<p dir="ltr">In this dissertation, I explore how human actions (climate change, road salt, land use change, species invasions) interact with and influence morphology, disease, and population dynamics in freshwater organisms (amphibians and aquatic crustaceans). First, I examined how the incidence and timing of disease epidemics in native species (<i>Daphnia dentifera</i>) caused by a generalist parasite (<i>Metschnikowia bicuspidata</i>) influenced the success and impact of an invasive species (<i>Daphnia lumholtzi</i>) in freshwater zooplankton (Chapter 1). In the following chapter, I explored how host-parasite interactions are affected by the interactive effects of multiple environmental stressors, focusing on American bullfrog tadpoles (<i>Lithobates catesbeianus</i>), two of their common parasites (<i>Batrachochytrium dendrobatidis</i> (<i>Bd</i>) and trematode parasites in the family Echinostomatidae), and two common stressors (fluctuating temperatures and sublethal road salt pollution; Chapter 2). Finally, I investigated how the combination of climate (temperature and precipitation) and land use (developed and/or forested area) change have influenced the body size of a common toad (Fowler’s toad, <i>Anaxyrus fowleri</i>) from 1930 – 2020 utilizing museum specimens (Chapter 3). Together, this research establishes how emerging and persistent anthropogenic environmental stressors will interact to affect morphology, disease, and population dynamics in vulnerable freshwater organisms.</p>

This thesis examines the parasite dynamics and the mechanisms affecting parasite load and transmission focalising on the role played by host and habitat heterogeneities. This study is based on the gastrointestinal nematode Heligmosomoides polygyrus and the small mammal yellow necked mouse and uses data gathered from experimental field manipulations of parasites intensities and data gathered from trapping monitoring. Initially the parasite community of yellow-necked mouse (Apodemus flavicollis) was explored in North-Eastern Italian Alps with the aim to describe the major patterns and identify the factors affecting parasite community structure. Despite the observed spatial variability it has been found that differences within the host population such age and secondly sex and breeding conditions, were the major factors acting on parasite occurrence and intensity. Habitat differences had a less apparent effect on parasite community structure. The consequences of H. polygyrus infection on other parasite species infections have been analysed, in specific the infestation of the tick Ixodes ricinus in populations of A. flavicollis. H. polygyrus load and tick infestation were monitored as well as were carried out field manipulations of H. polygyrus intensity and were monitored changes in tick infestation. It has been found that H. polygyrus load was negatively related to I. ricinus infestations. Host factors mediated the H. polygyrus-I. ricinus interaction such that young and non-breeding mice exhibited higher I. ricinus to H. polygyrus intensity respect breeding adults. The role of host sex on parasite abundance was then investigated carrying out a field experiment where the H. polygyrus intensity were manipulated in relation to mice gender. In specific, H. polygyrus was removed alternately from either sexes and the parasite load was analysed in the untreated sex. It was found that males mice were responsible to drive parasite transmission in the host population and this was observed in absence of sex-bias in parasite infection, suggesting that this pattern was not a mere consequence of quantitative differences in parasite loads between sexes. To disentangle the possible mechanism causing this sex bias in parasite transmission mathematical simulations based on parameters obtained for the field experiment were used. Two non mutually exclusive hypotheses causing sex bias in parasite transmission were tested: a- males immune response is less efficient and this causes the development of more successful parasite infective stages or b-males behaviours allow them to be more efficient is spreading in more exposed areas parasite infective stages. Multi-host models were developed and simulations were compared with field results. While it was not disentangled the most dominant mechanism causing sex bias in parasite transmission this study underlined the importance of host sexes in affecting parasite dynamics and host-parasite interaction. In conclusion this thesis highlighted the importance of considering host and environmental differences when investigating host parasite interactions. This finding could be extremely important when planning measured of disease control or to avoid disease outbreak. Controlling target group of individuals host could avoid economical losses and a more effective measure of intervention.

Fasioloides magna is a trematode parasitizing in the liver parenchyma of ruminants. Its life cycle is associated with the humid environment and includes intermediate freshwater snail hosts from family Lymnaeidae. According to the ability of host to form a certain type of a pseudocyst during fascioloidosis, they can be,divided in three groups, specific definitive hosts (red deers, fallow deers, roe deers), nonspecific definitive hosts (cattle, wild boars and elks) and atypical hosts (sheeps and goats). Beside the natural infections also the experimental infections of other potential host species has been realized (chamois, llama and bighorn sheep and traditional laboratory animals such as mice, guinea pigs, rats and rabbits). In the context of different diseases, many changes in infected organism can occur. These can be qualitatively and quantitatively evaluated. Similarly, during fascioloidosis the changes associated with the presence of the parasite in the host's body is possible to monitor, e.g. antibody production, increase in the number of eosinophils, release of eggs in faeces, internal bleeding, or the level liver damage. The liver damage is corresponding primarily to biochemical parameters of blood, not only the liver enzymes, but also other blood components, like blood proteins, lipids,...

A esquistossomose é uma doença importante em saúde pública. Dos genes selecionados como diferencialmente expressos em esquistossômulos a partir do transcriptoma do S. mansoni, 56% foram confirmados por RT-PCR em tempo real. Entre eles, a proteína Ly6.5, está presente no tegumento de esquistossômulos e vermes adultos por âncoras de GPI. Não foi detectada a função de inibir o sistema complemento, mas pode estar envolvido na manutenção do tegumento. O gene SmVal7 revelou transcritos nas glândulas esofágicas de vermes adultos por hibridização in situ, enquanto a localização da proteína não está definida. Anexina está associada ao tegumento de esquistossômulos e vermes adultos, de maneira dependente de cálcio. A supressão do gene por RNAi não resultou em alteração fenotípica significativa em esquistossômulos in vitro. Foi observada atividade parcial de inibição de coagulação e potencial atividade de endocitose de anticorpos ligados à superfície. A imunização com rLy6.5, rSmVal7, rAneI-II ou rAneII-III não levou a redução da carga parasitária após desafio. / Schistosomiasis is an important disease in public health. Genes selected from the S. mansoni transcriptome, 56% of them were confirmed as differentially expressed in schistosomula by real time RT-PCR. Among them, the protein Ly6.5 is present in the tegument of schistosomula and adult worms by GPI anchors. The function of inhibiting the complement system was not detected, but it may be involved in maintenance of the tegument. The gene SmVal7 revealed transcripts in the esophageal glands of adult worms by in situ hybridization, while the localization of the protein is not defined. Annexin is associated with the membranes of the schistosomula and adult worms tegument in a calcium-dependent manner. The suppression of the gene by RNAi did not resulted in a significant phenotypic change in schistosomula in vitro. Parcial inhibition of the coagulation activity and potential function of endocytosis of membrane-bound antibodies were observed. Immunization with the rLy6.5, rSmVal7, rAneI or rAneII-II-III did not show reduction in worm burden recovery after challenge.

Escherichia coli enteropatogênica (EPEC) é uma das principais causas de diarreia em crianças. Na carência de fosfato (Pi), um conjunto de genes conhecido como regulon PHO é induzido. Esse regulon é controlado pelo sistema Pst, que além de ser um transportador de Pi, reprime a expressão de PHO quando Pi é abundante, e pelo sistema de dois componentes PhoB/PhoR. A deleção de pst reduziu a adesão de EPEC à células epiteliais in vitro, pois diminuiu da expressão dos reguladores PerA/PerC, que por sua vez controlam a expressão de genes envolvidos na adesão. Este efeito foi exclusivo de pst e não devido a expressão constitutiva dos genes de PHO causada pela deleção de pst. A expressão da fímbria BFP, PerA e PerC também dependem da síntese de ppGpp, uma molécula de alarme envolvida na regulação de genes relacionados à carência nutricional. ppGpp regula positivamente a expressão de PerA e PerC. Entretanto, RpoS, o fator relacionado à resposta ao estresse, afetou negativamente o nível de adesão de EPEC e a expressão de BFP. / Enteropathogenic E. coli (EPEC) is one of the causes of diarrhea in children. Phosphate (Pi) shortage induces transcription of the genes known as the PHO regulon. These genes are controlled by the Pst system, that is also a high-affinity Pi transporter, and represses PHO expression under Pi-replete conditions. PHO is also controlled by the two-component system PhoB/PhoR. Deletion of the pst operon reduced the adhesion of EPEC to epithelial cells in vitro due to a decrease in the expression of the regulators PerA and PerC that in turn control the expression of genes related to adhesion. The constitutive expression of the PHO genes in the pst mutant was not the cause of adhesion inhibition. Expression of bfp and the regulators PerA and PerC was also dependent on ppGpp, an alarmone involved in the regulation of genes related to nutrient limitation. On the other hand, RpoS, the factor that controls the general stress response, negatively affected EPEC adhesion and bfpA expression.

L’histoire de l’ostréiculture française met en évidence la fragilité de cette production face à la surexploitation des stocks et l’apparition de maladies. En particulier, la production d’huître plate, Ostrea edulis, a fortement diminué suite à l’apparition de deux maladies parasitaires dont la bonamiose. Les moyens de lutte contre la bonamiose sont relativement restreints. Ils sont essentiellement basés sur la surveillance de la santé des huîtres afin de limiter la dissémination et la propagation de la maladie. Cependant l’utilisation de modèles prédictifs de l’évolution de la maladie en zone infectée permettrait d’optimiser la gestion des stocks et minimiser l’impact des agents pathogènes. De plus, le développement d’animaux résistants à l’infection pourrait permettre de relancer cette production. Ces différentes approches nécessitent des outils diagnostiques adaptés, une bonne connaissance du cycle de vie de l’agent pathogène, et, plus particulièrement des interactions du parasite avec son hôte. Dans ce contexte, l’objectif principal du travail de thèse proposé est de comprendre les interactions entre l’huître plate Ostrea edulis et son parasite Bonamia ostreae, et, plus particulièrement les bases moléculaires de la résistance au parasite. Dans un premier temps, la réalisation d’une banque soustractive d’ADNc a permis d’identifier des ESTs différentiellement exprimées chez des hémocytes en réponse au parasite. L’expression de certains gènes dont une galectine a été mesurée en PCR en temps réel dans le contexte d’infections in vitro. En complément, la réponse cellulaire a été étudiée par cytométrie en flux et l’infection contrôlée en microscopie. Ces expériences ont montré une multiplication parasitaire dans les hémocytes au cours du temps associée à une diminution de la production d’EOR et d’estérases. Dans un second temps, il a été entrepris une étude comparative entre une population d’huîtres plates résistantes à la bonamiose et une population naturelle. Les résultats obtenus tendent à montrer qu’une modulation de l’apoptose et une diminution de la phagocytose seraient impliquées dans les mécanismes liés à la résistance à la bonamiose. Ce travail est le premier à étudier la réponse des hémocytes d’huîtres plates à une infection par le parasite Bonamia ostreae au niveau cellulaire et moléculaire. / The history of the French oyster production highlights the fragility of this production against overexploitation and disease outbreaks. In particular, the production of flat oyster, Ostrea edulis, has decreased following the emergence of two parasitic diseases including bonamiosis. The means to fight against bonamiosis are relatively limited. They are mainly based on oyster health surveillance to limit the spread of the disease. However, the use of predictive models of disease progression in infected area would help to improve stock management and minimize the impact pathogens. Moreover the development of resistant animals could help to revive this production. These different approaches require appropriate diagnostic tools, a good knowledge of the life cycle of the pathogen, and the interactions between the parasite and its host. In this context, the main objective of the phD work is to understand the interactions between the flat oyster Ostrea edulis and the parasite Bonamia ostreae, and particularly the molecular basis of the resistance to the parasite. In a first step, a subtractive cDNA bank allowed the identification of ESTs differentially expressed in haemocytes in response to the parasite. Expression of some genes, among which a galectin, was measured by Real Time PCR in the context of in vitro infections. In addition, the cellular response was investigated by flow cytometry and the infection was checked by microscopy. These experiments showed a multiplication of the parasite inside haemocytes associated with a decreased of esterases and of the production of ROS. In a second step, a comparative approach was carried out between a population of oysters resistant to bonamiosis and a natural population. Results suggest that modulation of apopotosis and decrease of phagocytosis could be involved in mechanisms related to resistance to bonamiosis. This work is the first study on the response of haemocytes of flat oysters to an infection with the parasite Bonamia ostreae at the cellular and molecular levels.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-03-22T13:55:27Z No. of bitstreams: 1 2016 - Mariana Gomes Lima.pdf: 3236185 bytes, checksum: 3517b88022c7943663ae0274085cc96a (MD5) / Made available in DSpace on 2017-03-22T13:55:27Z (GMT). No. of bitstreams: 1 2016 - Mariana Gomes Lima.pdf: 3236185 bytes, checksum: 3517b88022c7943663ae0274085cc96a (MD5) Previous issue date: 2016-02-23 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The nematode Angiostrongylus cantonensis has gained notoriety under the public health point of view out of its endemic area in Asia, since in recent years cases of eosinophilic meningoencephalitis have been reported in other continents, such as in the Americas. In Brazil over thirty cases of this zoonotic disease have been recorded and there are reports of the presence of this nematode in 11 states. In its life cycle A. cantonensis has, along their development, need for intermediate and final host, involving predominantly molluscs and rodents, respectively, and different paratenic hosts, humans are accidental hosts. They can also infect a rich variety of terrestrial and aquatic molluscs and in this study, there were used specimens of neotropical planorbids transmitters of schistosomiasis, Biomphalaria straminea and Biomphalaria tenagophila, born and reared in the laboratoryfor experimental infection by A. cantonensis. In order to characterize the resulting physiological profile that host-parasite relationship, the host molluscs were separated into groups of infected and uninfected containing thirty molluscs per group, all made in triplicates. The investigation lasted 21 days, after the pre-patent period molluscs were dissected to collect hemolymph, removal of tissues and use the shell for calcium concentration check. The reproductive biology of both species was observed on the viability of the eggs, as well as (A) Number of eggs/ eggs mass; (B) Number of eggs/mollusc; (C) Number of eggs mass/mollusc and (D) Number of hatched molluscs/mollusc. In the hemolymph there were quantified total protein, glucose, uric acid, urea, the activity of transaminases AST and ALT, lactate dehydrogenase - LDH, and organic acids (oxalic, pyruvic, succinic and lactic). In the cephalopedal mass and digestive gland the glycogen content was measured and in the albumen gland, it was measured the galactogen. The results showed that the snails infected, of both species showed significant reduction in the viability of eggs, 50% for B. straminea and 10% for B. tenagophila. The metabolic status was also strongly affected, with mobilization of glycogen and hemolymph glucose reduction of 68% to B. straminea and 6.48% for B. tenagophila, followed by anincrease of LDH activity of 518.25% to 320.14% B. straminea and to B. tenagophila as well as the concentration of lactic acid, 5,7mM to B. straminea and 0.34 mM for B. tenagophila, and reduction in the concentration of pyruvic and succinic acids. The intense release of gluconeogenic amino acids from protein catabolism reversed the excretory pattern from uricotelic to ureotelic in both B. tenagophila and B. straminea. The tissue injury showed increase of 69% in the concentration of total proteins in the hemolymph of B. straminea and the intense activity of aminotransferases, ALT and AST, an increase of 241.26% and 360%, respectively, compared to the control group. The infection reduced by 51.57% the concentration of total circulating proteins to B. tenagophila, and increased ALT and AST activity at 310% and 280% compared to uninfected snails. The results of this study indicate that the energetic and structural metabolism of both species is strongly affected by the infection, showing the activation of an anaerobic to compensate for the depletion of glicidics stores as well as the change in the excretory pattern depending on the use of substrates protein for the production of ATP. Although studies on clinical, pathological and epidemiological neural angiostrongyliasis have been extensively carried xiii out, data on the metabolic and reproductive profile of snails infected by A. cantonensis are scarce. This pioneering study focused on species of host-parasite relationship certainly contribute to the knowledge of this interaction, as well as to the epidemiology of transmission A. cantonensis in addition to providing new subsidies which can be used in prevention and control of eosinophilic meningitis, zoonosis considered emerging in Brazil. / O nemat?deo Angiostrongylus cantonensis tem ganhado notoriedade sob o ponto de vista da sa?de p?blica fora de sua ?rea end?mica na ?sia, uma vez que nos ?ltimos anos casos de meningoencefal?te eosinof?lica v?m sendo reportados em outros continentes, como nas Am?ricas. No Brasil mais de trinta casos dessa zoonose j? foram registrados e h? relatos da presen?a desse nemat?deo em 11 estados. Em seu ciclo de vida heteroxeno A. cantonensis tem, ao longo do seu desenvolvimento, a necessidade de hospedeiro intermedi?rio e definitivo, envolvendo predominantemente, moluscos e roedores, respectivamente, al?m de v?rios hospedeiros parat?nicos, sendo o homem um hospedeiro acidental. Pode infectar uma rica variedade de moluscos terrestres e aqu?ticos e neste estudo, foram utilizadas para infec??o experimental popula??es nascidas e criadas em laborat?rio de duas esp?cies de planorb?deos neotropicais transmissores da esquistossomose mans?nica, Biomphalaria straminea e Biomphalaria tenagophila. Com a finalidade de caracterizar o perfil fisiol?gico resultante dessa rela??o hospedeiro-parasito, os moluscos hospedeiros foram separados em grupos de infectados e n?o infectados, contendo trinta moluscos por grupo, todos feitos em triplicatas. A investiga??o durou 21 dias, ao fim do per?odo pr?-patente os moluscos foram dissecados para a coleta de hemolinfa, retirada de tecidos e utiliza??o da concha para verifica??o de concentra??o de c?lcio. A oviposi??o dos moluscos de ambas as esp?cies foi observada quanto ? viabilidade dos ovos, assim como (A) N?mero de massas ov?geras/molusco; (B) N?mero de ovos/molusco; (C) N?mero de ovos/massa ov?gera e (D) N?mero de moluscos eclodidos/molusco. Na hemolinfa foram quantificadas as prote?nas totais, glicose, ?cido ?rico, ureia, a atividade das transaminases ALT e AST, lactato desidrogenase - LDH, e os ?cidos org?nicos (ox?lico, pir?vico, succ?nico e l?tico). Na massa cefalopediosa e gl?ndula digestiva, foram mensurados os conte?dos de glicog?nio e na gl?ndula de alb?men, foi mensurado o galactog?nio. Os resultados obtidos mostraram que os moluscos infectados, de ambas as esp?cies, apresentaram redu??o significativa na viabilidade dos ovos, de 50% para B. straminea e de 10% para B. tenagophila. O estado energ?tico tamb?m foi vigorosamente afetado, com mobiliza??o das reservas de glicog?nio, e queda da glicemia de 68% para B. straminea e de 6,48% para B. tenagophila, seguida de uma eleva??o na atividade da LDH de 518,25% para B. straminea e 320,14% para B. tenagophila, bem como na concentra??o do ?cido l?tico, 5,7mM para B. straminea e 0,34 mM para B. tenagophila, e redu??o na concentra??o dos ?cidos pir?vico e succ?nico. A intensa libera??o de amino?cidos gliconeog?nicos a partir do catabolismo proteico inverteu o padr?o excretor de uricot?lico para ureot?lico tanto em B. tenagophila quanto em B. straminea. A les?o tecidual mostrou eleva??o de 69% na concentra??o das prote?nas totais na hemolinfa de B. straminea, bem como a intensa atividade das aminotransferases, AST e ALT, com aumento de 241,26% e 360%, respectivamente, em rela??o ao grupo controle. A infec??o reduziu em 51,57% a concentra??o de prote?nas totais circulantes para B. tenagophila, e elevou a atividade de ALT e AST em 310% e xi 280% em compara??o com os moluscos n?o infectados. Os resultados observados neste estudo indicam que o metabolismo energ?tico e estrutural de ambas as esp?cies foi vigorosamente afetado pela infec??o, mostrando a ativa??o de uma via anaer?bia para compensar a exaust?o das reservas glic?dicas, assim como a mudan?a no padr?o excretor em fun??o da utiliza??o de substratos proteicos para produ??o de ATP. Embora, estudos sobre aspectos cl?nicos, patol?gicos e epidemiol?gicos da angiostrongil?ase neural t?m sido extensivamente realizados, dados acerca do perfil metab?lico e reprodutivo de moluscos infectados por A. cantonensis s?o escassos. Este estudo pioneiro na rela??o parasito-hospedeiro das esp?cies focadas seguramente contribuir? tanto para o conhecimento dessa intera??o, quanto da epidemiologia da transmiss?o de A. cantonensis, al?m de fornecer novos subs?dios que poder?o ser utilizados em medidas de preven??o e controle da meningite eosinof?lica, zoonose considerada emergente no Brasil

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-02T21:37:15Z No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5) / Made available in DSpace on 2017-05-02T21:37:15Z (GMT). No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5) Previous issue date: 2016-02-25 / CNPq / The interaction between intermediate hosts and helminths can cause metabolic and reproductive changes in the host snail, they start using their reserves to maintain its vital functions, immune system activation, repair of tissue damage and also to supply the necessary energy for the parasites development. Investigations of co-infection by different helminths can provide essential information about the biology of their co-existence. Our aims are investigate the reproductive changes, evaluate the activity of lactate dehydrogenase and the concentrations of glucose in the hemolymph and glycogen in the digestive gland complex and cephalopedal mass, and also verify histological and histochemical changes of Biomphalaria glabrata experimentally co-infected with Echinostoma paraensei and Angiostrongylus cantonensis. Five groups of snails i.e. uninfected, with single and double infections, either E. paraensei first (E+A) or A. cantonensis first (A+E) were followed; three times a week during four weeks the numbers of egg masses, eggs and hatched snails were counted. Histological and histochemical samples of the ovotestis, albumen gland, digestive gland complex and cephalopedal mass were collected after four weeks as well samples for the biochemical analysis. The number of egg masses/snail, eggs/snail and hatched snails showed significant differences comparing the control group to all infected groups, especially in group E + A, with the majority of values of parameters analyzed lower than 50% of those observed for control snails. A significant decrease in glucose levels and glycogen content, in contrast an increase of lactate dehydrogenase activity was observed in the infected snails. These results indicate intense energy demand that takes the snail increasing the anaerobic degradation of carbohydrates in the infected snails to obtain energy in an attempt to maintain homeostasis. The histological analysis showed that presence of both parasites in the all organs analyzed and the parasites were associated with cell disorganization. / A intera??o entre hospedeiros intermedi?rios e helmintos pode causar altera??es metab?licas e reprodutivas no molusco hospedeiro, que come?am a usar suas reservas para manter as fun??es vitais, na elabora??o de resposta imunol?gica e repara??o de danos teciduais e tamb?m para compensar a drenagem de nutrientes/energia pelos parasitos em desenvolvimento. Investiga??es sobre as co-infec??es por helmintos de esp?cies diferentes podem fornecer informa??es essenciais sobre a biologia da sua co-exist?ncia, refletindo aspectos mais pr?ximos ?queles que encontramos em condi??es naturais. Nossos objetivos foram investigar as altera??es reprodutivas, avaliar a atividade da lactato desidrogenase e as concentra??es de glicose na hemolinfa e glicog?nio no complexo gl?ndula digestiva-g?nada e massa cefalopediosa, e tamb?m verificar altera??es histol?gicas e histoqu?micas de Biomphalaria glabrata experimentalmente co-infectados com Echinostoma paraensei e Angiostrongylus cantonensis. Cinco grupos de moluscos (n?o infectados, com infec??es simples (Echinostoma paraensei ou Angiostrongylus cantonensis) e dupla, seja E. paraensei primeiro (E + A) ou A. cantonensis primeiro (A + E)) foram acompanhados; tr?s vezes por semana durante quatro semanas o n?mero de massas de ovos, n?mero de ovos e n?mero de moluscos eclodidos foram contados. Amostras para a an?lise histol?gica e histoqu?mica do ovoteste, gl?ndula do alb?men, complexo gl?ndula digestiva e massa cefalopediosa foram coletadas ap?s quatro semanas, assim como amostras para a an?lise bioqu?mica. O n?mero de massas de ovos/molusco, ovos/molusco e moluscos eclodidos dos moluscos infectados apresentaram diferen?as significativas quando comparados com o grupo controle, especialmente no grupo E + A, cuja a maioria dos valores dos par?metros analisados foi inferior a 50% dos valores observados para os moluscos controle. Um decr?scimo significativo nos n?veis de glicose e de glicog?nio, em contraste com um aumento de atividade da lactato desidrogenase foi observado nos exemplares infectados. Estes resultados indicam que a demanda de energia intensa leva o molusco infectado ao aumento da degrada??o anaer?bia de carboidratos para obten??o de energia buscando atender ao aumento da demanda energ?tica, numa tentativa de manter a homeostase glic?mica, por?m redu??es significativas s?o observadas. As an?lises histol?gicas e histoqu?micas mostraram a presen?a de ambos os parasitos nos ?rg?os analisados associados com intensa desorganiza??o celular.

Trygonorrhina fasciata (Rhinobatidae) specimens naturally infected by three monogenean species were captured and maintained in marine aquaria to promote a continuous parasite load. Monogenean eggs recovered from aquaria provided larvae for descriptions and life history experiments. I describe the adult, larva and post-larval development of a new species of hexabothriid, Branchotenthes octohamatus, from the gills. This is the first monogenean larva described with only eight hooklets. This character may be useful to help resolve problematic relationships within the Hexabothriidae and offers insight into more general hypotheses about relationships within the Monogenea. I also redescribe the adult of Calicotyle australis (Monocotylidae) from the cloaca and describe the larva. The number and arrangement of larval ciliated epidermal cells and sensilla was revealed using silver nitrate. I redescribe Pseudoleptobothrium aptychotremae (Microbothriidae) adults from the skin of T. fasciata, representing a new host and locality record. Larval anatomy and post-larval development are also documented. The presence of six needle-like spicules in the larval haptor is confirmed, supporting an earlier theory that spicules are ancestral vestiges. My studies revealed three different egg hatching, host finding strategies and larval ‘types’. Branchotenthes octohamatus has a ‘sit-and-wait’ strategy, entirely dependent on mechanical disturbance to stimulate eggs to hatch. Larvae are unciliated, cannot swim, lack pigmented eyespots and show no photo-response but may survive for more than two days after hatching at 22ºC. In contrast, eggs of C. australis hatch spontaneously with a strong diurnal rhythm in the first few hours of daylight when exposed to a LD12:12 illumination regime. Larvae are ciliated and can swim, have pigmented eyespots, are photo-positive and can remain active and survive for up to 24 h after hatching at 22ºC. Eggs of P. aptychotremae may have a ‘bet-hedging’ strategy. Some eggs hatch spontaneously and rhythmically in an LD12:12 regime during the last few hours of daylight but their low hatching success rate suggests that other eggs may require a different cue provided by the host. Larvae are ciliated, can swim, lack pigmented eyespots, show no photo-response and remain active for only a few hours at 22ºC. Experiments using the fluorescent dye, 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) revealed B. octohamatus on gills of T. fasciata within 30 min of exposure to the host. This provides strong evidence that larvae invade the gills directly via the host’s inhalant respiratory current and do not migrate after initial attachment elsewhere. Five rhinobatid species (Aptychotrema vincentiana, T. fasciata, Trygonorrhina sp. A, A. rostrata and Rhinobatos typus), with overlapping distributions spanning west, south and east Australian coastal waters were surveyed for monogeneans at four locations between Fremantle, Western Australia and Stradbroke Island, Queensland. Genetic homogeneity, using the mitochrondrial gene Cytochrome b (cytb) and the nuclear marker, Elongation factor-1 alpha (EF1a), was observed for all Branchotenthes and Calicotyle specimens irrespective of collection locality or rhinobatid species. Genetic homogeneity was observed for Pseudoleptobothrium specimens collected in western and southern Australia. However, local genetic heterogeneity was apparent among Pseudoleptobothrium specimens collected from two sympatric host species in New South Wales. Analyses revealed a highly divergent clade, indicating a morphologically cryptic, ancestral species. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1323070 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008

Ceratomyxa shasta infects salmonids in the Pacific Northwest (PNW) of North America, occasionally causing losses in wild and captive populations. Host-specific parasite genotypes (O, I, II, III) were previously characterized molecularly using markers in the ribosomal DNA and phenotypically by type host in the Klamath River, CA/OR. This thesis sough to re-examine the composition of the parasite population elsewhere in the PNW and to further evaluate the host specificity of each genotype. I surveyed salmonids native to the PNW primarily from the Fraser, the Columbia and the Sacramento River basins. I also conducted sentinel studies on the Willamette and Deschutes Rivers that exposed native and non-native salmonids to parasite populations above and below migration barriers. These studies expanded upon the known host range of each genotype: O was specific to rainbow/steelhead (Oncorhynchus mykiss) and coastal cutthroat trout (O. clarkii); I was specific with Chinook salmon (O. tshawytscha); II was non-specific, being detected in six species, but predominating in coho (O. kitsuch), chum (O. keta), and pink (O. gorbuscha) salmon; and III was also non-specific infecting many native and non-native salmonids. Hosts infected with genotype III were considered “adequate” if myxospore development occurred, and included brook char (Salvelinus fontinalis), brown (Salmo trutta), rainbow, cutthroat, and steelhead trout; but Chinook, kokanee/sockeye (O. nerka) and Atlantic salmon (Salmon salar) were less suitable. The distribution, life history, and phylogenetic traits of each salmonid host are factors that potentially explain the host specificity and the spatial and temporal patterns of each genotype. Data collected in this thesis provide evidence that host-specific C. shasta coevolved with Pacific salmonids, adapting unique host-parasite relationships over time. / Graduation date: 2012