The overall goal of this research was to verify the involvement of membrane progestin receptor alpha (mPRα) in mediating progestin-stimulated sperm hypermotility in the Atlantic croaker and southern flounder. Sperm motility in Atlantic croaker and southern flounder were tested with both the endogenous progestin, 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) or the selective mPRα agonist, 10-ethenyl-19-norprogesterone (Org OD 02-0). In croaker, the Pi3k/Akt/Pde and ErbB2/Mapk intracellular signaling pathways were examined. The role of mPRα in mediating sperm hypermotility and fertility in southern flounder was also studied. The effects of seasonal hypoxia on sperm motility in croaker were investigated in a field study in the northern Gulf of Mexico in the fall of 2010. Finally, the effects of acidified activator solution (simulating ocean acidification) were studied in the laboratory. In vitro, Org OD 02-0 mimicked the stimulatory actions of 20β-S in inducing sperm hypermotility and intracellular signaling cascades in croaker and flounder sperm, indicating that mPRα is the mediator of progestin signaling in the sperm of these species. In croaker sperm, both the Pi3k/Akt/Pde and ErbB2/Mapk intracellular signaling pathways were shown to be important mediators of progestin-induced sperm hypermotility, suggesting novel functions of G [subscript olf] βγ-subunits in teleost sperm. In flounder sperm, mPRα was shown to be important in mediating sperm hypermotility as only high motility sperm with high expression of mPRα were responsive to progestin stimulation, resulting in higher fertilization success compared to low motility sperm. A single LHRHa injection resulted in increased sperm motility and fertility, associated with an increase in mPRα expression in the sperm plasma membrane. The results also suggest that the mPRα/Acy/cAMP pathway first described in croaker sperm is present in flounder sperm. Field studies of male Atlantic croaker exposed to chronic seasonal hypoxia showed that hypoxia exposure resulted in smaller gonads, lower spermatogenesis, reduced testicular mPRα expression, and in some sites, reduced sperm motility. Studies with croaker sperm using acidified activator solution to simulate ocean acidification indicated that croaker sperm were sensitive to environmental insult. Furthermore, the results suggested that the progestin signaling mechanism is more sensitive to changes in ocean pH levels than the mechanism that controls sperm motility. / text

Made available in DSpace on 2014-06-11T19:35:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-17Bitstream added on 2014-06-13T20:06:41Z : No. of bitstreams: 1 zanetti_es_dr_jabo.pdf: 2115508 bytes, checksum: 9a9404d8a1fa80cf4752b8fd1ff6f56d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O conhecimento das técnicas de reprodução assistida nas espécies selvagens pode ser importante para futuros programas de conservação in situ e ex situ. Nosso objetivo foi estabelecer um protocolo de superovulação para o veado-catingueiro (Mazama gouazoubira). Para tanto, foram realizados dois experimentos: (1) No ano de 2007, seis fêmeas da espécie M. gouazoubira receberam um dispositivo intravaginal de progesterona (CIDR®) por 8 dias seguido por uma aplicação i.m. de 0,5mg de BE no momento da inserção (D-8) e de 265μg de cloprostenol (PGF2α) no momento da retirada (D0). Posteriormente foram divididas em três grupos (n=2): no primeiro os animais receberam 600UI de eCG (Tratamento A), no segundo 300UI de eCG (Tratamento B) e no terceiro 250UI de FSH+PVP. (Tratamento C), todos administrados no dia 4 (D-4). O segundo experimento (2) foi desenvolvido no ano de 2009 e utilizou outras seis fêmeas da mesma espécie, divididas em dois grupos (n=3): o primeiro recebeu CIDR® por 8 dias, seguido por uma aplicação i.m. de 0,25mg de BE no D-8, 700UI eCG no D-4 e 265μg PGF2α no D0 (Tratamento D) e o segundo recebeu CIDR® por 7,5 dias seguido por uma aplicação i.m. de 0,25mg de BE no D-7,5; 130mg de FSH em oito doses iguais (iniciando em D-3 e finalizando em D-0,5) e 265μg de PGF2α no D0 (Tratamento E). Em cada experimento os tratamentos foram delineados no esquema “cross-over”, com um intervalo de 44 dias após a remoção do CIDR®. Todas as fêmeas receberam uma aplicação i.m. de PGF2α 14 dias após a retirada do CIDR®. Um macho fértil foi utilizado para a detecção do estro e fertilização. A eficácia dos Tratamentos foi avaliada pelo comportamento reprodutivo, contagem de CLs e folículos (maiores que 3mm) e colheita de embriões por laparoscopia, sete dias após a primeira cópula. No Experimento 1, ainda... / Knowledge of assisted reproduction techniques for wild animals is useful for future in situ and ex situ conservation programs. The present study aimed to establish a superovulation protocol for brown brocket deer (Mazama gouazoubira) by evaluating the ovulation rate, the presence of functional corpora lutea (CL), the number of embryos and their development stages after superovulation using different treatments. For this, two experiments were realized: (1) in 2007, six female Mazama gouazoubira received an intravaginal device (CIDR) for 8 days, followed by 0.5mg i.m. injection of OB at the time of insertion (D-8) and 265μg of cloprostenol (PGF2α) at the time of removal (D0). Next, the hinds were divided into 3 groups (n=2): group 1 received an i.m. injection of 600IU eCG (Treatment A), group 2 300IU of eCG (Treatment B) and group 3 250IU of FSH+PVP (Treatment C), all on D4 (D-4). The second Experiment (2) was developed in 2009 and also used six hinds from the same species divided into 2 groups (n=3): the first received CIDR for 8 days, followed by 0.25mg i.m. injection of OB on D-8, 700IU of eCG on D-4 and 265μg of PGF2α on D0 (Treatment D) and the second received CIDR for 7.5 days followed by 0.25mg i.m. injection of OB on D-7.5, 130mg of FSH divided into eight equal doses [beginning on D-3 and ending D-0.5] and 265μg of PGF2α on D0 (Treatment E). In each experiment, the treatments were „crossed over‟ with 44 day intervals after CIDR removal. All the hinds received an i.m. injection of PGF2α 14 days after CIDR® removal. A fertile male was used for estrus detection and fertilization. Treatment efficacy was evaluated by reproductive behavior, observation of CL, unruptured follicles (over 3mm) and embryo collection via laparoscopy 7 days after the first copulation. In Experiment 1, feces were collected... (Complete abstract click electronic access below)

Tesis por compendio / [EN] The European eel (Anguilla anguilla, L., 1758) population is in dramatic decline, so much so that this species has been listed as "Critically Endangered" on the Red List of Threatened Species, by the International Union for Conservation of Nature (IUCN). The European eel has a complex life cycle, with sexual maturation blocked in the absence of the reproductive oceanic migration, and an inability to mature in captivity without the administration of hormonal treatments. Even though experimental maturation induces gamete production of both sexes, the fertilization results in infertile eggs, unviable embryos and larvae, which die within a few days of hatching. Therefore, understanding the eel reproductive physiology during maturation is very important if we want to recover the wild eel population. Furthermore, due to its phylogenetic position, representative of a basal group of teleosts, the Elopomorphs, the Anguilla species may provide insights into ancestral regulatory physiology processes of reproduction in teleosts, the largest group of vertebrates. In this thesis, characterization, phylogeny and synteny analyses have given us new insight into the evolutionary history of the reproductive process in vertebrates. The European eel possesses five membrane (mPRs) and two nuclear (nPR or pgrs) progestin receptors. Eel mPRs clustered in two major monophyletic groups. Phylogeny analysis of vertebrate nPRs and PLCz1 (sperm specific protein) places both eel PLCz1 and nPR sequences at the base of the teleost clade, which is consistent with the basal position of elopomorphs in the phylogeny of teleosts. To further resolve the origin of the duplicated eel nPRs, synteny analyses of the nPR neighboring genes in several vertebrate genomes were performed. Phylogeny and synteny analyses allowed us to propose the hypothesis that eel duplicated nPRs originated from the 3R. In order to gain a better understanding of the role of the genes implicated in eel reproduction, analyses of their regulation during experimental maturation were carried out. The change in salinity induced parallel increases in E2 plasma and nuclear estrogen receptor expression levels, revealing a stimulatory effect of salinity on the E2 signalling pathway along the BPG axis, leading to a control of spermatogonial stem cell renewal. Brain and pituitary estrogen receptors may then mediate the stimulation of androgens and steroidogenic enzymes linked to androgen synthesis. Androgen synthesis is not dependent on temperature, but further maturation requires higher temperatures to induce a change in the steroidogenic pathway towards estrogen and progestin synthesis. This is consistent with our studies on estrogen and progestin receptors. In the testis, progestin seems to regulate meiosis through membrane and nuclear progestin receptors, and final sperm maturation seems to be controlled by both estrogen and progestin through the estrogen and progestin membrane receptors. Finally, eel sperm-specific PLCz1 seems to have an important function in spermatozoa by inducing egg activation and temperature may play a role in its regulation, especially during the process of spermiogenesis. This thesis attempts to evaluate the physiological function of the genes involved in eel reproduction during spermatogenesis, and demonstrates that salinity and temperature play crucial roles in the sexual maturation of the male European eel. / [ES] La anguila europea (Anguilla anguilla, L., 1758) está sufriendo un declive dramático y ha sido incluida en la categoría de especies "En peligro crítico" en la Lista Roja de Especies Amenazadas, por la International Union for Conservation of Nature (IUCN). La anguila europea tiene un ciclo de vida complejo, con un bloqueo de la maduración sexual hasta que se produce la migración reproductiva, y no madura en cautividad sin la aplicación de tratamientos hormonales. La inducción de la maduración sexual conlleva la producción de gametos de ambos sexos, pero los resultados de la fertilización son huevos no fértiles, embriones no viables, o larvas que mueren pocos días después de la eclosión. Por tanto, la comprensión de la fisiología reproductiva de la anguila durante la maduración es imprescindible para recuperar sus poblaciones naturales. Además, dada su posición filogenética, como representantes de un grupo basal de los teleósteos, los elopomorfos, las especies del género Anguilla podrían proporcionar nuevas perspectivas sobre los procesos ancestrales de regulación de la fisiología de la reproducción de los teleósteos, el mayor grupo de vertebrados. En esta tesis, los resultados de caracterización, análisis de filogenia y sintenia ofrecen nuevas perspectivas de la historia evolutiva del proceso reproductivo de los vertebrados. La anguila europea posee cinco receptores de progestágenos de membrana (mPRs) y dos nucleares (nPR o pgrs). Los mPRs de la anguila se engloban en dos grandes grupos monofiléticos. Las filogenias de los nPRs y de la PLCz1 (una proteína específica del esperma) sitúan a las secuencias de la anguila de PLCz1 y de nPRs en la base del grupo de los teleósteos, lo que coincide con la posición basal de los elopomorfos en la filogenia de los teleósteos. Para resolver el origen de la duplicidad de los nPRs de anguila, se realizaron análisis de sintenia de los genes próximos a los nPRs. Los análisis de filogenia y sintenia nos permitieron formular la hipótesis de que los nPRs duplicados de la anguila se originaron en la 3° duplicación del genoma que se produjo en teleósteos. Para entender mejor el papel de los genes implicados en la reproducción de la anguila, se hicieron análisis de su regulación durante la maduración experimental. El cambio de salinidad indujo aumentos paralelos del nivel plasmático de E2 y de la expresión de los receptores nucleares de estrógenos, que refleja un efecto estimulador de la salinidad sobre la ruta de señalización del E2 dentro del eje cerebro-hipófisis-gónada, que conlleva el control de la renovación de las espermatogonias indiferenciadas. Los receptores de estrógeno en el eje cerebro-hipófisis-gónada podrían mediar la estimulación de la síntesis de andrógenos y de los enzimas esteroidogénicos unidos a ella. Esa síntesis de andrógenos no depende de la temperatura, pero la continuación del proceso de maduración requiere de temperaturas más altas para inducir un cambio en las rutas esteroidogénicas hacia la síntesis de estrógenos y progestágenos. Esto coincide con nuestros estudios sobre receptores de estrógenos y de progestágenos. En el testículo, los progestágenos parecen regular la meiosis mediante la participación de dos mPRs y un nPR, y la maduración final del esperma parece estar controlada tanto por estrógenos como por progestágenos mediante los receptores de estrógenos y de progestágenos de membrana. Finalmente, la PLCz1 de anguila podría tener una importante función en la activación del huevo inducida por el espermatozoide, y la temperatura podría jugar un papel en su regulación, especialmente durante el proceso de espermiogénesis. Esta tesis intentó evaluar la función fisiológica de los genes implicados en la reproducción de la anguila durante la espermatogénesis, y demuestra que la salinidad y la temperatura juegan papeles cruciales en la maduración sexual de los machos de anguila / [CA] La població d'anguila europea (Anguilla anguilla, L., 1758) està sofrint un declivi dramàtic, i aquesta espècie ha estat inclosa en la categoria d'espècies "En perill crític" en la Llista Roja d'Espècies Amenaçades per la International Union for Conservation of Nature (IUCN). L'anguila europea té un cicle de vida complex, amb un bloqueig de la maduració sexual que es manté fins que es produeix la migració reproductiva, i no madura en captivitat sense l'aplicació de tractaments hormonals. Però, fins i tot quan la inducció de la maduració sexual comporta la producció de gàmetes d'ambdós sexes, els resultats de la fertilització son ous no fèrtils, embrions no viables o larves que moren pocs dies després de l'eclosió. Per això, la comprensió de la fisiologia reproductiva de l'anguila durant la maduració és imprescindible per aconseguir la recuperació de les poblacions naturals d'anguila. A més, donada la seua posició filogenètica com a representant d'un grup basal de teleostis, els elopomorfos, les espècies del gènere Anguilla podrien proporcionar noves perspectives al voltant dels processos ancestrals de regulació de la fisiologia de la reproducció dels teleostis, el grup més nombrós dels vertebrats. En aquesta tesi, els resultats de caracterització i l'anàlisi de la filogènia i la sintènia ofereixen noves perspectives de la història evolutiva del procés reproductiu dels vertebrats. L'anguila europea posseeix cinc receptors de progestàgens de membrana (mPRs) i dos nuclears (nPR o pgrs). Els mPRs de l'anguila s'engloben en dos grans grups monofilètics. L'anàlisi filogenètic dels nPRs i de la PLCz1 (una proteïna específica de l'esperma) de l'anguila respecte a les de la resta de vertebrats situa a les seqüències d'aquestes proteïnes en la base dels grups dels teleostis, la qual cosa coincideix amb la posició basals dels elopomorfos en la filogènia dels teleostis. Per tal de resoldre l'origen de la duplicitat dels nPRs de l'anguila, es realitzaren anàlisis de sintènia dels gens pròxims als dels nPRs en els genomes de diversos vertebrats. Aquests anàlisis ens permeteren formular la hipòtesi de que els nPRS duplicats de l'anguila es van originar en la tercera duplicació del genoma que es va produir en teleostis. Per arribar a entendre millor el paper dels gens implicats en la reproducció de l'anguila, s'analitzà la seua regulació durant la maduració experimental. Els canvis en la salinitat induïren augments en paral·lel del nivell plasmàtic d'E2 i de l'expressió dels receptors nuclears d'estrògens, reflectint un efecte estimulador de la salinitat sobre la ruta de senyalització d'E2 en l'eix cervell-hipòfisi-gònada, que comportaria el control de la renovació dels espermatogonis indiferenciats. Els receptors d'estrògens en l'eix cervell-hipòfisi-gònada podrien, d'aquesta forma, intervindre en l'estimulació de la síntesi d'andrògens i dels enzims esteroidogènics units a la síntesi d'andrògens. Aquesta síntesi d'andrògens no depén de la temperatura, però la continuació del procés de maduració requereix de temperatures més altes per induir un canvi en les rutes esteroidogènics cap a la síntesi d'estrògens i progestàgens. En els testicles, els progestàgens pareixen regular la meiosi mitjançant la participació dels receptors de progestàgens de membrana i nuclears, i la maduració final de l'esperma sembla estar controlada tant pels estrògens com per progestàgens de membrana. Finalment, la PLCz1 específica de l'esperma de l'anguila podria tindre una funció de rellevància en l'activació dels ous induïda pels espermatozoides, i la temperatura podria tindre el seu paper en la regulació d'aquesta, especialment durant el procés de l'espermiogènesi. Aquesta tesi ha avaluat la funció fisiològica dels gens implicats en al reproducció de l'anguila durant l'espermatogènesi, i ha demostrat que la salinitat i la temperatura tenen papers clau / Morini, MAM. (2016). Molecular approaches related to the European eel (Anguilla anguilla) reproductive process [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/68513 / TESIS / Premios Extraordinarios de tesis doctorales / Compendio

Chez les vertébrés, la progestérone (P4) est un progestatif endogène agissant via les récepteurs nucléaires et membranaires de la progestérone pour médier des processus clés du développement et de la reproduction. De nombreuses molécules sont conçues pour mimer l’action de la P4. Ces progestatifs synthétiques et la P4 sont très utilisés en tant que substances pharmaceutiques et certains sont retrouvées dans l’environnement aquatique, posant un risque pour les organismes. Cependant, les informations sont insuffisantes pour évaluer les dangers et les risques de ces substances pour l’environnement. Dans ce travail de thèse, les mécanismes d’action et les effets biologiques des progestatifs sur le poisson zèbre ont été étudiés grâce à une combinaison d’outils in vitro et in vivo de type gène-rapporteur basés sur le mécanisme d’action des perturbateurs endocriniens. Nous avons caractérisé le profil toxicologique complexe de 26 progestatifs vis-à-vis de différents récepteurs nucléaires stéroïdiens humain et poisson zèbre et vis-à-vis de l’expression tissu-spécifique de gènes de la stéroïdogenèse (cyp19a1b dans le cerveau et cyp11c1 dans les interrénales). Nos résultats montrent que certains progestatifs peuvent perturber différentes voies de signalisation chez les larves en développement au niveau moléculaire, cellulaire et tissulaire ainsi qu’au niveau physiologique. Dans l’ensemble, les résultats de cette thèse permettent une meilleure caractérisation des dangers des progestatifs sur différentes cibles endocriniennes. / Among vertebrates, progesterone (P4) is an endogenous progestin acting through the nuclear and molecular progesterone receptor to mediate key processes of development and reproduction. Many molecules are designed to mimic the action of P4. These synthetic progestins and P4 are widely used as pharmaceuticals and some of them are found in this aquatic environment, thus posing risks to aquatic species. However, information is lacking to evaluate the dangers and risks of these substances for the environment. This thesis studied the mechanisms of action and the effects of progestins on zebrafish, using a combination of reporter-gene in vitro and in vivo tools based on the mechanism of action of endocrine disruptors. We characterized the toxicological profile of 26 progestins towards different human and zebrafish nuclear steroid receptors and towards the tissue-specific expression of steroidogenic genes (cyp19a1b in the brain and cyp11c1 in the interrenal cells). Our results show that some progestins can disrupt different signaling pathways in developing larvae at the molecular level, cellular and tissular level, and physiological level. Overall, the results of this thesis allow for a better characterization of the dangers of progestins on several endocrine targets.

Background: Estrogen has been reported to enhance well-being and quality of life during the climacteric phase. In women with an intact uterus estrogen treatment is always combined with progestins in order to protect the endometrium from hyperplasia and malignancies. However, in certain women the addition of progestins causes cyclicity in negative mood symptoms and physical symptoms similar to those encountered during ovulatory cycles in women with premenstrual dysphoric disorder (PMDD). The ovarian hormones estradiol and progesterone have profound effects on a number of neurotransmitter systems in the brain, such as the gamma aminobutyric acid (GABA) system and the serotonergic system. Progesterone metabolites, such as allopregnanolone and pregnanolone (also referred to as neurosteroids) modify the GABAA receptor in the central nervous system (CNS) and enhance GABAergic inhibitory transmission. Neurosteroid sensitivity in human studies can be studied by saccadic eye movement measurements using pharmacodynamic challenges with pregnanolone. Altered neurosteroid sensitivity has been suggested as a possible contributory factor to the progesterone/progestin-induced adverse mood effects of hormone replacement therapy (HRT). There is also evidence of estrogen treatment affecting the serotonergic system in postmenopausal women, although progestin addition has been less well studied. Aims and method: The aim was to investigate whether the negative mood symptoms experienced during the progestin or progesterone phase of HRT were associated with changes in neurosteroid sensitivity, or changes in platelet serotonin uptake site (transporter) and serotonin2A (5-HT2A) receptor binding. The intention was also to investigate whether hormonal changes during the normal menstrual cycle affect these peripheral serotonergic parameters. Postmenopausal women with climacteric symptoms were given HRT in two randomized, double-blinded, placebo-controlled crossover studies. The women received 2 mg estradiol (E2) continuously during 28- day cycles. Synthetic progestins or natural progesterone were added sequentially during the last 14 days, and compared to a placebo addition. Before treatment, as well as during the last week of each treatment cycle the pharmacodynamic response to pregnanolone was assessed using saccadic eye movement measurements. Throughout the studies daily symptom ratings were made. In the study regarding synthetic progestins, platelet serotonin transporter and 5-HT2A receptor binding were assayed before entering the study, as well as during the last week of each treatment cycle. In the study on reproductive women, blood samples were collected for analysis of platelet serotonin transporter and 5-HT2A receptor binding at six different points in time during the menstrual cycle. Results and conclusion: The addition of synthetic progestins to estrogen treatment increased negative mood symptoms and physical symptoms, whereas positive symptoms decreased. The addition of progestins also increased the sensitivity to pregnanolone. The addition of natural progesterone to estrogen treatment increased the sensitivity to pregnanolone. However, in this study the pregnanolone sensitivity was enhanced also during estrogen treatment. Women expressing cyclicity in negative mood symptoms were more sensitive to pregnanolone than women without symptom cyclicity. Thus, it is evident that mood deterioration during HRT is associated with altered neurosteroid sensitivity. Platelet serotonin transporter and 5-HT2A receptor binding did not change during the different treatment conditions in HRT. Thus, we were unable to explain the negative mood changes of HRT by use of these peripheral serotonergic parameters. In the study on reproductive women however, it was clear that the serotonergic variables did change during the menstrual cycle. Binding to the serotonin transporter was higher in the late follicular phase than in the ovulatory, early luteal or mid-luteal phases. Binding to the 5-HT2A receptor was higher in the early follicular phase and the early luteal phase than in the mid-luteal phase. These findings may provide a link between the ovarian steroids, and the GABAergic and serotonergic neurotransmitter systems, which in turn, could explain part of the specific vulnerability that women have for the development of adverse mood effects during HRT, mood and anxiety disorders and for the deterioration of mood so frequently seen during the luteal phase.

Submitted by DIEGO LOBON JIMENEZ FILHO null (jfilho2015@gmail.com) on 2016-11-29T16:57:43Z No. of bitstreams: 1 Tese_Diego_Lobon_Jimenez_Filho.pdf: 3364578 bytes, checksum: 8e70ceb974949881166a72cb68d4f112 (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-11-30T18:48:42Z (GMT) No. of bitstreams: 1 jimenezfilho_dl_dr_jabo.pdf: 3364578 bytes, checksum: 8e70ceb974949881166a72cb68d4f112 (MD5) / Made available in DSpace on 2016-11-30T18:48:42Z (GMT). No. of bitstreams: 1 jimenezfilho_dl_dr_jabo.pdf: 3364578 bytes, checksum: 8e70ceb974949881166a72cb68d4f112 (MD5) Previous issue date: 2016-11-18 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O objetivo geral do estudo foi avaliar os riscos sanitários associados ao uso de fontes exógenas de hormônios sexuais nos programas de sincronização e indução de ovulação “Inseminação Artificial em Tempo Fixo (IATF)”, “Transferência de Embriões em Tempo Fixo (TETF)”, "Superestimulação ovariana (SOV)” sobre a saúde coletiva e meio ambiente. Para isto, o trabalho foi dividido em quatro capítulos. O primeiro capítulo apresenta as considerações gerais sobre legislação, período de carência, equipamentos de proteção individual, grupos de risco e resíduos sólidos de saúde. No segundo capítulo o objetivo foi avaliar o conhecimento e a percepção de risco dos criadores de bovinos e médicos veterinários na utilização de hormônios e determinar os possíveis riscos para a saúde coletiva. Neste experimento foram entrevistados 65 criadores e 40 médicos veterinários que utilizavam hormônios nos programas de reprodução assistida em bovinos (IATF, TETF, SOV). A análise de correspondência múltipla foi processada com os fatores socioeconômicos (escolaridade, tempo na atividade) e a percepção de risco no uso de hormônios reprodutivos. A totalidade dos criadores entrevistados afirmaram que “os estabelecimentos que comercializam produtos veterinários não exigiram prescrição veterinária para a venda dos hormônios”. Os “períodos de carências” na carne e no leite eram desconhecidos por 69,2% dos criadores e por 65% dos médicos veterinários. Dos profissionais que afirmaram conhecer a informação, apenas dois a citaram corretamente. O “uso de equipamentos de proteção individual (EPI)” durante a manipulação dos hormônios foi declarado por 56,9% dos criadores e 92,5% dos médicos veterinários, sendo “as luvas de procedimentos” o EPI mais utilizado. Ao serem questionados sobre “o grupo de pessoas inaptas à manipulação hormonal”, 21,5% dos criadores e 62,5% dos médicos veterinários afirmaram conhecer o grupo de risco. Houve associação entre criadores com “ensino superior” e “tempo na atividade menor que 5 anos” com a variável “conhece o grupo de pessoas que não deve manipular hormônios” (p < 0,01). O fator socioeconômico dos médicos veterinários “mais de 11 anos na atividade” também apresentou associação com a variável “conhece o período de carência” (p < 0,05). O terceiro capítulo teve como objetivo avaliar o conhecimento dos criadores e médicos veterinários sobre o descarte dos resíduos sólidos de saúde (RSS) e avaliar a possível contaminação ambiental durante a lavagem dos dispositivos intravaginais de progesterona. Foram consideradas questões socioeconômicas e sobre o descarte dos RSS. Além das entrevistas, foram realizadas análises de progestinas na água utilizada na lavagem dos dispositivos intravaginais de progesterona. As progestinas foram extraídas por cromatografia líquida de alta eficiência (HPLC) e analisadas por espectrometria de massas sequencial (MS/MS). Dos 65 criadores, 18,5% disseram ter “recebido orientação de como promover o descarte dos RSS”, entretanto, apenas um criador entrevistado promove o “correto descarte desse material”. A prática de queima dos RSS é realizada por 86% dos criadores. A quantidade de progesterona que sobrou no resíduo da água utilizada na lavagem dos dispositivos foi de não detectável a 515 µg/kg. O quarto capítulo traz orientações para manuseio dos hormônios reprodutivos e orientações para o descarte dos resíduos sólidos de saúde gerados durante os programas de reprodução assistida em bovinos. Por fim, conclui-se que existe falta de informação entre criadores e médicos veterinários sobre os períodos de carências dos hormônios, a importância do uso de EPI na sua manipulação, o grupo de pessoas que não deve manipular os hormônios, a exigência da prescrição veterinária para compra dos fármacos e o modo de descarte dos RSS gerados durante os protocolos hormonais. / The overall objective of this study was to investigate health risks associated to manipulation of exogenous sources of sex hormones for ovulation induction and synchronization, such as through “fixed-time artificial insemination (FTAI)”, “fixed-time embryo transfer (FTET)”, "over-stimulation of the ovary (SOV)”; evaluating potential effects on public health and to the environment. For this purpose, we divided this study into four chapters. The first describes current legal considerations, withholding periods, personal protective equipment (PPE), risk groups and health solid wastes. In the second chapter, it was aimed to test the knowledge and perception of cattle raisers and veterinaries on the use of hormones, as well as determining potential risks against public health. In this part, hormone users, among 65 breeders and 40 vet doctors, from a bovine assisted-reproduction program were surveyed (using FTAI, FTET, and SOV methods). Multiple-correspondence analysis was performed on socioeconomic factors as education level and experience time, as well as on respondents’ perception of risk regarding manipulation of reproductive hormones. All of the surveyed farmers said that no requirement of prescriptions by vet facilities to the purchasing of hormones. Surprisingly, meat and milk withholding periods were unknown by 69.2% breeders and 65% veterinarians. Of the professionals who claimed to know the information, solely two of them specified it correctly. PPE use while handling hormones was stated by 56.9% raisers and 92.5% vets, among which surgical gloves was mostly used. When questioned about the inability of some people to manipulate hormones, 21.5% farmers and 62.5% doctors declared being aware of such risk group. Interestingly, one can note an association between higher-education breeders working less than 5 years in the activity to the variable "knowledge of the group risk for hormonal manipulation (p < 0.01). Veterinarians with more than 11 years in the activity were also associated to the variable "knowledge on withholding period" (p < 0.05). The third chapter aims at evaluating the knowledge of veterinarians and breeders on the disposal of health solid waste (HSW), as well as ascertaining potential environmental contamination during washing out of intravaginal progesterone devices. Furthermore, socioeconomic issues regarding the HSW disposal were also taken into consideration. In addition to the surveys, progestin analyses were made in water used in the washing of intravaginal progesterone devices. Progestin was extracted by high-performance liquid chromatography (HPLC) and analyzed by tandem mass spectrometry (MS/MS). From the 65 surveyed farmers, 18.5% declared they had "been oriented on how to dispose HSW"; however, only one performs it properly. Nearly 86% raisers perform the burning of HSW. Nonetheless, the amount of progesterone left in wastewater from washings was of an undetectable level (515 µg/kg). The fourth chapter come out with guidelines on HSW disposal during reproductive-hormone manipulation in bovine assisted-breeding programs. Finally, we may conclude that there is a lack of information among breeders and veterinarians on withholding periods for hormones, PPE use importance during manipulation, group of people unable to manipulate hormones, veterinarian prescription requirement for drug purchasing and disposal manner of HSWs generated during hormone protocols. / CNPq: 142282/2013-8

Orientador: Iveraldo dos Santos Dutra / Resumo: O objetivo geral do estudo foi avaliar os riscos sanitários associados ao uso de fontes exógenas de hormônios sexuais nos programas de sincronização e indução de ovulação “Inseminação Artificial em Tempo Fixo (IATF)”, “Transferência de Embriões em Tempo Fixo (TETF)”, "Superestimulação ovariana (SOV)” sobre a saúde coletiva e meio ambiente. Para isto, o trabalho foi dividido em quatro capítulos. O primeiro capítulo apresenta as considerações gerais sobre legislação, período de carência, equipamentos de proteção individual, grupos de risco e resíduos sólidos de saúde. No segundo capítulo o objetivo foi avaliar o conhecimento e a percepção de risco dos criadores de bovinos e médicos veterinários na utilização de hormônios e determinar os possíveis riscos para a saúde coletiva. Neste experimento foram entrevistados 65 criadores e 40 médicos veterinários que utilizavam hormônios nos programas de reprodução assistida em bovinos (IATF, TETF, SOV). A análise de correspondência múltipla foi processada com os fatores socioeconômicos (escolaridade, tempo na atividade) e a percepção de risco no uso de hormônios reprodutivos. A totalidade dos criadores entrevistados afirmaram que “os estabelecimentos que comercializam produtos veterinários não exigiram prescrição veterinária para a venda dos hormônios”. Os “períodos de carências” na carne e no leite eram desconhecidos por 69,2% dos criadores e por 65% dos médicos veterinários. Dos profissionais que afirmaram conhecer a informação, apenas ... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor