there were reductions in absolute (-8%) and relative (-17%) brain weights of LBW sheep (P<0.05) compared to controls. X-ray absorptiometry showed that the mature LBW sheep, compared to controls, had greater amounts of lean muscle (38.1 +/- 1.3 vs 35.3 +/- 0.5 kg, P<0.05) and tended to have more body fat (12.2 +/- 1.2 vs 9.6 +/- 0.9 kg; P=0.1); at autopsy abdominal fat mass was greater in LBW sheep (3.06 +/- 0.26 vs 2.20 +/- 0.25 kg, P<0.05). Plasma leptin concentrations were not different between groups. We conclude that, in sheep, LBW is associated with early postnatal catch-up in body weight, but body composition is permanently altered such that, relative to controls, adiposity is increased and brain weight is decreased.     

Pharmacokinetics of single intravenous and single and multiple dose oral administration of rifampin in mares

The disposition of rifampin in six healthy mares after single intravenous (i.v.) and oral (p.o.) doses and after seven oral doses of 10 mg/kg administered twice a day was investigated using a high performance liquid chromatographic (HPLC) method. Pharmacokinetic variables for rifampin determined using the HPLC method were comparable to variables reported from earlier studies utilizing a microbiological assay. Desascetylrifampin, a major metabolite of the parent compound, could not be detected in the serum but was detected at low concentrations in urine. Mean trough concentrations of rifampin increased from the first to the second dose of the multiple dose oral study and then remained unchanged through 72 h. At 84 h after the first dose (i.e. 12 h after the final dose) the rifampin concentration was significantly decreased ( P = 0.001). The harmonic mean of the half-life of rifampin decreased significantly from 13.3 h after a single oral dose of 7.99 h after the seventh oral dose. The mean serum protein binding of rifampin over the concentration range of 2–20 μg/ml was 78%. Mean trough serum concentrations of unbound rifampin after multiple oral doses ranged from 0.67 μg/ml at 24 h to 0.40 μg/ml at 72 h. The mean unbound serum rifampin concentration at 84 h (i.e., 12 h after the final dose) was 0.30 μg/ml. Trough concentrations and the 84-h sample concentration of unbound rifampin exceeded the minimum inhibitory concentration for most gram positive bacterial isolates from horses reported in this study. All organisms with minimum inhibitory concentrations less than 0.125 μg/ml were considered susceptible.
Based on the pharmacokinetics of rifampin after p.o. administration, we concur with the current dosage recommendation of 10 mg/kg twice a day by mouth. At this dose, most streptococci, Rhodococcus equi , and coagulase-positive staphylococci would be considered susceptible to rifampin.     

Physiological and pharmacological studies of oxytocin in virgin cattle III

Resting concentrations of oxytocin were measured in virgin cattle and it was determined if these animals would release oxytocin in response to milking associated stimuli. Half-life, clearance and entry rates for oxytocin were also quantified, to compare previous data from experiments carried out on lactating and nonlactating cows. Six Holstein virgin heifers received a two minute udder massage. Blood samples were taken before, at and after massage and analyzed for oxytocin by radioimmunoassay. Resting concentrations of oxytocin averaged 24.8 ±2.27 uU/ml (mean ± SE) and increased to 28 ±2.3 uU/ml at 2 min. The increase in oxytocin was clearly apparent after post-stimulation concentrations were corrected for basal (pre massage) levels. Serum oxytocin concentrations remained below baseline for 25 min post stimulation. No increase in oxytocin was seen prior to udder stimulation. Heifers were infused with 1.0 IU/min oxytocin to measure pharmacokinetic parameters. Oxytocin disappearance from sera was monitored by radioimmunoassay. A two compartment model for oxytocin elimination produced two half-life components. A rapid half-life component with a mean value of 3.07 ±.32 min and a slow half-life component with a mean of 24.3 ±2.2 min. The overall clearance rate for oxytocin was 9.4 ±.58 ml/kg min. The mean entry rate of oxytocin into the body was 231.96 ±25.96 uU/kg min. The sensitivity of the neuroendocrine reflex, eliciting oxytocin release, appears dependent upon the physiological state of the animal. Half-life and clearance measurements for oxytocin compare closely to those obtained in previous studies for cows in either early lactation, late lactation or undergoing mammary involution. It appears that the clearance and production of oxytocin are also related to the physiological state of the animal.