anti-GnRH Vaccine Research Articles

The On the importance of alpha behavior integrity in male Capybara Hydrochoerus hydrochaeris (Mammalia: Rodentia: Caviidae) following immuno-contraceptive treatment

As the human population continues to grow, habitat for wildlife shrinks, driving fauna either into extinction or into new habitats, which can create new problems. In Brazil, the Capybara Hydrochoerus hydrochaeris has become a pest by invading urban and agricultural areas. These mammals quickly multiply owing to abundant food supply and a lack of natural predators, and they can serve as amplifying hosts for Rickettsia rickettsii, the pathogen of potentially life-threatening Brazilian spotted fever. Species-specific population management strategies that respect public opinion and consider animal welfare are required for the effective mitigation of this tick-borne zoonotic disease. In order to control Capybara populations it is necessary to take into account their social dynamics, which are centered on polygynous dominant males with hormone-driven secondary sexual characteristics. To be a viable management tool, a contraceptive strategy targeting these males must preserve their social status to prevent other males from replacing them. As part of a larger research project on the efficiency of anti-Gonadotropin-releasing hormone (GnRH) vaccine treatment in free-ranging Capybaras, the aim of this study was to observe the impact of this treatment on alpha male and overall social group behavior. At the end of the 18-month-study, there were no recorded births involving the immunized animals, and alpha male sexual characteristics and group integrity were preserved. These results encourage the use of this anti-GnRH vaccine as an alternative population control tool in male Capybara.

Brazilian Spotted Fever Prevention through a Nonlethal Capybara Population Control Strategy.

Brazilian spotted fever (BSF), a lethal tick-borne Rickettsioses (2000 - 2018 >600 human deaths) involving synanthropic capybara as host. We introduced an alternative to mitigate human-capybara conflicts and epidemiologic concerns of BSF. Complex aspects like transmission dynamics, risk areas, host mobility, and birth rate control, were considered to develop a prevention strategy using an anti-GnRH vaccine. The propositioned immunocontraceptive potentially remove and prevent the spread of BSF from endemic areas. We propose the anti-GnRH vaccine as a BSF prevention strategy based on these favorable results.

Ovarian function following immunocontraceptive vaccination of mares using native porcine and recombinant zona pellucida vaccines formulated with a non-Freund's adjuvant and anti-GnRH vaccines

An important determinant in the selection of any contraceptive agent is the impact on ovarian function, both in the short and longer term. In this study, ovarian activity was monitored in mares immunised with one of the following vaccine formulations; native porcine zona pellucida (pZP), recombinant zona pellucida proteins ZP3 and ZP4 (reZP), pZP and reZP combined or a commercially available anti-GnRH vaccine. The ZP antigens were prepared in an adjuvant formulation consisting of 6% polymeric adjuvant (Montanide™ PetGel A, Seppic, France) and 500 μg polyinosinic-polycytidylic acid - TLR3-agonist (Poly(I:C) HMW VacciGrade™, Invivogen, USA). A vehicle-only control group was administered the adjuvant formulation without antigen. Ovarian activity was monitored using clinical observations (transrectal palpation and ultrasonography of the reproductive tract) in addition to blood sampling for serum progesterone and anti-Müllerian hormone (AMH) concentrations while employing a low sampling frequency. Treatments and measurements were initiated in December (southern hemisphere summer) and subsequent data collection was performed in January, February, March and May. Both reZP and anti-GnRH vaccination were associated with clinically evident ovarian suppression in the short term. Ovarian activity in mares administered a reZP or anti-GnRH vaccine was significantly different to adjuvant control and pZP treated mares. Serum AMH concentrations were different between pZP and anti-GnRH treated mares 3.5 months after the final vaccination. Serum AMH concentrations were significantly correlated with mare age, serum progesterone and ovarian volume.

Use of anti-gonadotropin-releasing hormone vaccines in African elephants (Loxodonta africana): A review

Background: Androgen-related aggressive behaviour and musth cause serious problems in captive African elephant bulls and often lead to human and animal injuries, and damage to property.Objectives: To review the work carried out with anti-gonadotropin-releasing hormone (GnRH) vaccines to control androgen-related behaviour and fertility in captive and free-ranging elephant bulls and the induction of anoestrus in elephant cows.Method: In the first study, an anti-GnRH vaccine from Pepscan was tested in six bulls (four captive and two free-ranging). Once the vaccine Improvac® became available, the effect on behaviour, the reproductive organs and semen quality was tested. Improvac® was also used to attempt induction of anoestrus in elephant cows.Results: The first study proved that aggressive behaviours are significantly associated with increased faecal androgen concentrations. Musth (n = 1) and aggressive behaviour (n = 2) were down regulated and correlated with a decline in faecal androgen concentrations. Aggression and musth could be controlled with Improvac® (600 µg), but were more consistent when the dose was increased to 1000 µg administered every five to six months. The same dose down regulated testicular function and bulls (n = 17) were rendered infertile within 12 months after commencement of treatment. Initial attempts to induce anoestrous with 600 µg in free-ranging elephant cows gave inconclusive results, but 1000 µg in captive cows delivered five-monthly was successful.Conclusion: The treatment of elephant bulls with Improvac® resulted in the successful down-regulation of androgen-related behaviour and sperm production in captive and wild elephant bulls of various ages (≤ 34 years). Preliminary studies to induce anoestrus in cows with Improvac® appear to be successful.

Immunocastration of goats using anti-gonadotrophin releasing hormone vaccine

The aim of this study was to evaluate the immuno-sterilizing action of anti-gonadotrophin-releasing hormone (anti-GnRH) vaccine in goats. Eighteen male goats were randomly distributed to receive three treatments: T1 (control) - whole animals, and T2 and T3 - application of 0.5 and 1.0 mL of anti-GnRH vaccine, respectively, with six replicates and one goat per experimental unit. Vaccine was administered at 8 months of age and 30 days after the first immunization. Testicular biometry was evaluated monthly, along with seminal collections, for the physical and morphological evaluation of semen. At the time of slaughter, the testicle were collected, and fragments were measured and removed for histological evaluation. The data were evaluated for normality by the Shapiro-Wilk test, followed by appropriate statistical tests for each variable. A reduction in width and length of the right and left testicles was observed and, consequently, the scrotal circumference of the immunized animals reduced after the second vaccine application (P < 0.05). Thirty-days after the first vaccine application, there was a negative effect on seminal production and quality; and 60 days after the second application, a pronounced reduction was observed in all seminal parameters in the vaccinated animals, including azoospermia (83.33% of animals; P < 0.05). Vaccine application reduced testicular weight, seminiferous tubule diameter, and gonadosomatic and tubulosomatic index (P < 0.05), but did not influence the proportion of testicular parenchyma components (P > 0.05). Two applications of the anti-GnRH conjugate are effective for the immunological castration of goats, and the 0.5 mL dose is recommended for use in crossbred goats.

Active immunization against GnRH in pre-pubertal domestic mammals: testicular morphometry, histopathology and endocrine responses in rabbits, guinea pigs and ram lambs

Effective tools for male contraception are important in the control of reproduction in animal populations. The aim of the present study was to evaluate the effects of active immunization against gonadotropin-releasing hormone (GnRH) on male reproductive function assessing testicular morphological changes and serum-gonadotropin levels in pre-pubertal rabbits, guinea pigs and ram lambs. An anti-GnRH vaccine was developed by linking a GnRH-homologous molecule to a tetanus clostridial toxoid (Al(OH)3 coadjuvant). After vaccination protocols testicular morphometry, histopathological alterations and endocrine responses (FSH, LH, testosterone and cortisol serum levels) were evaluated. Testicular volume was significantly reduced in vaccinated animals with respect to the control group in rabbits, guinea pigs and ram lambs (P<0.05 to P<0.001). The anti-GnRH vaccine generated a reduction in testicular volume of 15-, 27- and 11-fold, respectively. Tubule diameters decreased in the vaccinated group with respect to the control ~2.0-, 1.2- and 3.5-fold, respectively (P<0.001). Tubule, intertubular and lumen volumes significantly decreased in vaccinated rabbits (P<0.05), guinea pigs and ram lambs (P<0.01). Vaccinated animals of the three species showed significant reductions in spermatogonial numbers (10- to 40-fold; P<0.01). Sperm was absent in all seminiferous tubules of all rabbits, and most individuals of guinea pigs (80%) and ram lambs (60%). No significant differences were observed between vaccinated and control groups regarding FSH and LH during the experiments in the three experimental species/models used. Testosterone, however, was only significantly lower (~22-fold, P<0.01) in vaccinated rabbits. In conclusion, the present study demonstrated that pre-pubertal active immunization against GnRH leads to endocrine disruption and marked differences on testicular morphometry, development and activity among lagomorphs, hystricomorphs and ovine species with species-specific sensitivity regarding the anti-GnRH immune response.

101 Effects of Active Immunization Against GnRH in Oocyte Donors with Cystic Ovarian Disease

Cows intensively used as oocyte donors for in vitro embryo production (IVEP) are usually kept nonpregnant for prolonged intervals, exposed to successive hormonal treatments, and frequently become overweight. These are all risk factors for the development of endocrine unbalance and, consequently, cystic ovarian disease (COD). The aim of this study was to evaluate the effect of active immunization against gonadotropin-releasing hormone (GnRH) on (1) ovarian follicular population, and (2) development potential of oocytes used for IVEP. Nelore (Bos indicus) cows (n = 14), previously diagnosed with chronic COD (Faria et al. 2017 Anim. Reprod., in press), weighing 620.0 ± 12.8 kg and with body condition score of 4.1 ± 0.2, were assigned to control (n = 6) or treatment (n = 8) group. Cows in the treatment group received 2 SC injections of 1.0 mL of anti-GnRH vaccine (Bopriva, Zoetis, Brazil), 28 days apart (weeks 0 and 4), whereas cows in the control group received placebo on the same schedule. Transrectal ultrasonography was performed weekly from week 0 to evaluate the number and distribution of follicles among size classes, endometrial thickness, and clinical presence of mucometra. Immunization was considered effective (E-IM) when no follicles ≥5.0 mm were observed on the ovaries during a given examination. Cows having E-IM were then used as oocyte donors for IVEP. Cumulus-oocyte complexes (COC) were collected in 5 consecutive ovum pick-up weekly sessions. As a control for IVEP, oocytes from a slaughterhouse were used, with similar procedures performed on the same days and using the same semen batch. The MIXED procedure of SAS (SAS Institute Inc., Cary, NC, USA) with repeated-measures statement was used to evaluate the effects of treatment, time, and interactions on ovarian endpoints; and the GLM procedure was used to analyse embryo production data. Results are shown as mean ± SEM. There were time and time × treatment effects on ovarian parameters. Treated cows had a decrease (P &lt; 0.05) in the average diameter of the largest follicle and in the number of follicles ≥8 mm, and an increase (P &lt; 0.05) in follicular population after week 6. Nonetheless, individual response to treatment was variable: only 50% of the cows (4 of 8) were E-IM at week 8, whereas 25% (2 of 8) still had COD (largest follicle ≥18.0 mm) at this timepoint. Overall, a negative correlation was detected between follicular population and the diameter of the largest follicle (r = –0.60, P &lt; 0.0001) or the number of follicles ≥8 mm (r = –0.47, P &lt; 0.0001). There was no effect (P &gt; 0.05) of treatment on endometrial thickness or mucometra score. Cows with E-IM produced 22.2 ± 3.6 total and 12.9 ± 2.3 viable COC. Cleavage rate did not differ between E-IM and control (slaughterhouse) oocytes (70.8 ± 7.0 v. 75.1 ± 3.0%, respectively; P &gt; 0.05); however, blastocyst rate was greater in the E-IM group compared with controls (39.7 ± 5.5 v. 20.5 ± 4.7%, respectively; P &lt; 0.02). In summary, our results suggest that active immunization against GnRH leads to variable results in the distribution of the follicular population in cows with COD, but it does not negatively affect IVEP efficiency. This research was supported by Zoetis, CNPq, and CAPES.

Strategic use of anti-GnRH vaccine allowing selection of breeding boars without adverse effects on reproductive or production performances

Boar stations raise only entire male pigs for selection as reproductive boars, but the majority of them will fail the selection process, ending at slaughter with a high risk of boar tainted meat. The aim of this study was to investigate whether a single dose of Improvac given to 16-week old boars had a negative effect on their subsequent sperm numbers and motility in 16 artificial insemination boars. We also aimed to generate more knowledge on incidence of boar taint at slaughter among Finnish pigs, compare production performances as average daily gain, feed conversion ratio, and carcass and meat quality (lean meat percentage, back fat, pH, color, androstenone, and skatole) of immunocastrated boars (n = 248) with those of entire boars (n = 268). Moreover, we aimed also to explore whether a fat biopsy taken at 16 weeks of age could already reveal the presence of boar taint compounds and be predictive of boar taint development at slaughter age. We found that 32% of entire boars (Figen Landrace, Figen Large White, and their crossbreed) slaughtered at an age of 25 weeks presented levels of androstenone and/or skatole above the threshold for boar taint in their meat. These boars (control) had higher androstenone and skatole levels in the back fat samples at slaughter (0.77 ± 0.55 and 0.09 ± 0.06 μg/g, respectively, mean ± standard deviation) than those in the immuno group (0.20 ± 0.25 and 0.06 ± 0.03 μg/g, respectively, P < 0.001). A single dose of anti-GnRH vaccine, given at 16 weeks of age, did not affect future sperm numbers and motility of boars selected for artificial insemination. We found no difference in the levels of testosterone, anti-GnRH antibodies titers, testicle morphology, and sperm numbers and motility between the boars vaccinated once, at 16 weeks of age, with anti-GnRH vaccine and the control boars (no vaccination). There were no differences in average daily gain, feed conversion ratio, lean meat percentage, and back fat between the immunocastrated boars and entire boars. Meat from immunocastrated boars had a higher pH and better color than meat from entire boars (P < 0.001), suggesting slightly improved meat quality.

Effects of a GnRH vaccine on the movement and activity of free-living wild boar (Sus scrofa)

Context Fertility control is being promoted as a non-lethal means of managing wildlife populations. We recently evaluated a single-dose injectable immunocontraceptive vaccine (GonaCon™) on captive female wild boar for effectiveness and potential side effects; reproductive output was inhibited for 4–6 years, with no obvious detrimental effects on physiology and behaviour. Aims We injected individual free-living wild boar individuals with the fertility-control vaccine GonaCon™ to examine its effectiveness (measured as raised levels of GnRH antibodies) and looked for potential changes in movement and activity patterns. Methods We trapped, fitted telemetry devices to, and released wild boar individuals living in woodland in the West Midlands region of England between 2006 and 2010. We compared data on movements and activity among 10 adult females treated with the vaccine and 11 controls treated with saline only. We measured anti-GnRH antibody titres in six recaptured boar individuals as an indicator of the effectiveness of the vaccine. Key results Post-treatment GnRH antibody titres varied among the boar individuals; four of five treated sows resampled between 9 and 30 weeks post-injection had antibody titres high enough to block reproduction (detectable at 1 : 32 000–1 : 64 000 dilution). At least three treated females were pregnant at the time of vaccination; there was no subsequent evidence that the vaccine interfered with pregnancy. According to the distances moved per hour over a 24-h cycle and the daily activity cycle in relation to season, there were no differences in the behaviour of treated and control females that were likely to be biologically significant. The behaviour of two treated females monitored soon after vaccination and again 12 months later also showed no major differences. Conclusions Free-living wild boar responded to treatment with a 1.0-mL (1000 µg) dose of an anti-GnRH vaccine and no major adverse effects on activity and movement were subsequently detected. Implications Our results indicated that the vaccine could be more widely evaluated in the field against overabundant or nuisance populations. Such populations are increasingly found in urban areas and parks, where culling may not be an option. We suggest that further refinement of this approach for managing wild boar populations, including development of an oral vaccine, are warranted.

Immunization against GnRH in the male camel (Camelus dromedarius): Effects on sexual behavior, testicular volume, semen characteristics and serum testosterone concentrations

The effect of immunization against gonadotrophin releasing hormone (GnRH) on sexual behavior, total scrotal size, semen characteristics and serum concentrations of testosterone, was evaluated for 24 wks in sexually mature camels (Camelus dromedarius). Eight bull camels were randomly divided into a treatment and control group. Four male camels were immunized using 2 mg GnRH – tandem-dimer conjugated to ovalbumin, (Pepscan Systems, the Netherlands) administered subcutaneously, 4 wks apart. Control male camels received the same amount of saline solution. Significant decline in serum testosterone level was observed in three immunized camels out of four, whereas one camel showed no effect. The testosterone levels reached to <1.0 ng/mL serum by week 4 after booster injection and remained suppressed through the course of the study. The total testicular volume was not affected until the end of the experiment. In treated animals, the sexual behavior negatively affected after the booster injection. Anti-GnRH vaccine had a seriously detrimental effect on the acrosin amidase activity and normal acrosome percentages in treated male camels. It is concluded that the vaccine was effective in reducing serum testosterone levels and libido, and it had a serious harmful effect on the acrosin amidase activity and percentages of spermatozoa with normal acrosome. The immunogen did not affect the total testicular volume.

Immunisation against gonadotrophin-releasing hormone (GnRH) increases growth and reduces variability in group-housed boars

It is generally accepted that entire boars are leaner and more efficient than barrows but that they have poorer meat quality with an increased risk of boar taint. An additional issue in producing boars to meet market specifications is variation in growth performance, which may be exacerbated by high levels of aggression and mounting behaviour in group-housed animals. Immunisation against gonadotrophin-releasing hormone (GnRH), leading to immunological castration, offers a means of reducing aggression and mounting behaviour while maintaining most of the production efficiencies associated with entire boars. A total of 120 control boars, 120 immunised boars and 60 barrows (physically castrated at 5 days of age) were used in a study conducted in two replicates. Each replicate consisted of two pens each of boars, immunised boars and barrows, each pen containing 15 pigs, and two blocks of 15 individually-housed boars and immunised boars. The group-housed pigs had access to two single place electronic feeders per pen. Immunised boars were given the anti-GnRH vaccine (Improvac) at 14 and 18 weeks of age. In the 2 weeks immediately after the secondary immunisation at 18 weeks, average daily gain was greater in the immunised boars than the boars or barrows, particularly in those that were group housed, as indicated by the interaction (P = 0.035). From 18 to 23 weeks of age average daily gain was greater (P = 0.005) in immunised boars than in control boars and barrows and was greater (P &lt; 0.001) in individually- than in group-housed pigs. The average daily gain of group-housed immunised boars and individually-housed control boars were similar over the 5-week period after secondary immunisation (1090 vs 1099 g/day), being ~20 and 15% higher than the group-housed control boars and barrows, respectively. The average daily feed intake was greater (P = 0.011) in immunised boars than in control boars with barrows intermediate. The standard deviation of bodyweight increased with age but was lower (P from 0.032 to 0.09) in immunised boars than in control boars and barrows between 18 and 22 and 17 and 21 weeks of age. Carcass fighting damage and pork pH were higher (P &lt; 0.05) for control boars than for immunised boars or barrows. In conclusion, immunisation against GnRH increased average daily gain and average daily feed intake while decreasing variation in bodyweight and improving carcass and pork quality.

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.

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 μg 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.

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.

Cloning, expression, and purification of a highly immunogenic recombinant gonadotropin-releasing hormone (GnRH) chimeric peptide

To design an anti-gonadotropin-releasing hormone (GnRH) vaccine capable of eliciting strong immunogenicity, a gene fragment encoding a chimeric peptide was constructed using polymerase chain reaction and ligated into a novel expression vector for recombinant expression in a T7 RNA polymerase-based expression system. The chimeric peptide called GnRH3–hinge–MVP contained three linear repeats of GnRH (GnRH3), a fragment of the human IgG1 hinge region, and a T-cell epitope of measles virus protein (MVP). The expression plasmid contained the GnRH3–hinge–MVP construct ligated to its fusion partner (AnsB-C) via an unique acid labile Asp–Pro linker. The recombinant fusion protein was expressed in an inclusion body in Escherichia coli under IPTG or lactose induction and the target peptide was easily purified using washing of urea and ethanol precipitation. The target chimeric peptide was isolated from the fusion partner following acid hydrolysis and purified using DEAE–Sephacel chromatography. The purified GnRH3–hinge–MVP was determined to be highly homogeneous by IEF analysis and the N-terminal sequencing. Further, immunization of female mice with the recombinant chimeric peptide resulted in generation of high-titer antibodies specific for GnRH. The results showed that GnRH3–hinge–MVP could be considered as a candidate anti-GnRH vaccine.

Effects of immunization against GnRH, Melengestrol Acetate, and a trenbolene acetate/estradiol implant on growth and carcass characteristics of beef heifers

A 2 × 3 factorial experiment was conducted to determine the effects of an implant (trenbolene acetate/estradiol or no implant) and method of estrus suppression (immunization against GnRH, melengestrol acetate, or no suppression) on growth performance and carcass characteristics of heifers fed for slaughter. At the start of a 21 -d feed adaption phase, crossbred beef heifers (n = 144, 390 ± 2.8 kg) were given their first dose of an anti-GnRH vaccine or started on melengestrol acetate (MGA). Thereafter, heifers were fed a high-concentrate diet (78% barley grain) for 84 d (Days 0 to 83), received implants on Day 0, a second vaccination on Day 21, and were slaughtered on Days 84 or 85. Implanting increased average daily gain (1.72 vs 1.50 kg/d, P<0.01), feed efficiency (6.02 vs 6.75 kg dry matter intake/kg gain, P< 0.01), preslaughter weight (532 vs 513 kg, P < 0.01), carcass weight (301 vs 289 kg, P < 0.01), and ribeye area (88.6 vs 85.9 cm 2, P < 0.05), but had no affect (P > 0.05) on dry matter intake, grade fat thickness. marbling score, or lean yield. Compared to heifers fed MGA, those immunized against GnRH had a greater ribeye area (90.0 vs 84.6 cm 2) and lean yield (63 vs 61%), and had thinner grade fat (7.5 vs 8.6 mm; P < 0.05 for each). Furthermore, immunized heifers had lower (P < 0.001) plasma progesterone concentrations than control heifers on Days 42, 63 and 83. Heifers fed MGA had less estrus mounting activity (P < 0.05) and lower plasma progesterone concentrations (P < 0.001) than the remaining heifers. Method of estrus suppression did not affect (P > 0.05) preslaughter weight, average daily gain, dry matter intake, feed efficiency, carcass weight, or marbling score. In conclusion, implanting significantly increased growth performance and preslaughter and carcass weights. Compared to heifers fed MGA, immunization against GnRH significantly increased ribeye area and lean yield, and reduced grade fat thickness.

Anti-GnRH vaccine reduces testosterone levels

Anti-GnRH vaccine reduces testosterone levels

The effects of active immunization against gnRH on testicular development, feedlot performance, and carcass characteristics of beef bulls.

The objective was to determine the effects of a recombinant fusion protein anti-GnRH vaccine on testicular development, feedlot performance, and carcass quality of beef bulls. Crossbred beef bulls (n = 58, average weight 306 kg, 9 mo of age), were randomly allocated to two groups and received either an anti-GnRH vaccine (GnRH) or placebo (Control) by intramuscular injection on d 0, 56, and 112. There were group effects (P < 0.01; as a percentage of Control) on testicular weight (53%), daily sperm production (40%), and epididymal sperm reserves (16%). There were group x time interactions (P < 0.0001) for scrotal circumference and serum testosterone concentrations; at slaughter, bulls in the GnRH group had a smaller (P < 0.05) scrotal circumference (28.3 vs 33.9 cm) and lower (P < 0.05) serum testosterone concentrations (2.2 vs 8.6 ng/mL) than those in the Control group. Average daily gain, feed intake, and feed efficiency were not different between treatments during the backgrounding phase (d 0 to 84). During the finishing phase (d 98 to 182), ADG was greater (P < 0.05) for bulls in the Control group (1.69 vs 1.42 kg/d), as was carcass weight (6.9%; P < 0.01). However, GnRH bulls had numerically better feed efficiency (6.12 vs 7.08 kg DMI/kg gain; P < 0.23) and shear force values for ribeye that were 16% lower (P < 0.14) than Control bulls, warranting further investigation. Vaccinating bulls against GnRH suppressed testicular function, with growth and carcass characteristics similar to that expected with steers.

Pregnancy status and feedlot performance of beef heifers actively immunized against gonadotropin-releasing hormone.

The contraceptive effect of active immunization against GnRH was evaluated in beef heifers. Crossbred heifers were randomized by breed and weight (initial weight = 227.3 +/- 1.2 kg) and assigned to one of three treatment groups. Animals in Group IB (n = 48) were actively immunized against GnRH; heifers in Groups NB (n = 48) and NN (n = 49) did not receive the anti-GnRH vaccine. Sixteen weeks after primary immunization, bulls of proven fertility were introduced into pens containing Groups IB and NB. Bulls were maintained with heifers for 2 mo. Heifers in group NN were not intentionally exposed to fertile males. At the end of the breeding period, heifers received Synovex H implants and entered a commercial feedlot. Heifers were slaughtered after 116 d of feedlot confinement. Anti-GnRH titer was evident in all heifers (48 of 48) immunized against GnRH. Gravid uteri were present at slaughter in 40 of 48 (83.3%) NB heifers. In contrast, only four (8.3%) IB and two (4.1%) NN heifers carried gravid uteri at slaughter. Although ADG did not differ between groups during feedlot confinement, ADG during the breeding period was higher (P < .05) in IB heifers than in NN control animals. Dressing percentage and longissimus muscle area were decreased (P < .05) and marbling and quality grade were increased (P < .05) in NB heifers compared with NN control heifers. Carcass traits of IB heifers were intermediate between those of the NB and NN groups. Taken together, these data indicate that active immunization against GnRH reduces the fertility of terminal heifers. These observations suggest that immunoneutralization of GnRH may be an effective management tool that will reduce the incidence of unintended pregnancy in heifers destined for feedlots.