Deslorelin Implants in Pre‐pubertal Female Dogs: Short‐ and Long‐Term Effects on the Genital Tract

Deslorelin acetate is a GnRH agonist used for contraception in dogs. This study aimed to evaluate the treatment of pre‐pubertal female dogs with deslorelin acetate implants, to better investigate the primary stimulatory effect of the drug and the long‐term effects on the genital tract, throughout repeated treatments. Sicilian hound female dogs (24) were randomly assigned to treated group, control group 1 and control group 2. First group bitches were implanted at 4.5, 9.0 and 13.5 months and monitored clinically, ultrasonographically and endocrinologically, throughout the study period (13.5 months). Control group 1 bitches were not implanted and clinically monitored for the same period. At 18 months, the animals underwent ovariohysterectomy, thus allowing evaluation of the internal genitalia. Control group 2 bitches were ovariohysterectomized at the age of 4.5 months. The suppression of oestrus was obtained in the treated group despite the fact that the first implant caused a modest increase in plasmatic levels of 17‐beta estradiol and an evident cornification of the vaginal mucosa cells (50–80%). Estradiol and progesterone were at baseline levels for the remaining study period, in which no other oestrous manifestations were observed. The external genitalia maintained a juvenile appearance. The ovaries, ultrasonographically, showed no follicular structures and stayed the same size. At 18 months, the genital tract was still juvenile with inactive small ovaries and a thin filiform uterus. Deslorelin suppressed ovarian activity in pre‐pubertal bitches, and oestrous induction was not observed despite the presence of the primary stimulatory effect of the drug. Juvenile genitalia were an expected side effect of the treatment.     

Long‐Term Effect of Deslorelin Implant on Ovarian Pre‐Antral Follicles and Uterine Histology in Female Rats

The objective of the present study was to investigate the inhibitory effects of long‐term deslorelin implant administration on the ovarian and uterine structures of female rats. A total of 16 non‐pregnant female rats were randomly assigned to two groups, each consisting of eight animals. Animals in the implant group (DESL) received subcutaneously (s.c.) a single deslorelin implant (4.7 mg), an analogue of GnRH, while no treatment was applied to the control group (CON). A single adult male rat was introduced into the cages of both the DESL and CON females after 6 weeks of implant administration. After 1 year of implant administration, all animals were killed and follicular structures and volumes of ovaries and uterus were examined using stereological methods. Stereological observations showed that the mean ovarian total volume of the DESL group (0.28 ± 0.07 cm3) was lower than that of the CON group (1.55 ± 0.23 cm3) (p < 0.001). On the other hand, the total number of pre‐antral follicles in the ovaries of DESL (555.32 ± 151.47) females were significantly lower than the control group (1162.96 ± 189.19) (p < 0.001). In the DESL group, the mean volumes of epithelium, endometrium, myometrium and total volume of the uterus were significantly (p < 0.001) lower than in the control groups. In conclusion, these findings indicate that the long‐term deslorelin implant (i) interferes with the normal cyclicity of female rats and (ii) affects the pre‐antral follicle population. Further studies will be required to determine the effects of long‐term deslorelin treatment on the pre‐antral follicle numbers and future fertility in other species.     

Interovulatory intervals in mares receiving deslorelin implants in Ireland (2009 to 2010)

Deslorelin acetate implants, recently licensed in Ireland and the UK for ovulation induction in mares, have been associated with prolonged interovulatory intervals in USA studies, leading to the practice of removing implants postovulation. Trial data in Australia indicate a less pronounced effect on interovulatory intervals, suggesting possible geographical variation. Objectives of the current study were to assess the effect of deslorelin implants, with and without removal on oestrous cycle length in Irish- and UK-based Thoroughbred broodmares. Data were collected retrospectively from 88 oestrous cycles. A statistically significant difference (P=0.02) was found between interovulatory intervals in mares in which the deslorelin implant was not removed, compared with administration and removal of the implant or the use of human chorionic gonadotrophin. The results suggest that implant removal when possible is advisable. The delay in subsequent ovulations was less marked than that reported in some studies from the USA. This information is useful in deciding when to schedule subsequent breeding for mares which received a deslorelin implant during the previous oestrous period and provides evidence to counter-concerns that mares treated with deslorelin implants may experience a long delay in return to oestrus if the implant is not removed.     

Changes in the Aerobic Vaginal Bacterial Mucous Load after Treatment with Intravaginal Sponges in Anoestrous Ewes: Effect of Medroxiprogesterone Acetate and Antibiotic Treatment Use

Intravaginal sponges (IS) impregnated with progestagens are widely used for oestrous synchronization in ewes. As progestogens depress the immuno response, the first aim was to determine whether medroxiprogesterone acetate (MAP) content affects the vaginal bacteria number (VBN) in IS‐treated anoestrous ewes. The second aim was to compare the effectiveness of different antibiotic treatments to control the VBN increase caused by IS. In both experiments, IS were inserted during 14 days in anoestrous ewes. In the first, 11 ewes received commercial sponges (50 mg MAP), and 10 ewes received placebo sponges. For the second experiment, IS were inserted in three groups (n = 12/group), containing oxytetracycline im (20 mg/kg); injected into the sponge (0.02 mg), or control (no antibiotic). At sponge withdrawal, all ewes received 300 UI eCG. Mucous samples were collected from the vagina before sponge insertion, at sponge withdrawal, 24, 48 and 72 h later, and the VBN (colony‐forming units per ml; CFU/ml) was counted after 48‐h incubation. Medroxiprogesterone content did not affect VBN (log CFU/ml: 4.3 ± 0.2 vs 4.4 ± 0.2 with and without MAP, respectively). Bacterial number increased from 3.5 ± 0.2 at sponge insertion to 6.9 ± 0.1 at sponge withdrawal (p < 0.0001) and decreased the following day to 4.3 ± 0.2 (p < 0.0001). In the second experiment, VBN increased at sponge withdrawal (p < 0.0001) in all groups and decreased the following day (p < 0.0001). The CFU/ml at sponge withdrawal was lower in ewes treated with antibiotics (p < 0.0001), being even lower when local rather than systemic antibiotic was administered (log CFU/ml: 3.3 ± 1.8 vs 7.2 ± 1.8). The day of oestrous VBN was similar for all treatments and similar to that observed before sponge insertion. We concluded that MAP does not influence the increase in VBN, as the main effect is provoked by the sponge device itself, and local antibiotic treatment resulted in a lower bacterial growth than systemic treatments.     

Gonadotropin‐induced Puberty Does Not Impair Reproductive Performance of Gilts over Three Parities

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.     

Fertility in Adult Bitches Previously Treated with a 4.7 mg Subcutaneous Deslorelin Implant

The absence of fertility problems in male dogs after a single treatment with deslorelin acetate (Suprelorin^®) is well acknowledged. However, reports on the application of deslorelin in the bitch and information concerning fertility after implant treatment are still limited. In this retrospective study, data concerning induced and spontaneous oestruses of 39 bitches from 17 breeds, treated with deslorelin acetate implants (4.7 mg Suprelorin^®, Virbac, France), were retrieved to assess post‐treatment fertility (ovulation rate, pregnancy rate and litter size). Animals were grouped according to treatment characteristics: group 1 (Gr1) – females submitted to oestrus induction, showing natural oestruses afterwards (n = 19); group 2 (Gr2) – females re‐implanted with 4.7 mg deslorelin acetate to re‐induce oestrus, showing subsequent spontaneous post‐implant oestruses (n = 7); and group 3 (Gr3) – females submitted to a 4.7 mg deslorelin acetate implant for oestrus suppression, evaluated at subsequent spontaneous post‐implant oestruses (n = 13). Comparison of fertility traits between induced and post‐treatment spontaneous oestruses in Gr1 and Gr2 (short treatments), or between spontaneous oestruses after long‐treatment schedules (Gr 3) revealed a slightly better performance in spontaneous cycles compared with induced cycles: ovulation rate post‐treatment was 97.1%, 94.1% and 94.4% and the pregnancy rate post‐treatment was 91.2%, 88.9% and 84.6% for groups 1, 2 and 3, respectively. Nevertheless, fertility in induced and post‐treatment oestruses was considered normal. Moreover, the individual litter size did not differ within groups between induced and spontaneous cycles. From these findings, we concluded that treatment with 4.7 mg deslorelin implants did not compromise the bitches' fertility in subsequent oestruses.     

Dose–Response Studies for Pituitary and Testicular Function in Male Dogs Treated with the GnRH Superagonist, Deslorelin

We tested the effect of dose of GnRH superagonist on pituitary and testicular function in a study with four groups of four male dogs. The Controls received blank implants and the other three groups received implants containing 3, 6 or 12 mg deslorelin (d ‐Trp⁶‐Pro⁹‐des‐Gly¹⁰‐GnRH ethylamide). In all deslorelin‐treated groups, there was initially an acute increase in plasma concentrations of LH and testosterone, followed by declines such that both hormones became undetectable after approximately 12 days. There was a dose–response in some of these early aspects of the hormone profiles. With respect to long‐term effects of treatment, the 12‐mg dose had significantly greater effects than the smaller doses for the duration of minimum testicular volume [366 ± 77, mean ± SEM (3 mg), 472 ± 74 (6 mg), and 634 ± 59 (12 mg) days], absence of ejaculate [416 ± 88 (3 mg), 476 ± 83 (6 mg), and 644 ± 67 (12 mg) days], undetectable plasma concentrations of LH and testosterone [367 ± 64 (3 mg), 419 ± 72 (6 mg), and 607 ± 69 (12 mg) days], the delay until complete recovery of LH and testosterone secretion [394 ± 65 (3 mg), 484 ± 72 (6 mg) and 668 ± 47 (12 mg) days], and the delay until testes had regrown to normal volume [408 ± 77 (3 mg), 514 ± 74 (6 mg), 676 ± 59 (12 mg) days]. The time taken to restore full ejaculates was also longest for the 12‐mg dose: 716 ± 67 (12 mg) days vs 440 ± 66 (3 mg) and 538 ± 83 (6 mg) days after implantation. There was no correlation between delay to recovery of normal ejaculate quality and body mass. We conclude that the dose–response relationship with deslorelin implants is not expressed with respect to the degree of suppression of reproduction, but on the maximum duration of suppression and thus to delay until recovery.     

First Service Pregnancy Rates Following Post‐AI Use of hCG in Ovsynch and Heatsynch Programmes in Lactating Dairy Cows

Lactating dairy cows (n = 667) at random stages of the oestrous cycle were assigned to either ovsynch (O, n = 228), heatsynch (H, n = 252) or control (C, n = 187) groups. Cows in O and H groups received 100 μg of GnRH agonist, i.m. (day 0) starting at 44 ± 3 days in milk (DIM), and 500 μg of cloprostenol, i.m. (day 7). In O group, cows received 100 μg of GnRH (day 9) and were artificially inseminated without oestrus detection 16–20 h later. In H group, cows received 1 mg oestradiol benzoate (EB) i.m., 24 h after the cloprostenol injection and were artificially inseminated without oestrus detection 48–52 h after the EB injection. Cows in C group were inseminated at natural oestrus. On the day of artificial insemination (AI), cows in all groups were assigned to subgroups as follows: human Chorionic Gonadotrophin (O‐hCG) (n = 112), O‐saline (n = 116), H‐hCG (n = 123), H‐saline (n = 129), C‐hCG (n = 94) and C‐saline (n = 93) subgroups. Cows in hCG and saline subgroups received 3000 IU hCG i.m. and or 10 ml saline at day 5 post‐AI (day 15), respectively. Pregnancy status was assessed by palpation per rectum at days 40 to 45 after AI. The logistic regression model using just main effects of season (summer and winter), parity (primiparous and pluriparous), method₁ (O, H and C) and method₂ (hCG and saline) showed that all factors, except method₁, were significant. Significant effects of season (p < 0.01), hCG and parity (p < 0.01), and a trend of parity and season (p < 0.1) were detected. A clear negative effect of warm period on first service pregnancy rate was noted (p < 0.01). The pregnancy rate was the lowest in the H protocol during warm period (p < 0.05). Treatment with hCG 5 days after AI significantly improved pregnancy rates in those cows that were treated with the H protocol compared with saline treatments (41.5% vs 24.8%; p < 0.01). O and H were more effective in primiparous than in pluriparous cows (46.1% vs 29.9%; p < 0.1 and 43.6% vs 24.6%; p < 0.01). First service pregnancy rates were higher in primiparous hCG‐treated than in pluriparous hCG‐treated cows (57.9% vs 32.3%; p < 0.01). The pregnancy rate was higher for the hCG‐treated cows compared with saline‐treated cows during warm period (37.9% vs 23.6%; p < 0.001).     

Sustained-release deslorelin acetate implants disrupt oestrous cyclicity in the mare

There is a need for a safe, effective and practical method of oestrus suppression in the mare. The aim of this study was to monitor ovarian activity in mares exposed to either 9.4 or 28.2 mg deslorelin acetate, a GnRH agonist, in the form of a sustained-release implant. Following oestrus synchronisation, mares were randomly assigned to one of three groups (n = 4 per group) and administered either one (Des1 group; 9.4 mg) or three (Des3 group; 28.2 mg) implants of deslorelin acetate (Suprelorin-12, Virbac Australia) or one blank implant (Control group; Virbac Australia). Mares underwent weekly blood sampling for 12 weeks following implant placement (Day 0–Day 84), with transrectal palpation and ultrasonography of the reproductive tract at all sampling timepoints except Days 56, 70 and 77. All mares showed baseline serum progesterone concentrations (SPC; ≤1.3 nmol/L or 0.4 ng/ml) on Day 0. Cycling Control mares showed typical oestrous cyclicity characterised by peaks and troughs in SPC over time. Four of eight treated mares demonstrated a sustained elevation in SPC after the initial ovulation after implant placement; SPC declined to baseline levels (Des1 group; 2 mares) or remained elevated (Des3 group; 2 mares) at the final sampling timepoint on Day 84. Oestrous cyclicity was erratic in three of the remaining four treated mares. In total, 87.5% (7 of 8) of treated mares showed atypical oestrous cyclicity after implant placement. These results suggest that deslorelin acetate disrupts oestrous cyclicity in the mare, which warrants further research.     

Enhancement of Ovulatory Follicle Development in Maiden Sheep by Short-term Supplementation with Steam-flaked Corn

The effect of a short‐term nutritional supplementation with steam‐flaked corn on metabolism and folliculogenesis was evaluated in 14 maiden sheep. Oestrus was synchronized with two prostaglandin F_2α doses given 10 days apart. From day 11 to 15 of the oestrous cycle induced with prostaglandins, half of the ewes (group 2M) were supplemented with steam‐flaked corn, double the daily maintenance ration of the control sheep (group 1M). Body weight and condition remained unaffected, but the energetic supply increased plasma concentrations of glucose (3.6 ± 0.1 vs 4.3 ± 0.1 mmol/l, p < 0.0001) for the first 4 days and 3‐hydroxybutyrate (0.323 ± 0.58 vs 0.582 ± 0.04 mmol/l, p < 0.005) from day 2 to 4. The profile of insulin secretion was also affected by the treatment, increasing in group 2M to reach significant differences on days 13 and 14 (p < 0.05). From similar values at the start of the food supply, the treatment induced a higher follicular development in group 2M (1.1 ± 1.2 vs 7.4 ± 1.06 total follicles in day 15, p < 0.05), as evidenced by the lineal increase in the number of larger follicles (>4 mm, p < 0.005). Then, the number of follicles >4 mm in size in 2M was around 60% higher on day 16 (7.86 ± 0.45 vs 4.86 ± 0.63, p < 0.005). Thereafter, the mean number of corpora lutea per ewe was around 30% higher in group 2M (1.43 ± 0.2 vs 1.10 ± 0.1, although differences were not found to be statistically significant). These data suggest that the use of diets containing high starch sources, like the steam‐flaked corn, increases folliculogenesis and ovulation rate in sheep and can be applied in short‐term feeding practices.     

Luteal function and follicular growth following follicular aspiration during the peri-luteolysis period in Bos indicus and crossbred cattle

Follicular estradiol triggers luteolysis in cattle. Therefore, the control of follicle growth and steroidogenesis is expected to modulate luteal function and might be used as an anti‐luteolytic strategy to improve embryo survival. Objectives were to evaluate follicular dynamics, plasma concentrations of estradiol and luteal lifespan in Bos indicus and crossbred cows subjected to sequential follicular aspirations. From D13 to D25 of a synchronized cycle (ovulation = D1), Nelore or crossbred, non‐pregnant and non‐lactating cows were submitted to daily ultrasound‐guided aspiration of follicles >6 mm (n = 10) or to sham aspirations (n = 8). Diameter of the largest follicle on the day of luteolysis (7.4 ± 1.0 vs 9.7 ± 1.0 mm; mean ± SEM), number of days in which follicles >6 mm were present (2.3 ± 0.4 vs 4.6 ± 0.5 days) and daily mean diameter of the largest follicle between D15 and D19 (6.4 ± 0.2 vs 8.5 ± 0.3 mm) were smaller (p < 0.01) in the aspirated group compared with the control group, respectively. Aspiration tended to reduce (p < 0.10) plasma estradiol concentrations between D18 and D20 (2.95 ± 0.54 vs 4.30 ± 0.55 pg/ml). The luteal lifespan was similar (p > 0.10) between the groups (19.6 ± 0.4 days), whereas the oestrous cycle was longer (p < 0.01) in the aspirated group (31.4 ± 1.2 vs 21.2 ± 1.3 days). Hyperechogenic structures were present at the sites of aspiration and were associated with increase in concentration of progesterone between luteolysis and oestrus. It is concluded that follicular aspiration extended the oestrous cycle and decreased the average follicular diameter on the peri‐luteolysis period but failed to delay luteolysis.     

The Effect of a Chronic Stressor, Lameness, on Detailed Sexual Behaviour and Hormonal Profiles in Milk and Plasma of Dairy Cattle

The objectives of the present study were to quantify the effects of a biological chronic stressor (lameness) on the duration and frequency of different oestrous behaviours in parallel with milk hormone profiles. Dairy cows 51.8 ± 1.4 days postpartum (n = 59), including 18 non‐lame control cows, were scored for lameness and closely observed for signs of oestrus having had their follicular phases synchronized by administration of gonadotrophin‐releasing‐hormone (GnRH) followed by prostaglandin F_2α (PG) 7 days later. Lameness shortened the period when herd‐mates attempted to mount the lame cows (1.83 ± 0.69 h vs 5.20 ± 1.53 h; p = 0.042) but did not affect the overall duration of total behaviours (lame 12.3 ± 1.3 h vs non‐lame 15.2 ± 1.3 h). Lameness also lowered the intensity of oestrus [1417 ± 206 points (n = 18) vs 2260 ± 307 points (n = 15); p = 0.029]. Throughout the synchronized oestrous period, lame cows mounted the rear of herd‐mates less frequently (p = 0.020) and tended to chin rest less (p = 0.075). Around the period of maximum oestrous intensity, lameness also diminished the proportion of cows mounting the rear of another cow and chin resting (p = 0.048, p = 0.037, respectively). Furthermore, lame cows had lower progesterone values during the 6 days before oestrous (p ≤ 0.05). Fewer lame cows were observed in oestrus following PG (non‐lame 83%, lame 53%; p = 0.030); however, if prior progesterone concentrations were elevated, lame cows were just as likely to be observed in oestrus. In conclusion, following endogenous progesterone exposure, lameness shortens the period when herd‐mates attempt to mount lame cows but does not affect the incidence of oestrous. However, lame cows are mounted less frequently and express oestrus of lower intensity. This is associated with lower progesterone prior to oestrus but not with abnormal oestradiol or cortisol profiles in daily milk samples.     

The Treatment of Adrenal Cortical Disease in Ferrets with 4.7-mg Deslorelin Acetate Implants

Thirty pet ferrets with adrenocortical disease (ACD) of varying severity and duration were evaluated for response to a single administration of a slow release 4.7 mg deslorelin acetate implant. Clinical response to deslorelin was monitored via a physical examination performed every 3 to 4 months. Adrenal ultrasound measurements were taken every 3-4 months until clinical relapse. At clinical relapse, duration of symptom suppression and adrenal size and growth were determined. Administration of a single 4.7 mg implant of deslorelin acetate resulted in significant decreases in the clinical signs and hormonal concentrations associated with ACD. Within 14 days post-implant, vulvar swelling, pruritus, sexual behaviors and aggression decreased or disappeared. Hair re-growth was evident by 4-6 weeks post implant. Within two months post deslorelin implant, plasma concentrations of steroid hormones decreased: mean estradiol concentration decreased 28%; 17-hydroxyprogesterone levels decreased 89% and androstenedione levels decreased 88%. The response to a single 4.7 mg implant of deslorelin acetate was transitory. The mean ± SD time to recurrence of clinical signs was 17.6 ± 5.0 months (range, 8.0-30.0 months). Repeated ultrasound measurements revealed no statistical difference in size of the adrenals (right or left) before, during the months of deslorelin implant and at clinical relapse. Slow release 4.7 mg deslorelin implants can effectively be used to temporarily eliminate the clinical signs and reduce steroid hormone concentrations in ferrets with ACD. This dose of deslorelin does appear to influence adrenal tumor growth causing a decrease in adrenal size in some ferrets, and mild enlargement of adrenal glands in most ferrets with 2 of 30 implanted animals developing large tumors before clinical relapse. The long-term effect of treatment with deslorelin on adrenal tumor pathology requires additional investigation. At this time, surgical removal of the adrenal tumor remains the only curative treatment; however, 4.7 mg deslorelin implants are useful in the long-term management of ACD hormone-induced sequelae and may be as effective assurgical management.     

Ion,Protein, Phospholipid and Energy Substrate Content of Oviduct Fluid During the Oestrous Cycle of Buffalo (Bubalus bubalis)

The aim of this research was to analyse the composition of oviduct fluid (ODF) in buffalo cows at different oestrous cycle phases to fulfil the requirements of buffalo embryos in vitro. ODF was collected by chronic cannulation from three cows that were synchronized by administering a synthetic prostaglandin. Based on hormonal profiles, the pre‐ovulatory, ovulatory, post‐ovulatory and luteal phases of the oestrous cycle were defined. The volume of ODF produced (ml/24 h) was influenced by the oestrous cycle, with values (mean ± SE) around ovulation (1.0 ± 0.2) greater (p < 0.05) than in both the luteal (0.4 ± 0.1) and the post‐ovulatory phases (0.5 ± 0.1), but not different from the intermediate values in the pre‐ovulatory phase (0.8 ± 0.2). Among cycle phases, no differences were found in sodium, potassium, calcium and magnesium concentrations (130.0 ± 1.1, 5.1 ± 0.3, 2.8 ± 0.1 and 0.59 ± 0.04 mmol/l respectively). Interestingly, the chloride secretion (μm /24 h) was higher (p < 0.05) at ovulation (150.2 ± 16.5) than during both the luteal (73.7 ± 22.0) and the post‐ovulatory phases (63.7 ± 11.2), with intermediate values in the pre‐ovulatory phase (113.4 ± 23.5). Glucose concentration (mmol/l) was higher (p = 0.056) in the pre‐ovulatory phase (0.06 ± 0.02) than in the luteal (0.02 ± 0.01) and post‐ovulatory (0.02 ± 0.01) phases but not different from values in the ovulatory phase (0.04 ± 0.02). Concentrations of pyruvate and lactate among oestrous cycle phases were similar (0.08 ± 0.01 and 1.0 ± 0.1 mmol/l respectively). The total quantity of phospholipids (μmol/24 h) was greater (p < 0.05) at ovulation (0.21 ± 0.02) compared with the luteal, pre‐ovulatory and post‐ovulatory phases of the cycle (0.09 ± 0.02, 0.13 ± 0.02 and 0.09 ± 0.01 respectively). No differences were found in either the protein concentration (1.8 ± 0.3 mg/ml) or the quantity of proteins secreted in 24 h (1.8 ± 0.4 mg) among oestrous cycle phases. In conclusion, this study provides the first characterization of buffalo ODF during the oestrous cycle, showing species‐specific differences that may be useful for developing suitable media for buffalo in vitro embryo production.     

Attainment of Puberty in the European Mouflon (Ovis gmelini musimon) and the Domestic Manchega Ewe (Ovis aries)

The onset of puberty was assessed from plasma progesterone profiles for Mouflon (n = 8) and Manchega (n = 7) female lambs born in April. Four Mouflons exhibited their first ovulation at a mean date of 27 November ± 1.4 days, the mean age being 248.5 ± 3.9 days and the mean body weight 23.8 ± 0.6 kg. The remaining four Mouflons, which had a slower growth rate, attained puberty during the autumn of the following year. Five of seven Manchega lambs attained puberty in their first year at a mean date of 5 Oct ± 3.3, the mean age being 185.6 ± 2.6 days and the mean body weight 41.8 ± 2.0 kg. The mean live‐weight at the onset of puberty was 65.1 and 82% (p < 0.05) of adult body weights for Manchega and Mouflons, respectively. Overall, the results indicate that there is a threshold of body‐weight necessary for the attainment of puberty in the first breeding season for spring‐born lambs, which is very different between both Mouflon and Manchega. When body weight was below 23.8 ±0.6 and 41.8 ± 2.0 kg for Mouflon and Manchega lambs, respectively, first ovulation did not occur until the beginning of the next breeding season. The onset of puberty during the second year of life occurred within the specific reproductive seasons for Mouflon (October–April) and Manchega (July–March) ewes, despite minimal further growth.     

Influence of organic phosphorus on reproductive performance and metabolic profiles of anoestrous Farafra ewes in subtropics at the end of breeding season

The effect of organic phosphorus on metabolic, haematological and hormonal status, restoration of ovarian functions and conception rate in anoestrous Farafra ewes in subtropics were evaluated. Anoestrous Farafra ewes (n  = 24; 34.72 ± 0.52 kg body weight) were allocated into two equal groups: control and phosphorus groups. The ewes of phosphorus group were treated with sodium 4‐dimethylamino‐2‐methyl‐phenyl‐phosphonate as an organic bound phosphorous twice a week for successive 3 weeks. Ovarian follicle development and corpora lutea were checked three times a week till occurrence of oestrus using ultrasonography while pregnancy was confirmed at 30 days post‐service. Plasma metabolites, reproductive hormones, thyroid hormones and minerals were detected at weeks ?2, ?1, 0 (mating day) and + 4 weeks post‐oestrus. Phosphorus group had significantly (p  < .05) short interval to oestrous resumption if compared to control ewes (2.1 ± 0.8 weeks vs . 4.6 ± 1.1 weeks). In addition, phosphorous supplementation significantly (p  < .05) increased the number of antral follicles (developed and their sizes in addition to sizes of corpora lutea (8.72 ± 0.3 mm vs . 7.46 ± 0.9 mm) as well. Number of services per conception (2.6 vs . 1.4; p  < .01) was higher in control group than that of phosphorus group. Pregnancy rate (80 vs . 50%) was significantly (p  < .01) higher in phosphorus group when compared to control. White blood cells in treated ewes (10.8 ± 0.44; p  < .05) and monocytes (2.93 ± 0.13; p  < .01) were higher than that of control group (white blood cells; 9.53 ± 0.50 and monocytes; 2.24 ± 0.14). Metabolic parameters did not differ between phosphorus and control groups during different times of treatment. It could be concluded that phosphorous administration to anoestrous Farafra ewes in subtropics could improve reproductive performance and restore ovarian activity at the end of spring and early summer.     

Undernutrition During Foetal and Post‐Natal Life Affects Testicular Structure and Reduces the Number of Sertoli Cells in the Adult Rat

To test whether undernutrition during foetal to pre‐pubertal life would have long lasting effects on testicular histology in adult male offspring, eleven adult Sprague–Dawley pregnant rats were divided into two groups: Control group, n = 4, fed ad libitum, during gestation and lactation (until 25 day post‐partum). Underfed group pregnant females (n = 7) were kept in cages where only dams had access to food (standard rat chow, 33.5% of ad libitum intake of Control group pregnant dams). After parturition, litters were adjusted to either 14 (Underfed group) or eight (Control group) pups. Mothers were weighed weekly. At 25 day of age pups were weaned, housed at four animals per cage, fed ad libitum and weighed weekly until euthanized at 100 day of age. Testes were processed for standard histology and morphometrical evaluation. At weaning, mother weight was lower in Underfedthan in Control group (mean ± SD): 214.1 ± 26.2 g vs 361.9 ± 33.1 g. Body weight at 100 days of age (254 ± 26.9 g vs 342.4 ± 10.2 g, p ≤ 0.001), testicular weight (1.29 ± 0.16 g vs 1.45 ± 0.13 g, p = 0.03), number of Sertoli cells per seminiferous tubule cross section (18.2 ± 1.2 vs 20.2 ± 1.3, p ≤ 0.01) and per testis (30.5 ± 4.2×10⁶ vs 36.0 ± 5.4×10⁶) were lower (p < 0.05) in Underfed than in Control group. This is the first report stating that foetal to pubertal subnutrition is accompanied by changes in testicular structure and lower Sertoli cell numbers in adult life, strongly suggesting lower daily sperm production.     

Does Clinical Treatment with Phenylbutazone and Meloxicam in the Pre‐ovulatory Period Influence the Ovulation Rate in Mares?

The presence of anovulatory haemorrhagic follicles during the oestrous cycle of mares causes financial impacts, slowing conception and increasing the number of services per pregnancy. Non‐steroidal anti‐inflammatory drugs (NSAIDs) such as meloxicam and phenylbutazone are used in the treatment of several disorders in mares, and these drugs can impair the formation of prostaglandins (PGs) and consequently interfere with reproductive activity. This study aimed to evaluate the effects of treatment with NSAIDs on the development of pre‐ovulatory follicles in mares. In total, 11 mares were studied over three consecutive oestrous cycles, and gynaecological and ultrasound examinations were performed every 12 h. When 32‐mm‐diameter follicles were detected, 1 mg of deslorelin was administered to induce ovulation. The first cycle was used as a control, and the mares received only a dose of deslorelin. In the subsequent cycles, in addition to receiving the same dose of deslorelin, each mare was treated with NSAIDs. In the second cycle, 4.4 mg/kg of phenylbutazone was administered, and in the third cycle, 0.6 mg/kg of meloxicam was administered once a day until ovulation or the beginning of follicular haemorrhage. All of the mares ovulated between 36 and 48 h after the induction in the control cycle. In the meloxicam cycle, 10 mares (92%) did not ovulate, while in the phenylbutazone cycle, nine mares (83%) did not ovulate. In both treatments, intrafollicular hyperechoic spots indicative of haemorrhagic follicles were observed on ultrasound. Thus, our results suggested that treatment with meloxicam and phenylbutazone at therapeutic doses induced intrafollicular haemorrhage and luteinization of anovulatory follicles.