abstract: Differences between males and females can evolve through a variety of mechanisms, including sexual and ecological selection. Because coloration is evolutionarily labile, sexually dichromatic species are good models for understanding the evolution of sex differences. While many jumping spiders exhibit diverse and brilliant coloration, they have been notably absent from such studies. In the genus Habronattus, females are drab and cryptic while males are brilliantly colored, displaying some of these colors to females during elaborate courtship dances. Here I test multiple hypotheses for the control and function of male color. In the field, I found that Habronattus males indiscriminately court any female they encounter (including other species), so I first examined the role that colors play in species recognition. I manipulated male colors in H. pyrrithrix and found that while they are not required for species recognition, the presence of red facial coloration improves courtship success, but only if males are courting in the sun. Because light environment affects transmission of color signals, the multi-colored displays of males may facilitate communication in variable and unpredictable environments. Because these colors can be costly to produce and maintain, they also have the potential to signal reliable information about male quality to potential female mates. I found that both red facial and green leg coloration is condition dependent in H. pyrrithrix and thus has the potential to signal quality. Yet, surprisingly, this variation in male color does not appear to be important to females. Males of many Habronattus species also exhibit conspicuous markings on the dorsal surface of their abdomens that are not present in females and are oriented away from females during courtship. In the field, I found that these markings are paired with increased leg-waving behavior in a way that resembles the pattern and behavior of wasps; this may provide protection by exploiting the aversions of predators. My data also suggest that different activity levels between the sexes have placed different selection pressures on their dorsal color patterns. Overall, these findings challenge some of the traditional ways that we think about color signaling and provide novel insights into the evolution of animal coloration. / Dissertation/Thesis / Ph.D. Biology 2012

A full understanding of any biological trait requires investigation of its evolutionary origin. Primates inspire great curiosity amongst researchers due to the remarkable diversity across species in both anatomical and behavioural traits, including sociality, sexual behaviour, life histories, neuro-anatomy, cognitive abilities and behavioural repertoires. The study of primates has involved comparative approaches since its inception, however, the necessary tools for statistically investigating the macro-evolutionary processes responsible for current diversity in biological traits have been developed only in the last 30 years or so, namely phylogenetic reconstruction and phylogenetic comparative methods. Amongst a multitude of evolutionary questions that can be addressed by phylogenetic comparative analyses, this thesis attempts to address two in particular, concerning primates. First, chapters 3 and 4 use meta-analysis and phylogenetic comparative analyses to investigate the evolution of large, brightly coloured ‘exaggerated sexual swellings' in female Catarrhine (‘Old World') primates. Together, chapters 3 and 4 show that such swellings are signals of temporal fertility, and present evidence to suggest that swellings co-evolved with conditions favouring male mate choice and cryptic female choice, therefore shedding light on the general conditions under which female signals of temporal fertility should evolve. Second, chapters 5 and 6 use phylogenetic comparative analyses investigate the evolution of enlarged brain size in the primate order. Together, chapters 5 and 6 suggest that multiple selection pressures have contributed to diversity in brain size and cognitive traits across primates, including sociality, intra-sexual competition and extended life history. Further, analyses presented in chapter 6 suggest that reliance on learned behaviour is a self-reinforcing evolutionary process, favouring ‘runaway' increases in cognitive abilities and reliance on culture in some primate lineages, which parallels increases in brain size, cognitive ability and reliance on culture in human evolution.

Meliponiculture (or stingless beekeeping) is a powerful tool for sustainable economic development in the tropics. However, meliponiculture has many challenges as it lacks standardization in management and breeding practices. The aim of this thesis was to investigate stingless bee reproductive biology combined to management and breeding as background to meliponiculture improvement. In chapter 1, we performed a meta-analysis of heritability across the Hymenoptera in order to review current knowledge and discuss the challenges for bee breeding and conservation. In chapter 2, we assessed how sexual selection acts on male traits in the stingless bee Scaptotrigona aff. depilis and identified which traits may affect male fitness. In chapter 3, we estimated heritability and genetic correlations for the traits studied in the previous chapter, to understand how evolution shapes male traits and to identify important traits for breeding programs. Finally, in chapter 4 we investigated how the environment and long-term management influence colony productivity in Melipona subnitida (jandaíra), a commercially managed species in Northeastern Brazil. Our results revealed that morphological traits exhibit higher heritability estimates than fitness related traits, and in general, colony productivity traits showed potential for breeding. However, few studies are available for stingless bees yet. Male aggregations in S. aff depilis selected competitive males with higher quality sperm, indicating the importance of this mechanism in stingless bee mating biology. The studied male traits exhibited high heritability estimates, with exception of sperm length. Since aggregations selected males with shorter sperm, these results suggest selection for long-term sperm storage by queens and higher fertilization potential. The assessment of M. subnitida records revealed that honey production was affected by climate and management, and strategies to improve beekeeping practices were discussed. We believe this thesis provides important guidelines to establish successful stingless bee breeding programs / A meliponicultura, criação racional de abelhas sem ferrão, é uma poderosa ferramenta para o desenvolvimento econômico sustentável em regiões tropicais. No entanto, a prática da meliponicultura enfrenta diversos desafios, como a falta de padronização das técnicas de manejo e melhoramento genético. O objetivo desta tese foi investigar a biologia reprodutiva de abelhas sem ferrão aliada ao manejo e ao melhoramento genético, a fim de fornecer subsídios para o aprimoramento da meliponicultura. No capítulo 1, realizamos uma meta-análise sobre herdabilidade em Hymenoptera, a fim de compreender o conhecimento atual e os desafios associados ao melhoramento genético de abelhas e sua conservação. No capítulo 2, avaliamos como a seleção sexual atua em caracteres dos machos da espécie Scaptotrigona aff. depilis e identificamos quais caracteres podem influenciar o sucesso reprodutivo. No capítulo 3, estimamos a herdabilidade e correlações genéticas para os caracteres avaliados no capítulo anterior, a fim de entender como a evolução atua moldando os caracteres dos machos e quais desses caracteres podem ser utilizados em programas de melhoramento genético. Finalmente, no capítulo 4, investigamos o efeito do ambiente e do manejo na produtividade de colônias em Melipona subnitida (jandaíra), uma espécie manejada comercialmente no Nordeste brasileiro. Nossos resultados revelaram que caracteres morfológicos exibem estimativas de herdabilidade mais altas do que caracteres ligados ao sucesso reprodutivo. No entanto, poucos estudos com abelhas sem ferrão foram realizados até o momento. Agregações de machos em S. aff. depilis selecionaram machos mais competitivos que apresentaram maior qualidade espermática, indicando a importância desse mecanismo na biologia reprodutiva de abelhas sem ferrão. Os caracteres estudados apresentaram alta herdabilidade, com exceção do comprimento do espermatozoide. Como agregações selecionam machos com espermatozoides mais curtos, esses resultados sugerem seleção direcionada para um maior tempo de armazenamento do esperma pelas rainhas e maior potencial de fertilização. A avaliação dos registros de M. subnitida revelou que a produção de mel foi afetada pelo clima e pelo manejo, e estratégias a fim de melhorar as práticas da meliponicultura foram discutidas. Acreditamos que essa tese fornece importantes resultados para o estabelecimento de programas de melhoramento genético em abelhas sem ferrão

Orientador: Woodruff Whitman Benson / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-12T18:41:50Z (GMT). No. of bitstreams: 1 Peixoto_PauloEnriqueCardoso_D.pdf: 865304 bytes, checksum: 19540c783ae14daa3ba0b5f8dffc1d86 (MD5) Previous issue date: 2009 / Resumo: Brigas territoriais entre machos de insetos voadores são resolvidas das mais variadas formas. Em borboletas, existem evidencias de que tamanho, idade e motivação são importantes determinantes das chances de vitória. No entanto, as regras utilizadas nas brigas, o contexto biológico que afeta a intensidade dos comportamentos agonisticos e o papel funcional das características dos machos na geração de custos durante os confrontos ainda são pouco conhecidos. Neste trabalho investiguei uma serie de características que podem influenciar os custos e benefícios dos confrontos territoriais utilizando as espécies de satirineos Hermeuptychia hermes (Fabricius, 1775), Moneuptychia soter (Buttler, 1877) e Paryphthimoides phronius (Buttler, 1867) como modelos de estudo. No capitulo 1 investiguei se as disputas intra-especificas pela posse de territórios em machos de H. hermes e de M. soter ocorrem com contato físico e quais podem ser as características funcionalmente relevantes para a distinção entre machos territoriais e não territoriais. Para tanto, documentei as brigas entre machos utilizando filmagens de alta velocidade e realizei comparações morfológicas e fisiológicas entre machos possuidores de territórios (residentes) e machos que ocuparam territórios nos quais os residentes originais foram removidos (intrusos). Machos residentes de H. hermes apresentaram massa corporal e muscular similares as dos seus pares intrusos, porem, possuíram menor desgaste alar e maior quantidade de lipídeos. Machos residentes de M. soter, por outro lado, foram mais pesados que os machos intrusos que ocuparam seus territórios. Machos de H. hermes podem dividir a interação em vôos circulares, espirais ascendentes e perseguições lineares, enquanto machos de M. soter usam vôos circulares mais lentos, os quais são seguidos por perseguições lineares e emissões de som (estalos). Contatos físicos não foram registrados para nenhuma espécie. No capitulo 2 investiguei o papel da residência previa na resolução de conflitos territoriais entre machos de H. hermes. Ao induzir disputas entre dois machos que se comportaram como residentes (um residente original e um residente induzido) e compara-las com disputas naturais entre machos residentes e intrusos, foi possível demonstrar que a residência previa aumenta as chances de vitória em um combate. Massa corporal afetou positivamente as chances de vitória nas brigas induzidas, mas não nas brigas naturais. Finalmente no capitulo 3, investiguei a influencia da presença de recursos alimentares no estabelecimento de territórios de acasalamento por machos de P. phronius. Machos desta espécie defendem sítios de acasalamento localizados em manchas de sol sem recursos evidentes. No entanto, manchas de sol previamente desocupadas passaram a ser defendidas depois de receberem frutas fermentadas. Apesar de machos que se estabeleceram nestes locais aparentemente se alimentarem das frutas, experimentos de escolha de territórios sugerem que eles preferem defender sítios sem recursos e utilizam a defesa de áreas com alimento como tática alternativa de acasalamento. Este estudo representa a primeira investigação deste tipo de variação nas táticas de acasalamento em borboletas. Alem de auxiliar a compreensão da evolução de determinados sistemas de localização de parceiros, esse sistema permite testar qual a implicação da existência de mais de uma tática reprodutiva na intensidade dos confrontos territoriais entre machos / Abstract: Territorial conflicts in flying insects may be resolved through many different ways. In male butterflies, size, age and motivation are often reported as important determinants of the winning chances. However, the rules used when fighting, the biological context that affects the intensity of agonistic behaviors, and the functional role of male traits in the contests costs generation are poorly known. In this study I investigated a series of characteristics that may influence the costs and benefits of territorial conflicts using the satyrine species Hermeuptychia hermes (Fabricius, 1775), Moneuptychia soter (Buttler, 1877), and Paryphthimoides phronius (Buttler, 1867) as study organisms. In chapter 1, I used high-speed video imagery to investigate whether intraspecific disputes between male H. hermes and male M. soter occur with physical contact. Additionally, I removed resident males from their defended sites and the subsequent intruders that established on those areas after the resident removal to assess if wing wear, body mass, fat content and flight muscle ratio are important determinants of male residency status. Resident males of H. hermes were similar in body weight and thoracic muscle mass to their intruder rivals, but had fewer wing wear and higher fat content. Resident males of M. soter, on the other hand, were heavier than intruder males. Male H. hermes divided their interaction in up to three phases consisting of circular flights, ascending spirals and back and forth persecutions, whereas male M. soter used much slower circular flights that were often followed by linear persecutions and clicking sound emissions. Physical contacts were not observed for both species. In chapter 2, I investigated the role of previous residence on contest resolution in the butterfly H. hermes. By inducing territorial interactions between males that behaved as residents (one original resident and another resident-induced one) and comparing them with natural contests between resident and intruder males, it was possible to show that previous residence increases the chances of victory. Body mass positively affected the winning chances in the induced fights, but was unimportant among the natural ones. Finally in chapter 3, I investigated the role of feeding resources in the territory establishment by males of the butterfly P. phronius. Male of this species typically defend sunny clearings on the forest edge which do not contain any evident resource. However, previously undefended sunny clearings were occupied by territorial males after receiving soft fermenting fruit. Although males which established on sites containing feeding resources seem to forage on the fermenting fruit, territory selection experiments shows that males prefer to defend sites without resources. This type of variation in the mate locating tactics has never been previously investigated for butterflies. In addition of helping the understanding of the evolutionary processes leading to different mate-locating strategies, this system allows the testing of the influence of different reproductive behaviors on the intensity of territorial conflicts between males / Doutorado / Doutor em Ecologia

Machos dos opiliões goniosomatíneos Serracutisoma spelaeum e Serracutisoma guaricana, cujas fêmeas copulam com mais de um parceiro em um único período reprodutivo, apresentam uma grande variedade morfológica, especialmente relacionada a caracteres sexuais secundários. Considerando-se que machos menores frequentemente perdem disputas por fêmeas, esta tese objetivou verificar possíveis estratégias reprodutivas alternativas, resultantes de seleção sexual pré e pós-copulatória, adotadas por machos de pequeno porte dessas espécies assim como o papel da fêmea nesses mecanismos. Este trabalho foi, então, dividido em três capítulos, cada um com objetivos específicos. O primeiro deles refere-se à organização do trato reprodutor feminino e caracterização das espermatecas, órgãos de armazenamento de espermatozóides. Imagens de microscopia de luz e de microscopia eletrônica de varredura associadas às obtidas sob estereomicroscópio revelaram a existência de quatro espermatecas inseridas na musculatura da porção distal do ovipositor. Cada uma delas é constituída por quatro sáculos individuais de fundo cego e se comunica com a luz do ovipositor através de um único canal. A complexidade das espermatecas indica que a fêmea pode ser capaz de armazenar separadamente os espermatozóides dos diferentes machos com os quais vier a copular, enquanto a musculatura ao redor dessas estruturas sugere que ela possa determinar a paternidade de sua progênie através de contrações diferenciais. O capítulo 2 abrange observações comportamentais e características morfológicas e morfométricas do trato reprodutor masculino, objetivando identificar possíveis estratégias alternativas utilizadas por machos de pequeno porte na obtenção de sucesso reprodutivo. Para tal, foram efetuados acompanhamento do comportamento sexual e estudos morfométricos de genitália e da gônada. Machos de pequeno porte parecem adotar a estratégia de machos-satélites, copulando com as fêmeas quando machosguardiões estão com a atenção desviada. A ausência de diferenças estatísticas significativas na largura da placa ventral sugere que o pênis de machos grandes e de machos pequenos se ajustam à genitália feminina da mesma maneira e, portanto, sejam vii avaliados pelas fêmeas da mesma forma. Já a semelhança no comprimento da genitália e o menor tamanho corpóreo dos machos pequenos indicam que, durante a cópula, eles poderiam alcançar regiões mais profundas da espermateca, removendo ou deslocando os espermatozóides dos outros machos e, consequentemente, diminuindo o risco de competição espermática. A análise ultraestrutural dos espermatozóides (compostos por núcleo, citoplasma e acrossomo, além de diversas projeções na superfície) mostrou que, provavelmente, todos os machos possuam espermatozóides viáveis, visto que espermatozóides não-férteis, em geral, não possuem acrossomo. As similaridades entre machos de grande e de pequeno porte tanto na morfologia dos gametas como em seu comprimento indicam que tais características possam ter sido objeto de seleção sexual pós-copulatória. O terceiro capítulo propõe uma hipótese para a formação das projeções encontradas nos espermatozóides das duas espécies e que lhes confere uma morfologia peculiar. Para isso foi utilizada a associação de três técnicas de microscopia: microscopia eletrônica de transmissão, de varredura e confocal de varredura a laser. Os resultados obtidos evidenciam que essas estruturas são formadas ao longo do processo de espermatogênese. Uma rede de proteínas situada logo abaixo da membrana plasmática exerceria uma pressão sobre o citoplasma, que extravasaria pelos espaços existentes nessa espécie de malha. Dessa forma, as projeções seriam constituídas de citoplasma envolto por membrana plasmática. Em conclusão, os resultados obtidos na presente tese sugerem que a competição espermática e a escolha críptica da fêmea, componentes da seleção sexual póscopulatória, são pressões seletivas atuantes nessas duas espécies de opilião. / Males of the goniosomatine harvestmen Serracutisoma spelaeum and Serracutisoma guaricana, which females may copulate with more than one partner during the same reproductive season, show a large morphological variation, especially considering secondary sexual features. Since smaller males frequently lose fights against larger males for females, the present thesis aimed at verifying the occurrence of alternative reproductive strategies among these smaller males as a result of pre- and postcopulatory sexual selection, as well as the role of the female in these mechanisms. This study is composed of three chapters, each one with specific objectives, as described as follows. The first chapter describes the general organization of the female reproductive system and the spermathecae, structures for the storage of sperm. Ligth and scanning electronic microscopy images revealed the presence of four spermathecae inserted in the musculature of the distal portion of the ovipositor. Each spermatheca is composed of four individual dead-end sacs and is connected to the ovipositor lumen through a single channel. This complex spermathecae structure indicates that the female is able to store sperm from different males with which she copulates in different places, and that the musculature that involves these structures may be responsible for differential contractions which would result in different paternity for the females descendants. The second chapter includes behavioral observations and morphological and morphometric analysis of the male reproductive system (genitalic and gonadal features), aiming at identifying possible alternative reproductive strategies used by small males to achieve reproductive success. Small males seem to adopt a satellite-male strategy, copulating with females when larger guardian-males are not paying attention. The absence of significant statistic differences in the width of its ventral plate suggests that the penis of both large and small males may adjust to the female genitalia in the same way and, therefore, will be similarly evaluated by the female during copulation. In turn, the smaller length associated to the same sized penis of smaller males in comparison to the larger ones may indicate that they may reach deeper regions of the female genitalia during copulation, possibly removing or displacing the sperm of other males and, in consequence, reducing the risk of sperm competition. The ultrastructure analysis of the spermatozoa (which are composed of a nucleus, cytoplasm and an acrosome, in addition to several surface projections) showed that probably males of different sizes all have viable sperm, since generally non-fertile spermatozoa lack an acrosome. Similarities in the morphology and size of the gametes of both small and large males may indicate that these features have been subjected to postcopulatory sexual selection. The third chapter suggests a new hypothesis for the development of the projections found in the surface of the spermatozoa of the two species here studied. For this study, three microscope techniques were used: transmission electronic microscopy, scanning electronic microscopy, and confocal laser scanning microscopy. The results show that these projections are formed during the spermatogenesis. A net of proteins situated just below the plasmatic membrane restrains the cytoplasm, which flows through the spaces formed in the net, forming the projections, which are externally limited by plasmatic membrane. In conclusion, the results obtained in the present study suggest that sperm competition and female cryptic choice, which are components of the postcopulatory sexual selection process, are selection pressures working on the species studied herein.

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.

No description available.

Sex allocation and sexual selection have been heavily studied, but rarely linked. In this thesis I investigated the interface between them in the gregarious parasitoid wasp Nasonia vitripennis, both directly and through their interactions with the mating system and sexual conflict. Chapter 2 investigated sexual selection and mating at the natal site: earlier eclosing males mated more females independently of body size. Nasonia follows Local Mate Competition, which describes how a female laying eggs alone on a patch of resources (a so-called single-foundress) should lay an extremely female-biased brood to minimise competition between her sons, yet ensure all her daughters are fertilised. Based on this I predicted that males with with fewer brothers would be better inseminators. Despite finding significant among-strain variation in (1) single-foundress sex ratio, (2) mate competitiveness when alone and (3) when in competition, (4) sperm resources, but not (5) sperm-depletion (Chapters 3 & 4), I did not find the predicted relationship. Conversely males from strains with more brothers had a higher mating success under competition (Chapter 3) leading to the question: does mating success select on sex ratio or vice versa? Either way it is a result of an interaction between sexual selection and sex allocation. Chapter 5 investigated the role of male post-copulatory courtship on female re-mating, and found that among- strain variation in female re-mating was not associated with variation in the duration of the post-copulatory courtship. Chapter 6 reviewed sexual conflict in the Hymenoptera: their haplodiploid genetics, newly sequenced genomes and varied life- histories provides a base for future research to build on. Finally I highlight the novel links between sexual selection, sex allocation, sexual conflict and the mating system found during my studies that will hopefully prompt future research on this topic.

Abstract Divergence of behavioral traits (e.g. courtship rituals, habitat choice) has had a major impact on species formation and isolation. Species-specific courtship rituals preventing species hybridization may arise as a by-product of natural selection during spatial isolation or through direct action of natural selection to prevent species hybridization after a secondary contact. Coordination leading to the assumption of coevolution of signals and preferences is a prerequisite for effective courtship signaling between a male and a female of the same species. We found a reasonable amount of variation in the mate traits within the Drosophila montana and D. virilis species, but our findings did not reveal evidence of the coevolution between the male courtship signals and female preference for these signals. Variation also did not cause isolation within species. The form of female preference for carrier frequency of male song was found to be stabilizing even in changing environments, when female preference for the carrier frequency and pulse length of the male song was measured at ambient temperature. D. montana females always preferred males with high frequency songs, which probably advertises the male condition. The frequency of male song decreased due to male aging, although males with larger body size were able to maintain frequencies better. Ageing also had a deterioration effect on male reproductive success. The importance of male courtship song was not only highlighted in mate choice, but also in species-recognition. We were able to persuade D. montana females to copulate with alien D. lummei species by playing them simulated song resembling the song of conspecific males. Simulated courtship songs were demonstrated to be a practical tool in studies of preference between sexually isolated Drosophila species. / Tiivistelmä Kosintapiirteiden (esim. kosintarituaalit, habitaatin valinta) vaihtelevuudella on ollut suuri vaikutus lajien muodostumisessa ja eriytymisessä. Lajien välisiä risteytymisiä estäviä lajispesifisiä kosintarituaaleja voi syntyä luonnonvalinnan sivutuotteena spatiaalisen eristäytymisen aikana tai luonnonvalinnan suoralla vaikutuksella estämään lajiristeymät lajien uudelleen kohdatessa. Tehokkaan koiraan ja naaraan välisen kosintaviestinnän edellytyksenä on koordinointi, minkä vuoksi on oletettavaa, että signaalien ja vasteiden välillä on yhteisevoluutiota. Löysimme kohtuullisen määrän vaihtelevuutta kosintapiirteissä Drosophila montana – ja D. virilis – lajeilla, mutta emme löytäneet todisteita koiraan kosintasignaalien ja naaraan näihin signaaleihin kohdistamien vasteiden välisestä yhteisevoluutiosta. Vaihtelu ei myöskään aiheuttanut isolaatiota lajien välille. Naaraan koiraan kosintalaulun kantofrekvenssiin osoittaman vasteen funktion muodon havaittiin olevan tasapainottava jopa vaihtelevissa ympäristöissä, kun naaraan kantotaajuuteen osoittamaa vastetta mitattiin eri lämpötiloissa. D. montana naaraat suosivat koiraita, joilla oli korkeataajuinen kosintalaulu. Laulun korkea taajuus kertoo luultavasti naaraalle koiraan fyysisestä kunnosta. Koiraan laulun frekvenssin havaittiin laskevan koiraan ikääntyessä, mutta isommat koiraat pystyivät paremmin säilyttämään korkean taajuuden laulussaan. Ikääntyminen heikensi myös koiraan lisääntymismenestystä. Koiraan kosintalaulun tärkeys ei korostunut pelkästään parinvalinnassa, vaan myös lajintunnistuksessa. Pystyimme suostuttelemaan D. montana –naaraat parittelemaan vieraan D. lummei –lajin kanssa käyttämällä simuloituja kosintalauluja, jotka osoittautuivat käytännöllisiksi välineiksi tutkittaessa naaraan vasteita seksuaalisesti eriytyneillä Drosophila-lajeilla.

Cuticular hydrocarbons (CHCs) are found on the outer cuticle of all terrestrial arthropods. Although their primary function is in desiccation prevention, these compounds have also been shown to play a variety of roles in insect chemical communication, from species and sex recognition to providing cues of dominance and attractiveness. However, despite growing evidence of their versatility as cues, our knowledge of how CHCs are used in mating interactions is limited to Drosophila and field crickets. In this thesis I investigate the roles CHCs play in interactions at each stage of the mating process in the broad-horned flour beetle Gnatocerus cornutus. I assess the relative importance of CHCs in influencing male reproductive success and examine the complex interplay between different episodes of selection and the mechanisms of sexual selection acting on males. I use a combination of behavioural assays, experimental manipulations and gas chromatography. First, I identify the role of CHCs as cues of sperm competition risk and intensity, demonstrating how the presence of male-derived CHCs on the cuticles of virgin females elicits males to adjust their pre- and post-copulatory investment (chapter 2), by providing information on the state of their competitive environment. I then go on to look at the stability of CHCs as cues of sperm competition over time, finding that they are highly sensitive to environmental degradation (chapter 3) and do not persist in the habitat substrate of this species. Next, I investigate how male CHCs determine fighting and mating success. By estimating and comparing the strength and form of sexual selection imposed by male-male competition and female mate choice, I show that male CHCs are subject to strong antagonistic sexual selection (chapter 4). By experimentally manipulating male CHC profile, I then attempt to verify the selection gradients estimated for female choice 3 (chapter 5). However, my experimental manipulation fails to verify the importance of male CHCs for female mate choice. Finally, I explore the role of same-sex sexual behaviour (SSB) in determining male reproductive success (chapter 6). I find evidence to suggest that SSB may in fact be a form of aggression in its own right, and demonstrate that SSB and fighting may provide equivalent means for males to overcome female choice and secure a mating advantage. My results indicate that CHCs play key roles as chemical cues throughout the mating process and significantly impact male reproductive success. My thesis reveals the intricate nature of the relationships between mechanisms of sexual selection, alongside highlighting the need to consider both the social and physical environment when investigating the importance of chemical cues. I discuss the implications of these results for the evolution of male CHCs and how my findings can be used to further our knowledge of this field.