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Successful management of an invasive species requires in depth knowledge of the invader, the invaded ecosystem, and their interactions. The complexity of the species-system interactions can be reduced and represented in ecological models for better comprehension. In this study, a spatially explicit population model was created using the RAMAS software package to simulate the past and future invasion dynamics of the eastern grey squirrel (Sciurus carolinensis) in the fragmented habitat in case study areas in Ireland. This invasive squirrel species causes economic damage by bark stripping forest crops and is associated with the decline of its native congener (S. vulgaris). Three combinations of demographic and dispersal parameters, which best matched the distribution of the species shortly after introduction, were used to simulate invasion dynamics. Future population expansion was modeled under scenarios of no control and two different management strategies: fatal culls and immunocontraceptive vaccination programmes. In the absence of control, the grey squirrel range is predicted to expand to the south and southwest of Ireland endangering internationally important habitats, vulnerable forest crops, and the native red squirrel. The model revealed that region-wide intensive and coordinated culls would have the greatest impact on grey squirrel populations. Control strategies consisting solely of immunocontraceptive vaccines, often preferred by public interest groups, are predicted to be less effective. Complete eradication of the grey squirrel from Ireland is not economically feasible and strategic evidence-based management is required to limit further range expansion. Ecological models can be used to choose between informed management strategies based on predicted outcomes.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-10-18T12:27:24Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_livia_budziarek_eslabao.pdf: 1788001 bytes, checksum: c14f927ac7e6babdc2bd66cf80275556 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-10-23T11:12:08Z (GMT) No. of bitstreams: 2 dissertacao_livia_budziarek_eslabao.pdf: 1788001 bytes, checksum: c14f927ac7e6babdc2bd66cf80275556 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-10-23T11:12:21Z (GMT) No. of bitstreams: 2 dissertacao_livia_budziarek_eslabao.pdf: 1788001 bytes, checksum: c14f927ac7e6babdc2bd66cf80275556 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-10-23T11:12:30Z (GMT). No. of bitstreams: 2 dissertacao_livia_budziarek_eslabao.pdf: 1788001 bytes, checksum: c14f927ac7e6babdc2bd66cf80275556 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-03-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A imunocontracepção é reconhecida como um dos principais novos métodos contraceptivos para o controle e o manejo da fertilidade em diferentes espécies animais. Dentre os potenciais alvos utilizados em vacinas contraceptivas, o hormônio liberador de gonadotrofinas (GnRH) é considerado um dos mais atrativos. O GnRH é um decapeptídeo hipotalâmico que apresenta um papel central na reprodução de mamíferos. Entretanto, devido à sua baixa imunogenicidade, é necessário associar o GnRH com uma molécula carreadora capaz de estimular o sistema imune como, por exemplo, a subunidade B da enterotoxina termolábil de Escherichia coli (LTB). O presente estudo visou avaliar o potencial imunocontraceptivo de duas quimeras LTB/GnRH em camundongos machos da linhagem BALB/c. Os camundongos foram divididos aleatoriamente em oito grupos experimentais. O grupo LTB/GnRH-P recebeu a quimera expressa em P. pastoris. O grupo LTB/GnRH-P/A recebeu a quimera expressa em P. pastoris adsorvida em adjuvante oleoso. O grupo LTB/GnRH-E recebeu a quimera expressa em E. coli. O grupo LTB/GnRH-E/A recebeu a quimera expressa em E. coli adsorvida em adjuvante oleoso. O grupo LTB recebeu a proteína LTB expressa em E. coli. Os grupos Controle e Controle/A não receberam nenhum antígeno vacinal. O grupo Vacina comercial recebeu vacina comercial anti-GnRH utilizada em bovinos. A resposta imune humoral foi avaliada pela técnica de ELISA indireto e o potencial imunocontraceptivo foi avaliado por meio da quantificação de testosterona sérica e das análises histológicas das gônadas dos animais. Os grupos que receberam os antígenos LTB/GnRH (25 μg por vacina) apresentaram níveis de IgG total significativamente superiores quando comparados aos grupos controle, mostrando que ambos os antígenos são imunogênicos. Os grupos LTB/GnRH-P e LTB/GnRH-P/A apresentaram as maiores absorbâncias nos dias 28 (1,11 ± 0,09) e 42 (1,44 ± 0,04), respectivamente. Além disso, o grupo LTB/GnRH-P/A apresentou níveis de testosterona significativamente menores ao controle a partir do dia 28 (192 ng/dl ± 254,55). Os grupos LTB/GnRH-E e LTB/GnRH-E/A também obtiveram os maiores níveis de anticorpos nos dias 28 (0,33 ± 0,005) e 42 (1,44 ± 0,08), respectivamente. O grupo LTB/GnRH-E/A apresentou níveis de testosterona significativamente menores ao controle a partir do dia 28 (<12 ng/dl ± 0). Os resultados da histologia mostraram que ambos os antígenos causam alterações na espermatogênese, sendo que o antígeno expresso em E. coli foi relacionado com as maiores alterações nas gônadas. O presente estudo mostrou quimeras produzidas através da fusão da LTB com uma única molécula de GnRH é capaz de induzir a geração de resposta imune humoral, bloqueio das funções endócrinas relacionadas com a reprodução e alterações teciduais nas gônadas de camundongos machos. / Immunocontraception is recognized as one of the major new contraceptive methods for the control and management of fertility in different animal species. Among the potential targets used in contraceptive vaccines, gonadotropin-releasing hormone (GnRH) is considered one of the most attractive. GnRH is a hypothalamic decapeptide that presents a central role in mammalian reproduction. However, due to its low immunogenicity, it is necessary to associate the GnRH with a carrier molecule capable of stimulating the immune system such as, for example, the B subunit of Escherichia coli heat-labile enterotoxin (LTB). The present study aimed to evaluate the immunocontraceptive potential of two LTB/GnRH chimeras in male BALB/c mice. Mice were randomly divided into eight experimental groups. The LTB/GnRH-P group received the chimera expressed in P. pastoris. The LTB/GnRH-P/A group received the chimera expressed in P. pastoris adsorbed in oil adjuvant. The LTB/GnRH-E group received the chimera expressed in E. coli. The LTB/GnRH-E/A group received the chimera expressed in E. coli adsorbed in oil adjuvant. The Control and Control/A groups did not receive vaccine antigen. The Commercial vaccine group received an anti-GnRH commercial vaccine used in cattle. The humoral immune response was evaluated by indirect ELISA and the immunocontraceptive potential was assessed through the quantification of serum testosterone and the histological analysis of animal’s gonads. The groups that received LTB/GnRH antigens (25 μg per vaccine) presented total IgG levels significantly higher when compared to the control groups, showing that both antigens are immunogenic. The LTB/GnRH-P and LTB/GnRH-P/A groups showed higher absorbances at days 28 (1,11 ± 0,09) and 42 (1,44 ± 0,04), respectively. Furthermore, the LTB/GnRH-P/A group presented testosterone levels significantly lower to the control from day 28 (192 ng/dl ± 254,55). The LTB/GnRH-E and LTB/GnRH-E/A groups also obtained the highest antibody levels at days 28 (0,33 ± 0,005) and 42 (1,44 ± 0,08), respectively. The LTB/GnRH-E/A group presented testosterone levels significantly lower to the control from day 28 (<12 ng/dl ± 0). The results from histology showed that both antigens cause changes in spermatogenesis, wherein the antigen expressed in E. coli was associated with major changes in gonads. The present study showed that chimeras produced through the merge of LTB with a single GnRH molecule is capable to induce the generation of humoral immune response, block of the endocrine functions related to reproduction, and tissue alterations in the gonads of male mice.

The Bureau of Land Management (BLM) maintains a growing number of feral horses on public rangelands. With population growth rates as high as 22% annually, herds are exceeding their carrying capacity and millions of dollars are spent maintaining captured horses in holding facilities awaiting adoption. To manage the feral horse population, the BLM is seeking a contraceptive that is safe, can be remotely delivered, requires only a single administration and is effective for several years. Contraceptive strategies have been developed for feral horses that include hormone implants, chemical intrauterine devices, and immunocontraception. Porcine zona pellucida (pZP) immunocontraceptive vaccines have shown great potential for providing safe, long-term contraception in feral horses. ImmunoVaccine Technologies (Halifax, Nova Scotia, Canada) has developed a liposome encapsulated pZP formulation known as SpayVac™ (SpayVac), which after a single-dose provides multi-year contraceptive efficacy. In a continued effort to optimize the acceptability and efficacy of SpayVac, ImmunoVaccine Technologies developed alternative adjuvant preparations using either killed Mycobacterium butyricum (Modified Freund's Adjuvant; MFA) or a proprietary non-Mycobacterium based adjuvant (IVT) that are proposed to have less of the undesirable side-effects associated with Mycobacterium tuberculosis. Therefore, the objective of this research was to evaluate SpayVac in different adjuvant formulations for efficacy of contraception as measured by pZP titers and estrous cyclicity in treated mares. Domestic mares (n=28) were randomly assigned to four treatments (7 mares per treatment): adjuvant alone or saline (Control) or SpayVac vaccines in one of three adjuvant preparations: IVT or MFA in either an aqueous (MFA aq) or non-aqueous (MFA non-aq) suspension. Pre-immune blood samples were collected from each mare and mares were injected in the neck with a single injection of the Control or SpayVac. Subsequent blood samples were collected at weekly intervals for 26 weeks. Sera were analyzed for pZP titers and progesterone using ELISA. At the conclusion of the study, ovaries were recovered by ovariectomy (16 mares) or at necropsy (12 mares) for histologic analysis and collection of morphometric data and oocytes. Titers for pZP were greater (P<0.05) in IVT and MFA mares compared to Control mares and for MFA compared to IVT mares. Mares vaccinated with MFA aq had greater (P<0.05) pZP titers at 2 weeks post-injection compared to mares injected with IVT or MFA non-aq and at 3 weeks post-injection compared to mares injected with IVT. MFA non-aq mares had greater (P<0.05) pZP titers at 6 weeks post-injection compared to IVT mares and, although not significantly different, titers in MFA non-aq mares remained greater during weeks 8, 10, 14, 18 and 22 compared to IVT and MFA aq mares. Mean serum progesterone concentrations were greater (P<0.05) in Control compared to MFA non-aq mares. Mean ovarian weights, oocyte diameters, zona pellucida thicknesses and the number of horse sperm bound to oocytes recovered from vaccinated mares were greater (P<0.05) in Control mares compared to IVT and MFA mares. As judged by pZP titers and serum progesterone, these results suggest that SpayVac suspended in the MFA non-aqueous formulation exerted the greatest contraceptive effects in treated mares. This preparation of SpayVac may meet the criteria cited by the BLM for their most desirable immunocontraceptive. / Graduation date: 2012

Made available in DSpace on 2014-08-20T13:32:59Z (GMT). No. of bitstreams: 1 dissertacao_juliano_lacava_pereira.pdf: 1194037 bytes, checksum: 9990bc185909cc08288cd6230c0a9be6 (MD5) Previous issue date: 2011-07-18 / Several recombinant proteins have been studied as immunocontraception agents in mammals, among them the Gonadotropin-releasing hormone (GnRH). Nevertheless, due to its low molecular weight, GnRH needs to be conjugated with a carrier protein. Among the eukaryotic platforms the yeast Pichia pastoris, which are able of promoting post-translational modifications to expressed recombinant protein. The aim of this study was to clone and express the GnRH/LTB chimera in P. pastoris, and to test it as an immunocontraceptive. In order to accomplish this, sequences of P. pastoris codons usage were synthesized and cloned into the plasmid pPICZαB. The plasmid containing the sequence were propagated and transformed into yeast by electroporation. The transformed colonies were selected by resistance to antibiotics and confirmed by Colony Blotting. The selected clone was cultivated, induced and the protein expressed in supernatant. The protein was concentrated and used as a vaccine antigen. BALB/c males mice were vaccinated twice 15 apart with 100µg de LTB/GnRH adjuvanted with aluminum hydroxide. The immune response was evaluated by ELISA, showing seraconversion in vaccinated animals. The sperm concentration in the control group was 16.93 x 107 and 8.00 x 107 sperm/mL in the immunized group. Vaccination caused a reduction of spermatogenesis (p < 0,05), vacuolization and disorganization of the seminiferous tubules. We concluded that the yeast was able to express the recombinant protein LTB/GnRH, and it demonstrated immunocontraceptive potential in mice. / Diversas proteínas recombinantes estão sendo pesquisadas para utilização na imunocontracepção de mamíferos. Entre elas, o hormônio liberador de gonadotrofinas (GnRH), entretanto devido ao seu baixo peso molecular há necessidade de associá-lo a uma molécula carreadora. Dentre as plataformas eucarióticas, encontra-se a levedura Pichia pastoris que tem possibilitado expressão e modificações pós-traducionais das proteínas recombinantes. O objetivo deste trabalho foi clonar e expressar a quimera LTB/GnRH em P. pastoris, visando sua utilização como imunocontraceptivo. Para tanto, sequências com os códons preferenciais de P. pastoris da proteína foram sintetizadas e clonada no plasmídeo pPICZαB. O plasmídeo foi propagado e transformado na levedura por eletroporação. As colônias transformadas foram selecionadas por resistência antibiótica e confirmadas por Colony Blotting. O clone selecionado foi cultivado e após indução expressou a proteína no sobrenadante. A proteína foi concentrada e utilizada como antígeno vacinal. Camundongos BALB/c machos foram vacinados com duas doses de 100µg de LTB/GnRH adsorvidas em hidróxido de alumínio com intervalo de 15 dias. A resposta imune foi avaliada por ELISA, mostrando soroconversão nos animais imunizados. A concentração espermática no grupo controle foi 16,93 x 107 e no grupo imunizado 8,00 x 107 espermatozóides/mL. A imunização induziu a redução da espermatogênese (p < 0,05), vacuolização nos túbulos seminíferos e desorganização testicular. Concluiu-se que a levedura foi capaz de expressar a proteína LTB/GnRH recombinante e a mesma apresentou potencial imunocontraceptivo em camundongos.

Contraception through a vaccine has been a very attractive proposition and several attempts were made in the past. To achieve contraception through immunological means, several points need to be considered. First, the targeted antigen should be an important component of reproduction and interference in its actions should lead to infertility. Second, the antigen must be highly immunogenic and the antibodies elicited should be able to block the functions of the antigen. Third, the antibody titres should be effective and must sustain for longer periods. Gonadotropins fulfill all the above criteria and therefore, have been attractive targets for developing human contraceptive vaccines. The pituitary gonadotropins- Luteinizing hormone (LH) and the Follicle stimulating hormone (FSH) are the principal regulators of the reproduction process in all the mammalian species (McLachlan et al., 1995c; Moudgal et al., 1992b; Murty et al., 1979a; Selvaraj and Moudgal, 1994a; Weinbauer et al., 1991). In males, LH binds to its specific receptor-LHR, expressed on the Leydig cells and regulates the production of testosterone. This testosterone binds to the androgen receptors expressed in the Sertoli cells and along with FSH, which binds to the specific receptors present on the Sertoli cell membranes, regulate the testicular functions and the spermatogenesis (Simoni et al., 1997; Themmen and Huhtaniemi, 2000; Ulloa-Aguirre and Timossi, 1998). The well documented studies have unequivocally established that the specific immunoneutralization of either hormone by active or passive immunization, leads to disruption of the gonadal functions (Fraser et al., 1986a; Marathe et al., 1995; Moudgal et al., 1992b; Murty et al., 1979b; Shetty et al., 1996; Srinath et al., 1983b) and consequent infertility and this observation formed the basis of the human contraceptive vaccines (Moudgal et al., 1997b; Talwar et al., 2011a; Talwar et al., 2009a). Several studies using testosterone as the main male hormonal contraception method (Matsumoto et al., 1986; Matsumoto et al., 1983a) and anti-hCG vaccine as the female hormonal contraceptive vaccine reached Phase I and II clinical trials (Talwar, 1997; Talwar et al., 1994; Talwar et al., 1997) . However, these human contraceptive vaccines faced several limitations. There was a need to inhibit only particular segments of the entire reproduction process whereas others needed to remain completely unaffected. For example, in males, the FSH regulated functions, the sperm production and spermatogenesis needed to be inhibited whereas the LH/testosterone associated functions should be unaffected. Similarly in females, the functions of hCG alone, elaborated by the conceptus should be blocked without affecting either LH or FSH regulated functions, thus, maintaining the normal reproductive cycle. This however is a difficult task especially when the antigens share a large degree of homology and common subunits (Pierce and Parsons, 1981). Moreover, the issues relating to the development and sustenance of high titres of the bioneutralizing antibodies were major limitations of these human contraceptive vaccines. Therefore, despite reaching Phase I and II clinical trials, these studies did not progress further. However, the same concept of an immunocontraceptive vaccine involving the neutralization of the functions of the gonadotropins is an extremely attractive strategy for controlling the animal populations where the reproduction process could be inhibited in its entirety. The overgrowing populations of the stray animals such as dogs and cats pose problems unlike those experienced with the human overpopulation. Thus, there is an immediate need to develop the methods of controlling the populations of these animals both in the developed and the developing countries. Whereas, in countries like the US, the major emphasis is on the domestic animals, in countries like India, the populations of the stray animals need to be controlled. The current methods employed for reducing the numbers of these animals include either castration or culling of the animals. These methods are however, traumatic, unsafe and not widely accepted by the society. The animal contraceptive vaccines currently available are mostly GnRH vaccines which have high cost of production, are not safe for animal use and elicit unwanted side effects. Apart from these, the animals need multiple administrations of these vaccines to elicit high and effective antibody titres, mostly with the use of conventional but non-approved adjuvants (Boedeker et al., 2009; McCoy, 1994). As mentioned above, the gonadotropins, by virtue of their ability to control the mammalian reproduction process, are attractive targets for achieving contraception. Moreover, the ease of administration of this vaccine to neutralize the functions of the endogenous circulating hormones makes them ideal targets for developing animal immunocontraceptive vaccines. This method of neutralizing the functions of the gonadotropins is also humane and safe for the animals as opposed to the current methods which are employed to reduce their numbers. However, in case of animal contraception, particularly for strays such as dogs, where large numbers of animals need to be treated, the challenge is to develop a method to sustain the high levels of the bioneutralizing antibodies for prolonged periods preferably with a single administration of the immunogen and without the use of conventional adjuvants such as the Freund’s adjuvant. In the present study, an attempt has been made develop a strategy to achieve a sustained immune response to small quantities of the hormonal antigens, preferably with a single administration of the immunogen resulting in complete disruption of the gonadal function for prolonged periods. To achieve this goal, recent developments in the field of immunology and vaccinology have been employed. This involves targeting of the hormonal antigens to the dendritic cells. Targeting the antigens to the dendritic cells for vaccination is becoming an extremely fascinating strategy and is being used extensively to target the antigens involved in several diseases (Escudier et al., 2005; Frankel et al., 1998; Garcia et al., 2005; Nouri-Shirazi et al., 2000a; Nouri-Shirazi et al., 2000b; Steinman and Germain, 1998). Most antigens are targeted to the dendritic cells by coupling them to the antibodies specific for the receptors expressed on the dendritic cell surface. One such receptor is the DEC205, which is expressed on most of the dendritic cells (Jiang et al., 1995) and is being widely used to develop vaccines and vaccination strategies. Targeting the antigens to the dendritic cells provides advantages such as ability to induce hundred fold higher immune response to very low doses of antigen without the use of any conventional adjuvant (Bonifaz et al., 2004a). Therefore, in the present study, these features of the dendritic cells have been harnessed to target the hormonal antigens (hCG and hFSH) to the canine DEC205 receptor to induce a long-term immune response capable of disrupting the gonadal functions. Towards this goal of delivering hormonal antigens to the dendritic cells, a fragment of the canine DEC205 corresponding to the Cysteine Rich Fibronectin II domain (CR/FNII) was expressed and used to isolate several canine DEC205 specific recombinant antibodies in the form of single chain fragment variable (ScFvs) from the Tomlinson’s and the yeast human ScFv display libraries. From a pool of eight unique ScFvs screened from the Tomlinson’s libraries, three ScFvs namely B3, G10 and H4 were characterized. All these ScFvs could bind to the human DEC205 receptor but not to the mouse DEC205. Their inability to recognise the mouse DEC205 suggested that mouse could not be used as the model system for these studies and therefore, a surrogate model system was needed. As the canine CR/FNII shared a high degree of homology with the rabbit counterpart, adult rabbits have been used as the surrogate model for immunization studies after confirming the binding of the ScFvs to the rabbit dendritic cells. Since the goal of the study was to deliver the hormonal antigens to the dendritic cells, each ScFv was translationally fused to a core streptavidin fragment, thus creating bi-functional agents (ScFv-CS) capable of binding to the dendritic cells and also to any biotin-tagged antigen, thus delivering the antigen to the dendritic cells. Of the three ScFvs, the ScFv-CS-H4 which could bind to the canine CR/FNII with the KD of 25nM was used for demonstrating the ability of the ScFv-hormone complex to elicit the bioneutralizing antibody response. The ScFv-CS-H4-biotin-hCG or hFSH or both were administered to adult male rabbits along with poly IC: LC, a Toll-like receptor agonist and the antibody titres were monitored. It was possible to maintain high titres of the bioneutralizing antibodies for more than one year with a single administration of the immunogen. Testicular histology of the immunized animals showed extensive disruption of spermatogenesis with most of the germ cells being TUNEL positive undergoing apoptosis. There was complete absence of elongated spermatids and sperms in the testis indicating infertility caused by immunization with the gonadotropins. These data show that targeting the hormonal antigens to the dendritic cells leads to long-term infertility with minimal immunization. Although the ScFvs from the Tomlinson’s libraries were able to deliver the hormonal antigens to the dendritic cells and produce robust and sustained antibody response capable of disrupting the gonadal functions, the affinities of these ScFvs to DEC205 were moderate. It was felt that increasing the affinities of the ScFvs could enhance the effect with respect to the dose of the antigen that needs to be administered and the duration until which the high antibody titres could be maintained. Therefore, the yeast human ScFv display library offering higher diversity of the human ScFvs displayed, was screened for high affinity DEC205 specific binders. From a pool of several ScFvs, six unique ScFvs were characterized. The amino acid sequences of all ScFvs followed the Kabat's rules for identifying the complimentarity determining regions of the heavy and the light chains of the antibodies. All these ScFvs were unique in their amino acid sequences. The dissociation constants of all these antibodies for the canine CR/ FNII ranged from 10-9 to 10-11 M which was 20-300 fold higher than the ScFvs obtained from the Tomlinson’s libraries. The best ScFv obtained from this library was ScFv-92 with a KD value of 8 x10-11 M. All these ScFvs were able to deliver the payload antigen to both, the mouse DEC205 over-expressing cells and the bone marrow derived dendritic cells. Mice immunized with yeast display ScFvs also yielded antibody response to very small quantities of the immunogen with the highest antibody titres obtained with the ScFv-92. It was further demonstrated that all ScFvs also activated the cell-mediated immunity with significant increase in the antigen stimulated T cell proliferation. These ScFvs could also deliver the antigen to the human dendritic cells differentiated from the human monocytes in vitro, thus emphasising their utility in human vaccine development. An attempt was also made to develop nanoparticle (NP) based strategies of delivering the antigen to the dendritic cells. The PLGA-NPs, encapsulating hCG and coated with the DEC205 ScFv-92 was able to elicit high antibody response to very low doses of the antigen. This response could be sustained for 120 days and was higher than the response obtained with similar doses of hCG encapsulated NPs or hCG complexed to ScFv-92 alone. Targeting of the NPs also elicited antigen specific T cell response thus, potentiating their use in cell mediated immunity along with humoral immune responses. In conclusion, this approach of delivering the gonadotropins to the dendritic cells resulted in the production of bioneutralizing antibodies that could disrupt the gonadal functions for a prolonged period and can be effectively used in the fields for controlling the animal populations. This method fulfils all the criteria for any animal contraception. This strategy also elicits both T cell mediated and humoral immunity and can thus be used for producing vaccine against viral and parasitic infections. It can also be used for cancer immunotherapy. Another exciting feature of the strategy used in this study is the usage of ScFv-CS which allows the delivery of any biotin tagged antigen to the rodent and human dendritic cells. As discussed above, the methods for controlling the animal populations are expected to be effective, humane, safe, simple, non-surgical, single shot with long lasting effects, cheap, applicable in the fields and widely accepted by different societies. The methods presented in this study fulfill all these criteria and should be effective in controlling populations of different animal species.