Avaliação da toxicidade de florações naturais e de cultura de cianobactérias: efeitos sobre Ceriodaphnia silvestri (Crustacea, Cladocera)

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 (08º L e 39º W), in July of 2004 in Armando Ribeiro Gonçalves 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 (20µm.) 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 Ütermol 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 Laboratório de Ecologia e Toxicologia de Organismos Aquáticos (LETMA), in UFRN, and will be utilized in ecotoxicológical 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

Microcystin-LR in Brazilian Aquaculture Production Systems

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação dos efeitos de um extrato bruto de cianobactérias contendo microcistinas sobre o desempenho reprodutivo e a embriofetotoxicidade de ratos

The presence of cyanobacterial blooms in reservoirs intended for supply to the population can create public health problems for many species could produce potentially toxic compounds and these are not eliminated in the conventional procedures used in water treatment plants. So even in amounts less than the maximum allowable limit imposed by MS, cyanotoxins can be present in drinking water distributed to the population, creating a chronic exposure. There is little information about the long-term effects of oral exposure to cyanotoxins. This work aimed to show the exposure orally (v.o) of animals to a crude extract of cyanobacteria containing cyanotoxins to evaluate the reproductive performance of pregnant rats and their offspring and fertility of male rats. The presence of microcystins (MCs) in samples collected during the flowering processes in freshwater reservoirs in the Rio Grande do Norte, was analyzed by enzyme immunoassay and its variants have been identified and quantified by chromatographic methods. It was observed that by administration v.o. cyanobacterial extract containing MCs (40, 100 or 250 ng of MCs / kg / day) did not cause systemic toxicity in adult rats or effect on reproductive performance of male and female rats treated. It was also not observed any changes in skeletal study in the offspring of pregnant rats treated with the extract above. Because the solutions used contained MCs in a concentration equal to or greater than the tolerable daily intake for MCs, the results suggest, therefore, that the development of this work contributed to better assess public health risk as the oral exposure to cyanotoxins, increasing thus the credibility of the maximum allowable limit (LMP) of MCs in drinking water distributed to the population of several countries that use the LMP established by WHO in its legislation

Avaliação do potencial de toxicidade de florações de cianobactérias como subsídio para ações de gestão ambiental em açudes do semiárido potiguar

Blooms negatively compromise the aquatic environment, plants, animals and human health.Some species are toxin-producing, such as the neurotoxin saxitoxin and the hepatotoxin microcystin, which may contaminate water reservoirs, as those existing in the semiarid region of Rio Grande do Norte (Brazil) which are used to supply the population, fishing, aquiculture and recreational activities, thereby providing the risk of human exposure through water intake, dermal contact and respiratory tract. Thus, it is recommended a constant monitoring of the density of cyanobacteria with the quantification of cyanotoxins. One goal with this work was the monitoring of water in four reservoirs in semiarid of RN through the identification and enumeration of cyanobacteria and through the identification and quantification of cyanotoxins by ELISA. Furthermore, we intended to assess the environmental perception of farmers and artisanal fishers in reservoirs of semiarid of RN through semi-structured interviews with questions mostly related to water and eutrophication. Through these objectives the aim was the development of management strategies for aquaculture and prevention of risks to public health. The results showed that the highest values of microcystins were found in the rainy season. Standards for drinking water, according to the guidelines of Ministry of Health 2914/2011 and CONAMA 357/05, setting the maximum values for raw water density of cyanobacteria: 50,000 cel.mL-1; microcystin: 1 μg. L-1 and saxitoxin: 3 μg. L-1. The values found for microcystin ranged between 0.00227 μg. L-1 and 24.1954 μg. L-1. From 128 samples analyzed, 27% were above the limit. There was no clear seasonal pattern for saxitoxins and their values ranged between 0.003 μg. L-1 and 0.766 μg. L-1 with none of the values above the limit. Furthermore, 76% of the densities of cyanobacteria values were above the limit. About environmental perception, 52 interviews were conducted and the results show that the respondents recognize the main uses of water of the rervoirs, recognize the importance and have a positive view about the reservoirs. They also realize that the water has a poor quality and can cause health problems. The results provide data showing the persistence of cyanobacteria and cyanotoxins, many times over the limit, reinforcing the importance of constant monitoring. The assessment of environmental perception gives foundation for later proposed environmental education linked to public health management into the context of this particular population, making it more effective

Microcystin-producing genotypes from cyanobacteria in Brazilian reservoirs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos alelopáticos de microcistinas em rúcula (Eruca sativa Mill.)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ação alelopática de saxitoxinas e microcistinas em cianobactérias e microalgas verdes em condições controladas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biodegradacao da hepatotoxina(D-Leu1)-microcistina-LR por bactérias presentes em filtros biológicos de carvão

The persistence of MCs in aquatic environments and their difficult removal in the conventional water treatment is a challenge to companies of sanitation. However, the MCs are susceptible to degradation by bacteria present in water, sediment and sewage effluents. In this study, we investigated the biodegradation of MCs by microorganism present in carbon filters with biological activity (BAC) and their phylogenetic identification by sequencing gene 16S RNA. A study of water containing MCs was used, with different compositions, plus a filters BAC effluent. The results showed that of MCs were biodegraded by microorganism present in the biofilm. This study provides the ability to complete biodegradation of MCs by bacteria present in BAC filters and the possible use of these microorganisms as alternative of the removal of MCs in the treatment of drinking water

Efeito do extrato aquoso de folhas de guanandi (Calophyllum brasiliense Cambess) no controle do crescimento de Microcystis aeruginosa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Peptídeos cíclicos hepatotóxicos: MALDI-TOF como uma ferramenta analítica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de compostos inorgânicos, herbicidas e extratos foliares no desenvolvimento de microcystis aeruginosa e produção de microcistina

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Efeito de compostos inorgânicos, herbicidas e extratos foliares no desenvolvimento de microcystis aeruginosa e produção de microcistina

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Co-occurrence of microcystin and microginin congeners in Brazilian strains of Microcystis sp.

Species of Microcystis are the most common bloom-forming cyanobacteria in several countries. Despite extensive studies regarding the production of bioactive cyanopeptides in this genus, there are limited data on isolated strains from Brazil. Three Microcystis sp. strains were isolated from the Salto Grande Reservoir (LTPNA01, 08 and 09) and investigated for the presence of mcy genes, microcystins and other cyanopeptides. Microcystin and microginin production was confirmed in two isolates using high-resolution tandem mass spectrometry after electrospray ionization (ESI-Q-TOF), and the structures of two new microginin congeners were proposed (MG756 Ahda-Val-Leu-Hty-Tyr and MG770 MeAhda-Val-Leu-Hty-Tyr). The biosynthesis profile of the identified cyanopeptides was evaluated at different growth phases via a newly developed HPLC-UV method. Results demonstrated no substantial differences in the production of microcystins and microginins after data normalization to cell quota, suggesting a constitutive biosynthesis. This study represents the first confirmed co-production of microginins and microcystins in Brazilian strains of Microcystis sp. and highlights the potential of Brazilian cyanobacteria as a source of natural compounds with pharmaceutical interest.

Microcystin-producing genotypes from cyanobacteria in Brazilian reservoirs

The aim of this study was to evaluate the use of new oligonucleotide primers (mcyB-F/R, mcyB-F/R-A, and mcyB-F/R-B) designed from Brazilian cyanobacteria for the detection of microcystin-producing genotypes in 27 environmental samples from water reservoirs and 11 strains of Microcystis. Microcystins were found using HPLC in all 11 strains and 19 of the environmental samples. The new oligonucleotide primers amplified fragments of microcystin-producing genes, including the eight environmental samples in which no microcystins were detected by HPLC, but which presented amplified fragments, thereby demonstrating the existence of microcystin-producing genes. The new oligonucleotide primers exhibited better specificity when used with environmental samples and were more reliable in comparison with those described in the literature (mcyB-FAA/RAA and mcyA-Cd/FR), which generate false-negative results. The better performance of these new oligonucleotide primers underline the need for designing molecular markers that are well fitted to the regional biological diversity. As this is a fast predictive technique for determining the presence or absence of microcystins, it could be used either alone or in conjunction with other techniques, such as the screening of samples to be sent for quantitative toxicological analysis using HPLC, thereby reducing monitoring cost and time. (c) 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012.

Biomonitoring genotoxicity and cytotoxicity of Microcystis aeruginosa (Chroococcales, Cyanobacteria) using the Allium cepa test

Water pollution caused by toxic cyanobacteria is a problem worldwide, increasing with eutrophication. Due to its biological significance, genotoxicity should be a focus for biomonitoring pollution owing to the increasing complexity of the toxicological environment in which organisms are exposed. Cyanobacteria produce a large number of bioactive compounds, most of which lack toxicological data. Microcystins comprise a class of potent cyclic heptapeptide toxins produced mainly by Microcystis aeruginosa. Other natural products can also be synthesized by cyanobacteria, such as the protease inhibitor, aeruginosin. The hepatotoxicity of microcystins has been well documented, but information on the genotoxic effects of aeruginosins is relatively scarce. In this study, the genotoxicity and ecotoxicity of methanolic extracts from two strains of M. aeruginosa NPLJ-4, containing high levels of microcystin, and M. aeruginosa NPCD-1, with high levels of aeruginosin, were evaluated. Four endpoints, using plant assays in Allium cepa were applied: rootlet growth inhibition, chromosomal aberrations, mitotic divisions, and micronucleus assays. The microcystin content of M. aeruginosa NPLJ-4 was confirmed through ELISA, while M. aeruginosa NPCD-1 did not produce microcystins. The extracts of M. aeruginosa NPLJ-4 were diluted at 0.01, 0.1, 1 and 10 ppb of microcystins: the same procedure was used to dilute M. aeruginosa NPCD-1 used as a parameter for comparison, and water was used as the control. The results demonstrated that both strains inhibited root growth and induced rootlet abnormalities. The strain rich in aeruginosin was more genotoxic, altering the cell cycle, while microcystins were more mitogenic. These findings indicate the need for future research on non-microcystin producing cyanobacterial strains. Understanding the genotoxicity of M. aeruginosa extracts can help determine a possible link between contamination by aquatic cyanobacteria and high risk of primary liver cancer found in some areas as well as establish water level limits for compounds not yet studied. (C) 2012 Elsevier B.V. All rights reserved.