Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-02-05T23:00:04Z No. of bitstreams: 1 LiviaDeLourdesDeSousaPinto_DISSERT.pdf: 1563914 bytes, checksum: afff2cde0e6703ffa7b4fae03c2036ac (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-02-16T22:03:58Z (GMT) No. of bitstreams: 1 LiviaDeLourdesDeSousaPinto_DISSERT.pdf: 1563914 bytes, checksum: afff2cde0e6703ffa7b4fae03c2036ac (MD5) / Made available in DSpace on 2016-02-16T22:03:58Z (GMT). No. of bitstreams: 1 LiviaDeLourdesDeSousaPinto_DISSERT.pdf: 1563914 bytes, checksum: afff2cde0e6703ffa7b4fae03c2036ac (MD5) Previous issue date: 2015-03-13 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Condroitim sulfato (CS) ? um glicosaminoglicano natural presente na matriz extracelular de tecidos conectivos, podendo ser extra?do e purificado desses tecidos. O CS est? envolvido em diversas fun??es biol?gicas, o que pode estar relacionado ? variabilidade estrutural que possui, apesar da simplicidade da cadeia linear dessa mol?cula. Pesquisas na ?rea biotecnol?gica e farmacol?gica com rejeitos provenientes da aquicultura vem sendo desenvolvidas no Brasil. Nas ?ltimas d?cadas, a til?pia (Oreochromis niloticus), peixe nativo da ?frica, tem sido uma das esp?cies mais cultivadas em v?rias regi?es do mundo, inclusive no Brasil. A tilapicultura ? uma atividade economica cujo principal inconveniente ? a grande quantidade de res?duos que s?o descartados pelos produtores. Entende-se que o material descartado pode ser aproveitado em pesquisas como fonte de mol?culas com importante aplica??o biotecnol?gica, o que tamb?m contribui na redu??o de impactos ambientais e favorece o desenvolvimento dessa atividade de maneira ecologicamente correta. Dessa forma, v?sceras de til?pias nil?tica foram submetidas ? prote?lise, em seguida os glicosaminoglicanos foram complexados com resina de troca i?nica (Lewatit), fracionados com volumes crescentes de acetona e purificados atrav?s da cromatografia de troca i?nica DEAE-Sephacel. Na sequ?ncia, a fra??o foi analisada atrav?s de eletroforese em gel de agarose e resson?ncia magn?tica nuclear (RMN). O perfil eletrofor?tico do composto, a an?lise dos espectros 1H de RMN e a correla??o do HSQC permitem afirmar que o composto corresponde a uma mol?cula de condroitim sulfato. O ensaio de MTT foi utilizado para avalia??o da viabilidade celular na presen?a do CS isolado de til?pia e mostrou que o composto n?o ? citot?xico a c?lulas normais provenientes do fibroblasto de embri?o de camundongos (3T3). Ent?o, o composto foi testado quanto a capacidade de reduzir o influxo de leuc?citos em modelo de peritonite aguda (in vivo) induzida por tioglicolato de s?dio. Nesse contexto, foi realizada a contagem total e diferencial de leuc?citos do sangue e l?quido peritoneal coletadas, respectivamente, da veia cava e do lavado peritoneal de cada animal submetido ao experimento. O condroitim sulfato, pela primeira vez isolado de til?pia (CST ), foi capaz de reduzir a migra??o de leuc?citos ? cavidade peritoneal de camundongos inflamados em at? 80,4% na dose de 10?g/kg. Os resultados mostram ainda que houve redu??o significativa (p<0,001) da popula??o de polimorfonucleares do lavado peritoneal nas tr?s doses testadas (0,1?g/kg; 1?g/kg e 10?g/kg) quando comparado ao controle positivo (apenas tioglicolato). Portanto, uma vez que a estrutura e o mecanismo de a??o do CST tenham sido totalmente elucidados, esse composto pode apresentar potencial para uso terap?utico em doen?as inflamat?rias. / Chondroitin sulfate (CS) is a naturally glycosaminoglycan found in the extracellular matrix of connective tissues and it may be extracted and purified those tissues. CS is involved in various biological functions, which may be related to the having structural variability, despite the simplicity of the linear chain structure from this molecule. Researches in biotechnology and pharmaceutical field with wastes from aquaculture has been developed in Brazil. In recent decades, tilapia (Oreochromis niloticus), native fish from Africa, has been one of the most cultivated species in various regions of the world, including Brazil. The tilapia farming is a cost-effective activity, however, it generates large amount of wastes that are discarded by producers. It is understood that waste from tilapia can be used in research as a source of molecules with important biotechnological applications, which also helps in reducing environmental impacts and promote the development of an ecofriendly activity. Thus, nile tilapia viscera were subjected to proteolysis, then the glycosaminoglycans were complexed with ion exchange resin (Lewatit), it was fractionated with increasing volumes of acetone and purified by ion exchange chromatography DEAE-Sephacel. Further, the fraction was analyzed by agarose gel electrophoresis and nuclear magnetic resonance (NMR). The electrophoretic profile of the compound together the analysis of 1H NMR spectra and the HSQC correlation allow to affirm that the compound corresponds to a molecule like chondroitin sulfate. MTT assay was used to assess cell viability in the presence of CS tilapia isolated and showed that the compound is not cytotoxic to normal cells such as cells from the mouse embryo fibroblast (3T3). Then, this compound was tested for the ability to reduce the influx of leukocytes in model of acute peritonitis (in vivo) induced by sodium thioglycolate. In this context, it was done total and differential leukocytes counting in the blood and peritoneal fluid collected respectively from vena cava and the peritoneal cavity of the animals subjected to the experiment. The chondroitin sulfate for the first time isolated from tilapia (CST ) was able to reduce the migration of leukocytes to the peritoneal cavity of inflamed mice until 80.4 per cent at a dose 10?g/kg. The results also show that there was a significant reduction (p<0.001) of the population of polymorphonuclear leukocytes from peritoneal cavity in the three tested doses (0.1?g/kg; 1?g/kg and 10?g/kg) when it was compared to the positive control (just thioglycolate). Therefore, since the CST structure and mechanism of action has been completely elucidated, this compound may have potential for therapeutic use in inflammatory diseases

Made available in DSpace on 2014-12-17T14:01:59Z (GMT). No. of bitstreams: 1 EmersonESM.pdf: 931852 bytes, checksum: f768665fe7e8aa27e4023b4def16f98d (MD5) Previous issue date: 2009-03-27 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Considering their commercial importance, as these are the species of freshwater fish more commercialized in Brazil, their occurence in different kinds of aquatic environments (lakes, rivers and dams) and for being tolerant to a wide range of variation of various physical parameters and chemical water, the fish species Oreochromis niloticus, Cyprinus carpio and Colossoma macropomum were chosen for this study, furthermore, to test the toxicity we used the herbicide Roundup. The fingerlings of tilapia (Oreochromis niloticus), commun carp (Cyprinus carpio) and tambaqui (Colossoma macropomum) were submitted to the herbicide roundup in the following concentrations: 0.0 (control); 18,06; 19,10; 20,14; 21,18 and 22,22 mg.L-1, 0.0 (control); 13,89; 14,86; 15,83; 16,81 and 17,78 mg.L-1, and 0.0 (control); 18,06; 19,10; 20,14; 21,18 and 22,22 mg.L-1, respectively, three for 96 hours. The LC50 - 96h for O. niloticus, C. carpio and C. macropomum was 21,63, 15,33 and 20,06 mg.L-1 of the herbicide roundup, respectively. The results show that this herbicide is classified as slightly toxic to the three species. The values of dissolved oxygen, pH and temperature recorded in the aquarium control and aquarium experimental of the three fish species have remained without significant variations during the tests, which reduces the possibility of death caused by sudden variations of these parameters during the 96 hours the experiment. The values of LC50 between different species of fish were observed, noting that the species O.niloticus, C. carpio and C. macropomum showed no expressive differences. The values of environmental risk of Roundup were calculated to obtain more stringent parameters in assessing the dangerousness of those on nontargets. The risk of environmental contamination by Roundup for the Nile tilapia, common carp, and tambaqui are low for the lowest application rate (1 L.ha-1) and depths (1.5 and 2.0 m). The dilution of 100%, the highest recommended dose (5 L.ha-1) and depths (1.5 and 2.0 m) the risk is moderate for the three species. The values of the Risk Ratio (QR) were greater than 0,1, indicating that the values of the CAE and LC50 are above acceptable levels and there is a need, this study, a refinement in ecotoxicological tests / Considerando a import?ncia comercial, por se tratarem das esp?cies de ?gua-doce mais comercializadas no Brasil, por ocorrerem em diversos tipos de ambientes aqu?ticos (lagos, rios e a?udes) e ainda por serem tolerantes a um amplo espectro de varia??o de diversos par?metros f?sicos e qu?micos da ?gua, as esp?cies de peixes Oreochromis niloticus, Cyprinus carpio e Colossoma macropomum foram escolhidas para este estudo com a finalidade de testar a toxicidade do herbicida roundup. Diante do exposto, o presente trabalho teve como objetivo determinar a toxicidade aguda e avaliar o risco de contamina??o ambiental dessas tr?s esp?cies de peixes pelo herbicida Roundup. Os alevinos de Til?pia do Nilo (Oreochromis niloticus), Carpa-comum (Cyprinus carpio) e Tambaqui (Colossoma macropomum) foram submetidos ao herbicida roundup nas seguintes concentra??es: 0,0 (controle); 18,06; 19,10; 20,14; 21,18 e 22,22 mg.L-1, 0,0 (controle); 13,89; 14,86; 15,83; 16,81 e 17,78 mg.L-1 e 0,0 (controle); 18,06; 19,10; 20,14; 21,18 e 22,22 mg.L-1, respectivamente, os tr?s por 96 horas. A CL50 96h para O. niloticus, C. carpio e C. macropomum foi de 21,63, 15,33 e 20,06 mg.L-1 do herbicida roundup, respectivamente. Os resultados obtidos demonstram que esse herbicida est? classificado como levemente t?xico para as tr?s esp?cies. Os valores de oxig?nio dissolvido, pH e temperatura registrados no aqu?rio controle e nos aqu?rios experimentais das tr?s esp?cies estudadas mantiveram-se sem varia??es significativas ao longo dos testes, o que reduz a possibilidade de mortalidade causada por varia??es bruscas desses par?metros, durante as 96 horas de experimento. Os valores das CL50 entre as diferentes esp?cies de peixes foram observadas, constatando que as esp?cie Oreochromis niloticus, Colossoma macropomum e Cyprinus carpio n?o apresentaram diferen?as expressivas. Os valores de risco ambiental do Roundup foram calculados para a obten??o de par?metros mais rigorosos na avalia??o da periculosidade desses sobre organismos n?o-alvos. O risco de contamina??o ambiental por roundup para a til?pia do Nilo, carpa-comum e tambaqui s?o baixos para a menor dose de aplica??o (1 L.ha-1) e nas profundidades (1,5 e 2,0 m). Na dilui??o de 100%, na maior dose recomendada (5 L.ha-1) e nas profundidades (1,5 e 2,0 m) o risco ? moderado para as tr?s esp?cies. Os valores de Quociente de Risco (QR) foram maiores que 0,1, indicando que os valores da rela??o CAE e CL50 est?o acima dos n?veis aceit?veis, havendo a necessidade de um refinamento nas an?lises ecotoxicol?gicas

Made available in DSpace on 2014-12-17T14:33:02Z (GMT). No. of bitstreams: 1 MariaMLC.pdf: 1068178 bytes, checksum: 2211aebe12426e9f45d52a507df28c0f (MD5) Previous issue date: 2009-05-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The food chain theory predict that presence of omnivory prevent the trophic cascade and could be a strong stabilizing factor over resource and consumer community dynamics, and that the nutrient enrichment destabilize populations dynamics. Most of the freshwater tropical reservoirs are eutrophic, and strategies that seek improve the water quality through the control of phytoplankton biomass and nutrient input, become essential for the improvement and preservation of water quality. The aim of this study was test the zooplanktivory (when larvae) and omnivory (when young and adult) effects of Nile Tilapia over the structure and dynamics of plankton communities, in addition or absence of nutrients enrichment. For this, one field experiment was performed with a factorial design 2x3 resulting in six treatments: control, without fish and nutrient (C); with omnivorous fish (O); with zooplanktivorous fish (Z); without fish and with enrichment of nutrients (NP); with omnivorous fish and nutrients (ONP); and, with zooplanktivorous fish and nutrients (ZNP). The two planktivory types reduced the zooplankton biomass and increased the phytoplankton biomass, but the omnivory of filter-feeding fish attenuated the trophic cascade magnitude. The fertilization by nutrients increases the nutrient concentrations in water and the phytoplankton biomass, but the effect on zooplankton is dependent of the trophic structure. In a general way, the effects of the fish and nutrient addition were addictive, but significant interactions among those factors were observed in the answer of some zooplankton groups. The effects of omnivorous fish over the temporal variability of phytoplankton and zooplankton biomass were very variable, the increase or reduce in variability of the plankton depending of the level of nutrients and of the analyzed variable. With base in this study, we conclude that the planktivory type exercised by the fish and the concentrations of nutrients in the water affects the force of pelagic trophic cascades and probably the success of biomanipulation programs for the handling of water quality in lakes and tropical reservoirs / A teoria de cadeias alimentares prev? que a presen?a de onivoria pode atenuar os efeitos de cascata tr?fica e estabilizar as popula??es de aut?trofos e herb?voros, as quais s?o desestabilizadas pela fertiliza??o por nutrientes. Tendo em vista que muitos lagos e reservat?rios tropicais encontram-se eutrofizados, estrat?gias que visem o controle do aporte de nutrientes e a redu??o da biomassa fitoplanct?nica s?o essenciais para a melhoria da qualidade da ?gua desses ambientes. O presente estudo teve como objetivo avaliar o efeito da zooplanctivoria e onivoria por uma mesma esp?cie de peixe (Til?pia do Nilo), que quando larva ? zooplanct?vora e quando jovem e adulta torna-se filtradora on?vora, sobre a estrutura e din?mica das comunidades planct?nicas, na presen?a e aus?ncia de enriquecimento por nutrientes. Para tanto, foi realizado um experimento com desenho fatorial 2 x 3, resultando em 6 tratamentos: sem adi??o de peixes ou nutrientes (C); com adi??o de peixes zooplanct?voros (Z); com adi??o de peixes on?voros (O); com adi??o de nutrientes (NP); com adi??o de peixes zooplanct?voros e nutrientes (ZNP); e com adi??o de peixes on?voros e nutrientes (ONP). Os resultados mostram que os dois tipos de planct?voros reduziram a biomassa zooplanct?nica e aumentaram a biomassa fitoplanct?nica, mas que a onivoria dos peixes filtradores atenuou a magnitude das cascatas tr?ficas. Os resultados tamb?m mostram que a fertiliza??o por nutrientes aumenta as concentra??es de nutrientes na ?gua e a biomassa fitoplanct?nica, mas que o efeito sobre o zoopl?ncton depende da estrutura tr?fica. De um modo geral, os efeitos da adi??o de peixes e nutrientes foram aditivos, mas intera??es significativas entre esses fatores foram observadas na resposta de alguns grupos zooplanct?nicos. Os efeitos dos peixes on?voros sobre a variabilidade temporal da biomassa do fitopl?ncton e do zoopl?ncton foram muito vari?veis, aumentando ou reduzindo a variabilidade do pl?ncton dependendo do n?vel de nutrientes e da vari?vel analisada. ? poss?vel concluir, com base neste estudo, que o tipo de planctivoria exercido pelos peixes e as concentra??es de nutrientes na ?gua afetam a for?a das cascatas tr?ficas pel?gicas e provavelmente o sucesso dos programas de biomanipula??o para o manejo da qualidade da ?gua de lagos e reservat?rios tropicais