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Avalia??o da toxicidade de flora??es naturais e de cultura de cianobact?rias: efeitos sobre Ceriodaphnia silvestri (Crustacea, Cladocera)Mendon?a, Juska Milena dos Santos 30 July 2006 (has links)
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Previous issue date: 2006-07-30 / The incidence of toxic cyanobacterial blooms is one of the important consequences of eutrophication in aquatic ecosystems. It is a very common phenomenon in reservoirs and shrimp ponds in the State of Rio Grande do Norte (RN), Brazil. Cyanobacterias produce toxins which can affect aquatic organisms and men trough the food chain. Aiming to contribute to the studies of cyanobacterias in RN, we propose: a) to evaluate the toxicity of isolated cyanobacterias in important fresh-water environments; and b) to verify the effects of both natural and cultured blooms occurred in reservoirs for human supply and in the cladoceran Ceriodaphnia silvestrii. This study was carried out using samples of natural blooms occurred between March and October of 2004 in Gargalheiras Dam (08? L e 39? W), in July of 2004 in Armando Ribeiro Gon?alves Dam (06o S e 37o W) and in commercial shrimp ponds (Litopenaeus vannamei) located in fresh-water environments. The samples were collected with plankton net (20?m.) for identification, isolation and obtaining of phytoplanktonic biomass for liophilization and later toxicity bioassays. The toxicity of cultured samples and natural blooms was investigated through bioassays in Swiss mice. Quantification of cyanobacteria in samples was conducted following the ?termol method, with 300mL samples fixed with lugol. The toxicity test with Ceriodaphnia silvestrii followed ABNT, 2001 recommendations, and were accomplished with natural hepatotoxic bloom s samples and cultured samples of both non-toxic and neurotoxic C. raciborskii. In this test, five newborns, aged between 6 and 24 hours, were exposed to different concentrations (0 a 800 mg.L-1) of crude cyanobacterial extracts during 24 and 48 hours. Three replicates were used per treatment. The pH, temperature and dissolved oxygen at the beginning and after 24 and 48hours from the test were measured. We estimated the CL50 through the Trimmed Spearman-Karber method. The blooms were constituted by Microcystis panniformis, M. aeruginosa, Anabaena circinalis, Cylindrospermopsis raciborskii and Planktothrix agardhii, producers of mycrocistin-LR confirmed with HPLC analysis. Samples of hepatotoxic blooms registered toxinogenic potential for C. silvestrii, with CL50-24h value of 47.48 mg.L-1 and CL5048h of 38.15 mg.L-1 for GARG samples in march/2005; CL50-24h of 113,13 mg.L-1 and CL5048h of 88,24 mg.L-1 for ARG July/2004; CL50-24h of 300.39 mg.L-1 and CL50-48h of 149.89 mg.L-1 for GARG October/2005. For cultured samples, values of CL50-24h and CL50-48h for
C. raciborskii toxic strains were 228.05 and 120.28 mg.L-1, respectively. There was no mortality of C. silvestrii during the tests with non-toxic C. raciborskii strain. The toxicity test with C. silvestrii presented good sensitivity degree to cyanotoxins. The toxicity of natural hepatotoxic blooms samples (microcystins) and cultured neurotoxic saxitoxins producer samples analyzed in this study give us strong indications of that toxin s influence on the zooplanktonic community structure in tropical aquatic environments. Eleven cyanobacteria strains were isolated, representing 6 species: Anabaenopsis sp., Cylindrospermopsis raciborskii, Chroococcus sp., Microcystis panniformis, Geitlerinema unigranulatum e Planktothrix agardhii. None presented toxicity in Swiss mice. The strains were catalogued and deposited in the Laborat?rio de Ecologia e Toxicologia de Organismos Aqu?ticos (LETMA), in UFRN, and will be utilized in ecotoxicol?gical and ecophysiological studies, aiming to clarify the causes and control of cyanobacterial blooms in aquatic environments in RN. This state s reservoirs must receive broader attention from the authorities, considering the constant blooms occurring in waters used for human consumption / Incid?ncia de flora??es t?xicas de cianobact?rias ? uma das conseq??ncias da eutrofiza??o nos ecossistemas aqu?ticos, sendo comum a sua ocorr?ncia em reservat?rios e viveiros de cultivo de camar?o no Estado Rio Grande do Norte. Cianobact?rias podem produzir toxinas que podem afetar organismos aqu?ticos e o homem atrav?s da cadeia alimentar. Visando contribuir com o desenvolvimento de estudos ecotoxicol?gicos de cianobact?rias no RN, nos propomos avaliar a toxicidade de cianobact?rias isoladas de ambientes de ?guas doces e verificar o efeito de cultura e flora??es naturais ocorridas em reservat?rios para abastecimento humano, no crust?ceo clad?cero Ceriodaphnia silvestrii. O estudo foi conduzido com amostras de flora??es naturais ocorridas em mar?o e outubro de 2004 na Barragem Gargalheiras (08? L e 39? W), em julho de 2004 na Barragem Armando Ribeiro Gon?alves (06o S e 37o W) e em fazendas comerciais de cultivo de camar?o (Litopenaeus vannamei), localizadas em ambientes de ?gua doce do RN (6o 58 S e 38o 36 W). As amostras de flora??es foram coletadas com rede de pl?ncton (20?m) para identifica??o, isolamento e obten??o de biomassa fitoplanct?nica para liofiliza??o e posterior determina??o da toxicidade por bioensaios. A toxicidade das amostras de culturas e flora??es naturais foi investigada atrav?s de bioensaios em camundongos Swiss. A quantifica??o das cianobact?rias foi realizada pelo m?todo de ?termol, utlizando-se amostras (300mL) fixadas com lugol. O teste de toxicidade com Ceriodaphnia silvestrii seguiu as recomenda??es da ABNT (2001) e foram realizadas com amostras de flora??es naturais hepatot?xicas e culturas de C. raciborskii neurot?xica e n?o t?xica. No teste, cinco neonatos com idade entre 6 e 24 horas foram expostos a diferentes concentra??es (0 a 800 mg.L-1) de extrato bruto de cianobact?rias durante 24 e 48 horas. Tr?s replicatas por tratamentos foram usadas. Foram medidos o pH, a temperatura e o oxig?nio dissolvido no in?cio, com 24 e com 48 horas do teste. Atrav?s do m?todo Trimmed Spearman-Karber foi estimado a CL50. As flora??es eram mistas e compostas por Microcystis panniformis, M. aeruginosa, Anabaena circinalis, Cylindrospermopsis raciborskii e Planktothrix agardhii, produtoras de microcistina- LR, confirmada atrav?s de an?lise por HPLC. Amostras de flora??es hepatot?xicas registraram potencial toxinog?nico para o C. silvestrii, com valores de CL50-24h de 47.48 mg.L-1 e CL50-48h de 38.15 mg.L-1 para a amostra de GARG mar/05; CL50-24h de 113,13 mg.L-1 e CL50-48h de 88,24 mg.L-1 para ARG jul/04; CL50-24h de 300.39 mg.L-1 e CL50-48h de 149.89 mg.L-1 para GARG 10/05. Para as amostras de cultura valores de CL50-24h e CL50-48h para a cepa t?xica de C. raciborskii foram de 228.05 e 120.28 mg.L-1, respectivamente. N?o houve mortalidade dos C. silvestrii nos testes com a cepa de C. raciborskii n?o-t?xica. O teste de toxicidade com C. silvestrii apresentou um bom grau de sensibilidade ?s cianotoxinas. A toxicidade das amostras de flora??es naturais hepatot?xicas (microcistinas) e culturas neurot?xicas produtoras de saxitoxinas verificadas neste estudo fornece forte indicativos da influ?ncia dessas toxinas sobre a estrutura da comunidade zooplanct?nica em ambientes aqu?ticos tropicais. Foram isoladas 11 cepas de cianobact?rias de reservat?rios e viveiros do Estado, representadas por 6 esp?cies: Anabaenopsis sp., Cylindrospermopsis raciborskii, Chroococcus sp., Microcystis panniformis, Geitlerinema unigranulatum e Planktothrix agardhii. Nenhuma cepa apresentou toxicidade em camundongos Swiss. As cepas foram catalogadas e depositadas no Laborat?rio de Ecologia e Toxicologia de Organismos Aqu?ticos (LETMA) da UFRN e ser?o utilizadas em estudos ecotoxicol?gicos e ecofisiol?gicos visando esclarecer as causas e controle de flora??es de cianobact?rias nos ambientes aqu?ticos do Estado. Os reservat?rios do Rio Grande do Norte devem merecer maior aten??o perante os ?rg?os respons?veis, tendo em vista os constantes florescimentos t?xicos de cianobact?rias em ?guas utilizadas para consumo humano
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