Follicle size and reproductive hormone profiles during a post-weaning altrenogest treatment in primiparous sows.

This study investigated the endocrine background of follicle size changes during post-weaning altrenogest treatment. altrenogest-treated sows received a 20-mg dosage daily at 8.00 a.m. from Day -1 to Day 14 after weaning. On Day -1, only 3/13 altrenogest-treated sows showed LH pulses compared with 8/8 control sows (P=0.001). On Day 0, control sows showed a typical high frequency-low amplitude LH pattern, indicative for recruitment of oestrogenic follicles. In altrenogest-treated animals on Day 0, half of the sows showed high frequency-high amplitude pulses from 4-5h after weaning. In altrenogest-treated sows, average follicle size increased from 3.1±0.5 mm on Day 0 to 4.4±0.6mm on Day 5, then decreased to 3.7±0.5 mm on Day 7 and stabilised thereafter. FSH and oestradiol (E2) concentrations showed a distinct diurnal pattern; high at 7.00 a.m. and low at 3.00 p.m. E2 concentrations (7.00 a.m.) showed a 2.5-fold increase from Day -1 to Day 2, and subsequently a 2-fold decline to reach a plateau at Day 8. FSH concentrations reached maximum levels by Day 5 and slowly declined afterwards. In conclusion, once-daily administration of altrenogest starting one day before weaning delays the weaning-induced increase in LH pulses. Although FSH and follicle size increase until Day 5 after weaning, follicle E2 production already decreased from Day 2 after weaning. Post-weaning altrenogest treatment thus results in a follicular wave of follicles that lose oestrogenic competence at Day 2 after weaning, presumably related to the changed LH dynamics during altrenogest treatment.

The induction of a secondary corpus luteum on day 12 post-ovulation can delay the time of luteolysis in high-producing Holstein cows.

Luteolysis before the time of maternal recognition of pregnancy is one cause of low fertility in high-producing dairy cows. The objective of this study was to assess whether induction of a secondary corpus luteum (CL) late in the luteal phase would delay the time of luteolysis. Twenty high-producing Holstein cows were synchronized to ovulation (Day 0) with the Ovsynch protocol and received hCG (1500 IU im) on Day 12. Corpora lutea formation (as evaluated by ultrasonography) and plasma P4 concentrations were monitored from Days 4 to 36. hCG treatment induced the formation of one secondary CL (CL2) in 11 of 20 cows (55%) from the dominant follicle (mean diameter: 14.2 ± 0.9 mm) of two-wave (3/11) and three-wave (8/11) cycles. The maximal diameter of the CL2 (23.3 ± 1.9 mm) was reached approximately 6 days after hCG treatment and was correlated with its structural lifespan (p < 0.01). Cows that formed a CL2 after hCG had higher mean plasma P4 concentrations on Day 14 (+4.5 ng/ml) and Day 18 (+3.0 ng/ml) compared with cows without CL2 (p < 0.05). The structural regression of CL2 begun approximately 8 days after that of the CL1, and the median time at which the first drop in circulating P4 levels occurred was later in cows that formed a CL2 than in those that did not (Day 26 vs Day 18; p < 0.01). Thus, the induction of a CL2 by hCG on Day 12 might reduce the risk of premature luteolysis in high-producing dairy cows after insemination.

Induction of an LH surge and ovulation by buserelin (as Receptal) allows breeding of weaned sows with a single fixed-time insemination.

The aim of this study was to demonstrate successful breeding of sows with a single fixed-time insemination following ovulation induction by buserelin, a GnRH analogue. In a first step, the optimal dose of buserelin (6, 10, or 16 µg) injected at 77 hours after weaning was determined in weaned sows (N = 15, 11, and 12, respectively) using its ability to induce an LH surge of similar magnitude as in control sows (N = 15) and induce ovulation. In 29/38 treated sows (76%), ovulation was induced and synchronized between 32 and 44 hours after injection, and the proportion of females ovulating during this time window was similar between groups at 73%, 73%, and 83% (6, 10, or 16 µg, respectively). Interestingly, whereas ovulation of 100% multiparous sows was induced and synchronized in the 32 to 44 hours posttreatment time window, successful induction was achieved in a lower proportion of primiparous sows (50%, 50%, and 67% following 6, 10, or 16 µg, respectively), the dose effect being nonsignificant. The magnitude of the LH surge was similar between control and treated sows, irrespective of the buserelin dose injected. Neither ovulation rate nor the number of good embryos on Day 5 postovulation differed between groups. Interestingly, the frequency of follicular cysts at slaughter was significantly affected by treatment (P < 0.05), being minimal and maximal in sows treated with 10 or 6 µg buserelin, respectively. In a second step, 419 sows from commercial herds in Spain, Germany, and France were randomly allocated to a control or treated group. The control sows were inseminated twice 12 ± 4 hours apart once estrus was detected. Treated sows received 10 µg buserelin at 86 ± 3 hours after weaning and were inseminated once 30 to 33 hours later. Farrowing rate of treated sows (87%, 166/192) was similar to that of control sows (84.5%, 169/200). Litter size was also similar between treated and control sows (13.6 ± 3.8 vs. 13.7 ± 3.2). In multiparous sows, neither duration of lactation nor magnitude of the fat loss during lactation significantly affected treatment effects. It is concluded that ovulation of weaned multiparous sows can be tightly synchronized by buserelin (10 µg) administration at 86 hours postweaning. This allows breeding once at a fixed time following buserelin injection while maintaining reproductive performance at a level similar to that of sows bred twice during estrus.

Variation in LH pulsatility during 24h after a postweaning altrenogest treatment in relation to follicle development in primiparous sows.

This study assessed pulsatile release of LH during altrenogest treatment after weaning in primiparous sows and related this to follicle development, estrus and ovulation rate. Weaned sows (n=10) received altrenogest 20mg/day from D-1 to D13 (weaning=D0) at 0800 h. On D13, blood samples were collected every 12 min from 1000 until 1900 h (1st sampling period) and from 2300 h until 0800 h (2nd sampling period). During the 1st sampling period, LH concentrations remained low and no LH pulses were detected in 8/10 sows. During the 2nd sampling period, average and basal LH concentrations (P<0.04) and frequency of pulses (P<0.0001) were higher than during the 1st sampling period. Sows with short vs. long intervals to estrus (<5 days vs. ≥5 days) had higher basal and average LH concentrations during the 2nd sampling period (P≤0.004) and showed more follicular growth during treatment (P=0.007), generating larger follicles at D14 (P=0.005). Sows with high ovulation rate (≥25) displayed more LH pulses in total than sows with low (<25) ovulation rates (P=0.03). In conclusion, this study showed that altrenogest efficiently prevented LH pulsatility during the first bleeding period and that low frequency/high amplitude LH pulses were generally present during the second bleeding period. This variability in LH release in between two altrenogest administrations (24h) may explain why follicular growth progresses to 5mm during altrenogest treatments. LH pulsatility was related to length of the follicular phase and ovulation rate, which signifies its relevance.

Selection of impubertal gilts by ultrasonography optimizes their oestrus, ovulatory and fertility responses following puberty induction by PG600.

In a group of gilts, occurrence of puberty is spread over several weeks. The optimal time to apply puberty induction is therefore difficult to define, as treatment of puberal gilts is meaningless. Changes in uterine aspect around puberty can be detected by ultrasonography. Two experiments were carried out to assess the effect of PG600(®) (400 UI of eCG and 200 UI hCG) administration to 6 months old gilts shown to be impubertal by ultrasonography on cyclicity and reproductive performance. Impubertal Large White gilts (n=94) were treated with either PG600 or solvent (controls). Administration of PG600 to impubertal gilts increased significantly the proportion of females displaying pubertal uterine ultrasound images 3 days after treatment (100% versus 65% in controls). The number of days to puberty was significantly reduced in gilts injected with PG600 (3.3 days) versus controls (4.7 days). In gilts of the PG600 group, ovulation rate was higher at the 1st oestrus compared to the 2nd, while this did not happen in controls. Progesterone concentrations were higher at mid-luteal phase in the PG600 treated gilts compared to controls (significant treatment by time interaction). Similar proportions of gilts returned to oestrus (89% versus 74% for controls). Following insemination at the 2nd oestrus, pregnancy rate and number of live embryos were unaffected by treatment. The combination of ultrasonography and PG600 optimizes the use of exogenous hormones by targeting treatment to gilts that need it, therefore facilitating the introduction of gilts into all in/all out system.

Regulation of folliculogenesis and the determination of ovulation rate in ruminants.

The paper presents an update of our 1993 model of ovarian follicular development in ruminants, based on knowledge gained from the past 15 years of research. The model addresses the sequence of events from follicular formation in fetal life, through the successive waves of follicular growth and atresia, culminating with the emergence of ovulatory follicles during reproductive cycles. The original concept of five developmental classes of follicles, defined primarily by their responses to gonadotrophins, is retained: primordial, committed, gonadotrophin-responsive, gonadotrophin-dependent and ovulatory follicles. The updated model has more extensive integration of the morphological, molecular and cellular events during folliculogenesis with systemic events in the whole animal. It also incorporates knowledge on factors that influence oocyte quality and the critical roles of the oocyte in regulating follicular development and ovulation rate. The original hypothetical mechanisms determining ovulation rate are retained but with some refinements; the enhanced viability of gonadotrophin-dependent follicles and increases in the number of gonadotrophin-responsive follicles by increases in the throughput of follicles to this stage of growth. Finally, we reexamine how these two mechanisms, which are thought not to be mutually exclusive, appear to account for most of the known genetic and environmental effects on ovulation rate.

Effects of altrenogest treatments before and after weaning on follicular development, farrowing rate, and litter size in sows.

In a previous study, we showed that follicle size at weaning affects the response of a sow to a short-term altrenogest treatment after weaning. In this study, an attempt was made to prevent the growth of follicles into larger size categories before weaning by using different altrenogest treatments before weaning to improve reproductive performance after postweaning altrenogest treatments. Sows (87 primiparous and 130 multiparous) were assigned to the following treatments: control (no altrenogest treatment; n=59), RU0-20 (20 mg of altrenogest from d -1 to 6; weaning=d 0; n=53), RU40-20 (40 mg of altrenogest from d -3 to 0 and 20 mg of altrenogest from d 1 to 6; n=53), and RU20-20 (20 mg of altrenogest from d -3 to 6; n=52). Follicle size was assessed daily with transabdominal ultrasound. Follicle sizes on d -3 (3.6 ± 0.7 mm) and at weaning (4.0 ± 0.7 mm) were similar for all treatments. Altrenogest-treated sows had larger follicles at the beginning of the follicular phase than did control sows [5.4 ± 0.1 and 3.8 ± 0.2 mm (least squares means), respectively; P < 0.0001] and on d 4 of the follicular phase [8.0 ± 0.1 and 6.7 ± 0.2 mm (least squares means), respectively; P < 0.0001]. Multiparous sows had larger follicles than did primiparous sows at the beginning of the follicular phase [5.3 ± 0.1 and 4.7 ± 0.1 mm (least squares means), respectively; P < 0.01] and on d 4 of the follicular phase [8.0 ± 0.1 and 7.0 ± 0.1 mm (least squares means), respectively; P < 0.0001]. Farrowing rate and litter size (born alive + dead) were not affected by treatment or parity. However, in primiparous sows, when mummies were included in litter size, altrenogest-treated sows had larger litters than did control sows (13.4 ± 0.5 and 11.9 ± 0.7 piglets, respectively; P=0.02). In primiparous control sows, backfat depth at weaning and litter size were positively related (slope of the regression line=0.82; P < 0.05), which was not the case in primiparous sows receiving altrenogest. In conclusion, the different altrenogest treatments before weaning did not prevent the growth of follicles before weaning and similarly affected subsequent follicle development and fertility. In primiparous sows, altrenogest treatment after weaning increased the number of fetuses during pregnancy, but positive effects seemed limited by uterine capacity. Altrenogest treatment after weaning improved litter size in primiparous sows with reduced backfat depth at weaning, which suggests a specific positive effect of a recovery period after weaning in sows with reduced BCS at weaning.

The effect of different postweaning altrenogest treatments of primiparous sows on follicular development, pregnancy rates, and litter sizes.

This study investigated follicular development during and after postweaning altrenogest treatment of primiparous sows in relation to subsequent reproductive performance. Primiparous sows (n = 259) were randomly assigned at weaning (d 0) to 1 of 4 groups: control (no altrenogest, n = 71), RU4 (20 mg of altrenogest from d -1 to 2, n = 62), RU8 (20 mg of altrenogest from d -1 to 6, n = 65), or RU15 (20 mg of altrenogest from d -1 to 13, n = 61). Average follicular size (measured by ultrasound) increased during altrenogest treatment and resulted in larger follicles at the start of the follicular phase for RU4, RU8, and RU15 compared with controls (5.3 ± 0.9, 5.5 ± 1.3, 5.1 ± 1.2, and 3.4 ± 0.6 mm, respectively; P < 0.0001). Farrowing rate was greater in RU15 (95%) than in RU8 (76%; P = 0.04). The RU15 group also had more piglets (2 to 3 more piglets total born and born alive; P < 0.05) than the other treatment groups. Follicular development at weaning clearly affected reproductive performance. At weaning, average follicular size: small (<3.5 mm), medium (3.5 to 4.5 mm), or large (≥ 4.5 mm), was associated with farrowing rates of 86, 78, and 48%, respectively (P < 0.001). Sows with large follicles at weaning had low farrowing rates (71%) in RU4, very low farrowing rates (22%) in RU8, but normal farrowing rates in RU15 (83%). In conclusion, this study showed that 15 d of postweaning altrenogest treatment of primiparous sows may allow follicle turnover in sows that had large follicles at weaning and that this was associated with an improved reproductive performance. It also showed that shorter treatment with altrenogest (4 or 8 d) is beneficial for sows with small follicles at weaning, but is not recommendable for sows with large follicles at weaning.

Reproductive features and faecal progesterone metabolite profile in female ferrets.

Elevated post-partum progesterone metabolite (P(4) -met) levels have been recently postulated to occur in lactating lynxes. The aims of this study were to monitor reproductive features in female ferrets, changes in the faecal P(4) -met concentrations throughout the breeding season and ovarian activity in post-partum lactating and non-lactating (NL) female ferrets. Our results indicate that coinciding with the results described in the lynx, elevated faecal P4-met concentrations occur in lactating ferrets, furthermore, that the duration of elevated secretion of P(4) seems to be dependent on the duration of lactation (P4-met at delivery, n=47: <500 ng/g; 5-7 days after delivery, during lactation, n=47: ≥ 500-800 ng/g; in females weaned at delivery, n=4: baseline levels). Three days after ovariohysterectomy of lactating females, P(4) -met concentrations decreased to baseline levels. In lactating females, the ovarian stroma is more active than that in NL ones implicating that the ovary is at least in part responsible for the elevated P4-met concentrations. Ovaries of lactating females contained many luteinized cells either as luteinized granulose cells in the wall of late pre-antral/early antral follicles or as corpus luteum (CL)-like structures. Early resumption of the entire ovarian activity (developed follicles and oestrus) occurs in NL post-partum females, while final follicular development is blocked (follicles stalls at antral stage) in the lactating ones (however, occasionally lactational oestrus may occur). We suppose that the elevated faecal P4-met during lactation together with suckling and other hormonal effects may contribute to prevention of early returning to oestrus in nursing female ferrets.

Evaluation of the clinical efficacy of Gonazon implants in the treatment of reproductive pathologies, behavioral problems, and suppression of reproductive function in the male dog.

Efficacy of a slow-release gonadotropin-releasing hormone (GnRH)-agonist implant (Gonazon) was assessed in 53 male dogs presented with benign prostatic hyperplasia (BPH), hypersexuality, aggressive behavior (either alone or in combination), excessive micturition, or to suppress fertility. Changes in testosterone (T) and estradiol (E2) concentrations and size of testes and prostate were monitored on Weeks 0, +8, and +26 after implantation. Additional measurements during and after this period were performed in 35 dogs. Clinical signs were assessed by the owners. All implants except one were retained throughout the study. Full downregulation of testicular function (T<0.35 nmol/L) was achieved in 46 dogs, five dogs showed partial downregulation (T = 0.36 to 0.47 nmol/L), one dog did not respond, and another one displayed a transient downregulation on Week +18. On Week +8, mean T and E2 levels were reduced by 96% and 62%, respectively, and did not further decrease. Full downregulation (T<0.35 nmol/L) lasted between 6 to >22 mo in most dogs except two. Compared with pretreatment values, mean testicular and prostatic size was reduced (P<0.00001) by 54% and 52%, respectively, on Week +8 and by 68% and 64%, respectively, on Week +26. Relative reduction of prostatic size was more marked in dogs with BPH than in healthy ones on Week +8 (P<0.05) and Week +26 (P<0.02), and clinical signs of BPH disappeared rapidly after implantation. Dogs affected with BPH were significantly older (P<0.001) than nonaffected ones (9.7 vs. 2.5 yr). Hypersexuality was more common in dogs<3 yr of age, and treatment clearly improved clinical signs. Age significantly affected the response to treatment in aggressive dogs; 75% of the cases responded with an improvement. The only minor and possibly treatment-related events observed were a short-lasting exacerbation of clinical signs of BPH (two dogs), increased weight gain (three dogs), and anxiety (three dogs) with one of these dogs developing a blunt coat. These results demonstrate the clinical efficacy and overall safety of the Gonazon implants.

Comparison of four treatments to suppress ovarian activity in ferrets (Mustela putorius furo).

Twenty-five ferret jills were randomly allocated to five groups of five animals; they were treated either before the breeding season with 15 mg medroxyprogesterone acetate (MPA), with 40 mg proligestone or with a slow-releasing device containing 4.7 mg of the gonadotropin-releasing hormone (GnRH) agonist deslorelin acetate (srGnRH), or at spring oestrus with 100 iu human chorionic gonadotropin (hCG), or were left untreated and mated. All the ferrets were assessed for signs of oestrus and their ovarian response was monitored by individual faecal progesterone metabolite (P4-met) profiles. The mean (sd) durations of treatment-induced ovarian quiescence were 94 (18), 99 (40), 53 (9) and 698 (122) days in the group treated with MPA, proligestone, hCG and srGnRH, respectively (P<0.001). Treatment with hCG and srGnRH proved to be the safest, while MPA treatment was associated with most side effects. Both MPA and proligestone treatments caused alopecia in one ferret per group, and after the first return to oestrus and mating an MPA-treated jill had a premature delivery and developed a purulent vaginal discharge. At the first post-treatment mating, the fertility (expressed as the percentage of ferrets mated in the group that produced a litter) was 75 per cent in the MPA-treated group, 60 per cent in the proligestone-treated group, 75 per cent in the hCG-treated group and 0 per cent in the srGnRH-treated group; in the control group, fertility was 100 per cent at mating in spring and 60 per cent at mating in summer. Three srGnRH-treated jills conceived at the second post-treatment oestrus.

Induction and synchronization of ovulations of nulliparous and multiparous sows with an injection of gonadotropin-releasing hormone agonist (Receptal).

The objective of this study was to determine if administration of a set dose (10 microg) of a gonadotropin-releasing hormone agonist, buserelin (Receptal; Rc), at set times after altrenogest (Regumate; RU) treatment or after weaning was able to induce and synchronize ovulation in female swine (gilts and sows). The pubertal (n=187) gilts were allocated to four groups, all synchronized with RU. Group 1 (RU) was inseminated twice at detected estrus, Group 2 (RU+Rc120) and Group 4 (RU+Rc104) received 10 microg Rc at 120 or 104 h after the end of RU treatment, respectively, and Group 3 (RU+eCG+Rc104) was treated with 800 IU equine chorionic gonadotropin (eCG) at 24h and Rc 104 h after the end of RU treatment, respectively. Gilts were inseminated twice at predetermined times, namely 144 and 168 h (Group 2), 128 and 144 h (Group 3), and 144 and 152 h (Group 4) after the end of RU treatment, respectively. Pregnant gilts were slaughtered at 30 d. Administration of Rc 104 h after the end of RU feeding synchronized ovulation over a 24-h time window in 97.9% and 100% of the gilts of Groups 3 and 4, respectively, whereas Rc administration at 120 h (Group 2) only successfully synchronized 88.9% of the gilts over 24h. Ovulation rates of gilts of Groups 2 and 4 were similar to that of the control group. Pregnancy rates were numerically higher in Groups 2 and 3 (92% and 96%, respectively) compared with those of Groups 1 and 4 (84% and 81%, respectively). Combination of eCG with Rc administration at 104 h (Group 3) increased ovulation rate (+4 CL) but decreased embryo survival to 62% at Day 30. The weaned sow experiment involved 61 sows of a range of parities (2.7+/-0.9), allocated to two control groups (Control 104 group and Control 94 group) and two treated groups (Rc104 group and Rc94 group), which received 10 microg Rc at 104 and 94 h after weaning, respectively. The females were inseminated at detected estrus. All pregnant sows farrowed. After treatment with Rc 94 h after weaning, 100% of sows ovulated over a 24-h time window versus only 68.7% of controls. Farrowing rate and litter size of the sows treated with Rc at that time were unaffected compared with that of control sows. In contrast, Rc administration at 104 h after weaning may have been too late; only 66.7% of the treated sows ovulated during a 24-h period. This proportion was numerically lower but not significantly different than that for control sows. Farrowing rate and litter size of treated sows were not significantly different than that of controls. Administration of Rc at the dose and times selected in this study tightened synchrony of ovulation in gilts and in sows after weaning. It remains to be established if such a synchrony is suitable to obtain good fertility after a single artificial insemination at a predetermined time.

Effect of hCG on early luteal serum progesterone concentrations in PG600-treated gilts.

Gilt oestrus and ovulation responses to injection of a combination of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) (PG600) can be unpredictable, possibly reflecting inadequate circulating LH activity. The objective of this study was to determine the effect of PG600 followed by supplemental hCG on gilt ovarian responses. In experiment 1, 212 Hypor gilts (160 day of age) housed on two farms in Spain received intramuscular (i.m.) injections of PG600 (n = 47), or PG600 with an additional 200 IU hCG injected either concurrently (hCG-0; n = 39), or at 24 h (hCG-24; n = 41) or 48 h (hCG-48; n = 45) after PG600. A further 40 gilts served as non-injected controls. Ovulation responses were determined on the basis of initial blood progesterone concentrations being <1 ng/ml and achieving >5 ng / ml 10 d after the PG600 injection. The incidence of ovulating gilts having progesterone concentrations >30 ng/ml were recorded. During the study period, 10% of control gilts ovulated whereas 85-100% of hormone-treated gilts ovulated. There were no significant differences among hormone groups for proportions of gilts ovulating. The proportions of gilts having circulating progesterone concentrations >30 ng/ml were increased (p < or = 0.02) in all hCG treated groups compared with the PG600 group. In experiment 2, a total of 76 Hypor gilts at either 150 or 200 days of age were injected with PG600 (n = 18), 400 IU eCG followed by 200 IU hCG 24 h later (n = 20), PG600 followed by 100 IU hCG 24 h later (n = 17), or 400 IU eCG followed by 300 IU hCG 24 h later (n = 21). Blood samples were obtained 10 days later for progesterone assay. There were no effects of treatment or age on incidence of ovulation, but fewer 150-day-old gilts treated with PG600 or 400 IU eCG followed by 200 IU hCG had progesterone concentrations >30 ng / ml. We conclude that hCG treatment subsequent to PG600 treatment will generate a higher circulating progesterone concentration, although the effect is not evident in older, presumably peripubertal, gilts. The mechanism involved and implications for fertility remain to be determined.

Controlled delivery of a GnRH agonist by a silastic implant (Gonazon) results in long-term contraception in queens.

Many female cats are spayed to prevent problems associated with calling and unwanted pregnancies. This study describes the safety and efficacy of an alternative approach, using an azagly-nafarelin containing implant (Gonazon) inserted subcutaneously in the neck of six treated queens for 3 years. These six queens together with six controls were permanently housed with vasectomized tom cats, and changes in progesterone concentrations were used to document the contraceptive efficacy of Gonazon. All six control queens ovulated regularly throughout the treatment period (3 years), as shown by regular changes in progesterone concentration. Sixteen ovulatory cycles were observed in each control throughout the study. In Gonazon treated queens, during the week following implant placement, two queens displayed a treatment-induced rise in progesterone concentration. Later on, all treated queens continuously displayed low progesterone concentrations until 3 years post-implant insertion, with the exception of a single isolated episode (at approximately 2.5 years of treatment), of follicular luteinization in two queens. In all queens, azagly-nafarelin concentrations peaked in the week following implant insertion remained high for 1 month and later decreased slowly. After 2.5 years of treatment, azagly-nafarelin concentrations were still greater than 150 pg/ml in 3/6 queens. During a 6-month long extension of the study (36-42 months post-treatment), all queens (treated and controls) were run with intact tom cats. None of them conceived. Following autopsy, ovarian weight and diameter of the uterine horns of 3/6 treated queens were shown to be similar to those of the controls. In conclusion, this study demonstrated that Gonazon efficiently prevented ovulation in queens (100%) for 3 years. Return to fertile heat was not observed towards the end of treatment. However, in half of the treated queens, reversibility of the treatment induced effects on the genital tract was demonstrated.

Reversible downregulation of endocrine and germinative testicular function (hormonal castration) in the dog with the GnRH-agonist azagly-nafarelin as a removable implant "Gonazon"; a preclinical trial.

Downregulation of anterior pituitary GnRH-receptors by application of a slow release GnRH-implant offers an effective and reversible alternative to surgical castration of the male dog. Aim of the present study was to test the efficacy and the underlying mechanisms of a new non-biodegradable controlled-release device implant (Gonazon((R)), Intervet, containing 18.5mg of the GnRH-agonist Azagly-Nafarelin). Eight male beagle dogs were implanted s.c. at the para-umbilical region. In four dogs implant removal was after 180 days (group 1), in the other four dogs after 365 days (group 2). Eleven weeks after implantation availability of LH was reduced (p<0.0001) by 70%. After an initial increase lasting for about 4 days, testosterone (T) and estradiol (E2) concentrations decreased (p<0.0001) to basal levels within 17.5+/-8.4 days. Size of testes was decreased by about 82% after 17 weeks, size of prostate by about 46% after 5 weeks (p<0.0001). Five to 7 weeks after implantation all dogs were aspermic. Testosterone and estradiol concentrations, together with testicular and prostatic size remained suppressed in all dogs in group 1 and one dog of group 2 until implant removal. The other three dogs of group 2 escaped from down-regulation between 223 and 324 days. Effects on the availability of LH, T, E2 and on testicular and prostatic size were fully reversible after implant removal or escape from down-regulation. In six dogs semen quality was back to pre-treatment values after about 29 weeks, however, one dog developed oligozoospermia while another one stayed azoospermic, probably due to an obstruction within the epididymal duct.

Ovsynch synchronization and fixed-time insemination in goats.

This study assessed the efficacy of an Ovsynch protocol (vs. the classical cronolone containing vaginal sponge+eCG treatment) to generate fixed-time insemination in goats during the breeding season. Each regimen was applied to 24 Boer goat does. Onset and duration of estrus were determined with an aproned male and follicular development was monitored by ultrasonography. Ovulation and quality of the corpora lutea were established from progesterone concentrations. In 10-11 goats per group, LH concentrations were determined throughout the preovulatory period. Does were inseminated at pre-determined times (16 h after the second GnRH injection and 43 h after sponge removal). Estrus was identified in 96% of the Ovsynch-treated goats (at 49 h after prostaglandin injection) and in 100% of the goats synchronized with sponges (at 37 h after sponge removal). Low progesterone concentrations at the time of AI were observed in 21/24 and 24/24 goats synchronized by Ovsynch and sponges, respectively. Synchronization of the LH surge was tighter following Ovsynch compared to sponge treatment. Kidding rates (at 58 and 46% in the Ovsynch and sponge groups, respectively) and prolificacy (at 1.86 and 1.83 in the Ovsynch- and sponge-treated goats) were similar for both groups, as were the number of ovulations (2.9 and 3.3) and the proportion of does with premature corpus luteum regression (29 and 17%). When excluding does with premature luteal regression and those with low progesterone levels when receiving prostaglandins, kidding rate reached 87.5% (14/16) after Ovsynch. During the breeding season, the Ovsynch protocol may thus be an useful alternative to the sponge-eCG treatment.

Treatment with a subcutaneous GnRH agonist containing controlled release device reversibly prevents puberty in bitches.

In most species, continuous administration of GnRH agonists desensitizes the pituitary to GnRH, and blocks ovarian function. The aim of this study was to assess the effects of a novel controlled release device containing azagly-nafarelin (Gonazon) to prevent puberty in young Beagle bitches (mean age: 4.88 +/- 0.32 months). Gonazon containing 18.5 mg azagly-nafarelin (n = 10) or a placebo implant (n = 10) was administered subcutaneously. Throughout the 1-year treatment, estrus behaviour was monitored weekly. Plasma progesterone concentrations, as well as body weight and height, were measured monthly. Following implant removal, estrus detection and progesterone measurement were continued until occurrence of puberty in all bitches. Control bitches displayed puberty (estrus, followed by ovulation) at approximately 11.9 +/- 2.7 (range, 8-16) months of age. In contrast, none of the Gonazon treated bitches displayed puberty during the period when Gonazon was present. Following removal of Gonazon, resumption of estrus and ovulation naturally occurred (seven bitches) or was induced (three bitches) approximately 8.5 (1.2-14.3) months later. As a consequence, age of puberty of the Gonazon treated bitches was 25.5 +/- 5 (18-31) months. No clinically detectable side effects were noted in Gonazon treated bitches. Height at withers was unaffected by treatment. Changes in body weight with time were also unaffected by treatment. Implants were well tolerated and generally easy to remove. These data demonstrated that Gonazon safely, efficiently and reversibly prevents reproductive function for 1 year in prepubertal bitches.

Efficacy of two types of vaginal sponges to control onset of oestrus, time of preovulatory LH peak and kidding rate in goats inseminated with variable numbers of spermatozoa.

In small ruminants, progestagen-impregnated vaginal devices (sponges) are useful tools to manage reproduction irrespective of season and to the application of timed artificial insemination (AI). A novel progestagen releasing vaginal-controlled release device (Chronogest CR), loaded with less (20mg) cronolone using proprietary procedures, was developed and its efficacy (synchronising ability, fertility and prolificacy following sponge removal) evaluated versus the existing Chronogest sponge containing 45 mg of cronolone in goats. Females (n=199) were maintained in field conditions and inseminated with graded amounts of spermatozoa at two stages of the year (breeding and non-breeding seasons). The use of the new Chronogest CR sponge was associated with an earlier initiation of the LH surge (28.7h versus 30.8h following sponge removal, P<0.01). A similar degree of synchronisation of the LH surge was obtained with both types of sponges. In both treatment groups, a longer time interval between sponge removal and the LH surge was noted in females with high milk production. Fertility and prolificacy were high and unaffected by the type of sponge used or the amount of spermatozoa inseminated. It is concluded that the new Chronogest CR sponge allows a reduction of the progestagen load from 45 to 20mg without detrimental effects on synchronisation, fertility and prolificacy.

[Specificities of prepubertal follicles and oocytes]. / Particularités du follicule et de l'ovocyte prépubères.

Ovarian physiology of prepubertal and adult animals is different. Some characteristics as follicular dynamics (follicular waves and growth) are similar but total follicular population and number of growing follicles are higher in prepubertal ovary. Prepubertal oocytes represent a negative model for in vitro studies since they lead to lower cleavage and blastocyst rates when they are used to produce embryos. This reduced ability to support embryonic development is due to follicular and oocyte differences. Follicular fluid and granulosa cells proteins, and steroidogenic potential differ between prepubertal and adult animals. Moreover, experiments using nuclear transfer demonstrate that cytoplasmic maturation of prepubertal oocytes is incomplete. These deficient oocytes are smaller, contain lower levels of MPF and MAP Kinase and differences in metabolism and cytoplasmic organelles are observed.

Regulation of ovarian follicular dynamics in farm animals. Implications for manipulation of reproduction.

In this review, the main features of folliculogenesis are summarized and compared among species. In the past few years, ultrasonography has clarified follicle growth patterns, and our understanding of follicle maturation has improved considerably. As the follicles develop towards the ovulatory stage, three features appear to be highly conserved across all species: 1) the sequence of events (recruitment, selection and dominance); 2) the sequential need for gonadotropins (FSH for recruitment, LH for dominance) and 3) the large variability of numerical parameters (number of waves per cycle, number of follicles per wave) as well as temporal requirements (time of selection, duration of dominance). In addition, specific follicles may also have variable gonadotropin requirements (thresholds). When patterns of follicle development at different physiological states are compared across species, follicular waves were detected in cattle, sheep and horses and during the prepubertal period in swine, suggesting that ovaries of all species operate on a wave basis unless they are prevented from doing so. Efficient estrus control treatments should have the ability to affect 1) the wave pattern by preventing the development of persistent dominant follicles containing aging oocytes, and 2) the recruitment of the future ovulatory follicle whatever the stage of the wave at the time of treatment. This would allow synchronous ovulation of a growing dominant follicle. Manipulation of the luteal phase follicular waves after mating or AI may also optimize fertility. Superovulation is still an efficient technique to obtain progeny from genetically valuable females. Administration of exogenous gonadotropins acts to reveal the underlying ovarian variability. Ovarian response of each female depends on the number of gonado-sensitive follicles present at the time when treatment is initiated. Identification of the number of such follicles for each female would improve efficacy of superovulation, by allocating potential nonresponders to other techniques (OPU/FIV). One of the main components of the within female response to superovulation is the stage of the wave when gonadotropins are injected. Treatment in the absence of a dominant follicle ensures a response close to the female's specific maximum. The development of practical approaches to achieve this still requires further research.