Influence of dioxin and metal-contaminated sediment on phase I and II biotransformation enzymes in silver crucian carp

Several biochemical responses were measured in silver crucian carp (Carassius auratus gibelio) after exposure to sediments obtained from contaminated Ya-Er Lake, No, 1 pond, and an unpolluted reference site, Honglian Lake. After 1 week of exposure, a significant induction of the phase I biotransformation enzyme (ethoxylresorufin-o-deethylase, EROD) was found (83-fold of control), whereas the phase II biotransformation enzyme (glutathione S-transferase, GST) exhibited a slight, but significant induction (1,4-fold of control) after 4 weeks of exposure. The level of cellular glutathione in the liver was also slightly elevated after 4 weeks of exposure. The delayed response of GST to the contaminants indicates that the phase I and phase II biotransformation enzymes are regulated differently in fish. The results suggest that EROD is a sensitive bioindicator to assess the toxicity of dioxin-contamined sediment in the laboratory, (C) 1998 Academic Press.

Phase I and II biotransformation enzymes and polycyclic aromatic hydrocarbons in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected in front of an oil refinery

The aim of the present study was to investigate the responses of phase I and II biotransformation enzymes and levels of PAHs in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected from three sites at different distance from an oil refinery. Phase I enzyme activities as NAD(P)H-cyt c red, NADH ferry red, B(a)PMO and phase II as UDPGT. GST were measured in digestive gland while 16 PAHs (US-EPA) in whole soft tissue. An added value to the data obtained in the present study rely on the RDA analysis which showed close correlations between PAHs levels and phase I enzyme activities in mussels collected in front of the refinery. And again a significant spatial correlation between B(a)P levels and NADPH-cyt c red activities was observed using linear models. No differences among sites for B(a) PMO and phase II GST activities were observed, while the application of UDPGT as biomarkers requires further investigation. (C) 2012 Elsevier Ltd. All rights reserved.

Benzo(a)pyrene Metabolism and EROD and GST Biotransformation Activity in the Liver of Red- and White-Blooded Antarctic Fish.

Climate change and anthropogenic pollution are of increasing concern in remote areas such as Antarctica. The evolutionary adaptation of Antarctic notothenioid fish to the cold and stable Southern Ocean led to a low plasticity of their physiological functions, what may limit their capacity to deal with altered temperature regimes and pollution in the Antarctic environment. Using a biochemical approach, we aimed to assess the hepatic biotransformation capacities of Antarctic fish species by determining (i) the activities of ethoxyresorufin-O-deethylase (EROD) and glutathione-S-transferase (GST), and (ii) the metabolic clearance of benzo(a)pyrene by hepatic S9 supernatants. In addition, we determined the thermal sensitivity of the xenobiotic biotransformation enzymes. We investigated the xenobiotic metabolism of the red-blooded Gobionotothen gibberifrons and Notothenia rossii, the hemoglobin-less Chaenocephalus aceratus and Champsocephalus gunnari, and the rainbow trout Oncorhynchus mykiss as a reference. Our results revealed similar metabolic enzyme activities and metabolic clearance rates between red- and white-blooded Antarctic fish, but significantly lower rates in comparison to rainbow trout. Therefore, bioaccumulation factors for metabolizable lipophilic contaminants may be higher in Antarctic than in temperate fish. Likewise, the thermal adaptive capacities and flexibilities of the EROD and GST activities in Antarctic fish were significantly lower than in rainbow trout. As a consequence, increasing water temperatures in the Southern Ocean will additionally compromise the already low detoxification capacities of Antarctic fish.

Les répercussions de l’insuffisance rénale chronique sur le transport des médicaments

L’insuffisance rénale chronique (IRC) affecte 13 % de la population américaine et son incidence ne cesse d’augmenter. Malgré un ajustement des doses de médicaments administrés en fonction du taux de filtration glomérulaire du patient urémique, près de 40 % des patients reçoivent une dose trop élevée en raison de modifications de l’élimination extrarénale des médicaments chez ces patients. Il est connu que l’IRC affecte l’élimination métabolique des médicaments par les cytochromes P450 et les enzymes de biotransformation de phase II. Nous avons aussi démontré, chez le rat, que l’IRC affecte l’expression et l’activité de transporteurs de médicaments intestinaux entraînant une augmentation de la biodisponibilité de certains médicaments. On retrouve des transporteurs de médicaments dans de nombreux organes comme le foie, les reins et la barrière hématoencéphalique (BHE) où ils jouent des rôles importants dans les éliminations biliaire et rénale et la pénétration des médicaments au cerveau. Le but de ce travail était de mesurer, chez des rats néphrectomisés, les impacts de l’IRC sur l’expression protéique et génique et l’activité des transporteurs de médicaments hépatiques, rénaux et cérébraux. Les transporteurs étudiés sont de la famille des transporteurs ABC (P-glycoprotéine, multidrug-resistance related protein, breast cancer resistance protein) ou des solute carriers (organic anion transporter, organic anion transporting protein). Aussi, une étude réalisée chez l’humain visait à évaluer la pharmacocinétique de deux médicaments : la fexofénadine, un médicament majoritairement transporté, et le midazolam, un substrat du cytochrome P450 3A4, chez des sujets dialysés. Nos résultats montrent que, chez le rat, l’IRC entraîne des modulations de l’expression des transporteurs d’influx et d’efflux hépatiques pouvant entraîner des diminutions du métabolisme hépatique et de l’excrétion biliaire des médicaments. Dans le rein, nous avons démontré des modulations de l’expression des transporteurs de médicaments. Nous avons aussi démontré que l’IRC diminue l’élimination urinaire de la rhodamine 123 et favorise l’accumulation intrarénale de médicaments transportés comme la benzylpénicilline et la digoxine. À la BHE, nous avons démontré des diminutions de l’expression des transporteurs de médicaments. Toutefois, nous n’avons pas observé d’accumulation intracérébrale de trois substrats utilisés (digoxine, doxorubicine et vérapamil) et même une diminution de l’accumulation intracérébrale de la benzylpénicilline. Il semble donc que, malgré les modulations de l’expression des différents transporteurs de médicaments, l’intégrité et la fonction de la BHE soient conservées en IRC. Chez l’humain, nous avons démontré une augmentation de la surface sous la courbe de la fexofénadine chez les sujets dialysés, comparativement aux témoins, suggérant une altération des mécanismes de transport des médicaments chez ces patients. Nous n’avons, toutefois, pas observé de modification de la pharmacocinétique du midazolam chez les patients dialysés, suggérant une activité métabolique normale chez ces patients. Un ou des facteurs s’accumulant dans le sérum des sujets urémiques semblent responsables des modulations de l’expression et de l’activité des transporteurs de médicaments observées chez le rat et l’humain. Ces travaux mettent en évidence une nouvelle problématique chez les sujets urémiques. Nous devons maintenant identifier les mécanismes impliqués afin d’éventuellement développer des stratégies pour prévenir la toxicité et la morbidité chez ces patients.

Biochemical responses in armored catfish (Pterygoplichthys anisitsi) after short-term exposure to diesel oil, pure biodiesel and biodiesel blends

Biodiesel fuel is gradually replacing petroleum-based diesel oil use. Despite the biodiesel being considered friendlier to the environment, little is known about its effects in aquatic organisms. In this work we evaluated whether biodiesel exposure can affect oxidative stress parameters and biotransformation enzymes in armored catfish (Pterygoplichthys anisitsi, Loricariidae), a South American endemic species. Thus, fish were exposed for 2 and 7d to 0.01mLL-1 and 0.1mLL-1 of pure diesel, pure biodiesel (B100) and blends of diesel with 5% (B5) and 20% (B20) biodiesel. Lipid peroxidation (malondialdehyde) levels and the activities of the enzymes glutathione S-transferase, superoxide dismutase, catalase and glutathione peroxidase were measured in liver and gills. Also, DNA damage (8-oxo-7, 8-dihydro-2'-deoxyguanosine) levels in gills and 7-ethoxyresorufin-O-deethylase activity in liver were assessed. Pure diesel, B5 and B20 blends changed most of the enzymes tested and in some cases, B5 and B20 induced a higher enzyme activity than pure diesel. Antioxidant system activation in P. anisitsi was effective to counteract reactive oxygen species effects, since DNA damage and lipid peroxidation levels were maintained at basal levels after all treatments. However, fish gills exposed to B20 and B100 presented increased lipid peroxidation. Despite biodiesel being more biodegradable fuel that emits less greenhouse gases, the increased lipid peroxidation showed that biofuel and its blends also represent hazards to aquatic biota. © 2013 Elsevier Ltd.

Pollution-induced metabolic responses in hypoxia-tolerant freshwater turtles

The physiological control to support the absence of O2 for long periods of diving, and oxidative damage impact caused by the whole process of hypoxia/reperfusion in freshwater turtles is well known. However, effects of contaminants may act as co-varying stressors and cause biological damage, disrupting the hypoxia/reperfusion oxidative damage control. In order to investigate the action of environmental stressors present in domestic or industrial wastewater effluent, we performed a biochemical analysis of biotransformation enzymes, oxidative stress, as well as neuromuscular, physiological and morphological parameters in Phrynops geoffroanus, an hypoxic-tolerant freshwater turtle endemic of South America, using animals sampled in urban area, contaminated by sewage and industrial effluents and animals sampled in control area. Here we demonstrate the physiological and biochemical impact caused by pollution, and the effect that these changes cause in antioxidant activity. Animals from the urban area exhibited higher EROD (ethoxyresorufin-O-deethylase, CYP1A1), GST (glutathione S-transferase), G6PDH (glucose-6-phosphate deshydrogenase), AChE (acetilcholinesterase) activities and also TEAC (trolox-equivalent antioxidant capacity) and TBARS (thiobarbituric acid reactive substances) values. We examined whether two morphometric indices (K - condition factor and HIS - hepatosomatic index) which help in assessing the general condition and possible liver disease, respectively, were modified. The K of the urban animals was significantly decreased compared to the control animals, but the HIS value was increased in animals from the urban area, supporting the idea of an impact in physiology and life quality in the urban freshwater turtles. We propose that this freshwater turtle specie have the ability to enhance its antioxidants defenses in order to protect from tissue damage caused by hypoxia and reperfusion, but also that caused by environmental contamination and that the oxidative damage control in hypoxic conditions has resulted in an adaptive condition in hypoxic-tolerant freshwater turtle species, in order to better tolerate the release of contaminated effluents resulting from human activity. © 2013 Elsevier Inc.

Variação em biomarcadores bioquímicos de contaminação em Tilápias do Nilo (Oreochromis niloticus) de diferentes portes

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

Avaliação de biomarcadores bioquímicos em tilápias (Oreochromis niloticus) expostos a óleo diesel e biodiesel

Pós-graduação em Química - IBILCE

Avaliação dos efeitos deletérios do diuron e de seus metabólitos em lambaris (Astyanax sp.): testes de toxicidade, marcadores de estresse oxidativo, marcadores genotóxicos e enzimas de biotransformação

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

Estrógenos na bacia hidrográfica Tietê-Jacaré: ocorrência e avaliação do impacto ambiental

Pós-graduação em Química - IQ

Ecological relevance of sentinels' biomarker responses: A multi-level approach

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Bioaccumulation of butyltins and liver damage in the demersal fish Cathorops spixii (Siluriformes, Ariidae)

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

Cytotoxic, biochemical and genotoxic effects of biodiesel produced by different routes on ZFL cell line

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Enzimas de biotransformação em tilápia (Oreochromis niloticus) exposta ao hormônio natural 17b-estradiol

Among the sexual hormones the estrogens are receiving major attention due to their biological activity. Such biological response is atributed to the best conformation recognized by their receptors, resulting in maximum responses. The estrogens are also considered responsible for most of disruptor´s effects caused by their presence in aquatic ecosystems. The 17β-estradiol hormone is produced by vertebrates during the reproductive phase of their lives and its presence has been detected in superficial waters. The objective of this study was to evaluate the cause-effect of tilapia exposition to the hormone 17β-estradiol through the analysis of biotransformation enzymes in liver and gills, complemented with the quantification of 17β-estradiol and estrone in water samples collected from fish ponds integrated to swine production. The present study was conducted under laboratory conditions, in a randomized experimental design with three levels of 17β-estradiol (E2) (0, 5, 15 µg L-1), with three replicates. After 7 days of exposure time, liver and gills were extracted to analyze three isoforms of cytochrome P450: EROD, BROD, PROD and the activity of Glutathione S-Transferase (GST). The results showed that the EROD activity (CYP1A), normally induced by the metabolism of aromatic compounds, did not present statistical differences among the treatments exposed to E2, what means that the hormone did not induce isoform 1A in fish under these particular experimental conditions. PROD activity was significantly altered in both concentrations, by means of 5 and 15 µg L-1, when compared to control. This result can indicate an important role of PROD on the metabolism of E2 present in water. Regarding to the BROD activity, it could be observed differences statistically significant between control and both groups of treatments. Two or more CYP isoforms can contribute to the metabolism of the same compound, what makes BROD a candidate as a next bioindicator of the exposure to E2 in aquatic ecosystems. Analysis of variance could confirm the effect of E2 statistically significant on the GST activity in liver tissues with >90% of significance (Prob>F = 0.0753). Furthermore, it was possible to observe that the values of GSTs activities in liver and gills in both, control and treatments, follow a tendency, that means, enzymatic activity in gills increase as the increasing of the activity in the liver tissues. In this study, the 17β–estradiol was found in measurable concentrations in three sampled points, and these values were similar to the findings of other authors at different locations in Brazil. In addition, those values are much higher than the minimum concentration that presented observable effects (10 ng L-1).

Biochemical responses, morphometric changes, genotoxic effects and CYP1A expression in the armored catfish Pterygoplichthys anisitsi after 15 days of exposure to mineral diesel and biodiesel

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