Investigation of glucagon-like peptide-1 response to six oral carbohydrates in ponies.

Glucagon-like peptide-1 (GLP-1), the principal incretin in horses, may play a role in the pathophysiology of insulin dysregulation (ID). This study aimed to describe its concentration in response to three preserved forages and four dynamic tests for ID in ponies. Twelve adult ponies of mixed ID status were given a meal of hay, soaked hay or haylage, an in-feed oral glucose test (OGT), oral sugar test (OST), an oral test using a proprietary breakfast cereal (WEET) or a combined glucose-insulin tolerance test (CGIT) weekly in a randomised cross-over study. Glucose, insulin and GLP-1 concentrations were measured before and following each intervention. Ponies were designated ID or non-ID and insulin resistant (IR) or non-IR according to OGT and CGIT results, respectively. All interventions apart from the CGIT provoked a GLP-1 response within 30 min. The OGT and WEET interventions, (containing the greatest dose of non-structural carbohydrate, 1.06 and 1 g/kg BW, respectively), resulted in a greater area under the curve (AUC) for GLP-1 compared to all other interventions (P < 0.001). No difference in GLP-1 response was detected according to ID or IR status, despite there being strong positive correlations (rs [95 % CI]) between GLP-1 and insulin concentrations measured at individual time points (0.67 [0.62 - 0.71]; P < 0.001) and as AUC (0.66 [0.49-0.79], P < 0.001). These data do not support of the use of GLP-1 as an adjunctive diagnostic test for ID or IR, as defined by conventional intravenous or oral dynamic tests.

Comparison of one novel and four established diagnostic tests for insulin dysregulation in ponies.

Several tests have been advocated for diagnosis of insulin dysregulation (ID). Tests using simple sugars may not reflect the response to naturally ingested carbohydrates. This study aimed to evaluate agreement between the oral glucose test (OGT), the oral sugar test (OST), a novel oral test using a proprietary cereal (WEET), the IV combined glucose-insulin tolerance test (CGIT) and fasted basal insulin (FI) for diagnosis of ID. Each of the five tests above was performed on a group of six normal and six insulin dysregulated mixed-breed ponies in a randomised crossover study. Area under the curve (AUC) and maximum concentration of insulin from OGT, OST and WEET showed strong to very strong bivariate correlations (r = 0.85-0.94, and r = 0.87-0.92, respectively; P ≤ 0.001) and were significantly different between tests (associated with dose of carbohydrate) and between CGIT-positive and -negative ponies. Dichotomous results showed substantial agreement between OST and both WEET (κ = 0.65; P = 0.02) and OGT (κ = 0.67; P = 0.01) and between CGIT and both OST (κ = 0.63; P = 0.03) and OGT (κ = 0.67; P = 0.01), and no agreement between FI, which had low sensitivity, and all other tests (κ = 0.15 - 0.31; P > 0. 05). Palatability of WEET was variable, resulting in one pony being excluded for analysis of WEET data. Further work on development of an oral test using a more palatable feedstuff and appropriate cut-offs or diagnostic thresholds for tests of ID is warranted.

Effects of lameness, subclinical mastitis and loss of body condition on the reproductive performance of dairy cows.

A total of 318 cows were monitored in the pre-breeding postpartum period for the presence of three production stressors: lameness, subclinical mastitis and body condition score (BCS) loss. For each stressor, cows were given a classification of severely, moderately or non-affected based on mobility scores, somatic cell counts and BCS change. The number of days from calving to onset of the first luteal phase was greater in cows that had one severe production stressor (median 44 days) or two moderate production stressors (41 days) compared with cows that had no stressors (31 days) (P=0.02 and P=0.04, respectively). More than one severe stressor increased the interval further. There was no difference between cows with one moderate stressor (median 38 days) and those with none (P=0.13). The delay to the first luteal phase was significantly longer in cows with two moderate stressors if the onset of one stressor occurred at the time when resumption of ovarian activity was expected. The presence of these production stressors in early lactation had no effect on the interval from calving to establishment of the next pregnancy or the number of inseminations required despite the negative effect on the onset of the luteal phase.

Height measurement in horses and ponies: optimising standard protocols.

Standard and modified measuring sticks were used to record height at the withers and a 'non-contact' laser was used to measure withers and loin heights. Sixty horses and ponies, ranging in height (115 to 155 cm) and body condition score (BCS) (moderate to obese) were measured by each method at 10-minute intervals for 40 minutes. Measurement series were repeated over three successive days. Unique regression models were constructed for method-specific data. Coefficients of variation were similar for stick and laser methods (0.002 to 0.004 per cent). Models were not influenced by day of measurement or BCS. Withers height decreased significantly (0.48 cm, 95 per cent confidence intervals -0.61 to -0.36 cm, P<0.001) over the first 20 minutes. In living animals, laser-derived measurements of withers height at T(20) exceeded stick measurements by approximately 1 cm (P<0.001). Loin height remained similar across time. Some alteration in relaxed withers height is an inevitable consequence of changes in muscle tone at the scapulothoracic synsarcosis.

The effect of intra-uterine devices on the reproductive physiology and behaviour of pony mares.

Spherical (35 mm diameter) glass intra-uterine devices (IUDs) are commonly understood to modify sexual behaviour in performance mares. The effect of IUDs on the endocrinology (luteinising hormone, oestradiol, progesterone, testosterone), ovarian and uterine function, sexual and social behaviours associated with the ovarian cycle were evaluated during the natural breeding season in nine, nuliparous pony mares (4.6±0.5 years old, 230±12 kg bodyweight). Age and weight-matched groups were allocated to sham-treatment (n=5) or IUD-implantation (n=4) groups. Fifty complete ovarian cycles were evaluated, with 26 ovarian cycles monitored for the nine mares prior to treatment and a further 12 cycles were evaluated for mares in each of the groups post-treatment. Detailed characterisation of cyclic reproductive changes in the pony may be of value for controlled breeding programmes. The presence of an IUD was well-tolerated, but failed to modify any recorded aspect of reproductive physiology or behaviour.

Changes in plasma gonadotrophin and prolactin concentrations following castration of the pony stallion.

Concentrations of gonadotrophins and prolactin were recorded in pony stallions castrated during the early breeding season, to examine the regulatory role of the gonad at a time when testosterone has been postulated to exert positive feedback on LH secretion. Further, gonadotrophin concentrations in geldings are reported to return to values within the normal range of the entire stallion. In an attempt to characterize this species-specific reversal, the gonadotrophin concentrations of 6 male ponies castrated on 25 March were monitored for 4 months, and 4 stallions were used to generate control data. Blood samples were collected daily, from 3 d before to 10 d after castration (Day 0), and weekly thereafter until Day 122. The pituitary response to castration was immediate. Castration resulted in a previously unreported, dramatic (13-fold) but transient (3 d) surge in circulating concentrations of LH. Concentrations of LH and FSH increased in a logarithmically scaled (LH, R2 = 0.77; FSH, R2 = 0.93) manner over the subsequent 5 wk, during which temporal changes in concentrations of both hormones were strongly correlated (R2 = 0.97). The ratio of plasma gonadotrophin concentrations was consistent throughout (LH:FSH, 1.43 +/- 0.04). Maximal concentrations of LH (20.58 +/- 1.97 ng/mL, Day 34.8 +/- 3.2) were attained approximately 2 wk before the peak in FSH (16.99 +/- 1.97 ng/mL, Day 49.7 +/- 3.0). Plasma gonadotrophin concentrations exceeded those of entire stallions throughout the study. The equine testes inhibited LH secretion during the early breeding season, and no chronic decrease in plasma gonadotrophin concentrations was recorded. However, the LH surge evident for 3 d immediately afer castration, may be related to the dynamic seasonal interaction between gonadal steroids and the regulation of pituitary gonadotrophin release.

Superovulation in red deer (Cervus elaphus) and Père David's deer (Elaphurus davidianus), and fertilization rates following artificial insemination with Père David's deer semen.

Two comparative studies were undertaken using adult, female red and Père David's deer to examine the ovulatory response of these animals to a superovulation regimen and fertilization rates following inter- and intraspecific laparoscopic insemination. In Expt 1 six Père David's deer and 12 red deer hinds were treated during the breeding season with an intravaginal progesterone-impregnated controlled internal drug release device (CIDR) for 14 days, with 200 iu pregnant mares' serum gonadotrophin (PMSG) administered 72 h before the device was withdrawn and eight injections of ovine FSH given at 12 h intervals starting at the time of PMSG administration. Oestrous behaviour began one day after CIDR device withdrawal (Père David's deer: 24.00 +/- 2.32 h; red deer: 24.60 +/- 2.23 h). The duration of oestrus was greater in Père David's deer than in red deer (17.50 +/- 1.43 h and 8.25 +/- 3.25 h, respectively, P < 0.001). The peak LH surge of Père David's deer was 68.65 +/- 4.74 ng ml-1 occurring 29.00 +/- 2.41 h after removal of the CIDR devices. In comparison, the peak LH surge in red deer was 17.09 +/- 3.64 ng ml-1 (P < 0.001), occurring 24.00 +/- 0.00 h after CIDR device withdrawal. Pre-surge concentrations of LH were also greater (P < 0.001) in Père David's deer (1.37 +/- 0.11 ng ml-1) than in red deer hinds (0.41 +/- 0.02 ng ml-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Successful embryo transfer following artificial insemination of superovulated fallow deer (Dama dama).

Thirty-four European fallow deer (Dama dama dama) were randomly allocated into embryo donor (n = 12) or embryo recipient (n = 22) groups. All does were treated with controlled internal drug release (CIDR) devices for 14 days. Animals in the embryo donor group were further treated with 200 I.U. pregnant mare serum gonadotrophin (PMSG) and 0.5 units ovine follicle-stimulating hormone (FSH). PMSG was administered 72 h before withdrawal of CIDR devices and FSH was given in eight 0.063 unit injections at 12-hourly intervals starting at the time of PMSG administration. All embryo donor animals were inseminated, by laparoscopy in both uterine horns, 36 h after withdrawal of CIDR devices with 25 x 10(6) fresh spermatozoa collected from Mesopotamian fallow deer (Dama dama mesopotamica). Embryos were recovered by laparotomy on Day 3 (n = 6) or Day 6 (n = 6) after withdrawal of CIDR devices and the ovarian response was determined. In total, 22 embryos were transferred into the oviduct (2-4-cell stage, n = 14) or uterine horn (morula stage, n = 8) on Day 3 or Day 6 after withdrawal of CIDR devices respectively. The overall means (+/- s.e.m.) of total follicular response and corpora lutea were 24.2 +/- 3.5 and 14.1 +/- 3.6 respectively. The mean number of large unruptured follicles was higher on Day 6 than on Day 3 (13.5 +/- 2.9 v. 6.7 +/- 1.3, P < 0.05). The overall embryo recovery rate was 45.8%. There was no difference in pregnancy rate following transfer of embryos on either Day 3 or Day 6 (7/14 v. 5/8 respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of age and time of day on the timing of the surge in luteinizing hormone, behavioural oestrus and mating in red deer hinds (Cervus elaphus).

The oestrous cycles of fourteen red deer hinds (six yearling; eight more than 2 years old) were synchronized during the early breeding season by removal of a progesterone-containing intravaginal device and blood samples were taken at intervals of 3 h commencing 13 or 25 h later and continued for 54 h. The controlled internal drug release devices (CIDRs) were removed at 08:00 h (group 1; three yearlings and four adults) or 12 h later at 20:00 h (group 2; three yearlings and four adults). There was no significant effect of time of removal of CIDR on the interval to the onset of oestrus (group 1, 34.5 +/- 4.05 h; group 2, 42.14 +/- 7.8 h) on the time of peak concentration (group 1, 41.81 +/- 5.69 h; group 2, 41.71 +/- 7.81 h) or on duration of the luteinizing hormone (LH) surge (group 1, 15.00 +/- 0.95 h; group 2, 14.57 +/- 0.78 h). The six yearling animals exhibited oestrus and LH surge significantly later than the adults (55 +/- 4.2 versus 32 +/- 6.3 h for the LH surge for yearling and adult females, respectively). In a further experiment, 20 hinds were synchronized during the breeding season by removal of CIDR at two times of day 12 h apart and placed with a stag. Mating took place at a mean time of 42.1 +/- 2.4 h and 37.0 +/- 1.3 h later in the two groups. There was no significant effect of time of removal of CIDR upon time to onset of oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of melatonin on pulsatile release of luteinizing hormone in female lambs.

This study investigated the effect of melatonin treatment of ewe lambs on LH pulsatility in an attempt to examine the mechanism whereby melatonin advances the onset of puberty. Six ewe lambs were given intravaginal melatonin implants at 12.8 weeks of age. Another six lambs received empty implants. All lambs were serially blood sampled every 15 minutes for six hours on several occasions prior to the onset of puberty. One week after implantation LH pulse frequency and mean LH levels were higher in treated lambs than the control lambs (pulse frequency 0.13/h vs 0.03/h; mean LH levels 2.0 +/- 0.2 ng/ml vs 1.3 +/- 0.1 ng/ml; p less than 0.05). Melatonin treatment failed to alter pulse frequency after the initial increase. Puberty was advanced by 3 weeks in the treated group. In the second experiment six lambs received melatonin implants at 13 weeks of age and another six lambs served as control. In this experiment blood samples were taken intensively during the first few weeks after treatment. Results of this study show that mean plasma LH levels and LH pulse frequency were again higher during the first week after implantation. This transient increase in LH release may be part of the mechanism initiating the eventual advancement of puberty although the significance of this increase is questionable. In both experiments the LH response to estradiol injection was monitored at various times after treatment, but no effects of melatonin were found, although the magnitude of the response increased with age.

Effect of oral melatonin treatment on the seasonal physiology of pony stallions.

This study tested the role of melatonin in the regulation of seasonal physiological change in the pony stallion. Four 3-year-old, Welsh Mountain pony stallions were housed initially under the prevailing short-day photoperiod in December (8 of light [L]:16 h of darkness [D]) before being transferred to long days (16L:8D) on 13 January for the remaining 22 weeks of the study. On Day 76 (11 weeks later) the stallions began an 11-week period of daily melatonin treatment (20 mg orally, 8 h after lights on). Marked changes in mean plasma testosterone, beta-endorphin and cortisol concentrations occurred in response to long days and to subsequent melatonin treatment. Photostimulation produced a sharp rise in overall mean daily testosterone to a peak of 6.74 nmol/litre by Day 30. Values then fell to a nadir (3.17 nmol/litre) by Day 85, suggesting a role for melatonin in the termination of breeding activity in the horse. Cortisol and beta-endorphin values remained low throughout the first 11 weeks, but by Day 105 (Day 30 of melatonin treatment) concentrations had risen sharply, attaining a peak on Day 125 (510 pg beta-endorphin/ml, 50 ng cortisol/ml). Concentrations of both hormones had fallen by Day 77 of melatonin treatment (Day 152), perhaps as a result of refractoriness. Parallelism between beta-endorphin and cortisol suggests a pituitary origin for peripheral beta-endorphin. Diurnal variation in cortisol was observed under long days but no change in beta-endorphin was detected. Long days and melatonin treatment stimulated shedding of the winter and summer coats respectively, whereas growth rate was increased (2.03 kg/week) during the period of melatonin treatment relative to that of long days only (0.37 kg/week). The study provides evidence that the diurnal pattern of melatonin secretion mediates the reproductive and non-sexual responses to photoperiodic change in pony stallions.

Fertility of ewes given either melatonin or progestogen sponges.

The administration of melatonin via intravaginal sponges is an effective method of advancing the breeding season in ewes. In the present study the fertility of melatonin-treated ewes has been compared with that of ewes induced to ovulate by conventional treatment. On June 25, 15, 15-month-old ewes (group 1) were given intravaginal implants containing melatonin in silastic tubing. On August 8, 13 similar ewes (group 2) were given Veramix sponges which were removed 12 days later, when they were given 500 iu pregnant mare's serum gonadotrophin (PMSG). Two intact raddled rams were introduced to the combined groups on August 21. The mean date of mating was September 3 +/- 1.5 for group 1 and August 21 +/- 0.2 for group 2 ewes. All the ewes in group 1 and 10 in group 2 (77 per cent) were mated. All the ewes were slaughtered approximately 50 days after mating and their reproductive tracts removed. The mean ovulation rates were 2.1 and 2.3 in groups 1 and 2, respectively. The results indicate that conception rates of 87 per cent and 61.5 per cent of ewes put to the ram were obtained in the melatonin-treated and PMSG-treated groups, respectively. At slaughter the melatonin-treated group were found to have a mean of 1.47 live fetuses per ewe put to the ram and the PMSG-treated group a mean of 1.08. It can therefore be concluded that melatonin implantation is an effective method for the advancement of seasonal breeding in anoestrous sheep, and that the fertility achieved is at least as good as that given by conventional progestogen-PMSG treatment.