anti-GnRH Vaccine Research Articles

Effect of anti-GnRH vaccine on testicular tissues in stallions

Anti-GnRH Vaccine is known as an alternative to surgical castration. The main advantage is the reversibility of immunization in 90% of the cases (Bruyas et al. Reprod Dom Anim, p.55: https://doi.org/10.1111/rda.12800). Anti-GnRH immunization reduces fertility by inhibition of gonadotropin synthesis involved in steroid production and spermatogenesis. Only few papers report information on the recovery of sperm production after prime immunization (twice 4 weeks apart) without booster in some stallions. We aimed to assess the effect of the vaccination protocol (Primo immunization or/and boosters) and the delay between the last injection of anti-GnRH vaccine and surgical castration on testicular histology. Twelve unvaccinated (control) and 28 immunized Lusitanian stallions were castrated at different time points after the last immunization (range 0.5-14 months). Vaccination protocol was a primo vaccination by intramuscular injection of 1 mL IMPROVAC® twice 4 weeks apart, followed by booster(s) when stallion behavior returned or scrotal size re-increased (range 1-10 injections). Testes were fixed in formaldehyde (10%) and stained by HES (Hematoxylin-Eosin-Saffron) and Androgen Receptor immunohistochemistry. Testicular size and spermatogenesis were evaluated by 7 parameters: Johnsen scores, seminiferous tubule diameter, number and density of Sertoli cells, number and density of germ cells, and the ratio of germ to Sertoli cells.Statistical analysis was performed by ANOVA. Values are given as mean ±standard deviation. Testis size was larger for unvaccinated controls (88±5 mm; P<0.001) compared to vaccinated stallions (70 ±8 mm). The Johnsen scores were higher in control (9.3 ±0.3; P<0.0001) compared to vaccinated stallions (8.0 ±1.2). Seminiferous tubule diameters, number and density of germ cells, and the ratio of germ to Sertoli cells were larger in control stallions: respectively 173 ±10 µm (P<0.001), 180 ±34 germ cells per tubule (P<0.001), 6330 ±897 germ cells per mm² of tubule (P<0.001), and 8.2 ±1.5 (P<0.001) germ cells per Sertoli cell compared to vaccinated stallions: 135 ±22 µm, 83 ±47 germ cells per tubule, 4404 ±1454 germ cells per mm² of tubule, and 3.3 ±1.9 germ cells per Sertoli cell. Conversely, Sertoli cell numbers and density were larger in vaccinated: 27 ±4 Sertoli cells per tubule (P=0.005) and 1720 ±582 Sertoli cells per mm² of tubule (P<0.001) compared to 23 ±4 Sertoli cells per tubule and 817 ±165 Sertoli cells per mm² of tubule in controls. There was no significant difference among the vaccination protocols (e.g., number of boosters, delay between last injection and gonadectomy) on these parameters. In conclusion, multiple rounds of anti-GnRH immunization in stallions: (1) induced testicular hypoplasia is more dependent on stallion rather the vaccination protocol, and (2) had no reversible effects on testes (according to Johnsen scores).

Efficiency of immunocastration with an anti-gonadotropin-releasing hormone vaccine on cryptorchid bulls.

Cryptorchid bulls have low economic value owing to the effects of masculinization. Moreover, surgical removal of an ectopic testis is difficult in certain clinical cases. Recently, immunocastration has garnered popularity as a nonsurgical castration method in pig farming; however, the effects of immunocastration on cryptorchid bulls are yet to be yet. Herein, we investigated endocrine changes due to immunocastration in cryptorchid bulls and studied its effectiveness. This study included 13 Holstein bulls diagnosed with cryptorchidism and classified into two groups based on pubertal period: <8 months of age (pregroup) and ≥8 months of age (postgroup). Antigonadotropin-releasing hormone (GnRH) vaccine was used for immunocastration, and two vaccine doses were administered. Blood testosterone and anti-Müllerian hormone (AMH) levels were measured and analyzed for endocrine evaluation. The testosterone levels significantly decreased following the start of immunocastration in both groups, thereby confirming the efficacy of antiGnRH vaccination in cryptorchid bulls. The AMH levels significantly increased in the pregroup with two antiGnRH vaccination, suggesting a compensatory response via the neutralization of GnRH antibodies. The AMH levels did not significantly change in the postgroup, indicating the partial suppression of AMH secretion in Sertoli cells during sexual maturation and failure of Sertoli cell maturation. Thus, we successfully restrained the serum testosterone levels in cryptorchid bulls using antiGnRH vaccine. The testosterone levels are a useful indicator of the immunocastration effect on cryptorchid bulls. Hereafter, a vaccine program that can sustain the castration effect on cryptorchid bulls is necessary.

Evaluation of testicular echotexture with Ecotext as a diagnostic method of testicular dysfunction in stallions

This study aimed to assess if Ecotext, a new software for evaluation of testicular echotexture, is a good method for diagnosis of stallions with testicular dysfunction (TD). Relationships between Ecotext parameters and sperm motility and production, testicular volume, and testicular blood flow were also studied. Ecotext provides a total of six echotexture parameters: Ecotext 1 (black pixels), 2 (white pixels) and 3 (grey pixels), and another 3 parameters related to hypoechogenic areas: Ecotext tubular density (ETD), Ecotext tubular diameter (ETd), and Ecotext tubular area (ETA). Stallions (n = 33) were assessed using proven diagnostic techniques (spermiogram, B-mode and Pulse Doppler ultrasound), and subsequent analysis with Ecotext. Animals were classified as “control stallions” (n:21, acceptable semen quality), and “stallions with TD” (n:12, poor semen quality (TM < 60%, PM < 45% and total nº of sperm with PM < 2000 × 106 spz), that were subdivided into “induced TD group” (immunized, anti-GnRH vaccine) and “acquired TD group”. The acquired TD group showed differences in all Ecotext parameters in relation to controls (Ecotext 1:0.11 ± 0.17 vs 2.82 ± 2.52, Ecotext 2:1584.0 ± 575.8 vs 388 ± 368.2, Ecotext 3:134.2 ± 9.26; ETA: 2.14 ± 0.59 vs 5.40 ± 1.90; ETd: 65.66 ± 6.27 vs 86.93 ± 10.65 and ETD: 92.35 ± 11.24 vs 132.10 ± 16.35, p ≤ 0.001). Results suggest acquired TD stallions were suffering testicular degeneration with loss of architecture and function as all Ecotext parameters were altered in relation to controls. Induced TD horses only showed a reduction in ETD (116.2 ± 8.59 vs 132.10 ± 16.35, p ≤ 0.001), despite all sperm parameters being worse. These findings suggested immunized stallions probably only experience an acute loss of testicular functionality and parenchyma architecture is likely not affected since differences in Ecotext parameters with control stallions were not detected. ETD was the best parameter to identify animals with TD (AUC: 0.84, optimal cut-off value of 124.3 seminiferous tubules/cm2). Correlations were found between ETD and Doppler indices (PI: 0.60; RI: 0.47 p ≤ 0.001), total testicular volume (r: 0.48; p ≤ 0.05) and sperm motility (TM:0.51; and PM:0.54; p ≤ 0.001) and production (r:0.51; p ≤ 0.001). In summary, Ecotext could identify changes in testicular echotexture of stallions with TD. Results open the possibility for new research focused on establishing the relationship between Ecotext parameters and histomorphometry features in stallion testes.

Testicular morphometry as a tool to evaluate the efficiency of immunocastration in lambs.

This study aimed to evaluate the efficiency of immunocastration in lambs using testicular morphometry. Thirty lambs were randomly divided into two treatments (subcutaneous administration of 1.0 mL and 0.5 mL of an anti-GnRH vaccine) and a control group (1.0 mL saline solution). The animals were vaccinated at four months of age, received a second dose 30 days later, and were slaughtered 90 days after the first vaccine dose. After slaughter, testicles were collected, and samples were removed for histological processing and evaluation of testicular morphometric parameters. Analysis of variance, Tukey’s test, and Kruskal–Wallis test were performed, with a 5% level of significance. There was a reduction in testicular weight, gonadosomatic index, seminiferous tubule diameter, germinal epithelium height, leydigosomatic index, and total tubule length. The total length per testicular gram increased in the immunocastrated group. Intrinsic spermatogenesis yield, Sertoli cell indices, and estimates of sperm and Sertoli cell production were reduced in the immunized groups (P < 0.05). The anti-GnRH vaccine in lambs at doses of 1.0 mL and 0.5 mL is sufficient to promote immunocastration, verified through severe changes in testicular morphometry from animals.

Immunization against Gonadotropin-Releasing Hormone in Female Beef Calves to Avoid Pregnancy at Time of Slaughter.

Simple SummaryPrecociousness of heifers kept in mixed beef herds with young and adult bulls leads to unwanted pregnancies. Inbreeding, premature calving followed by dystocia and a high stillbirth rate are the consequences. As an alternative, such heifers are slaughtered during the resulting pregnancy. The slaughtering of pregnant animals is an ethical problem, which is strongly criticized by consumers and animal welfare organizations. Therefore, the aim of this study was to postpone puberty in female beef calves housed in Swiss mixed herds to avoid pregnancy until scheduled slaughter at 11 months of age. We used a vaccine (Improvac®) that induces antibody production against sexual hormones, thereby suppressing the reproductive cycle. Monthly progesterone analysis in blood was performed to decide whether the cycle had already started. The results proved, that vaccinated female beef were not coming in heat until 11 months of age compared to the unvaccinated control group, which came in heat earlier. In conclusion, vaccination with Improvac® is an animal-friendly, non-invasive and reliable method to avoid early pregnancy in heifers as well as the slaughter of pregnant cattle.Precocious puberty in beef heifers can result in unwanted pregnancies due to accidental breeding by farm bulls. Inbreeding, premature calving followed by dystocia and a high stillbirth rate or slaughtering of pregnant heifers are the consequences of this behaviour. The aim of the study was to postpone puberty by using Improvac®, an anti-GnRH vaccine. Therefore, n = 25 calves were twice vaccinated, once at the age of 5 and then at 6.5 months. n = 24 calves served as unvaccinated case controls. The onset of puberty was assigned if progesterone analysis in the blood exceeded 1 ng/mL. Progesterone values were excluded if the corresponding serum cortisol levels were ≥60 nmol/L. Our target was met, as in the vaccinated group none of the calves exceeded a progesterone value >1 ng/mL until the scheduled age of slaughter at 11 months and only 12.5% of the animals exceeded a progesterone value of 1 ng/mL over the whole measuring period (>400 days) compared with 56.5% of the calves in the control group. In conclusion, the favourable results from our study using the vaccine Improvac® represent an animal-friendly, non-invasive and reliable way to avoid early pregnancy in heifers as well as the slaughter of pregnant cattle.

Efficiency of the anti-GnRH vaccine for castration of lambs intended for finishing in pastures

It was aimed to evaluate anti-GnRH vaccine efficiency on productive characteristics, seminal performance, size, and histology of testicles of lambs submitted to immunocastration. Twenty contemporary Texel lambs were evaluated, being 10 animals received two doses of 1 mL of anti-GnRH vaccine and 10 received two doses of 1 mL of saline, with an interval of 30 days. Seminal characteristics, weight and histological cut for the testicles, productive performance, and blood parameters were analyzed. Andrological monitoring was carried out every 30 days. All animals were kept on pasture of Urochloa spp. receiving daily supplementation protein-energetic by 90 days. There was effect of immunocastration on slaughter weight (44.3 vs. 48.3 kg), total gain (9.9 vs. 10.3 kg), daily gain (104.5 vs. 108.9 g/day), and feed conversion (5.83 vs. 5.97) respectively to immunocastrated and intact lambs. There was immunocastration effect on testicle weight (0.09 vs. 0.35 g), motility (4.5 vs. 61.0%), vigor (0.40 vs. 3.00), volume (0.09 vs. 0.74 mL) and swirling (0.20 vs. 2.70) in the third semen collection. Immunocastration through two doses of 1 mL of anti-GnRH vaccine with a 30-day interval between doses, causes infertility in sheep and can be adopted for lambs.

Active immunization against GnRH as an alternative therapeutic approach for the management of Bos indicus oocyte donors diagnosed with chronic cystic ovarian disease

The aim of this study was to evaluate the effect of active immunization against GnRH in Nelore (Bos indicus) cows repeatedly used as oocyte donors that developed chronic cystic ovarian disease (COD). In the first experiment, ovarian and uterine features were first compared between COD cows (n = 15) and healthy cows (n = 22, cycling control group) from the same breed and herd. Cows with COD had a greater number of large (P < 0.0001) and medium follicles (P < 0.01) but lesser small follicles (P < 0.05) than cycling controls. Mucometra was diagnosed in 73.3% of COD cows, but in none of the controls. No difference in average thickness of the endometrium was detected between groups; however, endometrial thickness and mucometra score were negatively correlated (R = −0.73, P = 0.0029) in COD cows. In the second experiment, COD cows were randomly allocated into two experimental groups, which received two 1.0 mL SC injections of either an anti-GnRH vaccine (COD immunized group, n = 8) or saline (COD control group, n = 7), given 28 days apart. Cows were examined weekly by transrectal ultrasonography during nine consecutive weeks after the first injection to evaluate the number and distribution of follicles among size classes, endometrial thickness, and presence of clinical mucometra. Vaccination against GnRH resulted in a progressive suppression of follicle growth and a reduction in the average size of the largest follicle, as well as in the number of large follicles (P < 0.01) in COD immunized cows compared with COD controls from week 7 onwards. Conversely, the number of small follicles in the COD immunized group increased after week 5 and was greater (P = 0.0023) than controls on week 9. Endometrial thickness and mucometra score were not affected (P > 0.05) by immunization against GnRH. In the third experiment, the COD immunized cows with effective suppression of follicle growth four weeks after the 2nd injection (n = 6) were submitted to three consecutive ovum pick-up (OPU) sessions (weeks 10, 11, and 12) for in vitro embryo production (IVEP). Cumulus-oocyte complexes (COC) collected from slaughterhouse ovaries were used as controls for IVEP. COD cows with produced 25.0 ± 3.8 COC per OPU session with no apparent detrimental effect of anti-GnRH vaccine on oocyte developmental potential in vitro, i.e., we observed similar cleavage rate (P = 0.5914) and greater blastocyst rate (P = 0.0177) in immunized cows compared with COC from slaughterhouse controls. Finally, in the fourth experiment wave emergence and follicular dynamics after follicle ablation were compared between COD immunized cows with effective suppression of follicle growth and a subset (n = 6) of the cycling, control group. No follicles grew beyond 4 mm diameter after follicle ablation in the COD immunized group, whereas a normal follicular wave emergence occurred in cycling controls. Antral follicle count was similar between cycling controls and COD immunized groups at 24 h and 96 h post-follicle ablation (P > 0.05), but greater in cycling controls at 48 h and 72 h post-follicle ablation (P < 0.05). In summary, our results suggest that active immunization against GnRH is effective to induce the regression of follicular cysts as well as increase the number of small follicles growing on the ovaries, in oocyte donors diagnosed with chronic COD, with no apparent negative effect on oocyte developmental potential in vitro.

Effectiveness of a New Recombinant antiGnRH Vaccine for Immunocastration in Bulls.

Simple SummaryCastration of males is a common procedure in cattle production. Surgical procedures are most commonly used, but there is an increasing interest in non-invasive alternatives to avoid risk of infection, bleeding, pain, stress and to improve animal welfare. Immunization against gonadotropin-releasing hormone is currently being used in livestock, but there is only one commercially available vaccine for cattle and results regarding the number of doses needed to maintain castration are variable. The efficacy, safety, and production parameters of a new antigen for immunocastration in bulls was assessed. Results showed that two doses of the vaccine to 40 10-month-old bulls achieved testosterone suppression below productive performance. Live weight at slaughter and carcass yield was greater in immunized animals than in surgically castrated cattle. Castration effects of the vaccine were maintained until the end of the trial at 24 weeks.Castration by surgical techniques is common in livestock; however, post-surgery complications and concerns for animal wellbeing have created a need for new non-invasive alternatives. The objective of this study was to evaluate immunocastration in bulls using antigen GnRX G/Q; a recombinant peptide proved to be effective in laboratory and companion animals. A nine-month trial with 80 9-month-old Normand x Hereford bulls, kept in a pastured system, was conducted. The herd was divided in half with 40 bulls surgically castrated (SC) and 40 castrated by immunization against GnRH (IC). The antigen was injected on days 0 and 40 of the experiment. After the second dose, the IC group had elevated GnRH antibodies and decreased testosterone levels (below 5 ng/mL) that were maintained for 23 weeks. At slaughter on day 190, the immunocastrated group obtained a higher weight, hot carcass, and dressing percentage than the SC group. There was no difference in pH, color of meat, fat coverage, cooking loss, or tenderness between groups. The bulls showed no inflammatory reaction at the injection site or adverse side effects from the vaccine. Our results demonstrate that immunocastration with GnRX G/Q is an efficient and safe alternative to surgical castration in livestock. Additional work evaluating antigen effects over a longer period is needed to validate commercial viability.

Short Term Safety, Immunogenicity, and Reproductive Effects of Combined Vaccination With Anti-GnRH (Gonacon) and Rabies Vaccines in Female Feral Cats.

Overpopulation of free-roaming cats is a major problem leading to negative impacts on animal health and welfare, public nuisance, transmission of zoonotic diseases, and well-documented harm to wildlife. Surgical sterilization had failed to provide a practical solution to free-roaming cats' overpopulation under field conditions; therefore, efficient and safe non-surgical immunocontraception methods are aspired. Rabies is a deadly virus that may infect people and animals. However, the safety and efficacy of combined vaccination with anti-GnRH and rabies vaccines in feral cats, which often suffer from disrupted health conditions and experienced high stress level, has never been studied. Therefore, our objective was to examine the short-term safety and efficacy of anti-GnRH vaccine (Gonacon), in combination with rabies vaccine in female feral cats. Mature feral female cats were captured and divided into the following groups: (I) GonaconX1-Rabies: queens vaccinated with both Gonacon and rabies (n = 5); (II) GonaconX2-Rabies: queens vaccinated twice with Gonacon (3 weeks apart) and with Rabies (n = 4); (III) OVx-Rabies: queens ovariohysterectomized and vaccinated with rabies (n = 4); (IV) Intact-Rabies: queens vaccinated against rabies and remained intact (n = 3). Comprehensive veterinary examinations and blood tests were performed every 2 weeks for 14 weeks. Data were analyzed by Repeated-Measures-ANOVA or Fisher-Exact-Test. There were neither systemic nor local adverse reactions at the vaccination sites. Blood count (PCV, TS, RBC, HGB, HCT, WBC) and chemistry (Total protein, Total globulin, Albumin, Urea, Creatinine, Creatine kinase, Bilirubin, GGT, ALT, AST) analyses revealed no differences among groups. There were no differences in serum rabies antibodies titers among groups, and queens kept a protective titer (>0.5 IU/mL) starting at 2–4 weeks after vaccination. Anti-GnRH antibodies were detected in all Gonacon-vaccinated queens, excluding one queen (GonaconX2-Rabies group). Anti-müllerian hormone serum concentrations reduced significantly after ovariohysterectomy, as well as gradually following vaccination with Gonacon, but it remained high in intact queens. Evaluation of vaginal cytology and ovarian histology suggested that reproductive cyclicity was suppressed in Gonacon-vaccinated queens. Our results support the conclusion that in the short term, the combined vaccination with Gonacon and rabies is safe and effective in female feral cats. However, further long-term studies are warranted to test this immunologic regimen in feral cats.

Effect of a single or two doses of an anti-GnRH vaccine on testicle morpho-functional characteristics in Nelore bulls

The aim of this study was to compare testicle morpho-functional characteristics in bulls undergoing a single or two immunizations against GnRH. Nelore (Bos taurus indicus) bulls were randomly allocated into three experimental groups: G1 (n=12), a single 400 μg dose of anti-GnRH vaccine on day 0; G2 (n=11), a first 400 μg dose of anti-GnRH vaccine on day 0 followed by a second (boost) dose 30 days later; and control group (CG, n=12), 1 mL saline 0.9% at day 0. Every 30 days, from day 0 until slaughter at day 90, the bulls were weighed and underwent testicular biometry, semen collection and analysis, and blood sample collection for testosterone measurement. Immediately after slaughter, the testicles were removed and transport at 15°C to the laboratory for histopathological analysis. There was a decrease in testicular height (P=0.0476), width (P=0.0021), and in scrotal circumference (P=0.0001), after either a single (G1) or two (G2) immunizations against GnRH. Both G1 and G2 had lower testosterone concentrations than CG from day 60 on (P<0.01), but in G2, it was also lower than in G1 at day 90 (P=0.0006). All sperm parameters were affected by active immunization against GnRH (P<0.05), and in G2, averages were lesser (P<0.05) than in G1 from day 60 on. No signs of seminiferous tubule degeneration were found in any sample from the CG, contrasting with 75.0% and 100.0% of the samples from G1 and G2, respectively. In summary, immunocastration affected testicle morpho-functional characteristics in bulls in a time- and dose-dependent way.

Vaccination against GnRH as a prelude to surgical castration of horses.

Surgical castration is one of the most frequently performed surgeries in horses and is associated with various post-surgical complications ranging in severity. We investigated the effects of administering an anti-GnRH vaccine to colts for reduction of testis size on the incidence of complications associated with their subsequent surgical castration. Randomised open controlled trial. Nineteen colts were randomly assigned to one of the three groups. Treatment Groups V1-57 and V2-100 received two treatments of Improvac® (Zoetis, South Africa) before their castration in the field on either Day 57 or Day 100 following first treatment, respectively. Controls similarly received placebo treatments followed by castration on Day 57. Serial measurements of testicular dimensions and serum samples for measuring serum testosterone concentrations (STCs) and anti-GnRH antibody titres (ABTs) were obtained pre- and post-vaccination and on the day of castration. Clinical data recorded for 10days determined post-surgical complications. All vaccinated colts showed a baseline STC concentration at castration with a strong ABT response. Mean testicular volume of Groups V1-57 and V2-100 reduced by 49.7% and 30.8%, respectively, on Day 57 and this further reduced in Group V2-100 by 63.9% at castration on Day 100. Testis size was significantly correlated with post-surgical preputial (P=0.001) and scrotal (P=0.025) swelling. A study population of young light horse breed colts and a relatively small sample size limited this pilot study. Administering two doses of Improvac® in colts 28days apart prior to castration effectively reduced their testicular volumes with associated significant post-surgical improvement in both the incidence and degree of local swelling. These results may inform mitigation of potential post-surgical complications associated with castration in the field.

Reproductive parameters of lambs immunocastrated with anti-GnRH vaccine.

The objective of this study was to evaluate the testicular biometric, seminal, and plasma testosterone levels in lambs subjected to an anti-GnRH vaccine as a method of castration. Thirty entire, crossbred Santa Inês male lambs were randomly distributed into three treatment (T): T1 was the control group, with the administration of 1 mL of saline solution subcutaneously (SC); 1.0 and 0.5 mL of an anti-GnRH vaccine were administered SC in T2 and T3, respectively. Testicular biometric variables, physical and morphological variables of semen, and plasma testosterone concentrations were evaluated. At D60, there was a reduction in testicular length, width, thickness, and scrotal circumference of the immunocastrated animals regardless of the vaccine dose used (P < 0.05). A reduction in semen physical variables at both dosages (P < 0.05) was observed, with azoospermia, in 80% and 70% of animals in the T2 and T3 groups, respectively. At D60, the immunocastrated animals also showed an increase in spermatozoa defects (P < 0.05), whereas plasma testosterone concentration decreased (P < 0.05). Immunocastration of lambs using the Bopriva vaccine at doses of 1.0 and 0.5 mL is efficient in inducing azoospermia in up to 80% of animals, although two doses in a 30-day interval are necessary for it to be an effective and safe method. Efficacy was demonstrated through a reduction in serum testosterone levels, testicular biometry, and seminal fluid analysis. Considering the efficacy of both doses in this study, we recommend using the lower dose (0.5 mL), which will allow for a 50% reduction in vaccine costs.

Shortening the immunocastration-to-slaughter interval from 3 to 2 weeks reduced carcass backfat and weight, increased sexual behaviour and carcass skin blemishes in group-housed male pigs sold at fixed age

Context Pig producers require strategies to temporarily slow growth rate, particularly when markets impose penalties on over-weight and fat carcasses. Immunocastrated pigs usually have higher feed intakes (e.g. during the 3–5-week immunocastration-to-slaughter interval, which is commonly used in Australian pig industry), resulting in higher carcass weight and backfat thickness. Shortening the immunocastration-to-slaughter interval by delaying the second vaccination by a week may be a fine-tuned strategy to reduce carcass backfat and weight of male pigs. However, postponing the age receiving immunocastration as a method to achieve a shorter immunocastration-to-slaughter interval may increase aggressive and sexual behaviours of pigs. Aims The experiment aimed to quantify the effects of shortening the interval between the second dose of immunocastration and slaughter from 3 weeks to 2 weeks on growth performance, carcass traits, aggressive and sexual behaviours, and carcass skin blemishes in group-housed male finisher pigs slaughtered at a fixed age. Methods Pigs (n = 58 pens; 45 pigs per pen) were selected into the experiment at 16 weeks of age. Half of the pens received the second anti-GnRH vaccine (V2) as a means of immunocastration at 19 weeks (a common immunocastration age used in Australian pig industry; control group), and the remaining half of the pens received V2 at 20 weeks of age. All the pigs were slaughtered at 22 weeks of age. Growth performance was recorded between 16 and 22 weeks of age. Behavioural observation was conducted at 20 and 21 weeks of age. Carcass weight, backfat thickness and severity of skin blemishes on carcasses were recorded in the abattoir. Key results Results showed that shortening the immunocastration-to-slaughter interval from 3 to 2 weeks reduced average daily feed intake by 9%, growth rate by 8%, carcass weight by 3%, and backfat thickness by 7% (all P &lt; 0.05), but the percentage of pigs that displayed sexual behaviour and carcasses with &gt;25 lesions both increased (both P &lt; 0.05). Conclusions Shortening the immunocastration-to-slaughter interval from 3 to 2 weeks by delaying V2 is an effective strategy to reduce feed intake, growth rate, carcass weight and backfat in group-housed male pigs, but at a risk of increasing carcass skin blemishes. Implications The increased sexual behaviour and carcass skin blemishes will need to be managed when using a shortened immunocastration-to-slaughter interval to temporarily reduce carcass weight and backfat.

Anti-GnRH vaccination of stallions shedding equine arteritis virus in their semen: a field study

Stallions are natural reservoirs of equine arteritis virus (EAV) in their semen, representing a potential source of outbreaks. The carrier-state is testosterone-dependent, and clears spontaneously in 4 to 40% stallions. Reduction of testosterone secretion may be obtained with the anti-GnRH vaccine Equity. In this report, 16 naturally infected stallions excreting EAV in their semen were vaccinated twice with the vaccine EquityTM and monitored irregularly under field conditions for EAV viral load in their semen and plasmatic testosterone concentration. The results are indicated in months (M) after the first vaccine injection. Testosterone concentrations decreased from 1.7 to 0.2 ng/mL (P&lt;0.002) after 3M. The EAV viral load decreased from 3.2×109 to 1.1×106 RNA copy/mL of semen (P&lt;0.001) after 5M. One stallion died at 7M for other reason. At M3-10, 12/15 stallions ceased to shed the virus in their semen. At M5-10, 9/15 stallions had plasmatic testosterone concentrations of ≥ 0.5 ng/mL but the 6 others showed a persistently low testosterone concentration (≤0.3 ng/mL). Of the 14 stallions that were expected to recover their reproductive activity at the time of the next breeding season (&lt;M12), 8 were EAV negative and produced foals, and 6 were not usable (4 for reproductive deficiency and 2 for EAV positivity). All the stallions were EAV negative at M22, with one stallion being vaccinated a third time at M15. These results suggest that the anti-GnRH vaccination could help to clear EAV shedding in stallions, without a significant effect on reproductive capacity for most of them, but some present a long lasting reduced testosterone secretion.

Testicular features of semi-confined Nellore bulls subjected to immunocastration

This study aimed to evaluate the effect of immunocastration on the scrotal circumference, as well as in the macroscopic and microscopic testicular features in Nellore bulls bred under a semi-confining system for 91 days. Sixty animals were divided into two groups: 30 intact animals (non-vaccinated) and 30 immunocastrated animals. The immunocastrated animals were treated with two anti-GnRH vaccine doses (BoprivaTM - Zoetis Australia Pty Ltd). The scrotal circumference (SC) was measured on days 0 and 56. Testicular parameters: biometry traits (length, width, and height – cm; volume – cm3 ) and weight (g) was measured on day 91. Three testicles from each group were histologically processed to find the seminiferous tubule diameter and epithelium thickness (µm). Data were analyzed in the Minitab® 19 statistical software. The macroscopic features (SC, and testes weight, volume, length, width, and height) and the microscopic features (seminiferous tubule diameter and epithelium thickness) were compared between the two groups through Student’s t-test and Mann-Whitney test. All the values were lower (p&lt;0.05) in animals immunized against GnRH, except for the right testis width and seminiferous tubule diameter. Results indicate that immunization against GnRH affected testicular development.

Effect of vaccination with a novel GnRH-based immunocontraceptive on immune responses and fertility in rats

1. As human-wildlife conflicts increase worldwide, novel methods are required for mitigating these conflicts. Fertility control, based on immunocontraceptives, has emerged as an alternative option to lethal methods for managing wildlife.2. Immunocontraceptives are vaccines that generate an immune response to key components of an animal's reproductive system. Some of these vaccines target the gonadotropin-releasing hormone (GnRH) and have been used successfully as contraceptives for many wildlife species. However, the need to capture animals for treatment limits the field applications of injectable vaccines. The availability of orally delivered immunocontraceptives would increase the breadth of applications of fertility control for wildlife management.3. This study explored a new approach to developing an oral immunocontraceptive, exploiting the bioadhesive and immunologically active properties of killed Mycobacterium avium cell wall fragments (MAF). The MAF was conjugated to a GnRH recombinant protein called IMX294, used as a GnRH-specific immunogen.4. An initial trial using the MAF-IMX294 conjugate provided the first evidence that an orally delivered immunocontraceptive vaccine could generate anti-GnRH antibody titres in laboratory rats.5. Increasing the dose and frequency of vaccine administered to rats, in a second trial, enhanced the immune response, eliciting titres that reduced the proportion of females giving birth. This provided the first evidence of the contraceptive effect of an oral anti-GnRH vaccine.6. Future work is required to further increase the immunogenic effect of the oral vaccine and to establish a dosing schedule that is effective for practical field applications.

Oestrus suppression in a dairy herd by means of anti-GnRH vaccination Improvac®: A prospective field study

The aim of this study was to evaluate the duration of estrus suppression after a double administration of the anti-GnRH vaccine Improvac® (Zoetis Schweiz GmbH, 2800 Delémont) in cows. Furthermore, it should be investigated, if a third administration could prolong the effect of the cycle suppression. A total of 21 cows (more than four weeks post partum) were vaccinated twice, at least 35 days apart, with 2 ml Improvac® (0.4 mg of a GnRH-analogon) subcutaneously on one side of the neck. Over a period of 368 days and in the course of 18farm visits these cows were examined gynecologically and re-vaccinated if they showed signs of estrus behaviour or ovarian activity. After the second vaccination the cycle of the cows was suppressed for an average period of 114 days (59-175 days) and the effect could be prolonged by a booster of the vaccine for another 127days in three cows. Estrus behaviour was absent for a longer period than ovarian activity was. The vaccine was tolerated well: apart from slight swelling at the injection site, no side effects were observed. Our results demonstrate that two immunizations with Improvac® are an easily applicable method for the suppression of cyclic activity in cows for a mean period of 114 days. The duration of cycle suppression was prolonged by a booster of the vaccination.

306 Welfare implications of husbandry procedures impair piglets’ welfare and production, and farms’ economic

Abstract Piglets commonly undergo a set of invasive procedures, which may include surgical castration, tail docking and teeth clipping. These procedures potentially result in pain and stress, which may negatively affect the animals’ health, welfare and production. Our objective was to examine welfare and production parameters of pigs from birth to slaughter. Litters (n = 32 sows; 329 piglets; 3 days after farrowing) were allocated randomly into 4 groups; G1: Surgical castration, tail docking and teeth clipping, without environmental enrichment; G2: same as G1, but meaningful environmental enrichment was provided; G3: Immunocastration (Improvac®), tail docking, teeth clipping, with environmental enrichment; G4: none of the invasive procedures were performed, piglet were vaccinated (Improvac®), and environmental enrichment was provided. Mixed-effects Linear Regression model revealed that standardized slaughter weight significantly increased when invasive procedures were avoided and environmental enrichment was provided (G1: 99.2±1.07, G2: 99.9±1.4, G3: 103.6±.1.58, G4: 106.5±1.6 Kg; P &amp;lt; 0.05). The odds ratio to be weak, dead or injured in the conventional, non-enriched G1, was 89% higher than in G4 (P &amp;lt; 0.05). Hair Cortisol on weaning day, as a marker for chronic stress, significantly decreased when welfare implications were avoided (G3: -32.18%, P = 0.014; G4: -24.5%, P = 0.102). Saliva Cortisol, from weaning to slaughter, as a marker for acute stress, was significantly lower in all enriched groups as compared to the conventional, non-enriched Group1 (Group2: -0.74±0.37, Group3: -1.1±0.35, Group4: -0.98±0.36; Linear Regression). Anti-GnRH vaccine was effective in reducing serum and hair testosterone, and hair DHEA, similar to surgical castration. An economic model based on this study suggests an expected rise of 1.48 to 1.92 billion USD if applied to the US pig industry. In conclusion, replacing surgical castration by anti-GnRH vaccine, avoiding tail docking and teeth clipping, and providing environmental enrichment, are better alternatives that would substantially benefit both the animals and farmers.

Field-testing a single-dose immunocontraceptive in free-ranging male capybara (Hydrochoerus hydrochaeris): Evaluation of effects on reproductive physiology, secondary sexual characteristics, and agonistic behavior

Controlling wildlife populations to mitigate human-wildlife conflicts and the spread of zoonotic diseases is an ever-growing necessity. The objective of this study was to evaluate a single-dose anti-gonadotropin-releasing hormone vaccine (GonaCon, USDA/NWRC, Fort Collins, CO, USA) as a non-lethal alternative for population control in free-ranging, synanthropic male capybara. In addition to infertility efficacy of this treatment, potential effects on the alpha male’s secondary sexual characteristics and agonist behavior need to be assessed because any alterations in these factors could lead to population management failure. The treatment group (n = 3) received 1 mL of the anti-GnRH vaccine, intramuscularly, and the control group (n = 2) a 1 mL sham vaccine. Reproductive behavior and social group dynamics were monitored for 30 days prior to inoculation (June 2017) with continuous observations occurring during the study period. Antifertility effects were assessed by conducting exams of testicular morphology, semen characteristics, and histological analysis (after 270 days via hemi-gonadectomy). Compared to the control group, the testicles of the treated males had severe atrophy (P < 0.05), oligozoospermia and greater numbers of sperm cells in a static developmental phase. Courtship and agonistic alpha male behavior were not altered, and the group's social integrity was maintained. Results indicate there was 100% infertility in capybara males, observed throughout the study period of 18 months, and equally important, the male’s alpha characteristics were not affected by the treatment, which is imperative for successful capybara population control efforts.

Effects of an anti-gonadotrophin releasing hormone vaccine on the morphology, structure and function of bull testes

Castration reduces aggressive and sexual behaviour and provides better carcass quality in bull calves. Vaccination against gonadotrophin-releasing hormone (GnRH) is used as an alternative to surgical castration for the purposes of reducing pain and distress in the animals. Currently, no anti-GnRH vaccine has been authorized for use in cattle in the European Union. The aim of the present study was to assess the effect of an anti-GnRH swine-specific vaccine (Improvac®, Zoetis, USA) on the morphology, structure and function of bull testes. Animals were vaccinated at days 1, 21 and 104 of the experimental period and were classified based on their live weight into the following two groups: LIGHT (172.9 ± 30.00 kg) and HEAVY (323.8 ± 37.79 kg). The scrotal circumference was measured on day 1 and prior to slaughter (day 164). At slaughter, the sperm motility and concentration in the caudae epididymis were assessed. Testes were weighed, measured and examined using ultrasound, and then tissue samples were collected and fixed in formalin. Histological and immunohistochemical studies were performed on the testes to measure the diameter of the seminiferous tubules and assess the testicular cell populations. The results revealed that suppression of testicular development was associated with the use of the Improvac® vaccine, which resulted in a smaller size of the testes and impaired spermatid production. However, the effect of Improvac® was more pronounced and consistent in calves vaccinated at a low live weight than at a heavy live weight, which suggested that vaccination is more effective when calves are vaccinated before or early during puberty. However, testes from calves vaccinated at a low live weight were more prone to the development of intraluminal concretions in the seminiferous tubules.