Acidentes com peixes peçonhentos são muito freqüentes na região costeira do Brasil. Os bagres marinhos são os peixes peçonhentos que causam o maior número de acidentes no Brasil, especialmente no litoral sudeste, onde o bagre amarelo Cathorops spixii (Ariidae) é o mais abundante. O acidente é caracterizados por lesão dolorosa, edema intenso e ocasionalmente alguns sintomas sistêmicos. Embora estes sejam muito comuns, apenas nesta década, iniciaram-se os estudos sobre as ações dos venenos. Sendo assim, este trabalho objetivou caracterizar as principais atividades tóxicas (edema, nocicepção, hemorragia, necrose, proteólise, alterações na permeabilidade vascular e na microcirculação) induzidas pelos venenos do muco e do ferrão de C. spixii em modelo murino (Swiss, 20-22g); bem como caracterizar algumas de suas propriedades bioquímicas. Os venenos do muco e do ferrão e a mistura (muco+ferrão) foram testados. Nos ensaios de nocicepção e edema, os venenos(3; 10; 30 ou 60μg / 30μl PBS estéril) foram injetados na pata dos camundongos; para os testes de hemorragia e necrose, 50; 150 ou 300μg / 50μl PBS estéril foram injetados no dorso dos camundongos. Para a proteólise utilizou-se 3; 10; 30 ou 60μg sobre um substrato de caseína. Para a permeabilidade vascular, 10; 30 ou 60μg foram injetados via i.p. Enquanto que as alterações na microcirculação utilizouse 30μg. Ambos os venenos induziram nocicepção a partir de 3μg. Edema foi considerado significativo quando acima de 30% do controle. Todas as doses, para ambos os venenos foram capaz de induzi-lo, com pico após 2h da injeção e permanecendo por até 24h. Hemorragia e necrose não foram induzidas, mesmo após a injeção de altas doses. A permeabilidade vascular foi observada após 2h com todas as doses e venenos testados. As alterações na microcirculação foram observadas para ambos os venenos. Com 30μg observou-se o aumento do rolling de leucócitos (após10 min), extase sanguínea (após 30min, apenas no caso do muco) e contração miofibrilar (após40min). SDS-PAGE (12%) mostrou um perfil eletroforético bastante complexo, com proteínas majoritárias, acima de 97 kDa, com aproximadamente 97 kDa, entre 45 e 60 kDa e entre 20 e 30 kDa, no caso do veneno do muco. E na faixa dos 33 e acima dos 66 kDa no caso do veneno do ferrão. Diferenças são observadas entre as condições redutora e não redutora principalmente nos grupos acima de 97 kDa. A cromatografia em gel filtração não foi eficiente em separar os componentes protéicos de ambos os venenos, porém, evidencia diferenças entre os perfis. O veneno do muco contém proteínas com PI próximo a 5,0. E o ferrão deve ser composto de proteínas primordialmente aniônicas. Estes resultados mostram as importantes ações tóxicas induzidas pelos venenos do bagre marinho C. spixii, além de aspectos bioquímicos, que podem ser úteis no acompanhamento da evolução clínica e tratamento dos acidentados. / Accidents with venomous fishes are very frequent in costal waters of Brazil. Marine catfishes are the venomous fishes that cause the greatest number of accidents in Brazil, specially in southeast littoral, where the yellow-catfish Cathorops spixii (Ariidae) is the most abundant. The accidente is characterized for painful injury, intense edema and some eventul systemic symptoms; ande even these accidents are very commom,only in this decade, studies about its venoms had started. So this work intend to caracterize the main toxic activities (edema, nociception, hemorrhage, necrosis, proteolysis, alterations is vascular permeability and in microcirculation) induced by mucus and sting venom of C. spixii inmurine model (Swiss, 20-22g); besides it\'s biochemical characterization. Mucus, sting and mixture (mucus+sting) venoms were tested. In nociceptive and edematogenic assays venoms (3; 10; 30 or 60μg / 30μl sterile PBS) were injected in mice paw, whereas in hemorrhagic and necrosis assays, venoms (50; 150 or 300μg / 50μl sterile PBS) were injected in mice back. To proteolytic assay, 10; 30 or 60μg were utilized over a casein substrate. To vascular permeability assay 10; 30 or 60μg were injected by i.p. way. Whereas on microcirculation alterations 30μg were aplied over exposed cremaster muscle. All venoms induced nociception with 3 μg or more. Edema was considered significant when over 30% of control group and was induced by all venoms, too, with bottom after 2h, and continuing until 24h. Hemorrhage and necrosis weren\'t observed even so high doses were injected. Alterations on micocirculation were observed for bath venoms. 30μg increased leukocyte rolling (after 10min), blod flow stopped (after 30min, but only with mucus venom), and caused contraction on miofibres (after 40min). SDS-PAGE (12%) showed a complex pattern with several bands allover the gel. Skin and Sting venoms share some identical bands but there are important differences. Mucus venom majority bands are found between 20-30 kDa, 45-60 kDa and over 97 kDa. Sting venom major ones are between 33 kDa and 55-60 kDa. Gel- Filtration (Sephadex) showed 4 (Mucus venom) and 5 peaks (Sting venom). In this differential peak, there are molecules with high molecular weight. These results show that both venoms are able to induce important toxic activities induced by mucus and sting venoms of C. spixii. Besides, biochemical aspects that can be usefull to understand envenonming and propose better treatment.

Orientador: George Shigueki Yasui / Resumo: A triploidização é uma ferramenta interessante para produzir peixes estéreis. No Mandi amarelo, Pimelodus maculatus isso pode ser aplicada como uma ferramenta de reconstituição de espécies ameaçadas através de transplante de células germinativastronco. No capítulo I, objetivou-se estabelecer um protocolo eficiente para a triploidização da espécie P. maculatus empregando choques de térmicos. As temperaturas testadas foram de 37°C, 38°C e 39°C, 2 minutos pós-fertilização durante 2 minutos. Os embriões intactos serviram como grupo controle diplóide. A ploidia foi confirmada por citometria de fluxo, diâmetro nuclear dos eritrócitos e citogenética. Taxas de fertilização e sobrevivência foram verificadas nos principais estágios de desenvolvimento embrionário (clivagem, blástula, gástrula, segmentação e eclosão), assim como a porcentagem de larvas normais e anormais, e eficiência da triploidização. O choque térmico reduziu significativamente a sobrevivência no estágio de gástrula (P = 0,0178), somito (P = 0,0469) e incrementou o porcentual de larvas anormais (P = 0,0261). A menor sobrevivência foi observada para o tratamento a 39°C. Todos os tratamentos apresentaram altas porcentagens de indivíduos triploides, sendo o maior valor observado para o choque a 38°C (96,7%). Com base nos resultados acima, foi obtido um eficiente protocolo de triploidização em P. maculatus utilizado choque quente (38°C, 2 mpf e duração de 2 minutos). No capitulo II, foram avaliados aspectos relacionados ao... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Triploidization is an interesting tool to produce sterile fish. In the spotted catfish Pimelodus maculatus, this may be used as a tool for reconstitution of endangered species based on stem germ cell transplantation. In Chapter I, the study aimed to establish an efficient protocol for triploidization in the spotted catfish P. maculatus using temperature shock. The temperatures tested were 37°C, 38°C e 39°C, 2 min postfertilization during 2 minutes. Intact embryos served as diploid control. Ploidy status was confirmed by flow cytometry, nuclear diameter of erythrocytes and karyotyping. Fertilization and hatching rates were verified at the main embryo stages (cleavage, blastula, gastrula, somite stage and hatching), the percentage of normal and abnormal larvae and also the efficacy of triploidization. Heat shock decrease the survival at blastula stage (P= 0,0178), somite (P = 0,0469) and increased the percentage of abnormal larvae (P = 0,0261). The lowest survival was observed at 39 °C. All treatments presented high percentages of triploid individuals, and the highest values were observed for heat shock at 38°C (96,7%). Based on results above, it was obtained an efficient protocol for triploidization in P. maculatus using heat shock (38°C, 2 min postfertilization during 2 minutes). In Chapter II, aspects related to the growth and reproductive performance of diploid (2n) and triploid (3n) of P. maculatus were evaluated. The objective was to evaluate growth according with the plo... (Complete abstract click electronic access below) / Mestre

As populações de Heptapterus mustelinus do rio Uruguai, sistema da Laguna dos Patos e os rios dos sistemas costeiros do sul do Brasil e Uruguai foram revisados com dados morfológicos e moleculares. Duas novas espécies de Heptapterus foram reconhecidas. Heptapterus sp.n.A com distribuição restrita para o rio Pelotas na bacia do alto rio Uruguai. Heptapterus sp.n.B endêmica de tributários do rio Ibicuí na bacia do baixo rio Uruguai. As novas espécies distinguem-se do seu congênere mais próximo H. mustelinus pelo menor número de vértebras. Dados morfométricos e moleculares são congruentes no reconhecimento das duas espécies novas. / The populations of Heptapterus mustelinus from the Uruguay River, laguna dos Patos system and the coastal streams of southern Brazil were reviewed with morphological and molecular data. Two new species of Heptapterus were recognized. Heptapterus sp.n.A with a restricted distribution to rio Pelotas, in the upper Uruguay River basin. Heptapterus sp.n.B endemic to tributaries of rio Ibicuí, in the lower Uruguay River basin. The new species are distinguished from its closest congener H. mustelinus by the lower number of vertebrae. Morphometric and molecular data are congruent to recognize the two new species.

Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-04-20Bitstream added on 2014-06-13T18:41:14Z : No. of bitstreams: 1 junqueira_mep_dr_botfm.pdf: 683998 bytes, checksum: e97592029abff7e2b2e4961aed2d337c (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / Gap / Nossos estudos objetivaram caracterizar as respostas imune inata e específica induzidas pelas peçonhas do muco e do ferrão do bagre Cathorops agassizii . A coleta dos espécimes foi realizada no complexo Baía-Estuário de Santos e São Vicente, localizado no litoral sul do Estado de São Paulo. As peçonhas (do Muco e do Ferrão) apresentaram perfil eletroforético similar entre si. Induzida a inflamação aguda em um modelo experimental murino, as peçonhas apresentaram igualmente a capacidade de induzir aumento da permeabilidade vascular e também edema de pata. A detecção de Leucotrieno B4 e Prostaglandina E2 no lavado da cavidade peritoneal dos camundongos injetados, com ambas as peçonhas, corroboram esta hipótese. Nossos resultados através da microscopia intravital mostraram que as peçonhas induzem um grande número de leucócitos rolantes nas vênulas pós-capilares com focos de extravasamento leucocitário, principalmente de neutrófilos seguido pelo influxo de macrófagos. Além disso, a peçonha do Ferrão induziu uma resolução mais rápida do influxo leucocitário ao contrário da peçonha do Muco que manteve o infiltrado macrofágico por até 7 dias. De maneira interessante, somente a citocina IL-6 foi detectada no lavado peritoneal induzida principalmente pela peçonha do Muco e as quimiocinas KC e MCP-1, por ambas as peçonhas, expressando naquele momento, a participação destes mediadores no recrutamento de neutrófilos e macrófagos para o sítio da lesão. As peçonhas foram eficazes ao induzir uma produção primária e secundária de anticorpos das classes IgM e IgG anti-venenos. Observamos ainda, uma especificidade dos anticorpos produzidos para os componentes das próprias peçonhas e também uma reatividade antigênica cruzada entre elas. / Our studies aimed to characterize the innate and specific immune responses induced by poisons of the mucus and sting of the catfish Cathorops agassizii. The collection of specimens was accomplished in the complex Bay-Estuary System of Santos and São Vicente located in the south coast of the São Paulo State. Poisons of the Mucus and Sting presented similarities in their electrophorectical profiles. In basis of an Induced process of inflammation throughout an experimental murine model, both poisons equally showed the capacity to increase the vascular permeability and also paw edema. The Leucotrien B4'S and Prostaglandine E2 detection in washings of mice peritoneal cavity injected with both poisons corroborates this hypothesis. Our results through an intravital microscopy procedure also showed that the poisons induced a great number of rolling leukocytes in the post-capillary venules with focus on the leukocyte overflow mainly neutrophiles followed by macrophage influx. Besides, the sting poison Sting induced a faster resolution of the leukocyte influx, as well as, the Mucus poison unlike maintained a macrophage infiltrated for up to 7 days. In an interesting way, only the cytokine IL-6 was detected in peritoneal washings induced mainly by the Mucus poison although Quimiokinins KC and MCP-1 were for both poisons, expressing on that moment the participation of these mediators in the neutrophile and macrophage recruitment for the lesion site. The poisons were also effective in the induction of a primary and secondary production of IgM and IgG classes of antibodies. In this research we still observed a specificity of the antibodies produced for the components of the own poisons and also a crossed antigenic reactivity among them.

Made available in DSpace on 2017-07-10T14:38:31Z (GMT). No. of bitstreams: 1 Thais_ Sobjak.pdf: 1847698 bytes, checksum: 24153036e5d148ad4316a83db73814fc (MD5) Previous issue date: 2016-12-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The advancement of agricultural techniques has enabled improvement and greater productivity, but also made the use of agrochemicals and inadequate in promoting exaggerated acute effects to non-target species. The pesticides are agents of greatest potential in degradation of aquatic environments, since, through the flood of agricultural areas, flowing organic or inorganic substances, natural or synthetic. Among the pesticides most used currently, include the non-selective herbicide glyphosate and organophosphorus compounds. To understand and prevent the damage caused to the environment, researchers have used multiple biomarkers as an effective tool for evaluation of environmental contamination. This study was divided into two studies: a manipulative experiment and other natural. The first study aimed to investigate the neurotoxic effects and about the system in larvae of Rhamdia quelen exposed to sublethal concentration of glyphosate. In this study it was possible to conclude that, in spite of the animals exposed to glyphosate having highest percentage of survival, there was an early induction of cholinesterase activity and antioxidant system, followed by the difficulty of maintaining the activities of antioxidant system in later times, resulting in a greater injury at the cellular level. The stress that animals suffer when they are exposed to the herbicide glyphosate causes a change in your metabolism being characteristic of elastic resistance. In the second study, the objective was to evaluate changes in antioxidant system and neurotoxic R. branneri present in streams with different levels of environmental contamination during autumn and winter. In this analysis, the response of biomarkers is related to the temporal variation, and possibly the exposure of these animals to pesticides. With the results obtained showed that the local biota responded differently in each season, signaling that the fauna is in contact with oxidizing agents and cholinesterasic in the period of autumn, when there is increased frequency of planting and use of pesticides / O avanço das técnicas agrícolas possibilitou melhoria e maior produtividade de alimentos, mas também fez com que o uso exacerbado e inadequado de agroquímicos promovesse efeitos agudos a espécies não-alvo. Os agrotóxicos são os agentes de maior potencial na degradação dos ambientes aquáticos, já que, por meio do deflúvio superficial de áreas agrícolas, escoam substâncias orgânicas ou inorgânicas, naturais ou sintéticas. Dentre os agrotóxicos mais utilizados atualmente pode-se citar o herbicida não-seletivo glifosato e compostos organofosforados. Para se compreender e prevenir os danos causados ao ambiente, pesquisadores têm utilizado múltiplos biomarcadores como uma ferramenta eficaz para avaliação de contaminação ambiental. O presente estudo foi dividido em dois estudos: um experimento manipulativo e outro natural. O primeiro estudo teve por objetivo investigar os efeitos neurotóxicos e sobre o sistema em larvas de Rhamdia quelen expostas a concentração subletal de glifosato. Neste estudo foi possível concluir que, apesar dos animais expostos ao glifosato terem maior porcentagem de sobrevivência, ocorreu uma indução precoce da atividade colinesterásica e do sistema antioxidante, seguido da dificuldade de manutenção das atividades do sistema antioxidante nos horários posteriores, resultando em uma maior lesão ao nível celular. O estresse que os animais sofreram ao serem expostos ao herbicida glifosato provoca uma alteração no seu metabolismo sendo característica de resistência elástica. No segundo estudo, o objetivo proposto foi avaliar alterações neurotóxicas e do sistema antioxidante em R. branneri presentes em riachos com diferentes níveis de contaminação ambiental nos períodos de outono e inverno. Nesta análise, a resposta dos biomarcadores está relacionada à variação temporal, e possivelmente à exposição desses animais à agrotóxicos. Com os resultados obtidos observou-se que, a biota local respondeu de forma diferente em cada estação, sinalizando que a fauna está em contato com agentes oxidantes e colinesterásicos no período de outono, quando há maior frequência de plantio e uso de agrotóxicos

This study was conducted on Willard Bay Reservoir, Utah, during the summers of 1972 and 1973, to study the spawning biology of the channel catfish (Ictalurus Punctatus), and to evaluate potential reproduction for this species in the reservoir. Potential reproduction was evaluated through examination of available natural habitat and use of artificial habitat provided for this study. Channel catfish were found to mature by the end of their fourth year and spawn from mid-June through August. A total of 12 nesting channel catfish were observed along 15, 0.8 kilometer (0.5 mile) sections of dike. Ten of these 12 nests were along the dike-bottom interface. Numerous other areas along these sections of dike were considered to be favorable spawning habitat. Only four fish, two black bullheads (Ictalurus melas) and two channel catfish, were observed using artificial spawning habitat. All four fish utilized milk cans as spawning structures, rather than tire groups or plastic trash cans. Results indicated that there is suitable habitat available in the reservoir to accommodate a much larger population of channel catfish spawners. Length frequency analysis and capture of young-of-the-year show that channel catfish had spawned successfully each year since 1971.

Channel Catfish (Ictalurus punctatus, Rafinesque, 1818) leukocyte immune-type receptors (IpLITRs) are a family of proteins sharing structural and phylogenetic relationships with mammalian immune receptors. Based on their predicted signalling potential and ligand-binding properties, IpLITRs may be important in the control of immune cell effector responses in fish. The main objectives of this thesis were to determine how stimulatory IpLITRs activate cells and to develop assays for the screening of IpLITR ligands. Using cellular transfections, coimmunoprecipitation, and flow cytometry, I determined that stimulatory IpLITRs associate with specific adaptor molecules, which is required for their surface expression and signalling ability. These adaptors assemble with IpLITRs via their charged transmembrane regions and contain cytoplasmic tails encoding tyrosines that may initiate kinase pathways leading to immune cell activation. This study represents the first step towards elucidating how IpLITRs turn on immune cells. Combined with the development of assays to identify IpLITR ligands, my work sets the stage for further investigations into the functional characterization of these receptors. / Physiology, Cell and Developmental Biology

Thesis (Ph.D.)--Mississippi State University. in Veterinary Medical Sciences in the Department of Basic Sciences. / Title from title screen. Includes bibliographical references.

Disease outbreaks continue to be a major problem in the sustainable development of the aquaculture industry. Clinical outbreaks can negatively impact on the welfare of the fish and the economic gain derived from this industry. Jaundice observed as a yellow colouration in the abdominal skin, sclera of the eyes and fin bases is a significant health problem affecting the Vietnamese freshwater catfish industry. This study was designed to investigate jaundice of farmed catfish, Pangasianodon hypophthalmus using several complementary approaches. These included clinical investigations and identification of potential aetiological agents as well as epidemiological analyses to identify farm-based risk factors for this economically devastating condition occurring in the catfish farms of the Mekong Delta. The results of this survey demonstrated that the jaundice was not linked to a single geographical location as affected fish were found widely distributed throughout the five main production areas. Nor was any association found between any weight groupings, feed type or feeding regime applied in the affected farms. The highest prevalence occurred between June to October and fish mortalities ranged from 1 to 10% in the study sites. The duration of this condition was significantly correlated (P < 0.05) to mortality but not to total farm area, depth of pond, stocking density, or amount of water exchanged. The number of fish ponds affected was not as high in the large-scale farms compared to the small-scale farms. The results from the clinical description study showed that the affected fish were suffering a form of jaundice or icterus. Histological examination revealed a number of serious pathologies in the affected fish. Spleenomegaly was associated with the loss of cell structure and connective tissue and the haematopoietic tissue had large areas of necrosis. In the liver, histological changes consisted of vasculitis and multifocal to diffuse hepatocellular necrosis. The presence of haemosiderin was observed in melano-macrophage centres in the spleen and kidney of jaundiced fish. No single pathogen was identified in the jaundiced fish. Myxosporean infection was found in both apparently normal fish and jaundiced fish. However, there was a definite tendency for jaundiced fish to be more heavily infected. Histopathological examination found several changes that could not be ascribed to specific aetiological factors and presume that both groups (jaundiced alone and myxosporean-affected jaundiced fish) have similar lesions. The results of this study would suggest that the parasite identified as M. pangasii was not a primary pathogen associated with the haemolytic jaundice. Neither were the gills myxosporeans associated with the haemolytic jaundice and they may be considered more as a nuisance rather than as primary pathogens in farmed P. hypophthalmus in the Mekong Delta, Vietnam. Univariate analysis of the whole dataset showed several variables were significantly associated with the haemolytic jaundice. However, none of the variables achieved lasting statistical relevance in multivariable models. In conclusion, this study identified a haemolytic jaundice condition affecting farmed P. hypophthalmus in Vietnam, but no single aetiological agent or farm based risk factor was identified with this condition. Several priority areas for further work were identified and include a prospective, longitudinal cohort study to identify further the risk factors associated with the clinical jaundice condition.

No description available.