Intrauterine infusion of killed semen adversely affects uterine blood flow and endometrial gene expression of inflammatory cytokines in mares susceptible to persistent breeding-induced endometritis.

Persistent breeding-induced endometritis (PBIE) is a leading cause of infertility in mares. The objective of the study was to assess genital perfusion and endometrial gene expression of inflammatory cytokines in mares classified as susceptible (n = 5) or resistant (n = 5) to PBIE. Ten mares were examined daily during estrus until 6 d after hCG-induced ovulation for two estrous cycles. Twenty-four hours after application of 1500 IU hCG, 4 mL of killed (by repeated freezing in liquid nitrogen and thawing at 50 °C) deep-frozen semen or sterile saline was instilled into the uterine body and examinations were carried out immediately before and 3, 6, and 12 h after intrauterine infusion. Examinations included blood sampling to determine plasma progesterone (P4) concentrations, and transrectal ultrasonography in B- and color Doppler mode to determine follicular and luteal size and blood flow, the extent of intrauterine fluid, as well as time-averaged maximum velocity (TAMV), blood flow volume (BFV), and blood flow resistance (expressed as pulsatility index, PI) of the uterine arteries. Additionally, endometrial biopsies were obtained at 24 h before, and 2 and 7 d after infusion, and mRNA expressions of IL1B, IL6, IL8, IL10, TNF, CASP3, and COX2 were determined by qRT-PCR. Statistical analyses were performed with mixed models. Intrauterine fluid retention (diameter >20 mm for at least 3 d) was found after infusion of killed semen in five susceptible mares. There was no treatment effect (semen vs saline; P > 0.05) on genital blood flow, plasma P4 concentration, and endometrial gene expression. In comparison to resistant mares, susceptible mares had an increased (P = 0.04) BFV of the uterine arteries at 24 h before intrauterine infusion of killed semen, and an increased (P = 0.03) PI at 2 d after infusion. The TAMV, plasma P4 concentrations, and follicular and luteal size and blood flow did not differ (P > 0.05) between resistant and susceptible mares. Endometrial mRNA expression of IL1B increased (P = 0.05) at 2 d after the infusion of killed semen in the susceptible mares, and the expression of IL10 increased (P = 0.003) at 7 d after the infusion within the resistant mares. Interleukin 6 mRNA was increased (P = 0.05) in susceptible compared to resistant mares at 2 d after infusion. In summary, an intrauterine infusion of killed semen increases uterine blood flow resistance and alters endometrial gene expression of inflammatory cytokines for at least 7 d but does not affect ovarian blood supply and luteal function in mares susceptible to PBIE.

Effects of an extension of the equilibration period up to 96 hours on the characteristics of cryopreserved bull semen.

This study was designed to investigate the effects of an equilibration period up to 96 hours and three extenders (AndroMed, OPTIXcell, and Triladyl) on the quality of cryopreserved bull semen and to evaluate, whether an extension of the equilibration time to 72 hours does affect fertility in the field. One ejaculate of 17 bulls was collected and divided into three equal aliquots and diluted, respectively, with the three extenders. Each aliquot was again divided into five parts and equilibrated for 4, 24, 48, 72, and 96 hours before freezing in an automatic freezer. Sperm motility, plasma membrane and acrosome integrity (PMAI), and DNA fragmentation index (% DFI) were measured during equilibration. In addition to the parameters measured during equilibration, the percentage of viable sperm cells with high mitochondrial membrane potential (HMMP) was measured immediately after thawing, and after 3 hours of incubation at 37 °C. Sperm motility was assessed using CASA, and PMAI, HMMP, and % DFI were measured using flow cytometry. Equilibration time did affect all parameters before freezing (P < 0.01), and also the extender affected all parameters except HMMP (P < 0.05). After thawing, all parameters except HMMP immediately after thawing were influenced by the equilibration period (P < 0.001), whereas all parameters except % DFI immediately after thawing were influenced by the extender (P < 0.001). The changes of semen characteristics during 3 hours of incubation were also dependent on the equilibration time and the extender used in all parameters (P < 0.01). In the field study, semen of nine bulls was collected thrice weekly, processed using Triladyl egg yolk extender, and frozen in 0.25 mL straws with 15 × 106 spermatozoa per straw. In total, the nonreturn rates on Day 90 after insemination (NRR90) of 263,816 inseminations in two periods were evaluated. Whereas semen collected on Mondays and Wednesdays was equilibrated for 24 hours in both periods, semen collected on Fridays was equilibrated for 4 hours in period one and equilibrated for 72 hours in period 2. No differences in NRR90 could be found (P > 0.05). In conclusion, extension of the equilibration time from 4 hours to 24-72 hours can improve motility and viability of cryopreserved semen after thawing. The extent of improvement in semen quality is dependent on the extender used. Prolongation of the equilibration period from 4 hours to 72 hours had no effect on fertility in the field.

Immunization against gonadotropin-releasing hormone in dairy cattle: antibody titers, ovarian function, hormonal levels, and reversibility.

Suppression of cyclic activity in cattle is often desired in alpine farming and for feedlot cattle not intended for breeding. A cattle-specific anti-GnRH vaccination (Bopriva, Zoetis Australia Ltd., West Ryde, Australia) is approved for use in heifers and bulls in New Zealand, Australia, Mexico, Brazil, Argentina, Turkey, and Peru. Eleven healthy, cyclic Swiss Fleckvieh cows were included in the study and vaccinated twice with Bopriva 4wk apart. Injection site, rectal body temperature, and heart and respiratory rates were recorded before and 3d following each vaccination. Blood samples were taken weekly for progesterone and estrogen analysis and to determine GnRH antibody titer. Ovaries were examined weekly, using ultrasound to count the number of follicles and identify the presence of a corpus luteum. Thirty weeks after the first vaccination, the cows were subjected to a controlled internal drug-releasing device-based Select-Synch treatment. The GnRH antibody titers increased after the second vaccination and peaked 2wk later. Estrogen levels were not influenced by vaccination, and progesterone level decreased in 7 of 11 cows up to 3wk after the second vaccination and remained low for 10 to 15wk following the second vaccination. The number of class I follicles (diameter ≤5mm) was not influenced by vaccination, whereas the number of class II follicles (diameter 6-9mm) decreased between 7 and 16wk after the first vaccination. Class III follicles (diameter >9mm) were totally absent during this period in most cows. The median period until recurrence of class III follicles was 78d from the day of the second vaccination (95% confidence interval: 60-92d). After vaccination, all cows showed swelling and pain at the injection site, and these reactions subsided within 2wk. Body temperature and heart and respiratory rates increased after the first and second vaccinations and returned to normal values within 2d of each vaccination. The cows in our study were not observed to display estrus behavior until 30wk after the first vaccination. Therefore, a Select-Synch protocol was initiated at that time. Ten cows became pregnant after the first insemination (the remaining cow was reinseminated once until confirmed pregnancy). Bopriva induced a reliable and reversible suppression of reproductive cyclicity for more than 2mo. The best practical predictor for the length of the anestrus period was the absence of class III follicles.

[Castration in the bull calf and ram lamb]. / Die Kastration beim männlichen Kalb und Lamm.

This paper aims to provide an overview of the accepted techniques of pain relief and castration and guidelines of how to best perform these painful interventions in an animal-friendly way under Swiss conditions. Calves should be castrated at the age of 14 days or less, at least 10 minutes after local anesthesia with lidocaine, applying a single rubber ring. Concurrently, a NSAID should be administered intravenously (ketoprofen, 3 mg/kg of bodyweight) and Tetanus-serum subcutaneously (off label use). If possible, ketoprofen(4.5 mg/kg BW) should be orally administered for 3 - 5 days postoperatively. At 10 days after applying the rubber ring, the dried-off scrotum including the rubber ring should be removed with a clean knife or a scalpel. Local anesthesia is not necessary for this procedure. Ram lambs should be castrated at the age of 14 days or less, at least 10 minutes after local anesthesia with lidocaine, applying a rubber ring. The toxic dose of 4 mg lidocaine/kg BW (corresponds to 1 ml lidocaine 2 % per lamb of 5 kg BW) should not be exceeded. Concurrently, a NSAID (off label use) and Tetanus-serum should be administered systemically. Immunization against GnRH represents an animal-friendly and economically feasible alternative to rubber ring castration. With two immunizations at an interval of 3 - 4 weeks testicular development can be inhibited for at least 3 months and the onset of puberty clearly delayed. However, a specific vaccine for use in ruminants is currently registered neither in Switzerland nor in Europe.

Le présent travail donne un aperçu des méthodes praticables pour l'anesthésie et la castration ainsi que des recommandations pour une réalisation de cette intervention dans le respect du bien-être animal dans les conditions suisses. Les veaux devraient être castrés dans les 14 premiers jours de vie au moyen d'un élastique posé au plus tôt 10 minutes après la réalisation de l'anesthésie locale avec de la lidocaïne. Il convient d'administrer simultanément un AINS (kétoprofène, 3 mg/kg PC) en i/v lente ainsi qu'un sérum antitétanique (reconversion). Dans la mesure du possible, il convient d'administrer oralement du kétoprofène (4.5 mg/kg PC) par voie orale pendant les 3 à 5 jours suivants. Le scrotum desséché y compris l'élastique doit être supprimé sans anesthésie au moyen d'un couteau propre ou d'un scalpel stérile 10 jours après la pose de l'élastique. Les agneaux doivent être castrés dans leurs 14 premiers jours de vie, comme les veaux avec un élastique posé au minimum 10 minutes après la réalisation d'une anesthésie à la lidocaïne. On prendra garde à ne pas dépasser la dose maximale de 4 mg/kg de lidocaïne (ce qui représente 1 ml de lidocaïne 2 % pour un agneau de 5 kg). Simultanément à l'anesthésie, on appliquera un AINS (reconversion) et un sérum antitétanique. La vaccination anti-GnRH représente une alternative pratique et économique ménageant les animaux pour la castration des veaux et des agneaux mâles. Une double vaccination à 3 à 4 semaines d'intervalle permet de bloquer le développement testiculaire pendant au moins 3 mois et de retarder ainsi la maturité sexuelle. Toutefois il n'existe actuellement pas en Suisse ni en Europe de vaccin spécifique enregistré pour l'application chez les ruminants.

[Fertility after treatment with Eazi-breedTM CIDR G for 6 or 12 days outside the breeding season in Lacaune dairy sheep]. / Fruchtbarkeit nach 6- und 12-tägiger Behandlung mit Eazi-breed™ CIDR® G ausserhalb der Zuchtsaison beim Lacaune Milchschaf.

This study compares the fertility after short- and long-term synchronization using a progesterone intravaginal device in Lacaune dairy sheep outside the breeding season. For the experiment 108 Lacaune sheep were treated with Eazi-breed™ CIDR® G intravaginal devices (Pfizer Animal Health, Zürich) for 12 days (Group L, n = 60) or 6 days (Group K, n = 48) in combination with eCG (Group L) or with eCG and 125 µg Cloprostenol (Group K) at device removal. Thereafter the ewes were kept together with rams for 60 days, ewes in estrus were recorded and the fertility was assessed after lambing. Blood progesterone concentration was measured at device application, withdrawal and 14 days later. Results show that neither treatment nor parity had an influence on estrus rate (Group L 91.7 %, Group K 93.8 %, nulli- and pluriparous animals 96.9 % and 90.8 %, respectively). Group L showed a tendency towards a better first cycle lambing rate and a significantly (P < 0.05) higher overall lambing rate compared to sheep of Group K (71.7 % vs. 60.4 % and 83.3 % vs. 72.9 %). Pluriparous ewes had higher (P < 0.05) lambing rates and greater (P < 0.05) numbers of lambs born per synchronized ewe than nulliparous sheep for the first cycle (75.0 % vs. 46.9 % and 1.4 ± 1.0 vs. 0.9 ± 1.1) as well as for the overall service period (92.1 % vs. 46.9 % and 1.7 ± 0.8 vs. 0.9 ± 1.1). Fourteen days after insert withdrawal progesterone concentrations were higher (P < 0.05) in Group L than in Group K (7.7 ± 4.3 vs. 5.6 ± 2.7 ng/mL) and in nulli- compared to pluriparous (9.1 ± 5.6 vs. 5.7 ± 2.1 ng/mL) ewes. In conclusion, the overall lambing rate was higher after long-term compared to short progesterone treatment and nulliparous ewes were less suitable for estrus induction outside the breeding season.

Dans ce travail, on étudie la fertilité chez de brebis de race Lacaune lait après une synchronisation des chaleurs de courte et de longue durée au moyen d'un pessaire intra vaginal à la progestérone. Pour cela, 108 brebis Lacaune lait ont été traitées pendant 12 jours (groupe L, n = 60) ou 6 jours (groupe K, N = 48) avec un pessaire vaginal Eazi-breed™ CIDR® G (Pfizer Animal Health, Zürich) en combinaison avec 500 IE d'eCG (groupe L) respectivement 500 IE d'eCG et 125 µg de Cloprostenol (groupe K) au moment du retrait du pessaire. Par la suite, les brebis ont été détenues pendant 60 jours avec des béliers, les chaleurs ont été relevées ainsi que la fertilité après l'agnelage. Le taux sanguin de progestérone a été mesuré lors de la mise en place et du retrait du pessaire ainsi que 14 jours plus tard. Les résultats montrent que ni le traitement ni le nombre de gestations antérieures n'avaient d'influence sur le taux de chaleurs (groupe L 91.7 %, groupe K 93.8 %, brebis nulli- et pluripares 96.9 % respectivement 90.8 %). Les brebis du groupe L montraient, un taux de mise bas tendentiellement meilleur lors du premier cycle et au total significativement plus haut (P < 0.05) que celles du groupe K (71.7 % par rapport à. 60.4 % et 83.3 % par rapport à 72.9 %). Les pluripares avaient, lors du premier cycle et en général, des taux de mise-bas plus élevés que les nullipares (75.0 % contre 46.9 % respectivement 92.1 % contre 46.9 %, P < 0.05) ainsi qu'un nombre d'agneaux plus élevé par brebis synchronisée (1.4 ± 1.0 contre 0.9 ± 1.1 respectivement 1.7 ± 0.8 contre 0.9 ± 1.1, P < 0.05). Quatorze jours après le retrait du pessaire, les taux de progestérone étaient plus élevés dans le groupe L que dans le groupe K (7.7 ± 4.3 contre 5.6 ± 2.7 ng/mL) aussi bien chez les nulli- que chez les pluripares (9.1 ± 5.6 contre 5.7 ± 2.1 ng/mL). En résumé on constate que le taux de mise-bas était meilleur après un traitement long qu'après un traitement court et que les brebis nullipares étaient moins adaptées à la synchronisation des chaleurs hors de la saison de reproduction.

Vaccination against gonadotropin-releasing factor (GnRF) with Bopriva significantly decreases testicular development, serum testosterone levels and physical activity in pubertal bulls.

The aim of this study was to evaluate the effects of vaccination against gonadotropin-releasing factor (GnRF) on testicular development, testosterone secretion, and physical activity in pubertal bulls. The experiment was performed using 44 bulls aged between 6 and 7 mo. Twenty-three animals were vaccinated twice 4 wk apart with 1 mL of Bopriva (Pfizer, Animal Health, Parkville, Australia) and 21 bulls served as matched controls. Serum GnRF antibody titer and testosterone concentration as well as body weight and scrotal circumference were determined in all bulls for 24 wk from the first vaccination. In addition, physical activity was analyzed in 11 vaccinated and in 10 control animals using the ALPRO DeLaval activity meter system (DeLaval AG, Sursee, Switzerland). The results show that vaccination significantly (P < 0.05) influenced all parameters evaluated except body weight. Antibody titers to GnRF began to rise 2 wk after the first vaccination and reached peak values 2 wk after the second injection. Significant group differences in anti-GnRF titer were present for 22 wk following the first vaccination. Testosterone concentrations were significantly lower between weeks 6 to 24 after first vaccination in bulls with Bopriva compared with control animals. In vaccinated bulls testicular development was impaired after the second injection and scrotal circumference was significantly smaller between weeks 8 to 24 after first vaccination. Physical activity of vaccinated bulls was reduced after the booster injection with significant group differences for a continuous period of 106 days. In conclusion, vaccination against GnRF with Bopriva in pubertal bulls decreased testosterone levels in peripheral blood, testicular development, and physical activity but did not affect weight gain.

Effect of vaccination against gonadotropin-releasing factor (GnRF) with Bopriva® in the prepubertal bull calf.

The aim of this study was to evaluate the effect of immunization against gonadotropin-releasing factor (GnRF) with Bopriva(®) (Pfizer Animal Health, Parkville, Australia) in prepubertal bull calves. For the study, 6 calves were vaccinated at the age of 3 and 6 weeks with 1 mL Bopriva(®), and 6 animals served as matched controls. Concentrations of GnRF antibodies, testosterone and LH were determined in serum samples out to 30 weeks after the first immunization. Body weight and scrotal circumference were measured for 59 weeks. At slaughter, 65 weeks after the first immunization, the quality of epididymal sperm was evaluated. The results showed that vaccination against GnRF influenced (P<0.05) anti-GnRF titer, LH and testosterone concentrations as well as scrotal circumference. Antibody titers significantly (P<0.05) increased after the booster vaccination and reached peak values 2 weeks later. Compared to control animals, inhibition (P<0.05) of the prepubertal LH secretion was observed in vaccinated calves at weeks 10 and 12-14 after the first vaccination. In vaccinated calves testosterone concentrations decreased after the booster injection to values below 0.5 ng/mL serum and remained for at least 22 weeks at this low level. Animals vaccinated with Bopriva(®) showed a delay in testes growth and smaller scrotal circumference. Puberty occurred at the age between 46 and 55 weeks in vaccinated and between 38 and 52 weeks in control animals and body weight gain was similar in both groups. All vaccinated bulls attained spermatogenic capacity at slaughter when they were 68 weeks old.

[Effect of vaccination against GnRH (Bopriva®) in the male pubertal calf]. / Wirkung einer Impfung gegen GnRH (Bopriva®) beim männlichen pubertären Kalb.

The objective of this experiment was to investigate the effectiveness of a newly developed bovine anti-GnRH vaccine (Bopriva®, Pfizer Animal Health, Australia). A total of 12 peripubertal bull calves aged between 6 and 8 months were used, 2 randomly selected animals served as controls. Animals were vaccinated twice at an interval of 4 weeks with 1ml of Bopriva® (400 µg GnRH-protein-conjugate) subcutaneously in the neck and observed for a total of 36 weeks. Scrotal circumference was measured every week and blood samples were also taken weekly for the determination of testosterone and GnRH antibodies. Three months after the second injection (booster), 5 animals were slaughtered and their testes histologically examined. GnRH antibody titers rapidly began to rise after the second vaccination and reached peak values 3 weeks later. Testosterone concentrations decreased to values below 0.5 ng/ml serum 1 week after the booster and remained at this low level for at least 10 weeks. The following increase of testosterone occurred individually within 11 and 23 weeks after the booster injection. Histological examination of testes in vaccinated animals showed an incomplete spermatogenesis with impaired or no production of spermatids and a reduced diameter of seminiferous tubules. From our results we conclude that in the peripubertal bull two injections with the new bovine anti-GnRH vaccine 4 weeks apart is effective in suppressing testicular growth and testosterone secretion during at least 10 weeks after the booster injection.

[Suppression of reproductive cyclicity by active immunization against GnRH in the adult ewe]. / Unterdrückung der Fortpflanzungsaktivität durch aktive Immunisierung gegen GnRH beim adulten weiblichen Schaf.

The objective of the present study was to evaluate the effect of an anti-GnRH vaccine on cycling activity in the adult ewe during the breeding season. For the experiments 22 cycling White Alpine sheep, aged between 2 and 4 years, were randomly divided into a treatment and control group of 11 animals, each. Sheep of the treatment group were immunized twice at an interval of 4 weeks with 2 ml (400microg GnRH-protein conjugate) of Improvac (Pfizer Animal Health, Australia) subcutanously in the neck. Sheep of the control group received the same amount of saline solution. Blood progesterone concentrations were measured weekly from 3 weeks before to 32 weeks after first immunization and anti-GnRH titers were determined monthly. All vaccinated ewes ceased cycling within 2 - 8 weeks after first immunization. Plasma progesterone was suppressed for a minimum of 12 weeks (2 ewes) with individual variation of 14 (1 ewe), 25 (1 ewe) and more than 25 weeks (7 ewes). Four animals resumed cyclicity while 7 animals remained suppressed until the end of the study. Antibody titers peaked one month after the booster injection and thereafter continuously dropped until the end of the study (8 months after first immunization) to values between 10.9 and 40.8 % binding. From our results it can be concluded that two vaccinations with Improvac 4 weeks apart suppress cycling activity in adult ewes for at least 12 weeks. The inhibitory effect on ovarian activity, however, varies individually and may last more than 31 weeks.

Suppression of testicular function and sexual behavior by vaccination against GnRH (Equity) in the adult stallion.

The aim of this study was to evaluate the effects of an anti-GnRH vaccine on testosterone concentration, antibody titer, scrotal width, semen quality and sexual behavior in the stallion. Adverse reactions to the vaccine were also determined. Eight clinically healthy sexually experienced stallions aged between 6 and 15 years from the National Stud in Avenches (Switzerland) were used. Five stallions were immunized 3 times at an interval of 4 and 8 weeks, respectively, with 200 microg of a GnRH-protein conjugate (Equity, CSL Limited, Australia) intramuscularly in the neck and 3 control animals received an equivalent volume of saline solution. Plasma testosterone concentrations and GnRH antibody titers as well as semen quality and libido were determined weekly during 1 year (52 weeks). In addition, scrotal width was measured in all stallions before and 4, 8 as well as 12 months after first vaccination. Our results demonstrate that in 4 stallions plasma testosterone started to decrease after the second vaccination and remained suppressed for at least 6 months whereas one stallion showed no effect. Until the end of the experiment 2 stallions reached prevaccination testosterone values. Antibody titers varied individually in all 5 stallions and reached peak concentrations within 2 weeks after the third vaccination. Scrotal width was significantly (P<0.05) lower in vaccinated than in control stallions 8 months after first vaccination. Semen quality started to decreased after the second vaccination and improved towards the end of the experiment. In 4 stallions libido was clearly reduced after the second immunization but normalized in 2 animals before the end of the study while 2 stallions continued to show poor libido. From our results we conclude that three immunizations with Equity are well tolerated in the stallion and may effectively suppress testosterone secretion and reduce semen quality as well as sexual behavior. The inhibiting activity of Equity on these parameters is individually different and may last for a minimum of 6 months.

Capillary haemangiomas of the scrotum and testicle in boars.

In boars, scrotal haemangiomas have been described frequently, but intratesticular haemangiomas are rare. Of 12 boars with scrotal haemangiomas, three animals also had testicular tumours, as follow: testicular haemangioma (TH) (n=1); TH with intratubular germ cell tumour (ITGT) (n=1); TH with intratubular germ cell-like tumour (ITGLT) and Sertoli cell tumour (n=1). In the nine remaining boars, no testicular tumours were found. Immunohistochemical examination of scrotal and testicular haemangiomas revealed labelling of endothelial cells for vimentin and factor VIII-related antigen. Labelling of smooth muscle actin was occasionally observed in the walls of neoplastic vessels. In the ITGT, no labelling was produced by any of the antibodies used. The Sertoli cell tumour was strongly positive for S-100. Sperm granulomas and degeneration of seminiferous tubules were also observed.

Influence of immunization against GnRH on reproductive cyclicity and estrous behavior in the mare.

The aim of this study was to evaluate the effect of active immunization against GnRH on ovarian activity, plasma progesterone and estradiol concentrations and on estrous behavior in adult mares. Eighteen cyclic mares were randomly divided into a treatment and control group. Nine mares were immunized twice with 2 mL (400 microg GnRH-protein conjugate) of a GnRH-vaccine (Improvac, CSL Limited, Australia) administered intramuscularly, 4 weeks apart. Control mares received the same amount of saline solution. Ovaries and uterus of all mares were examined weekly by ultrasonography from 3 weeks before to 60 weeks after first immunization. Thereafter, vaccinated mares were evaluated monthly until 100 weeks after first vaccination. In addition, mares were teased with a stallion for assessment of estrous behavior and blood was collected for progesterone, estradiol-17beta and GnRH antibody titer determination. Results demonstrate that vaccination against GnRH significantly (P<0.05) influenced all parameters, except estradiol-17beta concentration. All vaccinated mares ceased reproductive cyclicity (plasma progesterone <1 ng/mL, follicles <3 cm) within 8 weeks after the first injection and ovarian activity remained suppressed for a minimum of 23 weeks. Five mares resumed cyclicity (follicles >3 cm, progesterone >1 ng/mL) while three mares showed only follicular activity (follicles >3 cm) and one mare remained completely suppressed for the entire duration of the study. In spite of ovarian suppression, four mares expressed sporadic and one mare continuous estrous behavior. In conclusion, reproductive cyclicity in adult mares can be successfully suppressed by immunization against GnRH but the timing of resumption of cyclicity is highly variable and estrous behavior may occur in spite of ovarian suppression.

Influence of estradiol on adrenal activity in ovariectomized cows during acute stress.

Stress-dependent activation of the hypothalamo-pituitary-adrenal axis (HPA) can compromise reproductive function in animals and humans. In addition, it has been shown that estrogens are also capable of influencing the activity of the adrenal cortex. The objective of this study was to evaluate the effect of estradiol (E2) on adrenocortical secretion of cortisol and progesterone as well as on pituitary LH-release in cows during stress. Five ovariectomized Brown Swiss cows were exposed to acute restraint stress (2-h immobilization in a trimming chute), either with or without E2 treatment. Blood samples were taken every 15 min during a 5-h period for determination of cortisol progesterone and LH. Our results demonstrate that during the 2-h stress period mean cortisol concentrations significantly (P < 0.05) increased in all cows independent of E2 treatment. Mean progesterone concentrations also increased during stress, but the effect was only significant in E2-untreated cows. In contrast to cortisol and progesterone, mean LH values decreased in all animals during stress, but the decline was not significant. However, significantly lower mean LH values were seen at the end of the stress period comparing to values before stress. In cows without stress, E2 treatment had no significant effect on mean values of all three hormones analyzed. From our results it can be concluded that in ovariectomized cows (a) acute stress increases cortisol and progesterone secretion but decreases LH release and (b) the stress induced adrenocortical and pituitary responses were clearly attenuated under the influence of estradiol.

Histomorphological and immunohistochemical findings in testes, bulbourethral glands and brain of immunologically castrated male piglets.

The aim of this study was the histological and immunohistochemical evaluation and comparison of testicular, bulbourethral and brain tissue in immunized and intact control boars. Fourteen male piglets, aged between 10 and 16 weeks, were vaccinated twice subcutaneously 4 to 5 weeks apart with Improvac, an anti-GnRH vaccine. The pigs were sacrificed 1 to 16 weeks following the second injection. Testicular weight was recorded and various tissue samples were collected and fixed in formalin and Bouin's fixative for histological examination. In addition, 2 boars were immunized five times and slaughtered 60 weeks after the last injection. Histological and immunohistological studies performed on testes and epididymes showed clear signs of atrophy in the immunized animals and a significant reduction in paired testes weight was seen in treated boars. Microscopically, the mean diameter of the seminiferous tubules was markedly reduced. Spermatogonia as well as few spermatocytes were visible between the Sertoli cells and Leydig cells were atrophic. None or only few spermatozoa were detected in the epididymis. The bulbourethral glands of immunocastrated pigs were smaller than in control pigs and showed histological evidence of atrophy. Immunohistological detection of LH and FSH in the pituitary gland of treated and control boars showed no quantifiable difference in the amount of these two gonadotropins and no lesions were visible in the hypothalamus and the pituitary gland. From our findings it can be concluded that the anti-GnRH vaccine Improvac induces severe atrophy of testes and bulbourethral glands in immunized pigs. This effect appears to be reversible, depending on the immune response of each animal and the time elapsed after the last booster injection.

[Body condition and frequency of ovarian cysts in Brown Swiss cows during the first 40 days of lactation]. / Körperkondition und Häufigkeit von Ovarialzysten bei Braunviehkühen während der ersten 40 Laktationstage.

For this investigation 50 Brown Swiss cows from 21 different farms were used. Twenty-five peripartal overconditioned cows (back fat thickness > 38 mm) were compared with 25 peripartal not overconditioned animals (back fat thickness < 38 mm). On days 20, 30 and 40 post partum the ovaries were examined sonographically and 10, 15, 20, 30 and 40 days after calving plasma concentrations of progesterone and 17-beta estradiol were determined. In peripartal overconditioned animals 12 ovarian cysts were found while only one cyst was present in not overconditioned cows (P < 0.05). At first examination all ovarian cysts were classified by ultrasound as follicle theca cysts (progesterone < 0.5 ng/ml plasma). Follow examinations resulted in 3 cysts which persisted as theca cysts while 8 cysts became luteinized and 2 cysts completely regressed. There was no indication of increased plasma progesterone and/or estradiol concentrations in overconditioned cows with higher fat deposit before of ovarian cysts had occurred.

[Suppression of cycling activity in sheep using parenteral progestagen treatment]. / Die Unterdrückung der Brunst mittels parenteraler Gestagenbehandlung beim Schaf.

The objective of this study was to evaluate the effect of two synthetic progestagen preparations Chlormadinone acetate (CAP, Chronosyn, Veterinaria AG Zürich) and Medroxyprogesterone acetate (MPA, Nadigest, G Streuli & Co. Uznach) on cycling activity and fertility in sheep. A flock of 28 non pregnant white alpine sheep was randomly divided into three groups, A (n = 10), B (n = 9) and C (n = 9). During a period of 4 weeks the cycling activity was confirmed by blood progesterone analysis. Thereafter, the animals of group A were treated with 50 mg CAP, those of group B with 140 mg MPA and those of group C with physiological saline solution. All injections were given intramuscularly. Suppression of endogenous progesterone secretion lasted from 28 to 49 days (mean = 39 days) in group A and from 42 to 70 days (mean = 50 days) in group B. The synchronization effect of both preparations was unsatisfactory as the occurrence of first estrus was distributed over a period of 3 weeks in group A and 4 weeks in group B. These findings could also be confirmed by the lambing period which lasted 52 days in group A and 36 days in group B. Control animals lambed within 9 days due to the synchronizing effect of the ram. The first fertile estrus was observed 36 days (group A) and 45 days (group B) after the treatment. In group A all 10 animals and in groups B and C 8 of 9 ewes each became pregnant. Parenteral progestagen application with CAP and MPA is a simple, safe and reversible method of estrus suppression in the sheep. The minimal suppressive duration of 4 (CAP) and 5 weeks (MPA) is not sufficient when a period of 3 months (alpine pasture period) is desired.

[The castration of male lambs by immunization against GnRH]. / Die Kastration männlicher Lämmer mittels Immunisierung gegen GnRH.

The objective of this study was to evaluate the effect of a GnRH-vaccine in the ram lamb. Experiments were performed using 20 male lambs, randomly divided into a test (GnRH-immunization) and control group (physiological NaCl-solution). At a body weight of 20 kg (age 2-3 months) and three weeks later, all animals of the test group received 2 ml of Improvac (CSL Limited, Parkville, Victoria, Australia). The body weight as well as the blood testosterone concentration were measured weekly for 16 weeks. Thereafter, blood samples for testosterone analysis were taken monthly in immunized lams only. After the booster injection testicular growth was suppressed and plasma testosterone remained at low values < 0.1 ng/ml for at least 12 weeks. The mean corresponding testosterone concentrations for the control lambs ranged between 0.1 and 0.9 ng/ml plasma. An increase of testosterone was observed in 8 of 10 immunized animals between 3 to 7 months after the booster dose. The control lambs showed a tendency for better growth rate than vaccinated animals, but the difference was not significant. Our results demonstrate that in prepubertal ram lambs two immunizations with Improvac, three weeks apart, can suppress testosterone secretion and testicular growth at least for three months after the booster injection. For a suppression of reproductive function longer than three months after the second vaccination, a third immunization is needed at this time or when testicular growth is beginning.

Semen characteristics after vasectomy in the ram.

The objective of this study was to monitor the changes in semen characteristics in vasectomized rams and to determine if infertility was present 14 days after vasectomy. Experiments were performed using five cross-breed rams, aged between 18 and 30 months. Semen was collected weekly by artificial vagina from 2 months before to 5 months after vasectomy. After sexual rest for 10 days, vasectomy was performed by the cranial midscrotal approach. In all ejaculates the volume, concentration, total sperm number, motility and morphology (normal spermatozoa, loose heads) were determined and sperm viability (SYBR-14/PI) was evaluated in all semen samples collected after vasectomy. In the first ejaculate obtained 14 days post vasectomy all rams showed a significant (P < 0.05) drop in mean volume (from 1.2 to 0.5 mL), total sperm count (from 5176.8 to 51.1 x 10(6)) and morphologically normal sperm (from 84.1 to 15.7%), when compared to the last prevasectomy collection. We could also demonstrate a positive correlation (r = 0.89) between the individual cumulative total number of spermatozoa after vasectomy and the scrotal circumference measured before vasectomy. Sperm motility and viability could never be demonstrated after vasectomy and normal spermatozoa continuously decreased concomitant with an increase in loose heads. On post mortem examination 5 months after surgery, spermatocele formation and multiple sperm granulomas were present in all five rams. Our results show that in the first ejaculate collected by artificial vagina 14 days after vasectomy, no motile and viable spermatozoa could be detected. Despite weekly collections during a 5-month period after sterilization, azoospermia could never be achieved.

[Gynecomastia in a goat buck]. / Gynäkomastie beim Ziegenbock.

A 3-year old horned buck of the Toggenburger breed was referred to our clinic with signs of gynecomastia. Cytogenetic abnormalities could not be detected in the karyogram and compared to control animals the buck had normal plasma concentrations of testosterone and estradiol but not prolactin. Hyperprolactinemia has been considered the cause of gynecomastia leaving the semen quality undisturbed.