Orientador: Alexandre Ninhaus Silveira / Resumo: Os peixes durante o período reprodutivo, mobilizam grande parte de suas reservas energéticas para o processo de maturação dos oócitos, por isso quando em cativeiro, é necessário o fornecimento de uma dieta nutricionalmente adequada, com cuidado com o manejo alimentar e a escolha da ração, aonde os reprodutores devem receber uma dieta com componentes de alta qualidade e digestibilidade. O objetivo foi verificar se a adição de 5% de óleo de peixe marinho a dieta Gymnocorymbus ternetzi promoverá uma incorporação de ácidos graxos essenciais no tecido ovariano e possível alteração no desenvolvimento gonadal. Para tal, foram estabelecidos dois sistemas compostos com espécimes adultos de G. ternetzi, sendo um denominado grupo controle - GC, em que os animais foram alimentados com ração comercial e o outro grupo óleo - GO que foram alimentados com a mesma ração com adição de 5% de óleo de peixe marinho, por um período de 12 meses. Mensalmente, foram realizadas coletas do tecido hepático e gonadal de fêmeas de G. ternetzi, para determinação dos índices hepatossomático (IHS) e gonadossomático (IGS), para quantificação e determinação do tamanho dos oócitos, para análise das fases reprodutivas e determinação quanto ao perfil dos ácidos graxos. Os dados de biometria dos animais, não diferiram estatisticamente entre os grupos testados. A determinação dos ácidos graxos dos tecidos hepático e gonadal, apresentou valores constantes e similares de ácidos graxos poli-insaturados - família ômega... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Fish during the reproductive period mobilize most of their energy reserves for the oocyte maturation process, so when in captivity, it is necessary to provide a nutritionally adequate diet, careful with food management and choice of feed, where breeders should receive a diet with high quality components and digestibility. The objective was to verify if the addition of 5% of marine fish oil to the Gymnocorymbus ternetzi diet will promote the incorporation of essential fatty acids in the ovarian tissue and possible alteration in gonadal development. For this purpose, two composite systems were established with adult G. ternetzi specimens, one called control group - GC, in which the animals were fed with commercial feed and the other oil - GO group that were fed with the same feed with the addition of 5% marine fish oil for a period of 12 months. The liver and gonadal tissue were collected monthly from G. ternetzi females to determine hepatosomal (IHS) and gonadosomatic indices (IGS), to quantify and determine oocyte size, to analyze the reproductive phases and to determine the profile. of fatty acids. Animal biometrics data did not differ statistically between the groups tested. The determination of fatty acids from hepatic and gonadal tissues presented constant and similar values of polyunsaturated fatty acids - omega 3 family (PUFAs n3) for both groups. The gonadosomatic index (IGS) did not differ statistically between the groups, but the hepatosomatic index (IHS), in April /... (Complete abstract click electronic access below) / Mestre

The overall aim of this thesis was to determine the size and age compositions, growth and reproductive biology of Western Blue Groper (Achoerodus gouldii), Blue Morwong (Nemadactylus valenciennesi) and Yellowtail Flathead (Platycepahlus endrachtensis) in south-western Australian waters, in which these three species have divergent life cycle characteristics. As A. gouldii and N. valenciennesi are commercially and recreationally important in coastal waters and P. endrachtensis is one of the most recreationally important species in the Swan River Estuary, these biological data were then used to produce estimates of mortality and spawning stock biomass per recruit for each of these species. The biological data and stock assessment parameters were finally employed comparatively to ascertain whether any of the three species possessed characteristics that would make them particularly susceptible to the effects of fishing and whether there was evidence that any of the species is fully or even overexploited. Achoerodus gouldii typically uses reefs in protected inshore waters along the coast and around neighbouring islands as a nursery habitat and then, as it increases in size, moves to deeper, offshore reefs, where it spawns between early winter and mid-spring. The maximum total length and age of A. gouldii were 1162 mm and 70 years, the latter being the greatest age by far yet recorded for any species of labrid. However, most growth occurs in the first 20 years of life. Histological and demographic analyses demonstrated that all individuals begin life as females and, after attaining maturity, many become males, i.e. A. gouldii is a monandric protogynous hermaphrodite. The L50 at maturity and sex change were 653 and 821 mm, respectively, which correspond to ages of c. 17 and 37 years, respectively. As sex change took place over a narrower range in lengths (650 to 900 mm) than in ages (15 to 49 years), that change is apparently related more to size than age. Since sex change is typically accompanied by a change from green to blue, body colour can be used as a proxy for determining the length (L50) at which females change to males. von Bertalanffy growth curves fitted to the lengths at age of individuals of each sex of this hermaphroditic species using a novel technique demonstrated that, with increasing age, the lengths of males became increasingly greater than those of females. Thus, at ages 15, 30 and 60 years, the estimated lengths at age of females were c. 600, 670 and 680 mm, respectively, whereas those of males were c. 695, 895 and 975 mm, respectively. As A. gouldii is very long-lived and sexual maturity, and even more particularly sex change, occur late, this labrid is potentially very susceptible to overfishing. Thus, because the mortality estimates and per recruit analyses indicated that, at present, this species is close to or fully exploited, fisheries managers will need to take a precautionary and watchful approach to managing and thus conserving the stocks of this species. As with A. gouldii, N. valenciennesi moves to deeper, offshore waters as it increases in size and then matures and spawns in those waters. Although N. valenciennesi has a maximum length of nearly 1 m and thus, like A. gouldii, is moderately large, it has a far shorter life span, i.e. 19 vs 70 years. While female N. valenciennesi does not grow to as large a size as its males (max. lengths = 846 and 984 mm, respectively), the maximum age of both sexes was 19 years. From the growth curves, the females by ages 3, 6 and 10 years havd attained, on average, lengths of 435, 587 and 662 mm, respectively, compared with 446, 633 and 752 mm, respectively, for males. Both sexes grew little after 10 years of age. Juvenile N. valenciennesi < 400 mm in total length were found exclusively in shallow, coastal waters on the south coast, whereas their adults were abundant in offshore waters of both the south and lower west coasts. The females and males typically mature in offshore waters of the south coast at lengths of c. 600-800 mm and ages of c. 7-9 years. In contrast, the vast majority of females caught in offshore waters of the lower west coast (where they were of a similar length and age range to those in offshore waters on the south coast) became mature at lengths of 400-600 mm and 3-4 years of age. The attainment of maturity by N. valenciennesi at far lesser lengths and ages on the lower west coast than south coast suggests that the former coast provides better environmental conditions for the gonadal maturation and spawning of this species. Furthermore, the contrast between the almost total absence of the juveniles of N. valenciennesi in nearshore waters on the lower west coast and their substantial numbers in comparable waters on the south coast indicates that the larvae of this species produced on the lower west coast are transported southwards to the south coast, where they become juveniles. As spawning occurs between mid-summer and late autumn, the larvae, which spend a protracted period in the plankton, would be exposed, on the lower west coast, to the influence of the southwards-flowing Leeuwin Current at the time when that current is at its strongest. Although N. valenciennesi is caught by recreational line fishing and commercial gillnet fishing when they are as young as 3-4 years old, they do not become fully vulnerable to these fisheries until they are about 9 years old. Consequently, the individuals of this species can potentially breed over a number of years before they become particularly prone to capture by fishers. Mortality estimates and per recruit analyses suggested that N. valenciennesi in south-western Australia is not currently overfished. A greater resilience to fishing by N. valenciennesi than A. gouldii presumably reflects, in part, its far shorter lifespan, earlier maturity and possession of gonochorism rather than hermaphroditism. Platycephalus endrachtensis spawns in the Swan River Estuary between late spring and early autumn and completes the whole of its life cycle in this system. Although females attain a far larger length (615 mm) than males (374 mm), females and males were present in each age class. These data, together with a detailed examination of histological sections of a wide size and age range of individuals, demonstrated that this species, unlike some of its relatives, is not a protandrous hermaphrodite, i.e. it does not change from male to female with increasing body size. The combination of the presence of females and males in all age classes and the observation that all of the large number of individuals between 374 and 615 mm were females shows that the far greater length attained by that sex is largely related to its faster growth rate. The fact that females outnumbered males in each age class of P. endrachtensis in which the sample size was substantial, i.e. > 25, with the overall sex ratio being 2.7 females: 1 male, indicate that there has been strong selection for egg production in this species. As the minimum legal length for retention of P. endrachtensis is 300 mm, and relatively few males exceeded this length, the recreational fishery which targets this species is based largely on its females. The estimates of mortality and results of per recruit analyses provided no evidence that P. endrachtensis is currently overfished. From a management point of view, it is advantageous that the current size limit for this species exceeds the average length at which its females (259 mm) attain maturity. Furthermore, this species appears to be resilient to capture and release. The data presented in this thesis demonstrate that A. gouldii possesses biological characteristics which make it potentially more prone to the effects of fishing than is the case with either N. valenciennesi or P. endrachtensis. This presumably accounts, at least in part, for the indications that A. gouldii is the only one of these three species that is likely to be close to or at full exploitation.

O complexo de espécies crípticas de Anastrepha fraterculus compreende oito morfotipos dos quais três ocorrem no Brasil. Anastrepha sp.1 aff. fraterculus é o morfotipo Brasil-1 e é uma espécie de ampla distribuição no planalto sudeste/sul do Brasil e no norte da Argentina. O comportamento reprodutivo dessa espécie é complexo, envolvendo uma série de movimentos desempenhados pelos machos para atrair as fêmeas para acasalamento, e ocorre, preferencialmente, nas primeiras horas do dia. Dois genes, period e fruitless, entre outros, são conhecidos por participar do controle do comportamento reprodutivo de vários organismos, incluindo algumas espécies de moscas-das-frutas. O presente trabalho buscou informações sobre a atuação desses genes no comportamento reprodutivo de A. sp.1, utilizando o silenciamento transitório desses genes pela metodologia de RNA interferente. Foram, primeiramente, desenhados iniciadores específicos para amplificar fragmentos do DNA genômico desses genes, sendo demonstrado que apresentaram uma similaridade entre 97 a 99% com os genes equivalentes de outras espécies de Anastrepha. A seguir, após padronização e adaptação de protocolos, foram sintetizados os RNA de dupla-fita (dsRNA) dos dois genes, que foram, então, utilizados nos experimentos de silenciamento. Análises, para verificação se os genes foram realmente silenciados foram feitas a partir a injeção dos dsRNAfru e/ou dsRNAper no abdomên de machos sexualmente maduros, tendo sido demonstrado que os genes estavam silenciados ao máximo, sete a oito dias após a injeção. Interferências no comportamento de machos sexualmente maduros, com um ou outro gene silenciado, foram avaliados por testes relacionados com dois parâmetros do comportamento reprodutivo: alterações nas atividades gerais (qualquer tipo de movimentação dos insetos) dos machos durante o ciclo circadiano (dia/noite) e mais especificamente, a atividade relacionada ao comportamento reprodutivo. Ao injetar dsRNAfru em machos adultos de A. sp.1 foi observado que não houve alteração significativa nas suas atividades gerais. Porém, foi observado que houve uma diminuição significativa no número de machos que realizavam atividades reprodutivas, sugerindo que o silenciamento de fruitless no macho adulto altera o funcionamento normal do comportamento sexual masculino. Em contrapartida, ao injetar dsRNAper em machos adultos de A. sp.1 não foi observada alteração significativa tanto nas suas atividades reprodutivas quanto nas atividades gerais. O silenciamento dos genes aparentemente não afeta a produção de espermatozóides. Apesar de controverso, esses dados corroboram com o que foi observado em D. melanogaster, em que o mutante nulo per04 não apresenta alteração significativa nos seus comportamentos em relação aos machos selavagens em um regime cíclico Dia/Noite. Dessa forma, análises adicionais serão necessárias com o objetivo de elucidar as implicações desses dois genes no comportamento locomotor e reprodutivo de Anastrepha sp.1 aff. fraterculus. / The complex of cryptic species Anastrepha fraterculus comprises eight morphotypes three of which occur in Brazil. Anastrepha sp.1 aff. fraterculus correspond to the morphotype Brazil-1 and is a species of wide distribution in the southeast/south plateau of Brazil and in north of Argentina. The reproductive behavior of this species is complex, involving a series of movements performed by males to attract females for mating, and occurs in the early hours of the day. Two genes, period and fruitless, among others, are known to participate in the control of reproductive behavior of various organisms, including some species of fruit flies. The present work aimed to get information about the presumable role of these genes in the reproductive behavior of A. sp.1, using transient silencing of these genes by interfering RNA methodology. Specific primers were first designed to amplify fragments of the genomic DNA of these genes, showing they have a similarity between 97 to 99% with the equivalent genes of other Anastrepha species. After standardization and adaptation of protocols, the double-stranded RNA (dsRNA) of the two genes were synthesized, and used in the silencing experiments. Tests to verify that the genes were actually silenced, were made after injection of the dsRNAfru and/or dsRNAper into the abdomens of sexually mature males. These tests showed that the genes were silenced to the maximum, seven to eight days after the injection. Interferences in the behavior of sexually mature males with one or other silenced gene were evaluated by tests related to two parameters of reproductive behavior: changes in the general activities (any type of movement of insects) of males during the circadian cycle (day/night) and more specifically, activity related to reproductive behavior. Injection of dsRNAfru in adult males of A. sp.1 showed that they did not cause significant alterations in general activities, but it was observed that they cause a significant decrease in the number of males that performed reproductive activities, suggesting that silencing of fruitless. alters the normal functioning of male sexual behavior. In contrast, when injecting dsRNAper. in adult males of A. sp.1, no significant alteration was observed neither in their reproductive activities nor in their general activities. Moreover, silencing of the genes seems not to affect the production of spermatozoa. Although controversial, the data are in line with observations in D. melanogaster, in which the null mutant per04 did not present significant alterations in behaviors relatives to the control males in a Day/Night cyclic regime. Thus, additional analyzes will be needed to elucidate the participation of the two genes in the coordination of behaviors in Anastrepha sp.1 aff fraterculus.

O complexo de espécies crípticas de Anastrepha fraterculus compreende oito morfotipos dos quais três ocorrem no Brasil. Anastrepha sp.1 aff. fraterculus é o morfotipo Brasil-1 e é uma espécie de ampla distribuição no planalto sudeste/sul do Brasil e no norte da Argentina. O comportamento reprodutivo dessa espécie é complexo, envolvendo uma série de movimentos desempenhados pelos machos para atrair as fêmeas para acasalamento, e ocorre, preferencialmente, nas primeiras horas do dia. Dois genes, period e fruitless, entre outros, são conhecidos por participar do controle do comportamento reprodutivo de vários organismos, incluindo algumas espécies de moscas-das-frutas. O presente trabalho buscou informações sobre a atuação desses genes no comportamento reprodutivo de A. sp.1, utilizando o silenciamento transitório desses genes pela metodologia de RNA interferente. Foram, primeiramente, desenhados iniciadores específicos para amplificar fragmentos do DNA genômico desses genes, sendo demonstrado que apresentaram uma similaridade entre 97 a 99% com os genes equivalentes de outras espécies de Anastrepha. A seguir, após padronização e adaptação de protocolos, foram sintetizados os RNA de dupla-fita (dsRNA) dos dois genes, que foram, então, utilizados nos experimentos de silenciamento. Análises, para verificação se os genes foram realmente silenciados foram feitas a partir a injeção dos dsRNAfru e/ou dsRNAper no abdomên de machos sexualmente maduros, tendo sido demonstrado que os genes estavam silenciados ao máximo, sete a oito dias após a injeção. Interferências no comportamento de machos sexualmente maduros, com um ou outro gene silenciado, foram avaliados por testes relacionados com dois parâmetros do comportamento reprodutivo: alterações nas atividades gerais (qualquer tipo de movimentação dos insetos) dos machos durante o ciclo circadiano (dia/noite) e mais especificamente, a atividade relacionada ao comportamento reprodutivo. Ao injetar dsRNAfru em machos adultos de A. sp.1 foi observado que não houve alteração significativa nas suas atividades gerais. Porém, foi observado que houve uma diminuição significativa no número de machos que realizavam atividades reprodutivas, sugerindo que o silenciamento de fruitless no macho adulto altera o funcionamento normal do comportamento sexual masculino. Em contrapartida, ao injetar dsRNAper em machos adultos de A. sp.1 não foi observada alteração significativa tanto nas suas atividades reprodutivas quanto nas atividades gerais. O silenciamento dos genes aparentemente não afeta a produção de espermatozóides. Apesar de controverso, esses dados corroboram com o que foi observado em D. melanogaster, em que o mutante nulo per04 não apresenta alteração significativa nos seus comportamentos em relação aos machos selavagens em um regime cíclico Dia/Noite. Dessa forma, análises adicionais serão necessárias com o objetivo de elucidar as implicações desses dois genes no comportamento locomotor e reprodutivo de Anastrepha sp.1 aff. fraterculus. / The complex of cryptic species Anastrepha fraterculus comprises eight morphotypes three of which occur in Brazil. Anastrepha sp.1 aff. fraterculus correspond to the morphotype Brazil-1 and is a species of wide distribution in the southeast/south plateau of Brazil and in north of Argentina. The reproductive behavior of this species is complex, involving a series of movements performed by males to attract females for mating, and occurs in the early hours of the day. Two genes, period and fruitless, among others, are known to participate in the control of reproductive behavior of various organisms, including some species of fruit flies. The present work aimed to get information about the presumable role of these genes in the reproductive behavior of A. sp.1, using transient silencing of these genes by interfering RNA methodology. Specific primers were first designed to amplify fragments of the genomic DNA of these genes, showing they have a similarity between 97 to 99% with the equivalent genes of other Anastrepha species. After standardization and adaptation of protocols, the double-stranded RNA (dsRNA) of the two genes were synthesized, and used in the silencing experiments. Tests to verify that the genes were actually silenced, were made after injection of the dsRNAfru and/or dsRNAper into the abdomens of sexually mature males. These tests showed that the genes were silenced to the maximum, seven to eight days after the injection. Interferences in the behavior of sexually mature males with one or other silenced gene were evaluated by tests related to two parameters of reproductive behavior: changes in the general activities (any type of movement of insects) of males during the circadian cycle (day/night) and more specifically, activity related to reproductive behavior. Injection of dsRNAfru in adult males of A. sp.1 showed that they did not cause significant alterations in general activities, but it was observed that they cause a significant decrease in the number of males that performed reproductive activities, suggesting that silencing of fruitless. alters the normal functioning of male sexual behavior. In contrast, when injecting dsRNAper. in adult males of A. sp.1, no significant alteration was observed neither in their reproductive activities nor in their general activities. Moreover, silencing of the genes seems not to affect the production of spermatozoa. Although controversial, the data are in line with observations in D. melanogaster, in which the null mutant per04 did not present significant alterations in behaviors relatives to the control males in a Day/Night cyclic regime. Thus, additional analyzes will be needed to elucidate the participation of the two genes in the coordination of behaviors in Anastrepha sp.1 aff fraterculus.