Immunological Approaches to Contraception
Barber, M.R. and R.A. Fayrer-Hosken (2000). Evaluation of somatic and reproductive immunotoxic effects of the porcine zona pellucida vaccination. The Journal of Experimental Zoology 286(6): 641-646. ISSN: 0022-104X.
NAL Call Number: 410 J825
Abstract: Immunological, immunocytochemical and fertility analyses were performed to determine the potential toxic side effects of porcine zona pellucida (pZP) vaccinations on target animals, including horses and dogs. The study was designed to determine the effect of antibodies, raised against highly purified pZP, on somatic tissues. Immunocytochemical studies performed with fixed tissues showed that rabbit anti-pZP antiserum did not crossreact with brain, heart, lung, kidney, liver, bladder, stomach, small intestine, large intestine, muscle, skin, spleen, pancreas, or lymph node of either the dog or horse. To determine the effect or oral intake on nontarget animals, female rabbits were fed a contraceptive vaccine containing pZP glycoproteins and the synthetic trehalose dicorynomycolate in drakeol (S-TDCM) adjuvant. Enzyme-linked immunoadsorbent assay (LISA) analyses showed that rabbits fed with the adjuvanted pZP proteins did not develop circulating anti-pZP IgG antibodies that crossreacted with pZP. Furthermore, fertility studies performed on rabbits fed with adjuvanted pZP revealed no significant differences in the number of embryos or stage of the embryos produced between the treated and control animals. Results of these studies suggest that the pZP vaccine delivered to dogs or horses in field studies have no recognizable somatic tissue effects. Moreover, there were no side effects on nontarget animals should they eat the vaccine. This substantiates field trials results about the safety of the pZP immunocontraceptive vaccine.
Descriptors: animals, antibody formation, immunologic contraception, dogs, egg proteins, enzyme-linked immunosorbent assay, horses, immunoenzyme techniques, immunoglobulin G analysis, membrane glycoproteins, rabbits, swine, vaccines, zona pellucida.
Barber, M.R. and R.A. Fayrer-Hosken (2000). Possible mechanisms of mammalian immunocontraception. Journal of Reproductive Immunology 46(2): 103-124. ISSN: 0165-0378.
NAL Call Number: QR180.J64
Abstract: Ecological and conservation programs in ecosystems around the world have experienced varied success in population management. One of the greatest problems is that human expansion has led to the shrinking of wildlife habitat and, as a result, the overpopulation of many different species has occurred. The pressures exerted by the increased number of animals has caused environmental damage. The humane and practical control of these populations has solicited the scientific community to arrive at a safe, effective, and cost-efficient means of population control. Immunocontraception using zona pellucida antigens, specifically porcine zona pellucida (pZP), has become one of the most promising population control tools in the world today, with notable successes in horses and elephants. A conundrum has risen where pZP, a single vaccine, successfully induces an immunocontraceptive effect in multiple species of mammals. This review describes the most current data pertaining to the mammalian zona pellucida and immunocontraception, and from these studies, we suggest several potential mechanisms of immunocontraception.
Descriptors: animals, immunologic contraception, egg proteins, female, glycosylation, human, lectins, male, membrane glycoproteins, oligosaccharides, ovarian diseases, sperm-ovum interactions, adverse effects of vaccinations, zona pellucida.
Barber, M.R., S.M. Lee, W.L. Steffens, M. Ard, and R.A. Fayrer-Hosken (2001). Immunolocalization of zona pellucida antigens in the ovarian follicle of dogs, cats, horses and elephants. Theriogenology 55(8): 1705-1717. ISSN: 0093-691X.
NAL Call Number: QP251.A1T5
Abstract: A comparative evaluation of the location of immunoreactive porcine zona pellucida (pZP) glycoproteins was performed with polyclonal rabbit anti-pZP antibodies on ovarian sections of the dog, cat, horse, and elephant. For this, formalin (light microscopy) and glutaraldehyde (transmission electron microscopy [TEM]) fixed ovarian sections were incubated with antibodies raised against highly purified pZP. Staining patterns were determined with diaminobenzidine (DAB) at the light level. The dog ZP had a distinct staining distribution that is characterized by intense staining around the periphery of the ZP and the oolemma and less dense staining throughout the width of the ZP. In dog follicles that contained multiple oocytes, there were oocytes of identical and dissimilar stages. Cat ovarian sections showed uniform staining of the ZP. Horse results showed uniform staining of ZP and ooplasm, and granulosa cells (GC). Elephant sections showed staining of the ZP with dense staining at the oolemma, as well as staining of the ooplasm. In all species the staining of the ZP was not evident until GC differentiation. In all cases there was no staining of ovarian tissue with control normal rabbit serum. Specific staining patterns of ZP were evaluated by TEM and immunogold staining. The immunogold-linked anti-pZP antibodies stained the ZP matrix in all species. There was staining of ooplasm organelles suggesting that ZP secretion originates from the oocyte of the dog and cat. In addition, follicular and ZP measurements were taken that allowed accurate characterization of follicle stage. These findings suggest that in all four species the ZP is recognized by anti-pZP antibodies and there is also evidence to suggest the possible origins of ZP glycoproteins.
Descriptors: cats, immunologic contraception, dogs, elephants, horses, immunoglobulin g, electron microscopy, ovary, species specificity, zona pellucida.
Courchamp, F. and S.J. Cornell (2000). Virus-vectored immunocontraception to control feral cats on islands: a mathematical model. The Journal of Applied Ecology 37(6): 903-913. ISSN: 0021-8901.
NAL Call Number: 410 J828
Descriptors: cats, wild animals, vertebrate pests, pest control, biological control, population control, integrated control, contraception, baits, vectors, viruses, genetic engineering, mathematical models, islands, genetically modified viral vectors.
Dunbar, B.S., G. Kaul, M. Prasad, and S.M. Skinner (2002). Molecular approaches for the evaluation of immune responses to zona pellucida (ZP) and development of second-generation ZP vaccines. Reproduction Supplement 60: 9-18. ISSN: 1477-0415.
NAL Call Number: QP251.J75 Suppl.
Abstract: It has long been established that there are major variations in both the immunogenicity and antigenicity of native zona pellucida (ZP) proteins. These differences appear to be more pronounced with respect to genetically engineered ZP proteins, which do not have native post-translational modifications (for example glycosylation and sulphation). As the number of animal species that are now included in population management programmes using native porcine zona pellucida (PZP) proteins expands, it is increasingly important to carry out studies to evaluate the immune response variations among different species as well as the individual variation within a species. In an attempt to compare these complex immune responses, we have evaluated antibodies from numerous species immunized with native, genetically engineered ZP and synthetic ZP peptides. Such an immunocontraceptive method could have great potential. These studies are critical not only for the development of predictable immune responses that result in permanent sterilization versus reversible contraceptive effects, but also for predicting which vaccinogens (native ZP protein versus genetically engineered ZP proteins) might have detrimental effects on animal and human populations.
Descriptors: vaccinogens, immunologic contraception, egg proteins, zona pellucida, dogs, membrane glycoproteins, contraceptive vaccines, population control.
Fayrer-Hosken, R.A., H.D. Dookwah, and C.I. Brandon (2000). Immunocontrol in dogs. Animal Reproduction Science 60-61: 365-373. ISSN: 0378-4320.
NAL Call Number: QP251.A5
Abstract: Population control in dogs and cats is an important goal for many groups. Control measures over the years has included surgery, hormonal therapy and more recently immunological control. The current presentation discusses dog population control with an emphasis on immunologic control. Specifically, vaccination with purified zona pellucida (ZP) glycoproteins leads initially to immunocontraception and then to the profound and irreversible changes of immunosterilization. The preliminary studies are extremely encouraging on developing a vaccine for lasting canine population control.
Descriptors: dogs, egg proteins, membrane glycoproteins, population control, methods of sterilization, vaccination.
Ferro, V.A., M.A.H. Khan, V.S. Latimer, D. Brown, H.F. Urbanski, and W.H. Stimson (2001). Immunoneutralisation of GnRH-I, without cross-reactivity to GnRH-II, in the development of a highly specific anti-fertility vaccine for clinical and veterinary use. Journal of Reproductive Immunology 51(2): 109-129. ISSN: 0165-0378.
NAL Call Number: QR180.J64
Descriptors: anti-fertility vaccine, neutralisation, GnRH, active immunization, rats, luteinizing hormone, immunocastration, toxicity.
Fraser, H.M., A. Gunn, R. Borthwick, and A.F. Fraser (1975). Letter: Sterilising by immunisation. The Veterinary Record 96(14): 323. ISSN: 0042-4900.
NAL Call Number: 41.8 V641
Descriptors: dogs, female, gonadorelin, immunization, male, methods of sterilization.
Fromme, B., P. Eftekhari, M. Van Regenmortel, J. Hoebeke, A. Katz, and R. Millar (2003). A novel retro-inverso gonadotropin-releasing hormone (GnRH) immunogen elicits antibodies that neutralize the activity of native GnRH. Endocrinology 144(7): 3262-3269. ISSN: 0013-7227.
NAL Call Number: 448.8 EN2
Descriptors: dependent cancers, vaccines, fertility, receptor, peptides, cells.
Gonzalez, A., A.F. Allen, K. Post, R.J. Mapletoft, and B.D. Murphy (1989). Immunological approaches to contraception in dogs. Journal of Reproduction and Fertility Supplement 39: 189-198. ISSN: 0449-3087.
NAL Call Number: 442.8 J8222 Suppl.
Abstract: The demand for safe, effective and cost-efficient means of pet population control has generated interest in the development of alternatives to surgical gonadectomy. The purpose of this review is to discuss the potential of immunological approaches to fertility control and to outline recent developments that may aid their design. Among the most promising candidates for immunoneutralization is gonadotrophin releasing hormone (GnRH). We have developed a reliable and reproducible GnRH-protein conjugate antigen and demonstrated that immunologically induced infertility is possible in dogs.
Descriptors: immunologic adjuvants, antibody formation, contraception, dogs, pituitary hormone releasing hormones, zona pellucida.
Gorman, S.P., J.K. Levy, A.L. Hampton, W.R. Collante, A.L. Harris, and R.G. Brown (2002). Evaluation of a porcine zona pellucida vaccine for the immunocontraception of domestic kittens (Felis catus). Theriogenology 58(1): 135-149. ISSN: 0093-691X.
NAL Call Number: QP251.A1T5
Abstract: With a seasonally polyestrus breeding structure, the unwanted domestic cat population has proven difficult to control. Various lethal methods have been used in an attempt to lower this population of cats. Recently, humane attempts to control "pest species," such as the feral cat, have focused on immunocontraception. SpayVac is a vaccine that uses antibodies raised against porcine (ZP) antigens to prevent fertilization of the ovum. SpayVac, delivered in a single dose, has been evaluated in fallow deer and several species of seals with greater than or equal to 90% reduction in fertility and no adverse reactions. This study evaluated the effectiveness of SpayVac in reducing fertility in domestic kittens. Thirty female kittens were treated with SpayVac containing either Freund's complete adjuvant (FCA) or alum, or with a control vehicle. Kittens were monitored for side effects, estrus cycling at maturity, and fecundity. Anti-porcine ZP antibodies were quantified by ELISA. Immunohistochemical assays measured the species specificity of the antibodies produced and IgG binding in vivo. Despite high anti-porcine ZP antibody titers, neither formulation of SpayVac prevented estrus cycling at maturity or reduced fecundity. Immunohistochemical assays indicated that antibodies produced by cats treated with SpayVac recognized porcine ZP, but not feline ZP.
Descriptors: cats, contraception, antibody formation, pigs, zona pellucida, vaccine development, vaccination, adjuvants, elisa, immunohistochemistry, ovaries.
Gupta, S.K., N. Srivastava, C.K. Govind, N. Sivapurapu, and G.K. Gahlay (2000). Comparative molecular biology and immunobiology of zona pellucida glycoproteins: fundamentals and applied aspects for contraception. Proceedings of the Indian National Science Academy, Part B: Biological Sciences 66(1): 33-48. ISSN: 0370-0097.
Descriptors: acrosome reaction, fertilization, immunocontraceptive vaccine, sperm-oocyte binding.
Hassan, T., R.E. Falvo, V. Chandrashekar, B.D. Schanbacher, and C. Awoniyi (1985). Active immunization against LHRH in the male mongrel dog. Biology of Reproduction 32(Suppl. 1): 222. ISSN: 0006-3363.
NAL Call Number: QL876.B5
Descriptors: LHRH, immunocontraception, dogs.
Horoz, H., C.S. Konuk, K. Gurbulak, G. Kasikci, M.E.C. Sonmez, and A. Gurel (2000). The effect of intrauterine device (IUD) administration on fertility, serum progesterone, oestradiol 17 beta and uterine endometrium during the induced estrus cycle in the bitch. Veteriner Fakultesi Dergisi Istanbul 26(2): 325-335. ISSN: 0378-2352.
Descriptors: bitches, clinical aspects, endometritis, endometrium, estradiol, female infertility, intrauterine devices, estrous cycle, oestrus, progesterone, pyometra, dogs.
Language of Text: Turkish.
Ivanova, M., M. Petrov, D. Klissourska, and M. Mollova (1995). Contraceptive potential of porcine zona pellucida in cats. Theriogenology 43(5): 969-981. ISSN: 0093-691X.
NAL Call Number: QP251.A1T5
Abstract: The contraceptive potential of solubilized porcine zona pellucida (spZP) was studied in 2 groups of cats after active immunization using slightly different protocols. Cats from Group 1 (n = 3) were immunized with a total of 300 8g spZP divided in 4 sc multisite injections (each of 37.5 8g) given at 10 day intervals followed by a booster 150 days after the initial immunization. Cats from Group 2 (n = 5) were immunized with a total of 400 8g spZP divided in 4 im injections (each of 50 8g) given at 2 wk intervals followed by a booster 92 days after initial immunization. Immunogen was emulsified in Complete Freund Adjuvant for the first dose and in Incomplete Freund Adjuvant for the following 3 doses. The respective controls were immunized in the same manner using only adjuvant and PBS.
Descriptors: cats, contraceptives, zona pellucida, pigs, immunization, antibody formation, binding proteins, immunofluorescence, oocytes, spermatozoa, Bulgaria.
Jewgenow, K., M. Rohleder, and I. Wegner (2000). Differences between antigenic determinants of pig and cat zona pellucida proteins. Journal of Reproduction and Fertility 119(1): 15-23. ISSN: 0022-4251.
Abstract: Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.
Descriptors: cats, contraception, egg proteins, cross reactions, enzyme linked immunosorbent assay methods, fertilization in vitro, immunohistochemistry, lions, rabbits, sperm-ovum interactions, immunology.
Jewgenow, K. and M. Rudolph (2001). Timing and location of zona pellucida synthesis during oogenesis in domestic cats--an ultrastructural immunohistological investigation. Journal of Reproduction and Fertility 57(Suppl.): 23-29. ISSN: 0449-3087.
NAL Call Number: 442.8 J8222 Suppl.
Abstract: The application of zona pellucida proteins for contraception of wildlife and feral animals, including stray cats, has been promoted since it was demonstrated to be effective for free-roaming feral mares. Active immunization with zona pellucida proteins leads to either reversible or irreversible infertility. Therefore, knowledge of the timing and location of zona pellucida synthesis during oogenesis in cats is a key aspect of designing an immunocontraceptive for felids. Domestic cat ovaries obtained after ovariohysterectomy were used to produce a specific rabbit antibody against feline zonae pellucidae. Ultrathin sections (70 nm) of cat ovaries were treated with the anti-zona antibody followed by incubation with gold-labelled anti-rabbit IgG for ultrastructural investigation. The gold label was related to the ultrastructure of oocytes and granulosa cells. Cat follicles at different stages of development were examined. The antibody reacted very specifically with zona pellucida proteins in fully grown oocytes with a compact zona pellucida. In secondary follicles, gold labels were found on the zona pellucida and inside granulosa cells in the vicinity of oocytes. Primary follicles were labelled inside their cubic granulosa cells and on fragments of zona pellucida in the cleft between granulosa cells and the oocyte. Some primordial follicles were characterized by labelling of the granulosa cells. In conclusion, these results indicate that cat zona pellucida is produced exclusively by granulosa cells and not by oocytes. Synthesis of zona pellucida takes place at every stage of follicular development.
Descriptors: antibodies, cats, egg proteins, granulosa cells, immunoglobulin g, immunohistochemistry, electron microscopy, oogenesis, rabbits, zona pellucida.
Lacoste, D., R. St-Arnaud, S. Caron, A. Belanger, and F. Labrie (1988). The rise in testicular androgens during the first days of treatment with an LHRH agonist in the dog can be blocked by aminoglutethimide or ketoconazole. Journal of Steroid Biochemistry 31(6): 963-970. ISSN: 0022-4731.
NAL Call Number: QD426.A1J6
Descriptors: luteinizing hormone releasing hormone, androgens, aminoglutethimide, ketoconazole, receptors, agonists, effects on, serum levels, inhibition, dogs.
Ladd, A., G. Prabhu, Y.Y. Tsong, T. Probst, W. Chung, and R.B. Thau (1988). Active immunization against gonadotropin-releasing hormone combined with androgen supplementation is a promising antifertility vaccine for males. American Journal of Reproductive Immunology and Microbiology 17(4): 121-127. ISSN: 8755-8920.
NAL Call Number: QR180.A53
Abstract: Male rats and rabbits were immunized against gonadotropin-releasing hormone (GnRH) conjugated to tetanus toxoid (GnRH10-TT) using only materials approved for humans. Testosterone (T)-releasing implants or the long-lasting T ester testosterone-17-trans-4-n-butyl-cyclohexane carboxylate (TE) was used as supplemental androgen for maintaining libido. Immunization against GnRH10-TT effectively suppressed fertility (spermatogenesis) in rats and rabbits. Neither T nor TE administration restored fertility. Both androgens were effective in maintaining normal libido in rats. TE, which is not hydrolyzed in rabbits, was less effective in maintaining normal ejaculatory behavior in this species. Active immunization against GnRH could be a convenient and cost-effective method of fertility control in males.
Descriptors: androgens, antibody formation, epididymis, fertility, follicle stimulating hormone, gonadorelin, luteinizing hormone, male, organ weight, prostate, rabbits, rats, seminal vesicles.
Ladd, A., Y.Y. Tsong, A.M. Walfield, and R. Thau (1994). Development of an antifertility vaccine for pets based on active immunization against luteinizing hormone-releasing hormone. Biology of Reproduction 51(6): 1076-1083. ISSN: 0006-3363.
NAL Call Number: QL876.B5
Abstract: Male dogs and cats were immunized against LHRH in order to evaluate the feasibility of an immunological approach to pet contraception. In the first study, dogs were immunized with 100, 500, or 2500 micrograms of LHRH conjugated to tetanus toxoid. A significant decline in serum testosterone (T) levels was observed in all immunized dogs, reaching castration levels in some animals by Week 4 and remaining suppressed in all the immunized dogs through the course of the study. Testicular histology suggested arrest of spermatogenesis (infertility). The effects of "immunological castration" were reversible (study 2): steroidogenesis suppressed by "immunological castration" was restored as antibody titers declined. Effective antibodies were rapidly reinduced in dogs by a single injection of LHRH1-TT. In contrast, the level of antibodies induced in male cats (study 3) was not sufficient for "immunological castration." The conclusion was that active immunization against LHRH could provide a cost-effective, nonsurgical, reversible means to control the fertility of companion animals.
Descriptors: pets, antifertility vaccine, immunologic contraception, gonadorelin, immunization against LHRH, reversible contraception.
Mahi-Brown, C.A., T.T. Huang Jr, and R. Yanagimachi (1982). Infertility in bitches induced by active immunization with porcine zonae pellucidae. The Journal of Experimental Zoology 222(1): 89-95. ISSN: 0022-104X.
NAL Call Number: 410 J825
Abstract: In a study designed to evaluate the contraceptive potential of anti-egg zona pellucida immunization, bitches were injected with isolated and solubilized zonae pellucidae of either the pig or the dog in saline and Freund's adjuvant or with saline and adjuvant alone (controls). They were boosted monthly, and serum samples were collected before the first injection and 10 days after each injection. The titers of anti-zona pellucida antibodies in each serum sample were measured by treating fresh canine oocytes with the serum, then evaluating antibody binding as indicated by indirect immunofluorescence, precipitation of the zona surface, and penetrability of the zonae by spermatozoa in vitro. The bitches were bred when they came into estrus. All three bitches immunized with porcine zonae developed high titers (1:10,000 or more by indirect immunofluorescence) of antibodies that cross-reacted with canine zonae to cause precipitation of the zona surface both in vivo and in vitro and that completely inhibited penetration of the zonae by spermatozoa in vitro. The two bitches immunized with canine zonae developed only low titers, and their sera had little or no effect on treated zonae. The two control bitches did not develop anti-zona antibody. None of the bitches immunized against porcine zonae became pregnant when bred, but one bitch immunized against canine zonae and one control did become pregnant. The bitches immunized with porcine zonae had somewhat abnormal cycles for unknown reasons. Thus, we could not establish with certainty whether the infertility resulted from specific interference with fertilization, as in vitro, or from alterations in ovarian function, or both.
Descriptors: antibodies, cattle, comparative study, dogs, estrus, female, fertility, immunization, oocytes, ovum, zona pellucida.
Mahi-Brown, C.A., R. Yanagimachi, J.C. Hoffman, and T.T. Huang Jr (1985). Fertility control in the bitch by active immunization with porcine zonae pellucidae: use of different adjuvants and patterns of estradiol and progesterone levels in estrous cycles. Biology of Reproduction 32(4): 761-772. ISSN: 0006-3363.
NAL Call Number: QL876.B5
Abstract: To determine the changes in patterns of 17 beta-estradiol and progesterone levels underlying abnormal cycles in bitches immunized with solubilized crude porcine zonae pellucidae (cPZP), to attempt to circumvent these problems by immunizing with a purified zona fraction (pPZP), and to test the effectiveness of different adjuvants, bitches were immunized with cPZP or pPZP 2-6 times with no adjuvant, Freund's adjuvant, alum adjuvant, or the adjuvant CP-20,961. The bitch immunized without adjuvant had a low titer with a normal cycle and fertility. Immunization with cPZP and adjuvant produced moderate to high titers of antizona antibodies and infertility. Bitches with high titers experienced abnormal estrous cycles. Estradiol rose during proestrus, but instead of falling sharply in early estrus as in controls, it remained elevated. Progesterone did not rise. The moderate-titered bitches had normal cycles and steroid patterns. Bitches immunized with pPZP had moderate titers. Cycles were normal after 3 injections, but after 6 injections one bitch had an abnormal cycle. One pPZP-immunized bitch remained fertile but the others were infertile. Alum was the mildest adjuvant, causing no injection site lesions, but the highest titers occurred with Freund's and CP-20,961 adjuvants. All three adjuvants induced titers sufficient to inhibit fertility. Infertility in bitches immunized with PZP may be due to prevention of zona penetration, because their antisera inhibited zona penetration of oocytes by spermatozoa in vitro. However, alterations in ovarian function preventing ovulation and luteinization could be involved in high-titered bitches.
Descriptors: immunologic adjuvants, antibodies, contraception, diamines, dogs, estradiol, blood, estrus, fertility, Freund's adjuvant, immunization, ovum, progesterone, zona pellucida.
Mahi-Brown, C.A., R. Yanagimachi, M.L. Nelson, H. Yanagimachi, and N. Palumbo (1988). Ovarian histopathology of bitches immunized with porcine zonae pellucidae. American Journal of Reproductive Immunology and Microbiology 18(3): 94-103. ISSN: 8755-8920.
NAL Call Number: QR180.A53
Abstract: The ovarian histopathology of bitches immunized with crude (cPZP) or partially purified (pPZP) porcine zona pellucida proteins was examined in order to determine the cause of abnormal estrous cycles. The majority of immunized bitches had ovarian cytes. Those immunized with cPZP had follicular cysts lined with a thin layer of granulosa cells, while in those immunized with pPZP, the cysts were lined by a basement membrane with a clump of luteinized cells. In two bitches immunized with cPZP, oocytes were present only in primordial follicles. Similar abnormalities were not found in a bitch immunized with human serum albumin or in 12 untreated bitches. Oocytes flushed from the oviducts of mated, immunized bitches were degenerating, which may have been a primary cause of infertility in such bitches. Ovaries studied 2-6 weeks after immunization showed no loss of gap junctional communication between oocytes and granulosa cells, nor was any inflammatory reaction seen. IgG was bound to the zona as revealed by fluoresceinated protein A staining of frozen sections of those ovaries. Abnormal estrous cycles in PZP-immunized bitches appear to result from follicular dysgenesis or cyst formation, but the etiology of these conditions is unresolved.
Descriptors: immunologic contraception, dogs, estrus, fertilization, immunization, ovary, ovulation, swine, uterus, zona pellucida.
Mahi, C.A. and R. Yanagimachi (1979). Prevention of in vitro fertilization of canine oocytes by anti-ovary antisera: a potential approach to fertility control in the bitch. The Journal of Experimental Zoology 210(1): 129-135. ISSN: 0022-104X.
NAL Call Number: 410 J825
Abstract: Antisera raised against canine ovaries were found to induce light scattering of the surface of the egg zona pellucida even when diluted 10,000 times, and to delay digestion of the zona by pronase. High concentrations of antiserum were required, however, to inhibit in vitro fertilization of the oocytes. Absorption of the antisera with canine ovaries removed these effects, whereas absorption with liver, uterus and serum did not. These results demonstrate the antigenicity of the canine ovary and suggest the plausibility of an anti-zona pellucida vaccine for bith control in the bitch.
Descriptors: contraception, dogs, female, fertilization in vitro, immune sera, oocytes, ovary, pronase, rabbits, sperm-ovum interactions, zona pellucida.
Mircu, C., H. Cernescu, G.H. Ghize, G.H. Bonca, V. Ardelean, and V. Igna (2001). The influence of immunization with porcine zona pellucida upon bitch ovaries. Acta Veterinaria 51(4): 235-244. ISSN: 0567-8315.
NAL Call Number: 41.8 V6447
Descriptors: immunocontraception, veterinary medicine, dogs, zona pellucida, meiotic competence, ovarian structure.
Navarrete G, Y.M. and C.C.F.d.M.V. Concepcion Univ. (1997). Castracion quimica en perros machos con Digluconato de Clorhexidina al 3% en Dimetil Sulfoxido al 50%. [Chemical castration in male dogs with clorhexidine digluconate at 3% in DMSO [Dymethil Sulfoxide] at 50%]. Dissertation, Concepcion University, Fac. de Medicina Veterinaria: Chillan, Chile. 79 p.
Descriptors: dogs, males, castration, chemicals, animal husbandry methods, canidae, carnivora, gonadectomy, mammals, sex, sterilization, surgical operations.
Language of Text: Spanish.
Sabeur, K., B.A. Ball, T.M. Nett, H.H. Ball, and I.K. Liu (2003). Effect of GnRH conjugated to pokeweed antiviral protein on reproductive function in adult male dogs. Reproduction 125(6): 801-806. ISSN: 1470-1626.
NAL Call Number: QP251.J75
Abstract: This study evaluated the effect of a GnRH analogue conjugated to the cytotoxin, pokeweed antiviral protein (PAP), on reproductive function in adult, male dogs. Four dogs received 0.0042 mg GnRH-PAP kg(-1) hourly for 36 h, and four other dogs received 0.1 mg GnRH-PAP kg(-1) as one bolus injection daily for three consecutive days. One dog received a single bolus (0.1 mg x kg(-1)). Three adult male dogs received GnRH without the PAP conjugate, as controls. Twenty-five weeks after the initial treatment, all treated dogs received 0.1 mg GnRH-PAP kg(-1) as a single administration, whereas dogs in the control group received 0.0045 mg kg(-1) of the GnRH analogue. Serum concentrations of testosterone and LH were determined by radioimmunoassay, and testis size was measured for 9 months after treatment. Stimulation tests (5 microg GnRH kg(-1)) were used to evaluate LH release (-15, 0, 30, 60, 90, 120 min), which was assessed by measuring area under the curve. Serum testosterone concentrations were significantly lower (P<0.05) after treatment in the bolus and hourly groups than in the control group. Testosterone concentrations fell to less than 50 pg x ml(-1) in three of four dogs in the bolus group and one of four dogs in the hourly group by week 8-9 after treatment. Basal LH was lower (P<0.05) in the bolus and hourly groups than in the control group between weeks 0 and 33 after treatment. Treatment with GnRH-PAP reduced (P<0.05) LH release after GnRH stimulation in the bolus and hourly groups compared with the control group. Testis volume was lower (P<0.05) in all treated versus control dogs. In conclusion, administration of the conjugate GnRH-PAP at a 25 week interval resulted in a major disruption of reproductive parameters in male dogs; this effect was maintained for 11-12 weeks after a second injection of GnRH-PAP.
Descriptors: contraception, dogs, gonadorelin, GnRH analogue, testosterone, testis volume.
Saxena, B.B., A. Clavio, M. Singh, P. Rathnam, E.Y. Bukharovich, T.J. Reimers Jr, A. Saxena, and S. Perkins (2003). Effect of immunization with bovine luteinizing hormone receptor on ovarian function in cats. American Journal of Veterinary Research 64(3): 292-298. ISSN: 0002-9645.
NAL Call Number: 41.8 Am3A
Abstract: OBJECTIVE: To determine the effect of immunization with bovine luteinizing hormone receptor (LH-R) on ovarian function of cats. ANIMALS: 9 adult female domestic cats. PROCEDURE: 7 cats were immunized with 0.5 mg of LH-R encapsulated in a silastic subdermal implant (3 x 10 mm); 2 served as control cats. Receptors had 80% specific binding to 125I-human chorionic gonadotropin with a binding capacity of 2,682 pM/mg. Cats received booster injections of LH-R. Cats were induced to ovulate with luteinizing hormone (LH) releasing hormone on day 345. Samples of venous blood and vaginal cells were collected through day 395. Observation of estrus behavior continued until day 516. Serum concentrations of estradiol, progesterone, thyroid gland hormones, LH, and LH-R antibody were determined. RESULTS: LH-R antibody was detected in the sera of immunized cats within 21 days after implantation. Detection of LH-R antibody was associated with suppression of serum progesterone to < or = 0.5 ng/mL during the study period, compared with concentrations of 5 to 10 ng/mL in control cats. Immunized cats did not display signs of estrus. Release of LH after administration of LH-releasing hormone indicated an intact hypothalamic-pituitary axis but poor corpus luteum function. Serum estradiol concentrations remained between 30 to 40 pg/mL in immunized and control cats. With the decrease antibody titers, hormone concentrations returned to a pattern consistent with that during fertility. CONCLUSIONS AND CLINICAL RELEVANCE: Active immunization with LH-R suppressed corpus luteum function in cats. The effect was reversible. An LH-R-based antifertility vaccine may have clinical application in other vertebrates.
Descriptors: cats, drug implants, estradiol, immunization, LH-R, physiology of ovulation, females.
Saxena, B.B., A. Clavio, M. Singh, P. Rathnam, Y. Bukharovich, T. Reimers Jr, A. Saxena, and S. Perkins (2002). Modulation of ovarian function in female dogs immunized with bovine luteinizing hormone receptor. Reproduction in Domestic Animals 37(1): 9-17. ISSN: 0936-6768.
NAL Call Number: SF105.A1Z3
Abstract: Adult female dogs were immunized with 0.5 mg bovine luteinizing hormone receptor (LH-R) encapsulated in a silastic subdermal implant and subsequently with four intramuscular booster injections of 0.1 mg LH-R each. Circulating LH-R antibody was detected in the sera 3 weeks post-implant. The appearance of LH-R antibody was associated with a decline in the serum progesterone concentrations to a range of 0-0.5 ng/ml until day 365 in the immunized dogs in comparison with a range of 5-10 ng in the control animals, suggesting a lack of ovulation and corpus luteum function in immunized dogs. The immunized dogs did not show signs of 'standing heat' and failed to ovulate when induced by LH-RH challenge. Serum oestradiol levels, however, remained in the range of 30-40 pg/ml in both the immunized and the control dogs. With the decline in the antibody titres, the hormonal profile and vaginal cytology returned to a fertile state and the dogs exhibited signs of 'standing heat', as well as vaginal bleeding. Dogs immunized with LH-R did not show any serious metabolic, local or systemic adverse effects. The hypothalamic--pituitary gonadal axis remained intact as indicated by little difference in pituitary LH levels between control and immunized animals, and by the release of LH by LH-RH challenge. These studies demonstrate that active immunization of female dogs with LH-R could immunomodulate ovarian function to cause a reversible state of infertility. It may be postulated that, due to extensive interspecies homology, a recombinant LH receptor-based immunocontraceptive vaccine may also be effective in other vertebrates.
Descriptors: immunologic adjuvants, antibodies, corpus luteum, dogs, drug implants, estradiol, gonadorelin, immunization, luteinizing hormone, ovary, progesterone, LH receptors.
Shafik, A. (1994). Prolactin injection, a new contraceptive method: experimental study. Contraception 50(2): 191-199. ISSN: 0010-7824.
NAL Call Number: RG136.A1C6
Abstract: "Prolactin injection" is presented as a new contraceptive method. The method was tested in dogs. The dogs in the test group were injected with prolactin (PRL) in a dose of 600 micrograms/kg of body weight weekly for 6 months. During this period, the testicles, semen, reproductive hormones, renal function, and serum sodium and potassium were examined periodically. Testicular biopsy was obtained after 3 and 6 months of PRL injection. These investigations were repeated during the 6 months following withdrawal of the drug. Sperm count decreased to azoospermia in 3 months after PRL administration with decrease of sperm motility and increase of abnormal forms. Testicular biopsy showed degenerated seminiferous tubules. Reproductive hormones, renal function, and serum sodium and potassium revealed insignificant change (P > 0.05). Dog mating during the period of PRL administration induced no pregnancy. After 3 months of drug withdrawal, the sperm count normalized and dog mating produced pregnancy; offsprings showed no anomalies. The study demonstrates that PRL administration has the potential to be developed as a reversible male contraceptive.
Descriptors: reversible male contraception, intramuscular injection, dogs, prolactin (PRL), antispermatogenic agents, reduction in sperm count and motility.
Shigeta, M., A. Hasegawa, Y. Hamada, and K. Koyama (2000). Analysis of B cell epitopes of a glycoprotein porcine zona pellucida (pZP1). Journal of Reproductive Immunology 47(2): 159-168. ISSN: 0165-0378.
NAL Call Number: QR180.J64
Abstract: The zona pellucida (ZP) of mammalian oocytes forms an extracellular matrix composed of three major glycoproteins and plays an important role in sperm-zona interactions. As ZP had a strong organ-specific but species-cross-reactive antigenicity and passive or active immunization with ZP antigens could impair fertilization, the possibility of developing a immunocontraceptive vaccine has been extensively studied. Studies on active immunization with porcine ZP (pZP) that contain B cell epitopes and T cell epitopes demonstrated that a temporary infertility could be induced along with the elevation of antibody titers, but it was always associated with ovarian failure. This could be due to the oophoritis by activation of pathogenic T cell immunity. It is the general consideration that any adverse effects by vaccination should be avoided for an immunocontraception. From this point of view, the analysis of B cell epitopes of pZP protein would be helpful for construction of a safe immunocontraceptive vaccine with zona antigens. We determined the amino acid sequence of the B epitope in the pZP1 protein by using a monoclonal antibody (MAb-5H4) that possesses a fertilization blocking ability. In addition, antiserum raised to the epitope sequence was revealed to block in vitro fertilization of homologous animal species.
Descriptors: amino acid sequence, monoclonal antibodies, B lymphocytes, molecular cloning, immunologic contraception, zona pellucida, membrane glycoproteins.
Shivers, C.A., P.M. Sieg, and H. Kitchen (1981). Pregnancy prevention in the dog: potential for an immunological approach. Journal of the American Animal Hospital Association 17(5): 823-828. ISSN: 0587-2871.
NAL Call Number: SF601.A5
Descriptors: immune regulation of fertility, immunocontraception, dogs, prevention of pregnancy, porcine zona pellucida.
Skinner, S., S. Prasad, T. Ndolo, and B. Dunbar (1996). Zona pellucida antigens: targets for contraceptive vaccines. American Journal of Reproductive Immunology 35(3): 163-174. ISSN: 8755-8920.
NAL Call Number: QR180.A53
Descriptors: zona pellucida antigens, contraceptive vaccines, mammals, ovaries, follicular development.
Srivastava, N., R. Santhanam, P. Sheela, S. Mukund, S.S. Thakral, B.S. Malik, and S.K. Gupta (2002). Evaluation of the immunocontraceptive potential of Escherichia coli-expressed recombinant dog ZP2 and ZP3 in a homologous animal model. Reproduction 123(6): 847-887. ISSN: 1470-1626.
NAL Call Number: QP251.J75
Abstract: Dog zona pellucida glycoprotein 2 (dZP2), excluding the N-terminal signal sequence and the C-terminal transmembrane-like domain, was cloned and expressed as a polyhistidine fusion protein in Escherichia coli to evaluate the immunocontraceptive efficacy of ZP glycoproteins. The recombinant dZP2 (rec-dZP2) revealed a 70 kDa band corresponding to the full length transcript, as well as several low molecular mass fragments in western blot analysis. In addition to rec-dZP2, E. coli expressed recombinant dog ZP glycoprotein 3 (rec-dZP3), which has also been evaluated for its efficacy to block fertility in a homologous system. Three groups of female dogs (n = 4 per group) were immunized with rec-dZP2 conjugated to diphtheria toxoid (rec-dZP2-DT), rec-dZP3 conjugated to DT (rec-dZP3-DT) and DT alone. Immunization of female dogs with rec-dZP2-DT and rec-dZP3-DT led to generation of antibodies against the respective ZP proteins as well as to DT. Subsequent to mating, the four female dogs immunized with rec-dZP2-DT all conceived, which is indicative of failure of the anti-rec-dZP2 antibodies to block fertility. In the group of dogs immunized with rec-dZP3-DT, three of four animals did not conceive when mated with males of proven fertility. The block in fertility was associated with anti-dZP3 antibody titres. Ovarian histopathology revealed that the block in fertility in the group immunized with rec-dZP3-DT is probably manifested by inhibition in the development of follicles and is due to atretic changes in the zona pellucida. These results, although preliminary, indicate that immunization with dZP3 may be a feasible proposition to control dog populations provided that adequate antibody titres are achieved.
Descriptors: immunologic contraception, dog population control, zona pellucida, dZP2, Escherichia coli, membrane glycoproteins.
Talwar, G.P. and R.K. Naz (1981). Immunological control of male fertility. Archives of Andrology 7(2): 177-185. ISSN: 0148-5016.
NAL Call Number: QP253.A54
Abstract: A notable feature of the male gametes is the presence in them of proteins that are "foreign" to the immune system of both male and the female. It is there that are considered responsible for the elicitation of auto- and iso-antibodies in certain natural infertility cases. By virtue of their dual application in both sexes the sperm antigens have interesting potential for exploration as possible agents for control of fertility.
Descriptors: fertility control, immunologic contraception, BCG vaccine, dogs, male, oligospermia, drug effects on sperm motility, sperm antigens.
Thompson, D.L.Jr. (2000). Immunization against GnRH in male species (comparative aspects). Animal Reproduction Science 60/61: 459-469. ISSN: 0378-4320.
NAL Call Number: QP251.A5
Descriptors: GnRH, immunization, livestock, animal behavior, fertility, neutralization, odours, testes, male animals, reproduction, aggressive behavior, immunocastration.
Verdier, Y., N. Rouet, G. Farr, and F. Bou (2001). Identification of antigenic fox spermatozoa surface proteins for use in a contraceptive vaccine. Journal of Andrology (Supplement): 158. ISSN: 0196-3635.
Descriptors: immunocontraception, contraception method, immunologic method, contraceptive vaccine development, fertility control.
Notes: Meeting Information: VIIth International Congress of Andrology, Montreal, Canada, June 15-19, 2001.
Verhage, H.G., A.T. Fazleabas, P.A. Mavrogianis, M.B. O'Day-Bowman, A. Schmidt, E.B. Arias, and R.C. Jaffe (1997). Characteristics of an oviductal glycoprotein and its potential role in fertility control. Journal of Reproduction and Fertility Supplement 51: 217-226. ISSN: 0449-3087.
NAL Call Number: 442.8 J8222 Suppl.
Abstract: At the time of ovulation the lining epithelium of the mammalian oviduct consists of columnar ciliated and secretory cells. These mature cells are dependent on ovarian steroids in carnivores. Oestradiol induces differentiation of these cells and maintains their mature functional state, and progesterone induces dedifferentiation. The secretory cells synthesize and secrete an oestrogen-dependent high molecular weight glycoprotein. The cDNAs encoding oviductal glycoproteins from several species have been sequenced and show high similarity. The human cDNA hybridized with a single message on northern blots of total oviduct RNA obtained from oestradiol-treated cats (about 2.3 kb) and dogs (about 2.1 kb). This glycoprotein is the major nonserum protein present in the oviductal lumen at the time of ovulation, fertilization and early embryonic development. The glycoproteins associate with the zona pellucida of oviductal eggs in all species studied to date. Recent studies suggest that the bovine glycoprotein facilitates sperm capacitation and significantly increases the ability of bovine spermatozoa to fertilize bovine oocytes in vitro, that the hamster glycoprotein increases the sperm penetration rate of the zona pellucida by three times and that the human glycoprotein increases sperm binding to the zona pellucida by three times. All of the evidence for a biological function for this glycoprotein is derived from studies performed in several different species at reproductive stages before fertilization. The biological actions of this glycoprotein suggest a potential role for the glycoprotein in fertility control. Specifically, purified or recombinant glycoprotein may improve success in IVF procedures by enhancing binding of spermatozoa to the zona pellucida and improving fertilization rates. The glycoprotein may also be a potential immunocontraceptive target since antibodies generated against the oviductal glycoprotein may prevent fertilization by preventing binding of spermatozoa to the zona pellucida.
Descriptors: fertility, glycoproteins, animals, species specificity, sperm-ovum interaction, zona pellucida, immunocontraception.
Wango, E.O. and S. Gombe (1995). The effect of immunising dogs against an androgen binding cauda epididymal antigen (caba). Discovery and Innovation 7(3): 265-281. ISSN: 1015-079X.
Descriptors: ammonium sulfate precipitation, antibody, fertility control, immunocontraceptive vaccine, protein, testosterone, domestic and wild animals, epididymal proteins.
Zhu, X. and R.K. Naz (1999). Comparison of ZP3 protein sequences among vertebrate species: to obtain a consensus sequence for immunocontraception. Frontiers in Bioscience: A Journal and Virtual Library 4: D212-D215. ISSN: 1093-4715.
Abstract: The deduced ZP3 amino acid (aa) sequences of 13 vertebrate species namely mouse, hamster, rabbit, pig, porcine, cow, dog, cat, human, bonnet, marmoset, carp, and frog were compared using the PILEUP and PRETTY alignment programs (GCG, Wisconsin, USA). The published aa sequences obtained from 13 vertebrate species indicated the overall evolutionarily conservation in the N-terminus, central region, and C-terminus of the ZP3 polypeptide. More variations of ZP3 polypeptide sequences were seen in the alignments of carp and frog from the 11 mammalian species making the leader sequence more prominent. The canonical furin proteolytic processing signal at the C-terminus was found in all the ZP3 polypeptide sequences except of carp and frog. In the central region, the ZP3 deduced aa sequences of all the 13 vertebrate species aligned well, and six relatively conserved sequences were found. There are 11 conserved cysteine residues in the central region across all species including carp and frog, indicating that these residues have longer evolutionary history. The ZP3 aa sequence similarities were examined using the GAP program (GCG). The highest aa similarities are observed between the members of the same order within the class mammalia, and also (95.4%) between pig (ungulata) and rabbit (lagomorpha). The deduced ZP3 aa sequences per se may not be enough to build a phylogenetic tree.
Descriptors: amino acid sequence, Callithrix, cats, cattle, consensus sequence, dogs, egg proteins, fishes, hamsters, human, Macaca radiata, membrane glycoproteins, mice, rabbits, cell surface receptors, swine, Xenopus, zona pellucida.