Onset of puberty and duration of seasonal cyclicity in Lipizzan fillies

REASONS FOR PERFORMING STUDY: The onset of puberty and duration of seasonal cyclicity in Lipizzan fillies and young mares have been determined mostly by empirical data, but very little is known about the hormonal changes at puberty and during the seasonal cyclicity in this breed. OBJECTIVES: To determine the onset of puberty and the course of the cyclicity in young Lipizzan mares from age 1-4 years. METHODS: Blood samples were collected twice a month for 2 years from 46 Lipizzan fillies age 1-4 years. Progesterone concentrations, indicating ovarian activity, were measured using an EIA kit. Values above 6.4 nmol/l were taken to indicate luteal phase and those below 3.4 nmol/l, oestrus. RESULTS: Fifty percent of Lipizzan fillies entered puberty during the first spring after their birth at mean +/- s.e. age 15.25 +/- 0.31 months and 50% during the second spring, at age 25.2 +/- 0.57 months. For several fillies of all ages, manifesting anoestrus during the winter months, the oestrus started in March and reached a peak in June to September. The duration of the oestrus activity was age-dependent, at 2.37 +/- 0.62 months in yearlings and 8.56 +/- 0.46 months in fillies age 4 years. In December, >50% of fillies age 2-4 years still exhibited oestrus activity and some entered anoestrus in January the following year. A small proportion of fillies age 3-4 years remained cyclic during the whole year, indicating that factors other than photoperiod can be involved in regulating seasonal cyclicity. CONCLUSIONS AND POTENTIAL RELEVANCE: Our results demonstrate the influence of seasonal factors on the onset of puberty and on the course of oestrus activity in the majority of Lipizzan fillies. However, certain exceptions were also demonstrated but reasons for the prolonged periods of oestrus activity in some Lipizzan fillies were not identified, and the role of the hormone leptin should be studied in future investigations.     

Field Trial on Progesterone Cycles, Metabolic Profiles, Body Condition Score and their Relation to Fertility in Estonian Holstein Dairy Cows

Resumption of luteal activity postpartum and fertility were investigated in an Estonian Holstein high milk production and good fertility dairy herd. Body condition was scored after every 10 days in 54 multiparous dairy cows (71 lactations) calving inside from December to March during 4-year period. Blood samples were taken 1–14 days before calving and 1–14, 28–42 and 63–77 days after calving: analytes estimated were serum aspartate aminotransferase (AST), glucose, ketone bodies, total cholesterol, non-esterified fatty acids and triglycerides. The general linear mixed model was used to compare the data for cows with different characteristics in luteal activity postpartum based on their milk progesterone profiles. Forty-five per cent of cases had abnormal profiles; delayed resumption of ovarian cyclicity postpartum (DC) was the most prevalent abnormality. There was no difference in body condition scores between the groups. The DC and prolonged luteal phase groups had higher serum AST activity (p < 0.01) 1–14 days postpartum compared with normal group. The DC group also had higher cholesterol and triglyceride values (p < 0.05) 28–42 days postpartum and higher milk fat/protein ratio (p < 0.01) on the first month of lactation compared with normal profile group. Despite long post-calving anoestrous period (71 ± 5.0 days; mean ± SEM) DC group had 64.7% first service pregnancy rate (normal group 48.6% and PLP group 37.5%). This study did not find any detrimental effect of prolonged anovulatory period postpartum on subsequent fertility.     

Effects of Short-Term High Carbohydrate or Fat Intakes on Leptin, Growth Hormone and Luteinizing Hormone Secretions in Prepubertal Fat-Tailed Tuj Lambs

The aim of the current study was to evaluate the effects of high carbohydrate or fat diets, fed for 15 days at the end of breeding season, on leptin, GH and LH secretions in prepubertal fat‐tailed Tuj lambs. For that purpose, 9‐month‐old ram‐lambs were divided into three groups as control group (fed with basal ration, n = 4), high carbohydrate (HC) group, basal ration plus barley, n = 4), or high fat (HF) group (basal ration plus by‐pass fat, n = 4). For the measurement of leptin and GH, blood plasma samples were collected on days 1, 4, 9 and 14 of the experiment. For the measurement of LH pulse frequency, serial blood samples were collected every 15 min for 6 h on day 14. Lambs were weighed and body condition scored (BCS) on days 1 and 15. Body weight and BCS increased towards the end of the study (p < 0.05). The BCS was higher in high energy groups at the end of the experiment (p < 0.05). Diet affected plasma leptin concentrations (p = 0.002) but time did not. The GH concentrations were not affected by diet or time. The LH pulse frequency appeared to be higher in HC and HF groups but there were no statistical difference between the groups. There was a significant positive relationship between overall BCS and corresponding leptin concentrations (R² = 0.263; p = 0.010) and between LH pulse frequency and leptin concentrations (R² = 0.594; p = 0.003). In conclusion, the present study suggests that rather than type of energy, amount of energy intake and body energy reserves are much important regulators of plasma leptin concentrations and LH pulse frequency in fat‐tailed Tuj lambs.     

Seasonal reproduction in the mare: possible role of plasma leptin, body weight and immune status

The primary objective of this study was to evaluate the possible role of leptin, body weight and immune status on reproductive activity throughout the transition period from cyclicity to seasonal anestrus, during anestrus and resumption of ovarian activity in Lusitano mares. Mares in good body condition were monthly monitored throughout 2 years (10 mares in each year) for evaluation of their reproductive status by sequential ultrasonography and plasma progesterone determinations. On the second year, all mares were weighed. Progesterone and leptin were assayed by radioimmunoassay (RIA). Parameters of the immune status (phagocytosis and oxidative burst of neutrophils, characterisation of circulating lymphocyte subsets) were also evaluated. Phagocytosis and oxidative burst in blood neutrophils were measured by flow cytometry using commercially available kits. Lymphocyte subsets were assessed by indirect immunofluorescence staining after incubation with monoclonal antibodies specific for CD2, CD19, CD4, CD8 cells markers by flow cytometry. Natural killer cells and B cells were estimated mathematically. No significant difference was found in phagocytosis, oxidative burst and circulating lymphocyte subsets at anestrus and at either phase of the estrous cycle (p>0.05), suggesting that the immune status of the mare was not influenced by the seasonal changes in ovarian activity. This study also suggests that body weight has a direct relationship with plasma leptin levels. Increased concentrations of this hormone in circulation might be associated with the restart or maintenance of ovarian cyclicity in Lusitano mares.     

Expression of the μ Opioid Receptor and Effects of the Opioid Antagonist Naloxone on In Vitro Maturation of Oocytes Recovered from Anoestrous Bitches

The μ-opioid receptor (MOR) is expressed in bovine, human, equine and canine oocytes, and in seasonal breeders, it is expressed with higher intensity during the anoestrous phase. Supplementation of in vitro maturation (IVM) medium with opioid agents, agonists or antagonists, was shown to affect oocyte maturation in several species such as rat, bovine and equine. This study reports the effects of supplementing IVM medium with naloxone (Nx), an opioid antagonist, on nuclear and cytoplasmic maturation rate of oocytes recovered from anoestrous bitches. Cytoplasmic maturation was examined in terms of mitochondrial (mt) distribution. In order to confirm the receptor-mediated action of Nx, in oocytes of anoestrous bitches, MOR expression was analyzed by Western blot. Cumulus–oocyte complexes, recovered from the ovaries of bitches in anoestrous, were cultured in vitro and Nx was added at the concentrations of 1 × 10⁻⁶, 1 × 10⁻⁸ and 1 × 10⁻¹⁰  m . The rate of oocytes resuming meiosis after culture in presence of 1 × 10⁻⁶  m Nx (29%) was significantly higher than that of oocytes of control group (12%; p < 0.05). However, treatment with Nx did not affect mt distribution pattern. In denuded oocytes and in corresponding cumulus cells, a doublet of 65 and 50 kDa was observed. We conclude that, in oocytes of anoestrous bitches, MOR is expressed and Nx significantly improves nuclear maturation rate. Further studies should be performed to elucidate the expression of other opioid receptors, such as δ and κ, and possible interactive effects of their antagonists on canine oocyte maturation.     

Seasonal and pulsatile dynamics of thyrotropin and leptin in mares maintained under a constant energy balance

The objective of this study was to determine if seasonal and/or pulsatile variations occur in plasma concentrations of thyrotropin (TSH) and leptin in mares while maintaining a constant energy balance. Blood samples were collected every 20 min during a 24 h period in winter and again in summer from six Quarter Horse type mares. Plasma concentrations of TSH, leptin, and T₄ were determined by radioimmunoassay. No differences were observed in body weight between winter (388.1 ± 12.5 kg) and summer (406.2 ± 12.5 kg; P = 0.11). Plasma concentrations of TSH were greater in the summer (2.80 ± 0.07 ng/ml) when compared to winter (0.97 ± 0.07 ng/ml; P < 0.001). Pulse frequency of TSH was not different between winter (6.17 ± 0.78 pulses/24 h) and summer (5.33 ± 0.78 pulses/24 h; P = 0.49). Mean TSH pulse amplitude, pulse area, and area under the curve were all greater in summer compared to winter (3.11 ± 0.10 ng/ml versus 1.20 ± 0.10 ng/ml, 24.86 ± 0.10 ng/ml min versus 13.46 ± 1.90 ng/ml min, 3936 ± 72.93 ng/ml versus 1284 ± 72.93 ng/ml, respectively; P < 0.01). Mean concentrations of leptin were greater in summer (2.48 ± 0.17 ng/ml) compared to winter (0.65 ± 0.17 ng/ml; P < 0.001). Pulsatile secretion patterns of leptin were not observed in any horses during experimentation. Mean concentrations of T₄ were greater in winter (20.3 ± 0.4 ng/ml) compared to summer (18.2 ± 0.4 ng/ml; P < 0.001). These seasonal differences between winter and summer provide evidence of possible seasonal regulation of TSH and leptin.     

Effect of Prepartum Energetic Supplementation on Productive and Reproductive Characteristics, and Metabolic and Hormonal Profiles in Dairy Cows under Grazing Conditions

The effect of cracked corn grain supplementation (3.5 kg/day) during 3 weeks before the expected calving date on milk production and composition, body condition score (BCS), metabolic and hormonal profiles and length of postpartum anoestrus was evaluated in multiparous Holstein dairy cows under grazing conditions (Energy supplemented group, n = 10; Control group, n = 10). Body condition score was weekly recorded during the peripartum period, from days −21 to +35 (parturition = day 0). Non-esterified fatty acids, β-hydroxybutyrate, cholesterol, urea, insulin, insulin-like growth factor I (IGF-I), leptin, thyroxine (T₄) and 3,3'5-triiodothyroinine (T₃) were weekly determined in plasma from days −21 to +35. The reinitiation of ovarian cyclicity was twice weekly determined by ovarian ultrasonography and confirmed by plasma progesterone concentrations. Cows fed energy concentrate prepartum had higher BCS during the prepartum and postpartum and produced more milk. Non-esterified fatty acids plasma concentrations were significantly higher in the energy group, while cholesterol was higher in the control group. Treated cows had higher levels of plasma insulin, IGF-I and leptin pre-calving. IGF-I, leptin and T₄ were diminished during the early postpartum period in both groups. Insulin levels were also diminished in the control group, but levels remained high in the energy-supplemented group. Treated cows ovulated sooner after parturition than controls. We conclude that Energetic supplementation prepartum in cows under grazing conditions increased milk production and reduced the reinitiation of ovarian activity, consistent with a better EB (BCS), higher prepartum levels of IGF-I, leptin and insulin, and higher insulin levels during early postpartum.     

Development of One vs Multiple Ovulatory Follicles and Associated Systemic Hormone Concentrations in Mares

Ablation of follicles ≥  6 mm in diameter and treatment with PGF2α 10 days after ovulation were used to induce the development of ovulatory waves. Comparisons were made between induced waves with one (33 waves, 72%) and multiple (13 waves, 28%) ovulatory follicles. Diameter deviation was defined as the separation of follicles into dominant and subordinate categories. Multiple ovulatory follicles were preceded by more (p < 0.001) follicles ≥ 20 mm at the beginning of deviation, higher LH preceding deviation (approached significance, p < 0.08), lower (p < 0.05) concentrations of FSH on the day of deviation and thereafter, and higher (p < 0.0003) oestradiol by 2 days after deviation. During the peri-ovulatory period, systemic hormone concentrations for waves with multiple ovulations involved higher oestradiol before ovulation (approached significance, p < 0.07), lower FSH (p < 0.04) before and after ovulation, and both higher progesterone (p < 0.05) and lower LH (p < 0.05) beginning the day after ovulation. Results indicated that by the beginning of deviation there were more follicles ≥  20 mm and subsequently greater oestradiol production in waves that led to the development of multiple ovulatory follicles, and during the peri-ovulatory period differences between one and multiple ovulations were consistent with the negative effects of the ovarian hormones on the gonadotropins.     

Effects of Exogenous Tumour Necrosis Factor-α on the Secretory Function of the Bovine Reproductive Tract Depend on Tumour Necrosis Factor-α Concentrations

The aim of study was to correlate tumour necrosis factor-α (TNF) infused doses used with the TNF concentrations achieved and with the secretory function of both the ovary and the uterus in cows. We evaluated the concentrations of progesterone (P4), prostaglandin (PG)F_2α, PGE₂ nitric oxide (NO) and TNF in the jugular vein and vena cava caudalis as parameters of exogenous TNF action on the female reproductive tract. Aortae abdominalis of cows (n = 18) were infused with saline or two doses of TNF (luteolytic – 1 μg or luteotrophic – 10 μg). In the peripheral blood, 1 μg TNF concentrations achieved within the range of 30–45 pg/ml, and 10 μg TNF provoked a sharp increase in achieved concentrations at a range of 250–450 pg/mL). The TNF concentrations achieved in vena cava caudalis were five to six times higher than that in peripheral blood (p < 0.001). One microgram TNF increased PGF_2α and NO (p < 0.001) and decreased P4 (p < 0.05). The higher TNF dose stimulated P4 and PGE₂ (p < 0.01). TNF infusion at luteolytic dose achieved its concentrations at the physiological range previously observed in cows. Luteotrophic TNF dose achieved the concentrations in vena cava caudalis that are much higher than physiological level and were previously noted in pathological circumstances (i.e. mastitis , metritis ).     

Seasonal Changes in Testicular Cell Morphology in Domestic Male Cats (Felis catus)

The aim of this study was to asses the variation in the morphology of the seminal epithelium in relation to natural photoperiod in male cats. Tom cats (n = 240) were castrated every other week throughout the year. Each testis was fixed in Bouin's solution and cut into sections. The percentage of tubules with round spermatids (RS), elongated spermatids (ES), tailed spermatids (TS), mature spermatids (MS) and the number of Sertoli cells (SC) and Leydig cells (LC) were recorded in each sample. Testicles from males during short days (SHD) had a higher percentage of tubules with RS and ES compared to testicles from males during long days (LHD, 31.3 ± 0.6 vs 2.1 ± 0.6%, p < 0.001; 30.9 ± 0.7 vs 11.0 ± 0.7%, p < 0.001). Conversely, testicles from males during SHD had a lower percentage of tubules with TS and MS compared to testicles from males during LHD (24.5 ± 0.8 vs 29.7 ± 0.8%, p < 0.01; 13.1 ± 1.2 vs 57.0 ± 1.2%, p < 0.01). Furthermore, testicles from males during SHD had a higher number of SC and lower number of LC compared to testicles from males during LHD (11.4 ± 0.1 vs 8.0 ± 0.1%, p < 0.01; 19.2 ± 1.0 vs 38.0 ± 1.0%, p < 0.01). In conclusion, there are seasonal changes in testis cell morphology in the tom which may be related to seasonal sperm production.     

Effect of Progesterone Prior to GnRH-PGF2α Treatment on Induction of Oestrus and Pregnancy in Anoestrous Awassi Ewes

An experiment was conducted to examine the effect of progesterone prior to a GnRH‐PGF_2α treatment on oestrus and pregnancy in seasonally anoestrous Awassi ewes. Twenty‐four ewes were randomly assigned to three groups to be pre‐treated with 60 mg medroxyprogesterone acetate sponges (group A), 600 mg progesterone sponges (group B) or blank sponges (group C) for 4 days. All ewes were injected with 100 μg of GnRH 24 h after sponge removal followed, 5 days later, by 20 mg PGF_2α injection. Ewes were exposed to three fertile rams at the time of PGF_2α injection (day 0, 0 h) and were checked for breeding marks at 6‐h intervals for 5 days. Blood samples were collected from all ewes 1 day (day ?10) prior to sponge insertion, at the time of sponge removal (day ?6), 1 day following sponge removal (day ?5, at the time of GnRH injection) and at the time of PGF_2α injection (day 0) for analysis of progesterone. Progesterone concentrations on days ?10 and ?5 were basal and averaged 0.2 ± 0.04 and 0.2 ± 0.2 ng/ml, respectively. Progesterone concentrations on day ?6 were elevated only in group B ewes and were higher (p < 0.0001) than those of groups A and C. Progesterone concentrations on day 0 were higher (p = 0.002) in groups A and B than group C. Oestrous responses occurred only in ewes of groups A and B (p > 0.05). Induced oestrus conception rate was greater (p < 0.01) in group A than groups B and C. Ewes returned to oestrus 17–20 days following day 0 were two of eight, six of eight and three of eight of groups A, B and C, respectively, all of which eventually lambed. The overall lambing rate was 82% in progesterone‐primed ewes compared with only 38% non‐progesterone‐primed ewes (p < 0.05). Progesterone priming apparently sensitizes GnRH‐PGF_2α‐treated seasonally anoestrous ewes and increases their response in oestrus and pregnancy rates.     

Induction of Ovulation of Ovulatory Size Non‐Ovulatory Follicles and Initiation of Ovarian Cyclicity in Summer Anoestrous Buffalo Heifers (Bubalus bubalis) Using Melatonin Implants

This study was conducted on summer anoestrous buffalo heifers to monitor the efficacy of melatonin for induction of ovulation and ovarian cyclicity. During pre‐treatment period of 24 days, the ovarian dynamics of five cycling and 10 summer anoestrous heifers was monitored on each alternate day using a transrectal ultrasound scanner. Thereafter, during treatment period, these 10 anoestrous heifers along with additional seven anoestrous heifers were randomly allocated into non‐implanted (n = 5) and implanted (n = 12, one melatonin implant/50 kg, 18 mg melatonin/implant) group. Non‐implanted heifers were monitored on each alternate day till the confirmation of second‐ovulation in implanted heifers. Pre‐treatment period revealed the presence of dominant follicles in anoestrous heifers which attained the diameter comparable with ovulatory follicles of cycling heifers but failed to ovulate and regressed. Between 6 and 36 days (15.3 ± 2.9 days) post‐treatment, all the implanted heifers (p < 0.05) exhibited ovulation of dominant follicles; however none of the non‐implanted heifers ovulated during the corresponding period. The first‐interovulatory period in implanted heifers ranged between 8 and 28 days (18.0 ± 1.8 days). The implanted heifers with short (≤16 days) interovulatory period had short‐lived corpus luteum (CL) that had smaller diameter and secreted less progesterone (p < 0.05). The diameter of CL was large (p < 0.05) and plasma progesterone was high (p < 0.05) following second‐ovulation compared with first‐ovulation in implanted heifers. In conclusion, using melatonin implants, ovulatory size nonovulatory follicles observed in summer anoestrous buffalo heifers can be successfully ovulated to initiate ovarian cyclicity.     

Effects of dietary chromium picolinate supplementation on performance and plasma concentrations of insulin and corticosterone in laying hens under low ambient temperature

This experiment was conducted to evaluate the effects of chromium (chromium picolinate) on performance and plasma concentrations of insulin and corticosterone of laying hens (Ross Brown) under a low ambient temperature (6.9 °C). One hundred and twenty laying hens (46 weeks old) were divided into four groups, 30 hens per group. The laying hens were fed either a control diet containing 710.3 p.p.b. chromium or the control diet supplemented with 100, 200 or 400 μg chromium/kg diet. Increasing supplemental chromium increased live weight change (p ≤ 0.001, linear) and egg production (p ≤ 0.001, linear) and also improved feed efficiency linearly (p ≤ 0.001). Live weight change and egg production also had quadratic responses (p ≤ 0.001) to increasing chromium supplementation. Plasma insulin concentration increased linearly (p ≤ 0.001), whereas corticosterone concentration decreased linearly (p ≤ 0.001) as dietary chromium supplementation increased. The results of this study indicate that supplemental dietary chromium, 200 p.p.b. in particular, had a positive effect on performance and increased the plasma insulin concentration of laying hens under cold stress.     

Reproductive Performance of Gilts with Similar Age but with Different Growth Rate at the Onset of Puberty Stimulation

The aim of this study was to evaluate the reproductive performance of gilts that had a similar age but different weights at the onset of puberty stimulation by boar exposure at 144 days. Gilts were divided into two groups according to their lifetime growth rate from birth to approximately 144 days of age. Mean growth rates at this moment were 577 and 724 g/day for group 1 (G1; n = 58) and group 2 (G2; n = 58), respectively. After selection, gilts were weighed at approximately 155, 165 and 175 days of age, on the insemination day and at slaughter. Gilts were inseminated, on average, at 193 days of age and were slaughtered 32 days after insemination, when the number of corpora lutea and embryos were recorded. Higher growth rate gilts (G2) reached puberty earlier (155.3 vs 164.1 days; p < 0.01). More gilts of G2 group attained puberty by 190 days of age (p = 0.004) than G1 gilts (95%; 55/58 vs 76%; 44/58). The anoestrous rate, until 60 days after the onset of boar exposure was higher (p < 0.01) in G1 (19.0%; 11/58) than in G2 (3.4%; 2/58) group. However, there were no differences in the pregnancy rate (90.7 vs 94.5), ovulation rate (15.9 vs 16.5), total embryos (12.9 vs 11.7), viable embryos (12.0 vs 11.1) and embryo survival (73.7% vs 68.5%), between G1 gilts and G2 gilts, respectively (p > 0.05). High growth rate gilts attain puberty earlier and have a lower anoestrous rate than low growth rate gilts.     

Effect of hCG in the Presence of hCG Antibodies on the Follicle, Hormone Concentrations, and Oocyte in Mares

Follicle blood flow, follicular-fluid and plasma hormone concentrations, and oocyte quality were studied 30 h after an ovulation-inducing hCG treatment when the pre-ovulatory follicle was 32 mm. Mares were grouped as positive (n = 16) and negative (n = 44) for hCG antibodies before the experimental hCG treatment. Percentage of the follicle wall with blood flow signals was less (p < 0.05) in the antibody positive group than in the negative group. The concentrations of follicular-fluid oestradiol and free IGF1, and plasma oestradiol were greater (p < 0.001), and follicular-fluid progesterone (p < 0.001) and plasma LH (p < 0.02) were less in the antibody-positive group than in the negative group. For recovered oocytes at 30 h (n = 37), the antibody-positive group had fewer (p < 0.001) mature (MII) oocytes than the antibody-negative group. Results were attributable to highly effective neutralization of the hCG in the antibody-positive group.     

Dynamics of follicle populations and gonadotropin concentrations in fillies age two to ten months

Follicle populations and concentrations of circulating gonadotropins were studied during age 2-10 months in 10 spring-born pony fillies. Blood sampling and ultrasound scanning were done every 4 days and daily for four 30 day periods. During 5-12 weeks, FSH concentrations were lower in 6 fillies with follicles > or = 6 mm (mean +/- s.e. 1.4 +/- 0.1 ng/ml) than in 4 fillies with follicles < 6 mm (2.8 +/- 0.3 ng/ml). The diameters and numbers of follicles and gonadotropin concentrations increased progressively during age 2-4 months. A plateau in follicle activity and reduced levels of gonadotropins occurred during 5-7 months. During 8-10 months, follicles grew to > 10 mm and gonadotropin concentrations increased. Waves of follicular growth were identified during the 30 day periods by significant increases in the diameter of the 10 largest follicles. The waves did not partition into dominant and subordinate follicles. Results indicated an initial postnatal period of negative ovarian feedback, temporally related changes in gonadotropins and follicles for months 3-10, and development of follicles in waves.     

Induction of ovulation with GnRH and PGF2alpha in lactating Bos taurus x Bos indicus cows

Induction of ovulation for timed artificial insemination (TAI) with the Ovsynch protocol was evaluated in 49 anoestrous and lactating Bos taurus x Bos indicus cows. Palpation per rectum and transrectal ultrasonography were used on Days -30, -20, -10 and 0 (start of treatment) to confirm anoestrus but with the presence of follicles > or = 10 mm, and every other day during treatment to determine ovarian activity. Cows were randomly assigned to: (1) Ovsynch (n = 24; Day 0, 200 microg GnRH; Day 7, 150 microg PGF2alpha; Day 9, 200 microg GnRH + TAI 16 to 20 h later) and (2) control (n = 25; no treatment). Rates of ovulation for the first GnRH injection, detection of a corpus luteum (CL) at PGF2alpha injection, pregnancy and induction of cyclicity were greater (P < 0.05) with Ovsynch. There was no effect of body condition score (P > 0.05). In conclusion, the Ovsynch protocol was not effective in obtaining acceptable pregnancy rate for TAI, but it was effective for induction of cyclicity in anoestrous and lactating Bos taurus x Bos indicus cows under tropical conditions.     

Effect of hCG Treatment on the Oestrous and Ovulation Responses to FSH in Prepubertal Gilts

To ensure sufficient numbers of pregnant females, particularly at hotter times of the year, hormonal induction of gilt oestrus may be necessary. However, the gilt oestrus and ovulation responses to gonadotrophin treatment have often proven unpredictable. The objective of this study was to examine possible reasons for this unpredictability. Prepubertal gilts (approximately 150 days of age, n = 63) were assigned to one of three treatments: injection of 300 IU hCG (n = 15); pre-treatment with 100 mg FSH in polyvinylpyrrolidinone administered as 2 × 50 mg injections 24 h apart, followed by 600 IU eCG at 24 h after the second FSH injection (n = 23); or FSH pre-treatment as above followed by 300 IU hCG at 24 h after the second FSH injection (n = 25). To facilitate oestrus detection, gilts were exposed to a mature boar for 15 min daily for 7 days. Blood samples were obtained on the day of eCG or hCG injection and again 10 days later and gilt ovulation responses determined based on elevated progesterone concentrations. The oestrus responses by 7 days were 6.7%, 17.5% and 64.0% for gilts treated with hCG, FSH + eCG and FSH + hCG, respectively (p < 0.001). The oestrous gilt receiving hCG alone and one oestrous FSH + hCG gilt did not ovulate, all other oestrous gilts ovulated. A further two anoestrous FSH + eCG-treated gilts ovulated. These data suggest that FSH pre-treatment facilitated the development of ovarian follicles to the point where they became responsive to hCG, but had little effect on the response to eCG.     

Role of Luteinizing Hormone in Primiparous Sow Responses to Split Weaning

In 45 primiparous sows, we examined endocrine, ovarian and reproductive responses to split-weaning or five injections per day of 800 ng GnRH from 18 to 21 days of lactation. There was no effect of treatment on absolute or changes in sow weight or backfat depth during lactation. Average piglet growth rates were similar among treatments except that piglets suckling split-weaned sows grew faster (p < 0.05) during days 18–21. On day 18, mean plasma LH concentrations and LH pulse frequency remained relatively stable in conventionally weaned sows but increased (p < 0.01) in response to split-weaning and GnRH. Prior to weaning on day 21, mean plasma LH concentrations remained elevated in GnRH-treated sows but had returned to control levels in split weaned sows. There was no treatment effect on preweaning LH pulse frequency noted on day 21. Weaning was associated with an increase in plasma LH concentrations in all the treatment groups. Mean plasma IGF-I remained relatively constant in conventionally weaned and GnRH sows, decreased in response to split weaning on day 18 (p < 0.02), but were elevated (p < 0.03) in split wean sows on day 21. On the day after weaning, split wean sows had more (p < 0.04) ovarian follicles ≥3 mm than conventionally weaned sows, with GnRH sows being intermediate. The wean-to-oestrus interval was reduced in split-wean sows compared with those conventionally weaned (p < 0.01), with GnRH sows being intermediate. There was no effect of treatment on ovulation rates, numbers of embryos, or embryonic survival rates. These data indicate that split-weaning of litters results in a more rapid return to oestrus after weaning and that this effect is associated with a transient acute increase in circulating gonadotrophins and earlier resumption of ovarian follicular development.     

Seasonal Profiles of Ovarian Activity in Iberian lynx (Lynx pardinus) Based on Urinary Hormone Metabolite Analyses

The Iberian Lynx Ex-Situ Conservation Programme is an essential part of a co-ordinated action plan to conserve the most endangered felid species of the world. Successful captive breeding demands reliable methods for reproduction monitoring including reliable non-invasive pregnancy diagnosis. During a 3-year study, urine samples from six captive Iberian lynx females were obtained (one non-pregnant, one pseudo-pregnant and 11 pregnant cycles). Progesterone, pregnanediol and oestradiol were determined in urinary extracts and relevant urinary oestrogen metabolites were characterized by high-performance liquid chromatography (HPLC). Urinary progestins did not follow the typical pregnancy-related course of felids. In the lynx, we failed to demonstrate an urinary progestin elevation during pregnancy. In contrast, urinary oestrogens increased from 3.8 ± 0.6 to 8.6 ± 0.5 ng/mg creatinine (p < 0.001) during the pregnancy. A comparison of pseudo-pregnant with pregnant cycles revealed a further increase of oestrogens caused by implantation (p < 0.05). In one female, which refused to mate, no difference was estimated between oestrogens levels during the breeding and non-breeding seasons. Almost 10-fold higher oestrogen concentrations were measured in urines of females that shared enclosures with males. HPLC analysis of oestrogens in urine samples collected from Iberian lynx during the pregnancy revealed that lynx urine is composed of two polar oestrogen metabolites in addition to oestrone and minor amounts of oestradiol. Oestrone was detectable in all urinary extracts (8–12% of metabolites), whereas oestradiol was elevated only during late pregnancy (18%). Thus, seasonal luteal activity in Iberian lynx can be monitored by urinary oestrogens. The increase of urinary oestradiol during late pregnancy might indicate an oestradiol secretion by the lynx placenta.