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A (Na+,K+)-ATPase presente no tecido branquial dos crustáceos osmorreguladores é um componente essencial do sistema de regulação iônica e osmótica e de excreção ativa de NH4 +. Dessa forma, a caracterização cinética da (Na+,K+)-ATPase branquial de Clibanarius vittatus pode fornecer dados importantes para a compreensão dos mecanismos bioquímicos desses processos. Nesse sentido, foram realizados ensaios cinéticos com a fração microsomal obtida do tecido branquial de C. vittatus. Os dados obtidos revelaram a presença de um único pico com atividade ATPase. Os dados obtidos a partir da hidrólise do substrato ATP revelaram a presença de dois sítios, um de alta afinidade que apresenta interação sítio-sítio (nH=1,9; K0,5= 63,8 ± 2,9 nM e V= 19,1 ± 0,8 U/mg) e outro de baixa afinidade, com características Michaelianas (nH=1,0; KM= 44,1 ± 2,6 mM e VM= 123,5 ± 6,1 U/mg). Além disso, foi observada interação do tipo sítio-sítio (nH= 1,8) para estimulação dos íons magnésio (V= 132,0 ± 5,3 U/mg e K0,5= 0,36 ± 0,02 mM). Por outro lado, as interações dos íons sódio (K0,5= 7,4 ± 0,4 mM), potássio (K0,5= 1,51 ± 0,05 mM) e amônio (K0,5= 4,5 ± 0,2 mM) obedeceram a uma cinética Michaeliana. Os dados obtidos mostraram que em condições saturantes de íons potássio, a atividade enzimática aumenta de maneira concentração-dependente quando na presença de íons amônio (V= 290,8 ± 14,4 U/mg e KM= 2,9 ± 0,1 mM). A presença da ouabaína no meio reacional acarretou 66% de inibição (KI= 117,3 ± 3,5 mM), indicando a presença de outras ATPases. A atividade K+-fosfatase, medida usando o substrato sintético PNFF revelou a presença de um único sítio de hidrólise (V= 47,4 ± 1,9 U/mg e KM= 1,0 ± 0,04 mM) com características Michaelianas (nH= 1,1). Entretanto, para os íons Mg2+ (nH= 2,6 e K0,5= 0,6 ± 0,1 mM), K+ (nH= 1,4 e K0,5= 3,7 ± 0,1 mM) e NH4 + (nH= 1,9 e K0,5= 19,3 ± 0,7 mM) foram observadas interações sítio-sítio. Em conjunto, os resultados cinéticos obtidos para a (Na+,K+)-ATPase branquial de C. vittatus, reforçam a possibilidade de que essa enzima contribui eficientemente com o mecanismo adaptativo desencadeado em resposta à exposição a altas concentrações externas de amônia. É importante ressaltar que o presente trabalho representa o primeiro estudo de caracterização cinética da enzima (Na+,K+)- ATPase no ermitão C. vittatus, um animal inserido em um microambiente constituído pela concha. Finalmente, este trabalho agrega informações relevantes relacionadas ao grupo dos Decapoda, em relação ao papel da (Na+,K+)-ATPase nos processos metabólicos e de osmorregulação nos crustáceos. / (Na+,K+)-ATPase is an essential component of osmotic and iono-regulatory system of crustacean gill and plays a relevant role in NH4 + excretion. Thus, the kinetic characterization of Clibanarius vittatus gill microsomal (Na+,K+)-ATPase, can provide crucial data to the understanding of osmoregulating and excretion processes of these animals in different environments, especially the shell microenvironment. In this way, kinetic assays were performed with gill microsomal fraction. The results showed a single protein peak with ATPase activity. The results from ATP hydrolysis revealed the presence of two different sites. One of them is related to the high-affinity with site-site interaction (nH=1.9; K0.5= 63.8 ± 2.9 nM and V= 19.1 ± 0.8 U/mg) and the other shows low-affinity, obeying Michaelis-Menten kinetics (nH=1.0; KM= 44.1 ± 2.6 mM and VM= 123.5 ± 6.1 U/mg). Moreover, site-site interaction (nH= 1.8) was observed to the magnesium ions stimulation (V= 132.0 ± 5.3 U/mg and K0.5= 0.36 ± 0.02 mM). On the other hand, the stimulations of Na+ (K0.5= 7.4 ± 0.4 mM), K+ (K0.5= 1.51 ± 0.05 mM) and NH4 + (K0.5= 4.5 ± 0.2 mM), followed Michaelis-Menten kinetics. At saturating concentrations of potassium ions, the enzyme activity was increased in a dependent-concentration manner with increasing ammonium ions concentration (V= 290.8 ± 14.4 U/mg and KM= 2.9 ± 0.1 mM). Ouabain inhibited 66% of (Na+,K+)-ATPase activity (KI= 117.3 ± 3.5 mM) of the gill microsomal enzyme, indicating the presence of other ATPase activities. The K+-fosfatase activity was also measured using PNPP, a synthetic substrate. PNPP hydrolysis resulted in single site hydrolysis site, obeying the Michaelis-Menten kinetics (nH= 1.1). However, it was observed site-site interactions for Mg2+ (nH= 2.6; K0.5= 0.6 ± 0.1 mM), K+ (nH= 1.4; K0.5= 3.7 ± 0.1 mM) and NH4 + (nH= 1.9; K0.5= 19.3 ± 0.7 mM) ion estimulations. Taken together, these results provide important informations on the kinetic properties of gill (Na+,K+)-ATPase of C. vittatus, emphasizing the synergiistic stimulation induced by K+ and NH4 +, which appear to contribute to the efficient adaptive mechanism triggered by exposure of high external ammonia concentrations. It is important to emphasize that this is the first study on the kinetic properties of (Na+,K+)-ATPase enzyme of the hermit crab C. vittatus. Finally, this study provide information concerning the (Na+,K+)-ATPase role in the metabolic and osmoregulating mechanisms of this Decapoda group

Orientador: Antonio Fernando Monteiro Camargo / Banca: Wagner Cotroni Valenti / Banca: Newton Castagnolli / Resumo: Esta dissertação de mestrado está organizada em três capítulos. O capítulo I apresenta uma introdução geral, onde são abordados os impactos ambientais provocados pela atividade da aqüicultura e o tratamento de efluente por wetlands construídas. O capítulo II e III são referentes à experimentos desenvolvidos no Setor de Carcinicultura do Centro de Aqüicultura da UNESP (CAUNESP), em Jaboticabal - SP. Os principais objetivos destes capítulos foram: (Capítulo II) determinar os possíveis impactos provocados pelo manejo utilizado para a manutenção de camarões reprodutores da espécie Macrobrachium rosenbergii na água utilizada no viveiro; (Capítulo III) avaliar a eficiência de uma wetland construída e povoada com duas espécies de macrófitas aquáticas flutuantes (Eichhornia crassipes e Salvinia molesta) com diferentes exigências nutricionais para otimizar o tratamento do efluente de viveiro de manutenção de reprodutores de M. rosenbergii. O resultado do Capítulo II permite concluir que o manejo realizado para a manutenção de camarões reprodutores da espécie Macrobrachium rosenbergii entre as biomassas de 403,18 e 547,90 g.m-2, durante o período de 50 dias, provoca o aumento dos valores de condutividade elétrica, pH, turbidez e MPS; das concentrações de NIT, NT, P-PO4, PD e PT e reduz a concentração de O2 dissolvido. No Capítulo III, concluiu-se que a wetland construída e povoada com S. molesta e com E. crassipes + S. molesta foram menos eficiente no tratamento do efluente de viveiro de manutenção de M. rosenbergii do que a wetland construída e povoada com E. crassipes. / Abstract: The world production of Malaysia shrimp, Macrobrachium rosenbergii, presented considerable growth in the last decades. However, shrimp farming activity can provoke impacts in water used in the vivarium. The aim of this work was to determine the possible impacts provoked by the management accomplished for the maintenance of reproductive shrimp of the species M. rosenbergii in water used in the vivarium during the period 50 day. In the beginning of the experiment (03/14/2007) the vivarium, located in the CAUNESP, in Jaboticabal (SP-Brazil), with 193 m2 of surface and 1.1 m was full of 1.263 reproductive shrimp with medium weight of 61.61 ± 16.55 g.indivíduo-1. Weekly, it was determined value of temperature, pH, dissolved oxygen, electric conductivity, total alkalinity, turbidity, particulate material in suspension (PMS), total inorganic nitrogen (TIN), total nitrogen (TN), orthophosphate (P-PO4), dissolved phosphorus (DP) and total phosohorus (TP) of supply water and of the effluent of the vivarium. The values of electric conductivity and pH were significantly (p<0,05) higher in the effluent relation to the ones of the supply water. The 25 turbidity and PMS values were significantly higher in the effluent of the vivarium in seven weeks. Except for the dissolved oxygen and of TN, the TIN, P-PO4, DP and TP concentrations were significantly higher in the effluent when compared to the supplu water of vivarium in all the weeks evaluated. Among the forms of phosphorus, the biggest increases were observed for DP and and TP concentrations with 82.76 μg.L-1 and 174.75 μg.L-1, respectively. Among nitrogeneous forms, the biggest increase was observed for TIN with 81.32 μg.L-1. The management accomplished for the reproductive maintenance of M. rosenbergii during the period 50 days caused impact in water used in the vivarium... (Complete abstract, click electronic access below) / Mestre

Made available in DSpace on 2014-12-17T15:36:32Z (GMT). No. of bitstreams: 1 JosePL.pdf: 882497 bytes, checksum: cb18f10b1c9406dc277552138c430082 (MD5) Previous issue date: 2007-12-21 / Reproductive behavior of Dendrocephalus brasiliensis Pesta, 1921(Crustacean: Anostracan). The reproductive behavior of fresh water Anostracan has been poorly investigated in carcinology specialized literature, specifically in relation to Dendrocephlaus brasiliensis Pesta, 1921, with abundant data just about the geographical distribution of that Anostracan. The objective of this work was to know the reproductive behavior of this Anostracan, in different seasons (dry and raine). For this, ripe individuals of both sexes were collected in four pond of the of Fish farming Station of Paulo Afonso (EPPA), through monthly captures in each ponds, from December 2004 to November 2005. The type of reproduction was observed after the placing in aquariums (a) of then with males and (b) other on individually, at the nauplii phase, where they stayed for 15 days until the reproductive age. The sexual proportion was calculated through the relative frequencies of males and females, every month, for the whole collection period during 10 days. the production of cysts was related to the size of the female. The male: female ratio in the study period was 1 male:1,07 female. The proportion male:female was from 48,25% : 51,75% along the year. Concerning the type of reproduction, it was observed that this specie is characteerized by sexual reproduction / O comportamento reprodutivo de anostr?ceos de ?gua doce tem sido pouco divulgado na literatura especializada em carcinologia, especificamente em rela??o a Dendrocephlaus brasiliensis Pesta, 1921, havendo apenas dados abundantes sobre a distribui??o geogr?fica desse anostr?ceo dulciaqu?cola. O objetivo deste trabalho foi conhecer o comportamento reprodutivo deste an?straceo, em diferentes per?odos (seco e chuvoso). Para isto, foram utilizados indiv?duos maduros de ambos os sexos coletados em quatro viveiros da Esta??o de Piscicultura de Paulo Afonso (EPPA), atrav?s capturas mensais em cada viveiro, durante o per?odo de dezembro de 2004 a novembro de 2005. O tipo de reprodu??o foi observado atrav?s da coloca??o simult?nea em aqu?rios de algumas f?meas e machos, e outras individualmente a partir da fase de n?uplios, onde permaneceram por 15 dias at? a idade reprodutiva. A propor??o sexual dos indiv?duos foi calculada atrav?s das freq??ncias relativas de machos e de f?meas, a cada m?s, para todo o per?odo de coleta. Aos 10 dias, ? poss?vel a produ??o de cistos e o n?mero destes est? relacionado ao tamanho da f?mea. A rela??o macho:f?mea no per?odo de estudo foi de 1 macho para 1,07 f?mea. A propor??o macho: f?mea foi de 51,75% de f?mea para 48,25% de machos ao longo do ano. Quanto ao tipo de reprodu??o, foi observado tratar-se de reprodu??o sexuada

A (Na+,K+)-ATPase presente no tecido branquial dos crustáceos osmorreguladores é um componente essencial do sistema de regulação iônica e osmótica e de excreção ativa de NH4 +. Dessa forma, a caracterização cinética da (Na+,K+)-ATPase branquial de Clibanarius vittatus pode fornecer dados importantes para a compreensão dos mecanismos bioquímicos desses processos. Nesse sentido, foram realizados ensaios cinéticos com a fração microsomal obtida do tecido branquial de C. vittatus. Os dados obtidos revelaram a presença de um único pico com atividade ATPase. Os dados obtidos a partir da hidrólise do substrato ATP revelaram a presença de dois sítios, um de alta afinidade que apresenta interação sítio-sítio (nH=1,9; K0,5= 63,8 ± 2,9 nM e V= 19,1 ± 0,8 U/mg) e outro de baixa afinidade, com características Michaelianas (nH=1,0; KM= 44,1 ± 2,6 mM e VM= 123,5 ± 6,1 U/mg). Além disso, foi observada interação do tipo sítio-sítio (nH= 1,8) para estimulação dos íons magnésio (V= 132,0 ± 5,3 U/mg e K0,5= 0,36 ± 0,02 mM). Por outro lado, as interações dos íons sódio (K0,5= 7,4 ± 0,4 mM), potássio (K0,5= 1,51 ± 0,05 mM) e amônio (K0,5= 4,5 ± 0,2 mM) obedeceram a uma cinética Michaeliana. Os dados obtidos mostraram que em condições saturantes de íons potássio, a atividade enzimática aumenta de maneira concentração-dependente quando na presença de íons amônio (V= 290,8 ± 14,4 U/mg e KM= 2,9 ± 0,1 mM). A presença da ouabaína no meio reacional acarretou 66% de inibição (KI= 117,3 ± 3,5 mM), indicando a presença de outras ATPases. A atividade K+-fosfatase, medida usando o substrato sintético PNFF revelou a presença de um único sítio de hidrólise (V= 47,4 ± 1,9 U/mg e KM= 1,0 ± 0,04 mM) com características Michaelianas (nH= 1,1). Entretanto, para os íons Mg2+ (nH= 2,6 e K0,5= 0,6 ± 0,1 mM), K+ (nH= 1,4 e K0,5= 3,7 ± 0,1 mM) e NH4 + (nH= 1,9 e K0,5= 19,3 ± 0,7 mM) foram observadas interações sítio-sítio. Em conjunto, os resultados cinéticos obtidos para a (Na+,K+)-ATPase branquial de C. vittatus, reforçam a possibilidade de que essa enzima contribui eficientemente com o mecanismo adaptativo desencadeado em resposta à exposição a altas concentrações externas de amônia. É importante ressaltar que o presente trabalho representa o primeiro estudo de caracterização cinética da enzima (Na+,K+)- ATPase no ermitão C. vittatus, um animal inserido em um microambiente constituído pela concha. Finalmente, este trabalho agrega informações relevantes relacionadas ao grupo dos Decapoda, em relação ao papel da (Na+,K+)-ATPase nos processos metabólicos e de osmorregulação nos crustáceos. / (Na+,K+)-ATPase is an essential component of osmotic and iono-regulatory system of crustacean gill and plays a relevant role in NH4 + excretion. Thus, the kinetic characterization of Clibanarius vittatus gill microsomal (Na+,K+)-ATPase, can provide crucial data to the understanding of osmoregulating and excretion processes of these animals in different environments, especially the shell microenvironment. In this way, kinetic assays were performed with gill microsomal fraction. The results showed a single protein peak with ATPase activity. The results from ATP hydrolysis revealed the presence of two different sites. One of them is related to the high-affinity with site-site interaction (nH=1.9; K0.5= 63.8 ± 2.9 nM and V= 19.1 ± 0.8 U/mg) and the other shows low-affinity, obeying Michaelis-Menten kinetics (nH=1.0; KM= 44.1 ± 2.6 mM and VM= 123.5 ± 6.1 U/mg). Moreover, site-site interaction (nH= 1.8) was observed to the magnesium ions stimulation (V= 132.0 ± 5.3 U/mg and K0.5= 0.36 ± 0.02 mM). On the other hand, the stimulations of Na+ (K0.5= 7.4 ± 0.4 mM), K+ (K0.5= 1.51 ± 0.05 mM) and NH4 + (K0.5= 4.5 ± 0.2 mM), followed Michaelis-Menten kinetics. At saturating concentrations of potassium ions, the enzyme activity was increased in a dependent-concentration manner with increasing ammonium ions concentration (V= 290.8 ± 14.4 U/mg and KM= 2.9 ± 0.1 mM). Ouabain inhibited 66% of (Na+,K+)-ATPase activity (KI= 117.3 ± 3.5 mM) of the gill microsomal enzyme, indicating the presence of other ATPase activities. The K+-fosfatase activity was also measured using PNPP, a synthetic substrate. PNPP hydrolysis resulted in single site hydrolysis site, obeying the Michaelis-Menten kinetics (nH= 1.1). However, it was observed site-site interactions for Mg2+ (nH= 2.6; K0.5= 0.6 ± 0.1 mM), K+ (nH= 1.4; K0.5= 3.7 ± 0.1 mM) and NH4 + (nH= 1.9; K0.5= 19.3 ± 0.7 mM) ion estimulations. Taken together, these results provide important informations on the kinetic properties of gill (Na+,K+)-ATPase of C. vittatus, emphasizing the synergiistic stimulation induced by K+ and NH4 +, which appear to contribute to the efficient adaptive mechanism triggered by exposure of high external ammonia concentrations. It is important to emphasize that this is the first study on the kinetic properties of (Na+,K+)-ATPase enzyme of the hermit crab C. vittatus. Finally, this study provide information concerning the (Na+,K+)-ATPase role in the metabolic and osmoregulating mechanisms of this Decapoda group

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Understanding the population dynamic processes is fundamental to access its viability (health). Great part of these processes is closely related to the organism densities in a studied population. These data can reveal how the population is arranged in the environment and, in adverse conditions, it is possible to forecast the occurrence of dispersion/migration. This study aims to investigate some ecological parameters of an Aegla longirostri population, a freshwater anomuran from the southern Brazil: (i) to estimate the population density, (ii) the substrate preference, (iii) the home range and (iv) the possible dispersion patterns. The study site was the first order stream Sanga do Caranguejo, located in the central region of Rio Grande do Sul State, during 2010 and 2011. For population density and dispersion investigation covo traps were used, distributed through the stream, in different times in the year. The study of the substrate preference used PVC gutters covered by mesh and arranged on the stream substrate, with controlled density population; a surber was used for active search in the stream. The home range analysis used radio telemetry techniques. The population size was estimated in 950 individuals in spring and 210 in autumn (estimated according to the Bayesian Method). With respect to the substrate preference, we found that there is an ontogenetic association regarding the choice. The juveniles, in the first development stages, prefer sandy substrates, while adults (males and females) prefer more complex substrates. The estimates of home range of eglids vary between 8,41 to approximately 45,49 linear meters of stream, and these crustaceans showed the local dislocation, once they perform their activities around a specific point in the stream. To finalize, we identified that the juveniles and adults of eglids are active walkers, not being carried by the flow in a passive way as occurs in larvae stage in other crustaceans. Its activities are related to the environment temperature and density of adult males. We believe that the knowledge about the group has increased, however, as it is a complex group, with a rich evolutionary history, much remain to be investigated about these freshwater crustaceans. / A compreensão dos processos de dinâmica populacional é fundamental para se avaliar a viabilidade (saúde) de uma população. Grande parte desses processos estão estreitamente relacionados com a densidade de organismos da população estudada. Estes dados podem revelar como a população está distribuída no ambiente e, em condições de adversidade ambiental, é possível prever a ocorrência de dispersão/migração. Neste estudo, alicerçado nos fatores ecológicos e biológicos do aeglídeo Aegla longirostri foram investigados: (i) estimativa da densidade populacional, (ii) as preferências de substrato, (iii) a área de vida e (iv) os possíveis padrões de dispersão. Este estudo foi conduzido em um riacho de primeira ordem, Sanga dos Caranguejos, que se localiza na região central do Rio Grande do Sul, durante os anos de 2010 e 2011. Nas investigações sobre densidade populacional e dispersão foram utilizados coletores do tipo covo espaçados pelo riacho, em diferentes estações climáticas do ano. A investigação de preferência de substrato foi conduzida com o uso de calhas de PVC cobertas por malha e dispostas no riacho para o experimento em condições de densidade controlada, e com surber, para busca ativa no riacho. A análise de área de vida foi realizada com a utilização da técnica de rádio telemetria. A população em questão foi estimada em 950 indivíduos na primavera e 210 indivíduos no outono (estimativas segundo o Método Bayesiano). Com relação à preferência de substrato, verificamos que há associação ontogenética quanto à escolha. Os juvenis, nos primeiros estágios de desenvolvimento, preferem substratos arenosos, enquanto que adultos (machos e fêmeas) preferem substratos mais complexos. Estimou-se também que a área de vida desses eglídeos varia desde 8,41 metros até, aproximadamente, 45,49 metros lineares de riacho, e que estes crustáceos apresentam comportamento de deslocamento local, já que realizam suas atividades em torno de um ponto específico do riacho. Observou-se ainda que os juvenis eglídeos, assim como os adultos, são caminhadores ativos, não sendo carregados pela correnteza de forma passiva como ocorre nas fases larvais de outros crustáceos. Suas atividades estão relacionadas com a temperatura do ambiente e com a densidade de adultos machos no riacho. Acreditamos que muito se avançou em termos de conhecimento da ecologia do grupo como um todo, e não apenas para a espécie estudada, servindo este estudo de base para estudo futuros sobre dispersão e área de vida principalmente.

Les fonctions physiologiques spécifiques de la voie de N-méthylation de la phosphatidyléthanolamine (PE), une des deux voies de biosynthèse de la phosphatidylcholine (PC), restent relativement énigmatiques. Il a été démontré chez les poissons euryhalins qu’un stress hyperosmotique induisait une activation de cette voie métabolique au niveau hépatique. L’objectif de notre travail était de vérifier si ce phénomène se produit aussi chez d’autres animaux euryhalins. Les études réalisées in vivo sur deux espèces de crâbes, Eriocheir sinensis et Carcinus maenas, nous ont permis de montrer que l’acclimatation en eau de mer de ces animaux active la synthèse de PC par N-méthylation de la PE dans l’hépatopancréas. Les marquages radioisotopiques montrent aussi que cette PC est échangée avec le plasma et que ce phénomène est amplifié chez les animaux en eau de mer. Ce pool de PC est utilisé comme précurseur de la bétaïne, un osmoeffecteur organique important chez ces animaux. Nous avons ensuite caractérisé la voie de N-méthylation de la PE chez un animal osmoconformeur, la moule Mytilus galloprovincialis. Les résultats, obtenus in vivo et in vitro sur les tissus isolés, démontrent qu’une activité de N-méthylation de la PE en PC est exprimée dans la glande digestive et les hémocytes circulant de M. galloprovincialis. La PC ainsi synthétisée dans ces tissus est échangée avec l’hémolymphe de l’animal. De l’ensemble de ces observations, nous pouvons conclure que la synthèse de PC par N-méthylation est largement exprimée chez les animaux marins euryhalins et qu’une des fonctions physiologiques de cette voie métabolique est de synthétisée des osmolytes organiques comme la bétaïne / The specific physiological functions of the N-methylation of phosphatidylethanolamine (PE), one of the two biosynthetic pathways of phosphatidylcholine (PC), remain relatively mysterious. It has been demonstrated in euryhaline fish that hyperosmotic stress induced activation of this pathway in the liver. The aim of our work was to verify whether this phenomenon also occurs in other euryhaline animals. In vivo studies on two species of crabs, Eriocheir sinensis and Carcinus maenas, showed that seawater acclimation activates PC synthesis by N-methylation of PE in the hepatopancreas. Radioisotopic labelling also showed that PC is exchanged with the plasma and that this phenomenon is amplified in animals in seawater. This pool of PC is used as a precursor of betaine, an important organic osmoeffector in these animals. We then characterized the process of PE N-methylation in an osmoconforming animal, the mussel Mytilus galloprovincialis. The results, obtained in vivo and in vitro on isolated tissues, show that N-methylation of PE to PC is expressed in the digestive gland and circulating haemocytes in M. galloprovincialis. The PC synthesized in these tissues is exchanged with hemolymph of the animal. From all these observations, we conclude that the synthesis of PC by N-methylation is widely expressed in marine euryhaline animals and that a physiological function of this pathway is to provide organic osmolytes such as betaine

Marine bioeroders, borers, and burrowers can have drastic effects to marine habitats and facilities. By physically altering the structure of marine habitats, these organisms may elicit ecosystem-level effects that cascade through the community. While borer damage is typically restricted to a few substratum types, burrowing isopods in the genus Sphaeroma attack a diversity of substrata in tropical and temperate systems. My dissertation examined how boring sphaeromatid isopods affect coastal habitats (saltmarshes, mangroves) and other estuarine substrata as well as marine structures. I used a combination of lab and mensurative field experiments to quantify the effects of boring by isopods and examine how select factors affect the colonization, hence burrowing damage by isopods. I explored these questions primarily using the temperate boring sphaeromatid, Sphaeroma quoianum, as a model organism. My initial lab experiments quantified the per capita erosion rates of S. quoianum in four commonly attacked estuarine substrata. I found marsh banks and Styrofoam substrata were the most affected per capita. I supplemented this lab experiment with a year-long mensurative field experiment examining how erosion rates differ between marshes infested and uninfested by boring isopods. Marshes infested with isopods eroded 300% faster than uninfested marshes. I further examined the boring effects on Styrofoam floats. I compiled surveys and observations and conducted a short experiment to describe how isopods affect Styrofoam floats used in floating docks. I observed dense colonies of isopods attacking floats and expelling millions of plastic particles in the ocean. The boring effects to simulated Styrofoam floats were also affected by seawater temperature. Burrowing effects in Styrofoam floats exhibited a curvilinear relationship with temperature and peaked around 18°C. These results suggest a 1-2°C increase in water temperature could increase boring effects 5-17% of populations of isopods in Oregon and California bays. To examine the small-scale factors that mediate colonization and boring, I conducted a series of binary choice experiments. I found the presence of conspecifics, biofilm, and shade were important factors influencing colonization. These small scale factors likely explain why isopod attack is focused in some substrata. Finally, to examine the boring effects of tropical isopods in mangroves, I examined the associations between burrowing by S. terebrans and mangrove performance and fecundity. I found negative relationships between boring effects and performance and fecundity in two mangrove species in a restored mangrove stand in Taiwan. Together, these studies elucidate the effects of bioerosive isopods on saltmarshes, mangroves, and marine structures. However, the similar mechanisms involved in bioerosion in other boring species suggest that these results can be used to infer similar effects of other borers. In addition, since many species of sphaeromatid isopods have been introduced, this research shows how the effects of a non-native bioeroder can damage marine facilities and degrade and alter marine habitats. Through biological erosion and thus changing the physical structure of a marine habitat these non-native species can have ecosystem-level effects that cascade throughout the local community.

Global climate change is increasing the number of hot days along the California coast as well as increasing the incidence of off-shore upwelling events that lower the pH of intertidal seawater; thus, intertidal organisms are experiencing an increase in more than one stress simultaneously. This study seeks to characterize the global protein response of the eurythermal porcelain crab Petrolisthes cinctipes to changes in thermal, pH, and tidal regime treatments, either combined or individually. The first experiment examined temperature stress alone and sought to determine the effect of chronic temperature acclimation on the acute heat shock response. We compared the proteomic response of cheliped muscle tissue following a month-long acclimation to either (1) constant 10°C, (2) daily fluctuation from 10-20°C, or (3) daily fluctuation from 10-30°C, all followed by either a 30°C acute heat shock or 10°C control. We found that ATP supply via the phosphagen system, changes in glycolytic enzymes, muscle fiber restructuring, respiratory protein fragmentation, and immunity were primarily affected by acclimation and subsequent heat shock. Acclimation to the “extreme” regimes (10°C and 10-30°C) resulted in the greatest proteomic changes, while acclimation to the moderate regime (10-20°C) resulted in a more mild response to heat shock (i.e., fewer adjustments to relative protein abundance). The second experiment sought to determine the proteomic response of gill tissue following a 17 d acclimation to daily changes in pH (ambient pH 8.1 vs low pH 7.6), tidal regime (constant immersion vs 6 h emersion), and temperature (ambient 11°C vs 22-31°C heat shock during emersion). Low pH alone reduced expression of molecular chaperones of the endoplasmic reticulum, lectins, and serine proteases involved in activating the prophenoloxidase cascade. It also increased the abundance of Na+/K+-ATPase, nitrogen metabolism enzymes, and induced changes in tubulin expression, all suggesting an increase in ammonium excretion. Addition of emersion during low pH reduced the abundance of several metabolic proteins including those involved in the proposed ammonium excretion mechanism, suggesting a decrease in metabolic function in part to prevent toxic accumulation of ammonium in the branchial chambers. Combined pH, emersion, and thermal stress increased the abundance of proteins involved in cuticle binding and crosslinking. These results indicate that the responses to pH, tidal cycle, and temperature are highly dependent on one another and that changes in ER protein maturation, ion transport, immunity, and cuticle structure are the primary biochemical systems impacted by these environmental stressors in crustacean gill.

Orientador: Lílian Castiglioni-Ruiz / Banca: Fabiano Gazzi Taddei / Banca: Hermione Elly Melara de Campos Bicudo / Resumo: Os camarões do gênero Macrobrachium pertencem à família Palaemonidae. Habitam as proximidades do litoral e, também, ambientes de água doce. Com mais de cem espécies amplamente distribuídas pelas regiões tropicais e subtropicais do mundo, os palemonídeos possuem hábitos crípticos e noturnos. Particularmente na região noroeste do Estado de São Paulo, podemos encontrar várias espécies com importância econômica e ecológica consideráveis. Entretanto, existem poucos estudos na literatura relacionados ao conhecimento da biologia desses crustáceos. Assim, o presente trabalho objetivou analisar a variabilidade genética intra e interpopulacional, de amostras de duas espécies de camarões palemonídeos do gênero Macrobrachium (M. amazonicum e M. jelskii), coletadas em diferentes localidades, utilizando-se a análise das seqüências dos genes mitocondriais COI e rRNA 16S e, ainda, devido à problemática taxonômica existente entre essas duas espécies, também foi analisada a variabilidade genética interespecífica, utilizando-se os mesmos genes.. O cálculo dos índices de divergência genética intrapopulacionais apresentou valores baixos, refletindo uma provável estruturação genética destas populações. Apenas as populações de MjME e do CAUNESP apresentaram valores de divergência genética mais elevados, sugerindo um desequilíbrio na estruturação genética das mesmas, provavelmente causado por interferência antrópica e situação de cativeiro, respectivamente. Nas comparações interpopulacionais, envolvendo M. amazonicum, as taxas de divergência genética apresentaram uma ampla variação, também com média elevada. Os valores elevados nas comparações envolvendo a população do CAUNESP, permitiram descartar a hipótese da provável origem comum das populações de Macrobrachium, a partir de espécimes trazidos do Pará. Da mesma forma, as comparações... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The genus Macrobrachium (Bate, 1868) belongs to the Palaemonidae family. Their species are widely distributed in lakes, floodplains and rivers in tropical and subtropical regions of South America, having cryptic and nights habits. This genus presents nearly 210 known species with ecological and economic importance. Particulary in the northwestern state of São Paulo, we can find several species with considerable importance. However, there are few studies related to knowledge of the biology of these crustaceans. The aim this work was to analyze the genetic variability interpopulation, and interspecific of two Macrobrachium species (M. amazonicum and M. jelskii), using the mitochondrial gene sequence COI and 16S rRNA. Also, due to taxonomic problems between these two species we analyzed interspecific genetic variability, using the same genes. The intrapopulation rates of genetic divergence values were low, reflecting a probable genetic structure of these populations. Only populations MjME and CAUNESP showed higher divergence values, suggesting changes in their genetic structure of the same, probably caused by human interference and situation of captivity, respectively. In comparisons inferences involving M. amazonicum, rates of genetic divergence showed a wide range, also with high average. High values in comparisons involving the population of CAUNESP, allowed to reject the hypothesis of a probable common origin of populations the Macrobrachium specimens brought from State of Para. Interpopulation comparisons involving M. jelskii also showed high values of divergence, especially for the population collected from the Mendonça dam, whose genetic alteration in population structure was attributed to the frequent introduction of exotic specimens. The values of interspecific genetic divergence in the samples collected in the same geographic location were low for populations Adolfo (0.3%) ...(Complete abstract click electronic access below) / Mestre