Análise toxicológica in vitro e in vivo de uma fucana antitrombótica da alga marrom Spatoglossum schdöederi

Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. They are extracted mainly from the extracellular matrix of brown algae and echinoderms. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans named A, B and C. Our research group have been extracted non anticoagulant heterofucan from S. schröederi which possess antithrombotic activity in vivo. However, their toxicity in vitro and in vivo has not yet been determined. For the results in toxicity in vitro, we observed that the fucan A at 20, 500 and 1000 μg/plate showed no mutagenic activity in Kado test (Microsuspension), when the bacterial strains TA97a, TA98, TA100 and TA102, with and without S9 were used. The comet assay showed that fucan A (from 20 to 1000 μg/mL) did not cause any genotoxic effect on CHO cells. There was no damage to the DNA of these cells, as evidenced by the tail length and tail moment, which were similar to that found for the negative control. The fucan A from S. schröederi was administered at 20 μg/g of rat (dose which it showed high antithrombotic activity) during two months. After that, the animals were killed and examined. The data showed that fucan A did not cause any change in biochemistry and hematological parameters, as well as, in the morphology and size of the rat s organs analyzed. In conclusion, this study indicates that fucan is a compound with potential pharmacological that has no toxicity

Análise da presença de agentes mutagênicos nas águas do Rio Pitimbú/RN

The Pitimbu River is located at the oriental portion of the State of Rio Grande do Norte, including three importants cities named Macaíba, Parnamirim e Natal. Although its high importance as a water source, which supplys great part of the South Zone of the Natal city, this river receives a large quantity of domestic and industrial waste water without treatment. The Pitimbu River headhas its river-head located in the city of Macaiba, goes through Parnamirim, then it flowing into at the Jiqui Lake in Natal. The aim of this study was to evaluate, qualitatively and quantitatively, the environmental quality of the Pitimbu River by genotoxicity bio-assays, which are important tools for genetics toxicological evaluation. In this work, five samples sites, distributed along the river, were used to collect water samples. Another point site, located near Jiqui lake, was used to collect drinkable water, which was treated by CAERN, the water treatment entreprise of Rio Grande do Norte. The following assays where used to evaluate the quality of these samples: Allium cepa assay; Comet assay; Micronuclei (MN) assay; and Ames test. For the Allium cepa assay, sixteen specimes where used for each water sample from the sample sites. In this assay both microscopic, like cytogenetic damage, and macroscopic aspects, as morphological variation were evaluated. Red blood cells from periferical blood of the Crenicichla menezesi native specie were used not only for the MN assay, but also for the Comet assay. These fishes were collected at different points on the Pitimbu River and the negative control was developed using fishes of the same species that were bring to the laboratory and maintained for 100 days in the optimal experimental conditions. For the Ames test, TA100, YG1042, TA98 and YG1041 strains were used in the directed method without metabolic activation. The results found by the Allium cepa assay showed that two water sample sites induced increase of mitotic index (IM). Additionally, compared to the control, all the water samples increased the chromossomal aberrations frequency and/or micronucleus. Among the sample sites, two also showed an abnormal growth rate in its root and two samples induced morphological alteration. With the MN test in red blood cells, a high frequence of MN was observed in tree sample. By comparing all the results obtained on the water sample points and with the negative control, a significant variation on the MN frequency was observed. Positive results were also observed for the same sample to water test by the Comet assay. These results allow concluding that the proposed specie Crenicichla menezesi has a good profile as a bio-indicator for the evaluation of environmental water quality and the MN and comet test can be usuful for in situ evaluation. By the Ames test, it was possible to detect the mutagenic activity on the waters from the Pitimbu River in different levels of mutagenicity. This result suggests that this river has several substances that induced changes directly to the DNA. The mechanisms involved to this phenomenon could be by both processes, by changing of the reading frame and by nucleotide substitution. These data set indicate the presence of mutagenic agents, which can represent in risk to biot and human beens

Avaliação da genotoxicidade das águas do lençol freático sob a Universidade Federal do Rio Grande do Norte

The groundwater represents the most important freshwater supply of planet. Dailly, in all world a great amount of toxic and genotoxic material reaches the aquatic systems, mainly the aquifers. The Barreiras aquifer through of five water wells is responsible for the supplying of Universidade Federal do Rio Grande do Norte (UFRN). All water wells are polluted with nitrate and some heavy metals, two of them were disabled. The genotoxicity of groundwater samples from Barreiras Aquifer in UFRN was assessed using the Allium cepa test, the Ames test and the Salmonella typhymurium microsuspension test (Kado test). For the Allium cepa test the influence of the groundwater samples collected on macroscopic (root length, colour and form) and microscopic (root tip mitotic index, chromosome aberrations and micronucleus) parameters was examined. All water samples caused a significant increase of the chromosome and mitotic aberration frequency and reduction on the rooth growth compared to negative control. Bridges and chromosome stickness were the most frequent kind of aberration in dividing cells. Furthermore, breaks were also observed. No significant increase in the number of micronuclei was found in relation to the negative controls. For Ames test were used the Salmonella typhymurium strains TA98 and TA100 without metabolic activation, applying the direct method. Prior to the Kado test, organic fractions from the water samples were obtained through XAD resin concentration. The mutagenicity organic extracts were evaluated by Kado test using TA98 and TA100 strains, in the absence and presence of S9 mix (metabolic activation). The concentrations of seven heavy metal ions were measured in water samples, but only Ni, Cu and Cr levels exceeded the permissible maximum concentration for the natural reservoirs. The results obtained for mutagenic activity using the Ames test were negative in all raw water samples analyzed. Positive results in XAD4 extracts of water samples were obtained for TA98 in the presence of S9 mix for two stations. Concentrations of heavy metals and nitrate can be correlated with the toxicity and genotoxicity of water analyzed. The mutagenic effect detected with TA98 strain suggested that organic compounds (after metabolization) are involved with the mutagenicity detected in the samples analyzed. The data set obtained in this work indicated the presence of at least two classes of mutagens: organic and inorganic compounds

Análise toxicológica in vitro e in vivo de uma fucana antitrombótica da alga marrom Spatoglossum schdöederi

Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. They are extracted mainly from the extracellular matrix of brown algae and echinoderms. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans named A, B and C. Our research group have been extracted non anticoagulant heterofucan from S. schröederi which possess antithrombotic activity in vivo. However, their toxicity in vitro and in vivo has not yet been determined. For the results in toxicity in vitro, we observed that the fucan A at 20, 500 and 1000 μg/plate showed no mutagenic activity in Kado test (Microsuspension), when the bacterial strains TA97a, TA98, TA100 and TA102, with and without S9 were used. The comet assay showed that fucan A (from 20 to 1000 μg/mL) did not cause any genotoxic effect on CHO cells. There was no damage to the DNA of these cells, as evidenced by the tail length and tail moment, which were similar to that found for the negative control. The fucan A from S. schröederi was administered at 20 μg/g of rat (dose which it showed high antithrombotic activity) during two months. After that, the animals were killed and examined. The data showed that fucan A did not cause any change in biochemistry and hematological parameters, as well as, in the morphology and size of the rat s organs analyzed. In conclusion, this study indicates that fucan is a compound with potential pharmacological that has no toxicity