Inbreeding can have profound negative effects on individuals by reducing fertility and viability. In populations, inbreeding depression can reduce growth rates and increases extinction risk. The aims of this thesis are to investigate inbreeding depression in male guppies (Poecilia reticulata) and to study the evolution of mechanisms for inbreeding avoidance in females, using guppies from a feral population in Queensland, Australia. Male guppies are highly polymorphic in their sexual ornamentation, indeed they show one of the most extreme polymorphisms observed in nature. Female guppies exhibit complex mate choice based on preferences for ornamentation, as well as social context. I aim is to examine how these factors of inbreeding avoidance alter sexual selection. In male guppies I found strong inbreeding depression in male sperm numbers, which is amplified under semi-natural compared to laboratory conditions (Chapter 2). Moreover, inbreeding depression results in low fertility under sperm competition: an experiment using artificial insemination techniques reveals that highly inbred males are heavily disadvantaged in gaining paternity (Chapter 3). On population level, inbreeding depression is manifest in reduced growth rates, predominantly in the early stages of inbreeding (Chapter 4). Population growth at inbreeding coefficients f=0.375-0.59 did not seem to lead to inbreeding depression, whereas lower levels of inbreeding reduced population growth. Although the growth rates in inbred populations appear normal, severe inbreeding depression is uncovered after outbred immigrants are added. Specifically, male immigrants are most efficient in short-term genetic rescue, probably due to insemination of large numbers of females whereas females are limited in the number of eggs they can produce (Chapter 4). Male ornamental traits show significant inbreeding depression in semi-natural conditions only (Chapters 2 & 3). Inbreeding avoidance mechanisms seem to have evolved in females: they prefer courtship displays of non-inbred males (Chapter 2), unfamiliar males (Chapter 5) and males with rare patterns (Chapter 6). This preference might increase the mating success of immigrants, and may have evolved to facilitate the avoidance of inbreeding. Together with context-independent preferences for ornament combinations (Chapter 6), it also offers an explanation for the maintenance of polymorphism in this species.

Inbreeding can have profound negative effects on individuals by reducing fertility and viability. In populations, inbreeding depression can reduce growth rates and increases extinction risk. The aims of this thesis are to investigate inbreeding depression in male guppies (Poecilia reticulata) and to study the evolution of mechanisms for inbreeding avoidance in females, using guppies from a feral population in Queensland, Australia. Male guppies are highly polymorphic in their sexual ornamentation, indeed they show one of the most extreme polymorphisms observed in nature. Female guppies exhibit complex mate choice based on preferences for ornamentation, as well as social context. I aim is to examine how these factors of inbreeding avoidance alter sexual selection. In male guppies I found strong inbreeding depression in male sperm numbers, which is amplified under semi-natural compared to laboratory conditions (Chapter 2). Moreover, inbreeding depression results in low fertility under sperm competition: an experiment using artificial insemination techniques reveals that highly inbred males are heavily disadvantaged in gaining paternity (Chapter 3). On population level, inbreeding depression is manifest in reduced growth rates, predominantly in the early stages of inbreeding (Chapter 4). Population growth at inbreeding coefficients f=0.375-0.59 did not seem to lead to inbreeding depression, whereas lower levels of inbreeding reduced population growth. Although the growth rates in inbred populations appear normal, severe inbreeding depression is uncovered after outbred immigrants are added. Specifically, male immigrants are most efficient in short-term genetic rescue, probably due to insemination of large numbers of females whereas females are limited in the number of eggs they can produce (Chapter 4). Male ornamental traits show significant inbreeding depression in semi-natural conditions only (Chapters 2 & 3). Inbreeding avoidance mechanisms seem to have evolved in females: they prefer courtship displays of non-inbred males (Chapter 2), unfamiliar males (Chapter 5) and males with rare patterns (Chapter 6). This preference might increase the mating success of immigrants, and may have evolved to facilitate the avoidance of inbreeding. Together with context-independent preferences for ornament combinations (Chapter 6), it also offers an explanation for the maintenance of polymorphism in this species.

The invasion of individuals into new habitats can pose a major threat to native species and to biodiversity itself. However, the consequences of invasions for native populations that are not fully reproductively isolated from their invaders are not yet well explored. Here I chose the Trinidadian guppy, Poecilia reticulata, to investigate how different population traits shaped the outcome of Haskins's introduction, a well-documented invasion of Guanapo river guppies into the Turure river. I especially concentrated on the importance of behaviour for invasive success. I investigated if the spread of Guanapo guppies is due to superiority in behaviour, life-history and/or genetics, or if the outcome of this translocation is due to chance. Despite the fact that by today the invasive front has passed the Turure's confluence with the River Quare many kilometres downstream of the introduction site, and the original genotype only survives in small percentages, as was revealed by genetic analysis in this and other studies, no obvious differences between invasive and native populations could be detected in any of the tested behavioural, life-history and genetic traits. When tested for mate choice, neither Guanapo nor Oropuche (Turure) males seemed to be able to distinguish between the population origin of females, but courted and mated at random. At the same time, females did not prefer to school with individuals of the same population over schooling with more distantly related females. The formation of mixed schools after an invasive event is therefore likely. Because female guppies showed a very low willingness to mate, even after having been separated from males for up to six months, sperm transfer through forced copulations will become more important. Taken together, these behaviours could increase the speed of population mixing after an invasion without the need for behavioural superiority of the invasive population. When tested for their schooling abilities, offspring of mixed parentage, in contrast to pure breds, displayed a large amount of variety in the time they spent schooling, a circumstance that can potentially influence survival rates and therefore the direction of gene pool mixing. Guanapo fish did not show reproductive superiority in a mesocosm experiment, where both populations were mixed in different proportions. On the contrary, in two out of three mixed treatments, the amount of Oropuche (Turure) alleles was significantly higher than expected from the proportion of initially stocked fish. The almost complete absence of distinguishable traits other than genetic variation between the examined populations that belong to different drainage systems, opposes the recent split of the guppy into two different species following drainage system borders, as is argued in this thesis. However, the successful invasion of the Turure by Guanapo guppies and the nearly entire disappearance of the original population can be explained in absence of differing population traits. Here I demonstrate how behavioural and genetic interactions between subspecies influence the outcome of biological invasions and second, how factors other than population traits, such as the geographic situation, can produce an advantageous situation for the invader even in the absence of population differences.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-08-25T14:53:55Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-08-28T17:33:16Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) / Made available in DSpace on 2017-08-28T17:33:16Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DesempenhoGuppyPoecilia.pdf: 4012755 bytes, checksum: e38444c9cd0f96e294f8f50246678089 (MD5) Previous issue date: 2016-09-23 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O uso de modelos animais em pesquisa experimental nas últimas décadas tem se mostrado mais diversificado do que o modelo clássico através do uso de roedores ou primatas. Isso se dá principalmente pelos avanços nos estudos moleculares, morfológicos e funcionais que revelaram uma grande homologia entre os vertebrados. Nesta perspectiva, o peixe Zebrafish (Danio rerio) tem se mostrado como o animal não-mamífero com maior ascensão como animal de estudo em ciências biológicas nas últimas décadas. Entretanto, outras espécies de peixes também se mostram promissoras como alternativas de uso como modelo animal. Este estudo utilizou o Guppy (Poecilia reticulata) como modelo para pesquisa em comportamento através de diferentes abordagens experimentais. No estudo I, guppies foram expostos e reexpostos em diferentes turnos (manhã, tarde, noite e madrugada) nos testes de campo aberto e preferência claro-escuro. Os resultados encontrados mostram que em ambos os testes, machos e fêmeas apresentam diferenças comportamentais, sendo sensíveis a reexposição, com capacidade de aprendizagem e controle do ciclo circadiano. No estudo II, foi desenvolvido um labirinto em cruz com rampa no qual se verificou a sensibilidade da espécie ao aparato, o perfil de resposta mediante reexposição e o efeito de drogas. Os resultados revelaram sensibilidade ao aparato para uma altura de coluna d’água de 8cm e 5 minutos de sessão e diferenças entre os sexos e aprendizagem por habituação ao longo das reexposições. O estudo farmacológico indica que neste aparato a espécie é sensível a drogas ansiolíticas e ansiogênicas. Ao final, pode-se concluir que o Guppy apresenta comportamento similar e respostas as drogas compatível com os dados descritos para Zebrafish. Tais similaridades reforçam o uso de peixes como uma alternativa ao uso de mamíferos na experimentação animal. / The use of animal models in experimental research in the last decades has shown to be more diversified than the classical model through the use of rodent or primate. This is mainly by advances in molecular, morphological and functional studies that revealed a big homology between the vertebrates. On this perspective, the Zebrafish (Danio rerio) has shown to be the non-mammal animal with biggest ascension as a study model in biological sciences in the latest decades. However, other species of fish also show to be promising as alternatives of use as animal model. This study used Guppy (Poecilia reticulata) as model for the research in behavior through different experimental approaches. In study I, guppies were exposed and re-exposed in different shifts (morning, afternoon, evening and night) in the open field test and light-dark preference. The found results show that in both tests, male and female present behavioral differences, being sensitive to re-exposure, with capacity of learning e controlling of the circadian cycle. In study II, was developed a plus-maze with ramp where it was verified the sensibility of the specie to the apparatus, the profile of response upon re-exposure and the drug effect. The results revealed sensibility to the apparatus with a height of water column at 8 cm and 5 minutes of session and differences between sex and learning by habituation along the re-expositions. The pharmacological study indicates that in this apparatus, the specie is sensitive to anxiolytic and anxiogenic drugs. Finally, we can conclude that Guppy presents similar behavior and drug responses compatible with the data described to the Zebrafish. These similarities reinforce the use of fishes as an alternative to the use of mammals in animal experimentation.

Made available in DSpace on 2016-06-02T19:29:38Z (GMT). No. of bitstreams: 1 3822.pdf: 3918575 bytes, checksum: d646b2b0a3b661d4dfa2d3c8a1ee8a69 (MD5) Previous issue date: 2011-03-28 / Universidade Federal de Sao Carlos / The Alto Jacaré-Guaçu Basin was evaluated regarding limnological and ecotoxicological aspects in four water and sediment sampling campaigns at 14 points distributed in the area. Toxicity tests with water, in the laboratory and in situ, showed chronic toxicity, with effects on the survival, growth and gill morphology of juveniles of both fish species examined (Danio rerio and Poecilia reticulata). The lowest survival percentages in the laboratory assays were observed for point L, and in the field tests for point J1. The analysis of the gills revealed first-stage alterations in the specimens of both species exposed at all the sample points, and there were second-stage alterations (aneurism and vascular congestion) at various points. The high concentrations of nutrients and metals in the basin s water and sediments reveal a system impacted by human activities in the surrounding region. The analyses of metals quantified Cd, Cr, Cu, Mn and Fe levels in the water above those permitted by the Brazilian National Environmental Council (CONAMA 357/05), and of Cd in the sediments above the threshold levels permitted both by national (CONAMA 344/04) and international regulations (CCME, 2003). Simple synthetic sediments were prepared composed of sand, clay and organic matter (obtained from the aquatic macrophyte Scirpus cubensis, at concentrations of 0, 0.25, 0.5 and 1%) to assess the toxicity of Cd. The synthetic sediments were viable for the growth and survival of Ceriodaphnia silvestrii, Chironomus xanthus and Danio rerio, and were suitable for use in spiked experiments. The evaluation of the acute toxicity of the sediments fortified with Cd indicated mean 48-h;EC50 for C. silvestrii of 3.09mg.Kg-1 in the sediments with 0% OM and 2.98mg.Kg-1 in the sediments with 1% OM. The mean 96-h;LC50 for C. xanthus was 18.67 mg.Kg-1 in the sediments with 0% OM and nearly four times lower in those with 1% OM (4.93 mg.Kg-1). A similar pattern was noted for D. rerio, a higher 48-h;LC50 in the sediments with 0% OM (114.88 mg.Kg-1) than in the sediments with 1% OM (75.31 mg.Kg-1). In subsequent experiments, the sediments were also spiked with 0.6mg.Kg-1 of Cd, a concentration considered safe for protection of aquatic life according to the applicable national and international regulations. In these tests, the survival of C. silvestrii was impaired in the Cd spiked sediments, so it was not possible to assess the effects on reproduction. There were also effects on the survival, biomass, mentum morphology and Cd concentration in the tissues of C. xanthus. For D. rerio, the survival and growth were compromised in the sediments with higher percentage of organic matter. Besides this, the organisms showed impaired gill morphology and higher Cd concentration in tissue in function of exposure to sediments spiked with the metal. Based on the results, it can be concluded that C. silvestrii was the most sensitive species to the sediments fortified with Cd, followed by C. xanthus and D. rerio, the least sensitive. These effects on the iv organisms at concentrations considered safe for the biota suggest more conservative thresholds of this metal should be established for tropical aquatic habitats. / A bacia do Alto Jacaré-Guaçu foi avaliada quanto aos aspectos limnológicos e ecotoxicológicos em quatro campanhas de amostragem de água e sedimento em 14 pontos distribuídos na área. Testes de toxicidade com água, tanto em laboratório como in situ, apresentaram toxicidade crônica com efeitos na sobrevivência, crescimento e morfologia de brânquias de juvenis de ambas as espécies de peixes avaliadas (Danio rerio e Poecilia reticulata). As menores porcentagens de sobrevivência nos testes em laboratório foram observadas no ponto L e, nos testes in situ, em J1. A análise das brânquias mostrou alterações de primeiro estágio nos organismos expostos em todos os pontos avaliados, e em diversos pontos foram verificadas alterações de segundo estágio (aneurisma e congestão vascular). As elevadas concentrações de nutrientes e metais na água e sedimentos da Bacia do Alto Jacaré-Guaçu revelam um sistema impactado pelas atividades humanas que se desenvolvem em seu entorno. As análises de metais quantificaram Cd, Cr, Cu, Mn e Fe acima da CONAMA 357/05 na água e Cd acima dos valores estabelecidos pela legislação nacional (CONAMA 344/04) e internacional (CCME, 2003) nos sedimentos. Foram desenvolvidos sedimentos sintéticos simples compostos de areia, argila e matéria orgânica (obtida a partir da macrófita aquática Scirpus cubensis, nas concentrações de 0; 0,25; 0,5 e 1%) para avaliar a toxicidade do Cd. Os sedimentos sintéticos se apresentaram viáveis para o crescimento e sobrevivência de Ceriodaphnia silvestrii, Chironomus xanthus e Danio rerio, sendo adequados para uso em experimentos de fortificação. A avaliação da toxicidade aguda dos sedimentos fortificados com Cd, indicaram CE50;48h média para C. silvestrii de 3,09mg.Kg-1 nos sedimentos sem MO e 2,98mg.Kg-1 nos sedimentos com 1% de MO. A CL50;96h média para C. xanthus foi de 18,67mg.Kg-1 nos sedimentos sem MO e, cerca de 4 vezes menor nos sedimentos com 1% de MO, 4,93mg.Kg-1. Para D. rerio também foi notada tendência semelhante, CL50;48h maior nos sedimentos sem MO, 114,88mg.Kg-1 e menor nos sedimentos com 1% de MO, 75,31mg.Kg-1. Os sedimentos também foram fortificados com 0,6mg.Kg-1 de Cd, concentração considerada segura para a proteção da vida aquática segundo legislações nacionais e internacionais, em experimentos posteriores. C. silvestrii teve a sobrevivência comprometida nos sedimentos fortificados com Cd, de modo que não foi possível avaliar os efeitos na reprodução. C. xanthus também apresentou efeitos na sobrevivência, biomassa, morfologia de mento e concentração de Cd nos tecidos. D. rerio teve a sobrevivência e crescimento comprometidos nos sedimentos com maior porcentagem de matéria orgânica. Além disso, os organismos apresentaram a morfologia de brânquias comprometida e concentração de Cd em ii seus tecidos elevada em função da exposição aos sedimentos fortificados com o metal. Com base nos resultados foi possível concluir que C. silvestrii foi a espécie mais sensível aos sedimentos fortificados com Cd, seguido de C. xanthus e, a menos sensível, D. rerio. Tais efeitos observados nos organismos em concentrações consideradas seguras para a biota sugerem a avaliação mais cuidadosa dos valores para espécies e climas tropicais.

nÃo hà / O dengue permanece como problema de saÃde pÃblica no Brasil. No nordeste brasileiro, os grandes depÃsitos domiciliares utilizados para acumular Ãgua sÃo importantes criadouros para reproduÃÃo do Aedes aegypti, o principal transmissor do dengue. O uso de alternativas ao controle quÃmico desse vetor vem sendo incentivado. Os objetivos desse trabalho foram avaliar se a presenÃa de peixes larvÃfagos altera o padrÃo de postura do A. aegypti, identificar a sobrevivÃncia de peixes larvÃfagos ao cloro e descrever a eficÃcia do peixe Betta splendens em condiÃÃes de campo. O padrÃo de postura foi avaliado em uma gaiola com 6 m3. Os peixes avaliados foram Poecilia reticulata e B. splendens. Na gaiola foram inseridos oito depÃsitos, sendo quatro com peixes, quatro sem peixes (controle) e 100 mosquitos. Em cada depÃsito tinha Ãgua e uma palheta de eucatex para postura dos ovos. Ao final de cada semana os ovos postos nessas palhetas foram contados. Os ensaios foram replicados por sete semanas consecutivas para cada espÃcie. A sobrevivÃncia do B. splendens e P. reticulata, ao cloro, foi avaliada para trÃs concentraÃÃes (1,0; 1,5 e 2,0 mg/L) utilizando tambores com 35 litros de Ãgua. Foram utilizados 105, 140 e 175 peixes para cada concentraÃÃo testada, na proporÃÃo de 6 depÃsitos com cloro para cada controle (sem cloro). A eficÃcia foi avaliada a partir de dados gerados pelo Programa Municipal de Controle do Dengue, na cidade de Fortaleza. Foi avaliada a permanÃncia dos peixes em depÃsitos domiciliares e a infestaÃÃo nesses depÃsitos com B. splendens e o larvicida Bacillus thuringiensis israelensis. A presenÃa do B. splendens inibiu a postura de ovos pelas fÃmeas de Aedes aegypti com um Ãndice de Atividade de OviposiÃÃo (IAO) de -0,627. O nÃmero mÃdio de ovos postos em depÃsitos com B. splendens (32,5/semana) foi menor que nos depÃsitos com o P. reticulata (200,5/semana) e os controles (186,5/semana; p < 0,0001). Todos os B. splendens sobreviveram a concentraÃÃo de cloro de 1,0 mg/L; 72,5 e 39,3% sobreviveram as concentraÃÃes de 1,5 e 2,0 mg/L. Por outro lado, apenas 4,4% do P. reticulata sobreviveram a concentraÃÃo mÃnima de 1,0 mg/L. Em campo foram encontrados trÃs depÃsitos com a presenÃa do B. splendens e larvas de mosquitos (1,6%), infestaÃÃo significativamente menor que nos depÃsitos com o Bti, onde essa infestaÃÃo foi de 10,9% (p < 0,001). Nos depÃsitos onde o peixe nÃo permaneceu a infestaÃÃo foi de 27,8%, maior que nos depÃsitos com Bti (p < 0,010). Nos depÃsitos onde o peixe permaneceu ele foi 85% mais eficaz que o larvicida. A permanÃncia dos peixes foi maior nos tanques de alvenaria (48,5%), localizados no peridomicÃlio (47,5%) e ao nÃvel do solo (53,3%). ConcluÃmos que o B. splendens pode ser apropriado para controle biolÃgico de larvas de Ae. aegypti em grandes reservatÃrios domiciliares, desde que possa ser atestada sua permanÃncia nesses depÃsitos. / Dengue fever remains an important public health problem in northeast Brazil. Large domestic containers used to store water are important breeding sites of Aedes aegypti, the main vector. The use of alternatives to chemical vector control has increased in the last years. The objectives of this study were: to evaluate the inhibition of oviposition by female Ae. aegypti in domestic containers with larvivorous fish; to describe the survival of larvivorous fish to different chlorine concentrations; and to describe the efficacy of Betta splendens fish under field conditions. Oviposition was assessed in a cage of 6 m3 of size. The fish species Poecilia reticulata and B. splendens were used in the laboratory assays. In the cage, eight water containers were placed - four with fish, four without fish (control), and 100 mosquitoes. In each container with 15 l water eucatex strips were placed to facilitate oviposition. At the end of each week, eggs laid on these strips were counted. For each species, the assays were repeated for seven consecutive weeks. Survival of B. splendens and P. reticulata to different concentrations of chlorine (1.0, 1.5 and 2.0 mg / L) was assessed in drums with 35 liters of water. We used 105, 140 and 175 fish for each concentration: six test containers with chlorine for each control without chlorine. Secondary data of the Municipal Dengue Control Program of the city of Fortaleza were analyzed to assess the efficacy of B. splendens under field conditions. The presence of fish in household containers was verified after several weeks. Infestation of containers with mosquito larvae was compared to containers with the larvicide Bacillus thuringiensis israelensis. The presence of B. splendens inhibited oviposition by Ae. aegypti with an activity oviposition index of -0.627. The mean number of eggs laid in deposits with B. splendens (32.5 / week) was lower than in deposits with P. reticulata (200.5 / week) and controls (186.5 / week, p <0.0001). All B. splendens specimens survived a chlorine concentration of 1.0 mg / L, and 72.5% and 39.3% survived concentrations of 1.5 and 2.0 mg / L, respectively. On the other hand, only 4.4% of P. reticulata survived a concentration of 1.0 mg / L. Under field conditions, three containers were encountered with B. splendens and mosquito larvae (1.6%), significantly less than infested deposits with Bti (10.9%; p <0.001). In containers where the fish died or disappeared, infestation was 27.8% higher than in deposits with Bti (p <0.010). In deposits where the fish remained, efficacy was 85% better than Bti. The permanence of fish was higher in concrete tanks (48.5%), located outside the house (47.5%) and at ground level (53.3%). We conclude that B. splendens may be suitable for biological control of Ae. aegypti larvae in large domestic water containers.