Cleoni dos Santos Carvalho

Biomarker responses as indication of contaminant effects in Oreochromis niloticus.

The current study investigated oxidative stress parameters (enzymes activities, metallothionein content and lipid peroxidation) in freshwater fish, Oreochromis niloticus, tilapia exposure to Monjolinho River (in 4 months of year: January, April, July and November). One critical site in Monjolinho River (site B) was assessed in comparison to a reference site (site A). Water pH and oxygen concentration was lower than that recommended by CONAMA (Brazilian National Environmental Committee), resolution 357/2005 for protection of aquatic communities, and ammonium and the metals Cu, Zn, Mn and Fe (on all months) concentrations were higher than the maximum concentration recommended. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significantly decreased in liver and muscle in tilapia from Monjolinho River, throughout the year, in relation to reference except in gills that SOD activity increased. Glutathione S-transferase (GST) activity was significantly increased in liver of the tilapia from Monjolinho River in all sites, in relation to reference except in gills that GST activity increased in July and decreased in November, suggesting that GST activity could be induced to neutralize the pollutants toxicity. On the other hand, GST activity was significantly decreased in white muscle indicating a toxic effect of pollutants, resulting in a decreased ability of tilapia to perform defense reactions associated to GSTs. The decrease of catalase (CAT) activity in gills of the O. niloticus together with the increase of SOD activity, could explain the increased lipid peroxidation (LPO) level in this organ. Metallothionein levels in liver and gills were significantly high in all sites. Results indicate that the exposure to metals caused severe damage to tissues; despite the consensually assumed antioxidant induction as a sign of exposure to contaminants the effects seem in part to be mediated by suppression of antioxidant system with SOD, CAT and GPx as potential candidates for tissues toxicity biomarkers of pollutants.

Effect of temperature acclimation on the liver antioxidant defence system of the Antarctic nototheniids Notothenia coriiceps and Notothenia rossii.

The aim of this study was to determine whether endemic Antarctic nototheniid fish are able to adjust their liver antioxidant defence system in response to the temperature increase. The activity of the superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutathione reductase (GR) enzymes as well as the content of non-enzymatic oxidative stress markers such as reduced glutathione (GSH), lipid peroxidation (LPO) and protein carbonyl (PC) were measured in the liver of two Antarctic fish species, Notothenia rossii and Notothenia coriiceps after 1, 3 and 6days of exposure to temperatures of 0°C and 8°C. The GST activity showed a downregulation in N. rossii after 6days of exposure to the increased temperature. The activity profiles of GST and GR in N. rossii and of GPx in N. coriiceps also changed as a consequence of heating to 8°C. The GSH content increased by heating to 8°C after 3days in N. coriiceps and after 6days in N. rossii. The content of malondialdehyde (MDA), a LPO marker, showed a negative modulation by the heating to 8°C in N. rossii after 3days of exposure to temperatures. Present results show that heating to 8°C influenced the levels and profiles of the antioxidant enzymes and defences over time in the nototheniid fish N. rossii and N. coriiceps.

Copper levels and changes in pH induce oxidative stress in the tissue of curimbata (Prochilodus lineatus).

We analyzed the effect of exposure to 25% 96 h-LC50 of copper at low (24.5 μg L(-1) Cu, pH 4.5), neutral (7.25 μg L(-1) Cu, pH 7.0) and high pH (4.0 μg L(-1) Cu, pH 8.0) at 20 °C on antioxidant defenses and oxidative stress in the liver, gills and white muscle of the fish Prochilodus lineatus. Water at pH 4.5 and 8.0 affected the enzymatic and non-enzymatic antioxidant systems of the liver and gills, but not of the white muscles of P. lineatus, when compared to water at pH 7.0. After Cu exposure, SOD (superoxide dismutase), GPx (glutathione peroxidase), GR (glutathione reductase) and GST (glutathione S-transferase) activities increased and CAT (catalase) activity decreased in the liver at water at pH 4.5 and 8.0. Meanwhile, the activities of SOD, CAT, GPx, GR and GST increased in the gills at these pHs. SOD and CAT activities increased in the white muscle after Cu exposure at pH 8.0 and GPx, GR and GST activities decreased after Cu exposure at pH 4.5 and 8.0. LPO levels decreased in the liver and gills of fish that were exposed to water at pH 4.5 and 8.0 and, after Cu exposure, the LPO level increased in the liver, gills and white muscle of fish that were exposed to water at pH 4.5 and 8.0, when compared to the control group at pH 7.0. The metallothionein (MT) concentration increased in the liver of fish in water at pH 4.5 and 8.0 and the gill of fish in water at pH 8.0. After Cu exposure, MT in the liver and gills was significantly elevated in fish exposed to water at pH 4.5 and 8.0, but remained at levels similar to the control group in the white muscle. These results indicate a differing sensitivity of fish organs and tissues to essential metals, such as copper, and that toxicity may be relevant at environmental concentrations. These results indicate that the effect of Cu on the response of antioxidant defense systems is determined by water pH.

Utilization of endogenous reserves and effects of starvation on the health of Prochilodus lineatus (Prochilodontidae)

Prochilodus lineatus Valenciennes (curimbatá) is an important migratory Neotropical fish. It does not feed during spawning migration, and often survives after spawning. The mobilization of energy reserves and some effects of starvation (zero to eight weeks) on fish health were experimentally evaluated. Hepatic glycogen and lipids from the perivisceral fat bodies were the main reserves mobilized during the first four weeks of fasting. During this period, somatic indices and blood parameters showed that fish health was not significantly affected. However, after five weeks of food deprivation, the main energy reserves were depleted and the fish became anaemic. The loss of muscle mass indicates that protein breakdown was an important energy source after the reduction of hepatic and perivisceral reserves. Mortality was increasingly observed from seven weeks of starvation. Prior accumulation of high amounts of reserves is essential to allow movements for long distances during spawning migrations in this species.

Negative impact of a cadmium concentration considered environmentally safe in Brazil on the cardiac performance of bullfrog tadpoles

A drastic amphibian decline has been observed worldwide, which can be attributed (among other factors) to exposure to pollutants. Considering that cadmium corresponds to the most rapidly increasing trace metal in the environment, the aim of this work was to evaluate whether the exposure (2 and 16 days) of bullfrog tadpoles to this trace metal, at the concentration currently considered environmentally safe (at 1ppb) in class 1 and 2 waters by the Brazilian Environmental Council, can affect the cardiac performance of these animals. The acute exposure (2 days) of tadpoles to cadmium resulted in a marked bradycardic response, which was correlated with an incomplete cardiac relaxation, without any compensation by improved cardiac twitch force (Fc) or contraction velocity (TPT), nor even by cardiac hypertrophy. Indeed, after 16 days of exposure, the cardiac function of tadpoles became even more depressed due to a marked decrease in Fc, a prolongation of TPT, and also incomplete relaxation (i.e. increases in the ventricle resting tension), without changes in ventricle relative mass. Altogether, the cardiodepressive effects of cadmium (especially after more prolonged exposure periods) impose negative alterations on a tadpole׳s development and also impede adequate homeostatic adjustments to respond appropriately to the exposure to cadmium with increase in energetic demand to counteract the deleterious effects of the xenobiotic. These disturbances can impair tadpoles׳ growth, development and reproduction. It is a fact that allows us to strongly suggest that cadmium concentrations, which are currently considered environmentally safe in Brazil, should be revised.

Synthesis of biogenic silver nanoparticles using Althaea officinalis as reducing agent: evaluation of toxicity and ecotoxicity

Silver nanoparticles (AgNPs) are known mainly because of their bactericidal properties. Among the different types of synthesis, there is the biogenic synthesis, which allows the synergy between the nanocomposites and substances from the organism employed for the synthesis. This study describes the synthesis of AgNPs using infusion of roots (AgNpR) and extract (AgNpE) of the plant Althaea officinalis. After the synthesis through reduction of silver nitrate with compounds of A. officinalis, physico-chemical analyzes were performed by UV-Vis spectroscopy, nanoparticles tracking analysis (NTA), dynamic light scattering (DLS) and scanning electron microscopy (SEM). Toxicity was evaluated through Allium cepa assay, comet test with cell lines, cell viability by mitochondrial activity and image cytometry and minimal inhibitory concentration on pathogenic microorganisms. Biochemical analyzes (CAT - catalase, GPx - glutathione peroxidase e GST - glutationa S-transferase) and genotoxicity evaluation in vivo on Zebrafish were also performed. AgNpE and AgNpR showed size of 157 ± 11 nm and 293 ± 12 nm, polydispersity of 0.47 ± 0.08 and 0.25 ± 0.01, and zeta potential of 20.4 ± 1.4 and 26.5 ± 1.2 mV, respectively. With regard to toxicity, the AgNpE were the most toxic when compared with AgNpR. Biochemical analyzes on fish showed increase of CAT activity in most of the organs, whereas GPx showed few changes and the activity of GST decreased. Also regarding to bactericidal activity, both nanoparticles were effective, however AgNpR showed greater activity. Althaea officinalis can be employed as reducing agent for the synthesis of silver nanoparticles, although it is necessary to consider its potential toxicity and ecotoxicity.

EXCREÇÃO URINÁRIA DE URÉIA, ÁCIDO ÚRICO E AMÔNIA EM PINGÜINS PYGOSCELLIDAE

Estudando o metabolismo nitrogenado de aves antarticas RODRIGUES et al. (Antarctic Record. 42(1):111119, 1998), constataram niveis elevados de ureia no sangue e de arginase em figado e rins de pinguins, semelhantes aos niveis encontrados em mamiferos. O presente trabalho objetivou verificar as concentracoes de acido urico, ureia e amonia na urina de pinguins Pygoscellidae da Regiao Antartica. Os exemplares de pinguins capturados proximo a Estacao Antartica Comandante Ferraz Ilha Rei George – Antartica, foram imobilizados e as amostras de urina colhidas na saida do ureter, como reflexo da situacao de stress. Somente amostras urinarias claras, sem contaminacao com fezes, foram utilizadas para dosagens de acido urico, ureia e amonia. Amostras de urina de Pygoscellis antarctica (n=2), P. Adeliae (n=16) e P. Papua (n=11) foram diluidas com solucao gelada de carbonato de sodio 0,2 % (p/v) e agitadas em banho de gelo ate que todo o precipitado branco fosse dissolvido. A concentracao de ureia foi determinada segundo o metodo da urease como descrito por BERGMEYER (Methods of Enzymatic Analysis. 3ed., Vol. 9. Florida VCH Publ. 449-453, 1985), o acido urico analisado pelo metodo da uricase como descrito por BARHAN e TRINDER (P. Analyst. 97:142, 1972) e amonia determinada pelo metodo colorimetrico do nitroprussiato. As concentracoes de acido urico, ureia e amonia na urina de pinguins Pygoscellidae em estado nativo estao apresentadas na tabela 1. O perfil excretorio dos compostos nitrogenados presentes na urina comparado aos niveis dos mesmos no sangue de P. antarctica, P. adeliae e P. papua, deixa clara a capacidade ureogenica e o metabolismo excretorio uricotelico caracteristico das aves.