A exibição do comportamento vocal em muitas espécies de anfíbios anuros é relacionada aos níveis de hormônios esteroides gonadais e interrenais. Esses hormônios poderiam mediar a relação entre intensidade de sinais e imunidade, pois estão envolvidos no desenvolvimento das características sexuais secundárias, comportamento de corte e mobilização de reservas energéticas durante a atividade reprodutiva, enquanto apresentam também efeitos imunomoduladores. Nesse sentido, o objetivo deste trabalho foi explorar as relações entre comportamento reprodutivo, imunidade e níveis plasmáticos de testosterona e corticosterona em machos do sapo do semiárido brasileiro, Rhinella granulosa, em atividade reprodutiva e após manipulação hormonal. A precipitação foi o principal determinante ambiental para o aumento dos níveis de testosterona e corticosterona circulantes em machos de R. granulosa, estimulando o comportamento de vocalização. As relações fisiológicas encontradas indicam que os altos níveis plasmáticos de testosterona nos primeiros dias após a chuva devem promover o início do turno vocal, porém a corticosterona deve modular o esforço de vocalização. De forma geral, a exacerbação do comportamento vocal de R. granulosa tem efeitos negativos sobre a imunocompetência, porém alguns indivíduos que apresentam maiores concentrações plasmáticas de corticosterona apresentam concomitantemente alto esforço vocal e alta imunidade. De acordo, a aplicação transdérmica de corticosterona promoveu elevação aguda dos níveis plasmáticos deste glicocorticoide, bem como um aumento da função imune. Assim, apesar de a atenção principal ser comumente colocada no papel da testosterona na mediação de sinais honestos, nossos resultados corroboram a importância da corticosterona na mediação da expressão do comportamento de corte e imunocompetência em machos R. granulosa / In most anuran species, vocal behavior is usually related to plasmatic levels of gonadal and interrenal steroid hormones. These hormones could mediate the relationship between signal intensity and immunocompetence, since they are involved in the development of sexual traits, courtship behavior and energetic mobilization during reproductive activity and also exert immunomodulatory effects. The aim of this study was to explore relations among vocal behavior, immunity and plasmatic levels of testosterone and corticosterone in males of a toad species from Brazilian semi-arid, Rhinella granulosa, during reproductive activity and after hormonal manipulation. Precipitation was the main environmental determinant to increase circulating levels of testosterone and corticosterone in R. granulosa males, stimulating vocal behavior. Physiological relations indicate that high circulating testosterone levels in the days right after rain may promote the initiation of vocal behavior, while corticosterone may modulate vocal effort. In general, exacerbation of vocal behavior in R. granulosa has negative effects on immunocompetence, however, some individuals with higher plasmatic levels of corticosterone can present both high vocal effort and good immunocompetence. Corroborating this interpretation, acute increase in glucocorticoid plasmatic levels after hormonal manipulation was accompanied by raised immune function. Despite the emphasis on androgens influence on sexual selected male traits, our results corroborate greater influence of corticosterone in the mediation of expression of courtship behavior and immunocompetence in R. granulosa

De acordo com o modelo de seleção intersexual mediado por parasitas, variações nas condições de ornamentos ou taxa de displays sexuais refletem diferentes graus de resistência ao parasitismo. Em coros naturais de anfíbios, vocalizações mais altas, longas e complexas e/ou emitidas a maiores taxas são, provavelmente, mais facilmente detectadas e atraem mais fêmeas. Entretanto, o esforço vocal está positivamente relacionado aos níveis plasmáticos de esteróides potencialmente imunossupressores, tais como testosterona e corticosterona. O objetivo deste trabalho foi avaliar as relações entre taxa de vocalização, imunocompetência e níveis plasmáticos de corticosterona em Hypsiboas albopunctatus, um anuro de médio porte que ocorre na América do Sul. Tais relações foram estudadas a partir de observação focal do comportamento vocal em coro natural, seguido de coleta de sangue para dosagem de corticosterona e avaliação de alguns parâmetros imunitários. O presente estudo demonstrou uma relação positiva entre a taxa de vocalização com níveis plasmáticos de corticosterona e negativa entre estes dois parâmetros com a imunocompetência mediada por células. Estes resultados apontam para a corticosterona como possível hormônio mediador do compromisso entre esforço vocal e imunidade mediada por células em anuros. / According to the model of intersexual selection mediated by parasites, changes in conditions of sexual ornaments or display rates reflect different degrees of resistance to parasitism. In natural choruses of amphibians, calls that are higher, longer, more complex and / or issued at higher rates are probably more easily detected and attract more females. However, the vocal effort is positively related to plasma levels of potentially immunosuppressive steroids, such as testosterone and corticosterone. The aim of this study was to evaluate the relationship between calling rates, immunocompetence and plasma levels of corticosterone in Hypsiboas albopunctatus, a midsize anuran occurring in South America. Such relationships were studied from focal observations of calling behavior, followed by blood collection for measurements of corticosterone plasma levels and evaluation of some immune parameters. The present study showed a positive relationship between the calling rate with plasma levels of corticosterone, and a negative correlation between these two parameters with cell-mediated immunocompetence. These results highlight the importance of the hormone corticosterone as a possible mediator of trade-off between vocal effort and cell-mediated immunity in anurans.

En utilisant comme cas d’étude le mouflon Méditerranéen (Ovis gmelini musimon × Ovis sp.), les objectifs de cette thèse étaient de mieux comprendre comment sont liés diversité génétique, comportement des individus, flux de gènes et dynamique parasitaire. Au travers d’approches de génétique des populations et de génétique du paysage, nous avons pu mettre en évidence que la structure génétique spatiale de la population étudiée était impactée par son histoire d’introduction, sa structure socio-spatiale et le paysage dans lequel elle évolue. Etant donné l’impact de ces divers éléments sur les flux de gènes des mouflons, nous nous attendions à ce qu’ils déterminent également les flux de parasites dans la population. Nos résultats ont, au contraire, révélé que les parasites circulent mieux que les gènes de mouflons dans la population. Enfin, nous avons montré que les capacités de résistance des hôtes face à leurs parasites dépendaient de la diversité génétique neutre et adaptative, notamment de l’hétérozygotie d’un gène lié à l’immunité. Les résultats de ce travail décrivent avec précision la distribution de la variabilité génétique et son lien avec les risques sanitaires dans la population d’étude, apportant ainsi des informations cruciales pour la mise en place de stratégies de gestion et de conservation des populations de mouflons dans le contexte actuel de changements globaux et de réémergences de maladies. / Using as a case study the Mediterranean mouflon (Ovis gmelini musimon × Ovis sp.), we aimed at better understanding how are linked genetic diversity, individual behaviour, gene flows and parasitic dynamic. Using population and landscape genetics approaches, we showed that the spatial genetic structure of the studied population was determined by its introduction history, its socio-spatial structure and the landscape in which it evolves. Given the impact of these elements on mouflon gene flow, we expected them to also determine parasite transmission in the population. Our results nevertheless evidenced that parasite are better dispersed than mouflon genes. Finally, we showed that host resistance to parasites depends on neutral and adaptive genetic diversity, and more specifically on heterozygosity at a immunity-linked locus. Our results precisely describe genetic variability spatial distribution and its link with sanitary risks in the studied population, bringing crucial information for wild sheep population management and conservation in the current context of global changes and disease reemergence.

Darwin's theory of sexual selection explains the existence of sexual dimorphism, or within-species sex differences in shape, color, size, and behavior. In some cases, sexually dimorphic traits, especially extravagant male ornaments, seem maladaptive and thus in opposition to natural selection. The crux of Darwin's theory was that sexual selection arises from individual differences in reproductive success that result from competition for mates. In this dissertation, I investigated several aspects of sexual selection and the evolution of female mating preferences and male ornaments in the great frigatebird (Fregata minor). Frigatebirds as a group (family Fregatidae) are the most ornamented of any seabirds, and are among the most ornamented of any animal group. Their most prominent ornament is a gular (throat) pouch which becomes red in males during the breeding season, and which is inflated and displayed to females during courtship. Male courtship display also includes a warble vocalization and extension and trembling of the wings. I investigated the following issues concerning sexual selection and ornamentation in great frigatebirds: 1) the source of ornamental coloration in male great frigatebird gular pouches. I determined that this was a carotenoid-based color display; 2) the relationship of male mating success to gular pouch size and coloration. I determined that mating success was not related to the size or color of this ornament; 3) the relationship between male vocal display traits and female preferences. Again, I found no relationship between vocal display traits and female preferences, and finally, 4) the role of a major histocompatibility complex (MHC) locus in female mate choice. The MHC is a highly polymorphic multi-gene family associated with immune defense and has been proposed to play a role in mate choice. I found a significant disassortative mating pattern amongst mated pairs compared to random pairings based on MHC genotypes. In summary, I found no evidence for female mating preferences based on visual or auditory display traits associated with male ornamentation. However, I did find evidence for female mating preferences based on genetic dissimilarity at an MHC locus.

This thesis investigates the impact of avian malaria (Haemosporidia) parasites on the outcome of interspecific competition between two closely related bird species, pied (Ficedula hypoleuca) and collared (F. albicollis) flycatchers. I further investigated how variation in timing of breeding, life history strategies and immune competence genes (MHC genes) modulate the fitness effects of malaria parasites in one of the two species i.e. collared flycatchers. Collared flycatchers colonized the Baltic island Öland in the late 1950-ties and has since then been expanding their breeding range while competitively excluding pied flycatchers from the favourable habitats (deciduous forests). I investigated the underlying mechanisms behind this exclusion by combining detailed long-term breeding data with modern molecular genetic techniques identifying both the presence/absence and lineage specificity of haemosporidian blood parasites. I found that the rapid decline of pied flycatchers can be explained by the combined effects of competition over nestling sites, hybridization and haemosporidian infections. Haemosporidian infections have a negative impact on survival of pied flycatcher females but no detectable effect on collared flycatchers’ longevity or reproductive success. This may be due to the fact that collared flycatchers carry (and are potentially exposed to) a higher diversity of parasites than pied flycatchers, which in turn may select for a higher diversity of MHC genes and hence a better overall protection from the negative impact of parasites. Indeed, functional MHC diversity correlates negatively with malaria prevalence among collared flycatchers from Gotland. Moreover, I found that both, malaria infection intensity and immunoglobulin level influences how infected collared flycatchers respond to increased nestling food-demands. The latter results mean that there is variation in allocation strategies (i.e. in resource allocation between reproductive effort and immune competence) within the collared flycatcher population. Hence, this population has the ability to respond to novel selection pressures in terms of optimal allocation of resources into immune functions. In summary, my results show that local parasites may facilitate the expansion of a new colonizer. This is important in the context of global climate change that will probably increase the colonization rate of southern species and lead to novel host-parasite interactions.

A exibição do comportamento vocal em muitas espécies de anfíbios anuros é relacionada aos níveis de hormônios esteroides gonadais e interrenais. Esses hormônios poderiam mediar a relação entre intensidade de sinais e imunidade, pois estão envolvidos no desenvolvimento das características sexuais secundárias, comportamento de corte e mobilização de reservas energéticas durante a atividade reprodutiva, enquanto apresentam também efeitos imunomoduladores. Nesse sentido, o objetivo deste trabalho foi explorar as relações entre comportamento reprodutivo, imunidade e níveis plasmáticos de testosterona e corticosterona em machos do sapo do semiárido brasileiro, Rhinella granulosa, em atividade reprodutiva e após manipulação hormonal. A precipitação foi o principal determinante ambiental para o aumento dos níveis de testosterona e corticosterona circulantes em machos de R. granulosa, estimulando o comportamento de vocalização. As relações fisiológicas encontradas indicam que os altos níveis plasmáticos de testosterona nos primeiros dias após a chuva devem promover o início do turno vocal, porém a corticosterona deve modular o esforço de vocalização. De forma geral, a exacerbação do comportamento vocal de R. granulosa tem efeitos negativos sobre a imunocompetência, porém alguns indivíduos que apresentam maiores concentrações plasmáticas de corticosterona apresentam concomitantemente alto esforço vocal e alta imunidade. De acordo, a aplicação transdérmica de corticosterona promoveu elevação aguda dos níveis plasmáticos deste glicocorticoide, bem como um aumento da função imune. Assim, apesar de a atenção principal ser comumente colocada no papel da testosterona na mediação de sinais honestos, nossos resultados corroboram a importância da corticosterona na mediação da expressão do comportamento de corte e imunocompetência em machos R. granulosa / In most anuran species, vocal behavior is usually related to plasmatic levels of gonadal and interrenal steroid hormones. These hormones could mediate the relationship between signal intensity and immunocompetence, since they are involved in the development of sexual traits, courtship behavior and energetic mobilization during reproductive activity and also exert immunomodulatory effects. The aim of this study was to explore relations among vocal behavior, immunity and plasmatic levels of testosterone and corticosterone in males of a toad species from Brazilian semi-arid, Rhinella granulosa, during reproductive activity and after hormonal manipulation. Precipitation was the main environmental determinant to increase circulating levels of testosterone and corticosterone in R. granulosa males, stimulating vocal behavior. Physiological relations indicate that high circulating testosterone levels in the days right after rain may promote the initiation of vocal behavior, while corticosterone may modulate vocal effort. In general, exacerbation of vocal behavior in R. granulosa has negative effects on immunocompetence, however, some individuals with higher plasmatic levels of corticosterone can present both high vocal effort and good immunocompetence. Corroborating this interpretation, acute increase in glucocorticoid plasmatic levels after hormonal manipulation was accompanied by raised immune function. Despite the emphasis on androgens influence on sexual selected male traits, our results corroborate greater influence of corticosterone in the mediation of expression of courtship behavior and immunocompetence in R. granulosa

De acordo com o modelo de seleção intersexual mediado por parasitas, variações nas condições de ornamentos ou taxa de displays sexuais refletem diferentes graus de resistência ao parasitismo. Em coros naturais de anfíbios, vocalizações mais altas, longas e complexas e/ou emitidas a maiores taxas são, provavelmente, mais facilmente detectadas e atraem mais fêmeas. Entretanto, o esforço vocal está positivamente relacionado aos níveis plasmáticos de esteróides potencialmente imunossupressores, tais como testosterona e corticosterona. O objetivo deste trabalho foi avaliar as relações entre taxa de vocalização, imunocompetência e níveis plasmáticos de corticosterona em Hypsiboas albopunctatus, um anuro de médio porte que ocorre na América do Sul. Tais relações foram estudadas a partir de observação focal do comportamento vocal em coro natural, seguido de coleta de sangue para dosagem de corticosterona e avaliação de alguns parâmetros imunitários. O presente estudo demonstrou uma relação positiva entre a taxa de vocalização com níveis plasmáticos de corticosterona e negativa entre estes dois parâmetros com a imunocompetência mediada por células. Estes resultados apontam para a corticosterona como possível hormônio mediador do compromisso entre esforço vocal e imunidade mediada por células em anuros. / According to the model of intersexual selection mediated by parasites, changes in conditions of sexual ornaments or display rates reflect different degrees of resistance to parasitism. In natural choruses of amphibians, calls that are higher, longer, more complex and / or issued at higher rates are probably more easily detected and attract more females. However, the vocal effort is positively related to plasma levels of potentially immunosuppressive steroids, such as testosterone and corticosterone. The aim of this study was to evaluate the relationship between calling rates, immunocompetence and plasma levels of corticosterone in Hypsiboas albopunctatus, a midsize anuran occurring in South America. Such relationships were studied from focal observations of calling behavior, followed by blood collection for measurements of corticosterone plasma levels and evaluation of some immune parameters. The present study showed a positive relationship between the calling rate with plasma levels of corticosterone, and a negative correlation between these two parameters with cell-mediated immunocompetence. These results highlight the importance of the hormone corticosterone as a possible mediator of trade-off between vocal effort and cell-mediated immunity in anurans.

The decline of bumblebee (Bombus spp.) populations in the UK and worldwide has been well reported. It has been generally assumed that such declines result in the genetic impoverishment of some species, potentially leading to reduced fitness and increased extinction risk. This study tested the fundamental assumption linking population fragmentation with fitness, in a model system of two Bombus species native to the UK. Bombus monticola has declined significantly in range across the UK in recent years and occupies fragmented upland areas, while Bombus pratorum has remained abundant and widespread over many habitat types. The effects of genetic diversity on fitness have been addressed in wild Bombus species, but this is the first study to explicitly compare data from species of differing levels of population connectivity and hence test the assumptions of traditional population genetic theory. As genetic diversity has often been linked with immunocompetence, aspects of the innate immune response were quantified, together with parasite load. These empirical measures of fitness showed lower than expected variability between the two study species, and no evidence was found to support the theory of lower fitness in fragmented populations. However, the considerable variability between sample sites in both species for all parameters measured raised interesting questions as to the underlying evolutionary processes; it is postulated that B. monticola populations may maintain a higher than expected Ne, despite their fragmented distribution. This study also provided methodological developments. An alternative method for the quantification of wing wear as a proxy for age was proposed, which could be easily applied to other Bombus species and possibly adapted for use in other flying insects. In addition possible sources of error in AFLP analysis were highlighted which have not been adequately discussed in the current literature, namely the effects of sample storage. Given the utility of AFLPs for non-model species, this is an important avenue for future research, and would be applicable to studies in other systems. Overall, the data presented here emphasise the challenges of studying fitness in wild populations, and underline the requirement for research into the fundamental principles underlying many assumptions made by conservation genetic theory.

Experiments were performed to assess the suitability of Rhabditis pellio, a nematode found in earthworms, as a challenge organism for use in development of a biomarker assay to determine the potential of chemicals to suppress the immunocompetence of the non-specific immune system. To accomplish this goal, information on the life cycle of R. pellio was determined; including effects of incubation time and temperature on growth rates; along with information on the immune response elicited in the earthworm, Lumbricus terrestris. Immune parameters measured were coelomocyte migration toward and attachment to R. pellio larvae and adults. Preliminary background information showed that R. pellio has potential as a challenge organism for development of a biomarker assay.

The goals of captive breeding programs for endangered species include preserving genetic diversity and avoiding inbreeding. Typically this is accomplished by minimizing population mean kinship; however, this approach becomes less effective when errors in the pedigree exist and may result in inbreeding depression, or reduced survival. Here, both pedigree- and DNA-based methods were used to assess inbreeding depression in the critically endangered Attwater’s prairie-chicken (Tympanuchus cupido attwateri). Less variation in the pedigree-based inbreeding coefficients and parental relatedness values were observed compared to DNA-based measures suggesting that errors exist in the pedigree. Further, chicks identified with high parental DNA-based relatedness exhibited decreased survival at both 14- and 50-days post-hatch. A similar pattern was observed in later life stages (> 50 days post-hatch) with birds released to the wild; however, the pattern varied depending on the time post-release. While DNA-based inbreeding coefficient was positively correlated with mortality to one month post-release, an opposite pattern was observed at nine months suggesting purging of deleterious alleles. I also investigated whether immunocompetence, or the ability to produce a normal immune response, was correlated with survival; however, no significant correlation was observed suggesting that inbreeding was a more important factor influencing survival. Pairing individuals for breeding by minimizing DNA-based parental relatedness values resulted in a significant increase in chick survival. This study highlights the importance of using DNA-based methods to avoid inbreeding depression when errors exist in the pedigree.