Biomedical subjects

M Pacheco

Publications and source records attributed to M Pacheco.

At least 19 recordsLinked to original sources

PubMed2006

Endocrine and metabolic responses of Anguilla anguilla L. caged in a freshwater-wetland (Pateira de Fermentelos--Portugal).

The present short-term in situ study was carried out in a freshwater-wetland--Pateira de Fermentelos--considered an enlargement of Cértima River, in the centre of Portugal. This ecosystem is an important fishing and recreation place, receiving effluents from different origins namely, electroplating industrial effluents containing heavy metals, domestic wastes, as well as pesticides and fertilizers resulting from agriculture activities. The aim of the present research work was to monitor the effects induced by the contaminants present in Pateira de Fermentelos water, using Anguilla anguilla L. (European eel) as a bioindicator. The eels were caged for 48 h at four Pateira de Fermentelos sites, differing in their distances to the main known pollution source (Cértima River): A (close to the lagoon entrance), B, C and D (the farthest from the Cértima River). A reference site was selected near the Cértima river spring. The following parameters were measured: liver ethoxyresorufin-O-deethylase (EROD) and alanine transaminase (ALT) activities, plasma levels of cortisol, 17beta-estradiol (E2), thyroid-stimulating-hormone (TSH), free thyroxine (T4), free triiodothyronine (T3), as well as glucose and lactate. The erythrocytic nuclear abnormalities (ENA) frequency was also scored as a genotoxicity indicator. The results revealed increased plasma cortisol and glucose concentrations at all exposure sites, displaying a similar response pattern. Plasma T3 showed a significant decrease only at site A when compared to reference site, whereas plasma E2 increased at sites B and D when compared to all the other sites, including reference site. The present results indicate the Pateira de Fermentelos water contamination, demonstrating the usefulness of the adopted strategy.

Alanine Transaminase↗

PubMed2006

Cytochrome P4501A, genotoxic and stress responses in golden grey mullet (Liza aurata) following short-term exposure to phenanthrene.

This study represents a first approach to short-term effects of phenanthrene (Phe) in fish. The teleost Liza aurata was exposed to 0.1-2.7microM Phe during 16h. CYP1A induction was assessed as liver ethoxyresorufin-O-deethylase (EROD) activity. Genotoxicity was evaluated in gill and liver as DNA integrity (by alkaline unwinding), whereas in blood the erythrocytic nuclear abnormalities (ENA) frequency was determined. Stress responses were determined as cortisol, glucose and lactate plasma levels. Liver EROD activity was significantly increased by Phe 0.3-2.7microM. Phe genotoxicity in gill was not found, whereas liver DNA integrity significantly decreased after exposure to Phe 0.1 and 0.9microM demonstrating its genotoxicity which did not correlate with liver CYP1A induction. Phe genotoxicity in blood was demonstrated by a significant ENA increase from 0.1 up to 2.7microM. In terms of stress responses, plasma cortisol was significantly increased by Phe 0.3-2.7microM, though plasma glucose was only significantly increased by Phe 0.9 and 2.7microM. The Phe observed effects on L. aurata detected at different levels demonstrate a physiological unbalance and a probable ecological risk to ichthyofauna.

Animals↗

PubMed2006

Oxidative stress and genotoxic effects in gill and kidney of Anguilla anguilla L. exposed to chromium with or without pre-exposure to beta-naphthoflavone.

Fish in the aquatic environment can be subjected to a multipollution state and the occurrence of sequential exposures is an important aspect of eco-toxicological research. In this context, a preceding exposure can affect a toxic response to a subsequent exposure. Therefore, the current study was based on sequential exposures, viz. to a PAH-like compound (beta-naphthoflavone, BNF) followed by a heavy metal (chromium, Cr), focusing on the assessment of oxidative stress responses and their role in induction of genotoxicity. Oxidative stress responses in gill and kidney were investigated in European eel (Anguilla anguilla L.), and measured as lipid peroxidation (LPO), glutathione peroxidase (GPX), catalase (CAT) and glutathione S-transferase (GST) activity, and reduced glutathione (GSH) concentration, whereas genotoxicity was measured as DNA strand breakage. Fish were exposed for 24 h to two Cr concentrations (100 microM, 1 mM), with or without pre-exposure to BNF (2.7 microM, 24 h). In gill, a GSH decrease was observed along with loss of DNA integrity at all exposure conditions except at the lowest Cr concentration, showing a crucial role of GSH over genotoxicity. Moreover, sporadic induction of antioxidant enzymes was not effective in the protection against genotoxicity. However, a different mechanism seems to occur in kidney, since the loss of DNA integrity detected for all exposed groups was not accompanied by alterations in antioxidant levels. With regards to peroxidative damage, both organs showed an LPO increase after sequential exposure to BNF and 100 microM Cr. However, no association between LPO induction and antioxidant responses could be established, showing that LPO is not predictable solely on the basis of antioxidant depletion. The interference of BNF pre-exposure with the response of organs to Cr showed a marked dependence on the Cr concentration. Gill showed synergistic effects on LPO and GPX increase, as well as on CAT and GSH decrease for the lowest Cr concentration. However, for the highest concentration an additive effect on decrease of DNA integrity and an antagonistic effect on the increase of GPX were observed. In kidney, synergistic effects were evident on LPO increase and GSH decrease for the lowest Cr concentration, as well as on CAT and GST decrease for the highest concentration. In contrast, an antagonistic action was observed on DNA integrity loss for both Cr concentrations. The current results are relevant in assessing the interactions of PAHs and metals and contribute to a better knowledge about oxidative stress and mechanisms of genotoxicity in fish.

Anguilla↗

PubMed2006

Anguilla anguilla L. Genotoxic responses after in situ exposure to freshwater wetland (Pateira de Fermentelos, Portugal).

Pateira de Fermentelos is a Cértima River enlargement, close to its river mouth (by the Agueda River), where the introduction of agricultural chemicals such as fertilisers and pesticides, domestic sewage, as well as heavy metals from electroplating industries, results in increased water pollution. The present research work concerns a 48 h in situ exposure of caged eels (Anguilla anguilla L.) at the Pateira de Fermentelos. Five exposure sites were selected, i.e., site A, site B, site C, site D and site E in order to study its water genotoxicity potential, measured in gill, blood, liver and kidney as DNA strand breaks. Eels were also exposed at a reference site by the Cértima River spring. Bottom water samples were collected for further physical-chemical analysis. Site A exposure, significantly decreased gill, blood and liver DNA integrity. Gill and liver DNA integrity was also significantly decreased at site B. At site C only blood DNA integrity was significantly decreased. The present field in situ study demonstrated that the three exposure sites close to the Pateira initial part, such as A, B and C are polluted by pro and/or genotoxic compounds. The genotoxic effects induced in A. anguilla L. suggest a different contamination of the exposure sites A, B and C, in genotoxic chemicals. Thus, according to its genotoxic potential the exposure sites A, B and C, may be ordered as follows site A>site B>site C. No genotoxic effects on A. anguilla L. were observed at site E as DNA strand breaks increase.

Anguilla↗

PubMed2006

Biotransformation, stress and genotoxic effects of 17beta-estradiol in juvenile sea bass (Dicentrarchus labrax L.).

The effects of 17beta-estradiol (E(2)) on fish became a matter of concern, since significant levels of this hormone were detected in the aquatic environment released mainly by domestic sewage treatment plants. In this perspective, the current study was focused on E(2) effects upon biotransformation, stress and genotoxic responses of juvenile Dicentrarchus labrax L. (sea bass). Fish were exposed to E(2) during 10 days in two different ways: water diluted (200 ng/L or 2,000 ng/L) and i.p. injected (0.5 mg/kg or 5 mg/kg). A battery of biological responses was evaluated: liver ethoxyresorufin-O-deethylase (EROD) and alanine transaminase (ALT) activities, liver somatic index (LSI), plasma cortisol, glucose and lactate concentrations, as well as erythrocytic nuclear abnormalities (ENA). All the exposure conditions induced endocrine disruption, measured as plasma cortisol decrease, and genotoxicity, measured as ENA increase. Thus, no differences were detected either between different exposure routes or tested concentrations. Concerning liver EROD and ALT activities, as well as plasma glucose and lactate concentrations no differences were found between treated and control groups. LSI was the only parameter to respond differently in the two exposure routes, as only E(2) water diluted induced a significant increase in this hepatic indicator.

Alanine Transaminase↗

PubMed2005

Responses of European eel (Anguilla anguilla L.) circulating phagocytes to an in situ closed pulp mill effluent exposure and its association with organ-specific peroxidative damage.

The effect of bleached kraft pulp mill effluent (BKPME) persistent compounds on phagocyte activities and its organ-specific influence in gill, kidney and liver was studied in European eel (Anguilla anguilla L.). Eels were caged and plunged at 3 different sites-50 m (site 1), 100 m (site 2) and 2,000 m (site 3) away from the end of the closed BKPME discharging channel for 8 and 48 h. Gill, head kidney and peritoneum phagocytes oxidative burst activity (OBA) was measured by the nitroblue tetrazolium reduction assay whereas lipid peroxidation (LPO) in eel gill, kidney and liver was measured by thiobarbituric acid reaction. A significant gill OBA induction was found at 8h on site 2, and on sites 1, 2 and 3 at 48 h exposure. However, in head kidney and peritoneal exudate phagocytes, OBA induction was significant only at sites 2 and 3 after 48 h exposure. In those particular sites, a significant increase in gill, kidney and liver LPO was measured that is assumed to result from OBA induction. Considering OBA and LPO, gill is the most affected tissue compared to kidney and liver. Gill vulnerability towards peroxidative damage was demonstrated at 8h on site 2 and at 48 h on sites 1, 2 and 3, whereas in kidney was observed at sites 2 and 3 only at 48 h. Liver LPO increased at site 2 only after 48 h exposure. Our results demonstrate that the OBA activation pattern in gill and kidney is associated with the induced peroxidative damage extent in those organ, together with water pollution the exposure route, resulting from previous BKPME effluent sediment contamination which may affect the activation pattern of circulating fish phagocytes.

Anguilla↗

PubMed2005

Sparus aurata L. liver EROD and GST activities, plasma cortisol, lactate, glucose and erythrocytic nuclear anomalies following short-term exposure either to 17beta-estradiol (E2) or E2 combined with 4-nonylphenol.

Immature Sparus aurata L. (gilthead seabream) were exposed to 17beta-estradiol (E(2)) 4000 ng/l and to the same E(2) concentration mixed with 50,000 ng/l 4-nonylphenol (E(2) + NP) during 4, 8, 12 and 16 h. E(2) availability and E(2) plasma level variations were assessed. Liver biotransformation capacity was measured as ethoxyresorufin-O-deethylase (EROD) and glutathione-S-transferase (GST) activities. Plasma cortisol, lactate and glucose were also determined. Genotoxicity was assessed through erythrocytic nuclear anomalies (ENA) frequency. Liver EROD activity significantly decreased during the whole experiment for both treatments, with the exception of 16 h exposure to E(2). Liver GST activity was significantly increased after 8 and 12 h of exposure either to E(2) or E(2) + NP. An endocrine disruption expressed as plasma cortisol decrease was observed after 16 h exposure under both tested conditions, concomitantly with a plasma lactate increase. No genotoxic responses, measured as ENA frequency, were detected. Analyzing the E(2) water concentration in aquaria without fish it was demonstrated an intense and fast E(2) loss, considerably reducing its availability to fish. In the presence of fish, E(2) water levels were drastically reduced after 4 h exposure, being this reduction more pronounced in E(2) aquarium when compared to E(2) + NP aquarium. In addition, it was demonstrated a rapid E(2) uptake from the water since the highest E(2) plasma concentrations were observed after 4 h exposure, followed by a continuous decrease, which became more pronounced between 8 and 12 h of exposure. Furthermore, during the first 8 h exposure to E(2) and E(2) + NP, seabream plasma E(2) concentrations were higher than the initial water exposure concentration. Comparing the E(2) plasma levels in both seabream-exposed groups, it was clear that its concentration is always higher in E(2) + NP-treated fish. Despite the previous results, no significant differences were found in the measured responses between E(2) and E(2) + NP.

Animals↗

PubMed2005

Physiological and genetic responses of European eel (Anguilla anguilla L.) to short-term chromium or copper exposure-Influence of preexposure to a PAH-like compound.

Anguilla anguilla L. (European eel) was exposed for 24 h to chromium (Cr-100 microM and 1 mM) or copper (Cu-1 and 2.5 microM), with or without a 24-h preexposure to beta-naphthoflavone (BNF-2.7 microM), a polycyclic aromatic hydrocarbon (PAH)-like compound, simulating sequential exposure to PAHs and heavy metals. Plasma cortisol, thyroid-stimulating hormone (TSH), free triiodothyronine (T3), and free thyroxine (T4) were determined in order to assess the effects on endocrine function. Plasma glucose and lactate also were measured. The frequency of erythrocytic nuclear abnormalities (ENA) was scored as a genotoxicity indicator. Plasma T4 decreased in eels when exposed to Cr only. The interference of BNF preexposure on Cr effects was observed as a significant plasma glucose increase. Single exposures to Cu elevated plasma cortisol and glucose (2.5 microM), as well as plasma lactate (1 microM), whereas a T4 decrease was found for both concentrations. BNF preexposure prevented plasma cortisol and lactate increases; however, a greater T4 decrease was observed in eels exposed to 2.5 microM Cu. Moreover, this pretreatment was crucial for genotoxicity expression because only BNF+2.5 microM Cu-exposed fish exhibited significant ENA induction. In general, plasma T4 was the most affected hormone, as it responded to all Cr and Cu exposure conditions.

Anguilla↗

PubMed2005

Morphological criteria to distinguish cell death induced by apoptotic and necrotic treatments.

We present a comparative study of apoptotic and necrotic morphology (light and scanning electron microscopy), induced by well known experimental conditions (photodynamic treatments, etoposide, hydrogen peroxide, freezing-thawing and serum deprivation) on cell cultures. Our results indicate that morphological criteria (apoptotic cell rounding and shrinkage, and appearance of membrane bubbles in early necrosis) allow to distinguish these cell death mechanisms, and also show that, independently of the damaging agents, the necrotic process occurs in a characteristic sequence (coalescence of membrane bubbles in a single big one that detaches from cells remaining on the substrate).

Apoptosis↗

PubMed2005

Biotransformation and genotoxic biomarkers in mullet species (Liza sp.) from a contaminated coastal lagoon (Ria de Aveiro, Portugal).

The aim of the present study was to assess liver ethoxyresorufin-O-deethylase (EROD) activity and erythrocytic nuclear abnormalities (ENA) frequency in three mullet species, as biotransformation and genotoxicity biomarkers, respectively, for monitoring aquatic ecosystems. Specimens of Liza aurata, Liza ramada and Liza saliens were caught bimonthly, between May 2000 and March 2001, at nine stations in the Ria de Aveiro (Portugal), a polluted coastal lagoon. The survey revealed two critical areas (Vagos and Carregal) where it was observed elevated EROD activity and ENA frequency, comparing to the reference sites. The results often reflect a contamination by a mixture of pollutants, from point and non-point sources. Nevertheless, the responses observed at Vagos are probably attributable to high polycyclic aromatic hydrocarbon (PAH) water levels measured at that station. All species were able to detect inter-site differences; however L. aurata seems to be the most appropriate bio-indicator due to its larger geographical distribution and abundance during the whole year. The adopted biomarkers, applied to mullet species, can be successfully employed for biomonitoring anthropogenic contamination in both Atlantic and Mediterranean European coastal waters.

Animals↗

PubMed2005

Endocrine and metabolic changes in Anguilla anguilla L. following exposure to beta-naphthoflavone--a microsomal enzyme inducer.

Anguilla anguilla L. were exposed during 24 and 48 h to 2.7 muM beta-naphthoflavone (BNF), a known microsomal enzyme inducer. The BNF effects on thyroid-stimulating hormone (TSH), free triiodothyronine (T3), free thyroxine (T4) and cortisol plasma levels were investigated. Alterations on plasma glucose and lactate levels were also measured as an indication of energy-mobilizing hormones alterations. BNF showed to be able to decrease significantly A. anguilla plasma T4 levels, whereas TSH, T3 and cortisol plasma remained constant. However, plasma glucose levels were significantly increased, demonstrating that intermediary metabolism has been affected. These results demonstrate that BNF a PAH-like compound alters the normal functioning of the hypothalamo-pituitary-thyroid (HPT) axis in A. anguilla.

Anguilla↗

PubMed2004

Juvenile sea bass biotransformation, genotoxic and endocrine responses to beta-naphthoflavone, 4-nonylphenol and 17 beta-estradiol individual and combined exposures.

Juvenile sea bass, Dicentrarchus labrax L., were exposed during 2, 4, 8, and 24 h to 0.9 microM beta-naphthoflavone (BNF), 131 nM 17 beta-estradiol (E(2)), 4.05 microM 4-nonylphenol (NP), as well as to BNF combined either to E(2) or NP (maintaining the previous concentrations). Liver cytochrome P450 content (P450), ethoxyresorufin-O-deethylase (EROD), and glutathione S-transferase (GST) activities were measured in order to evaluate biotransformation responses. Genotoxicity was assessed as erythrocytic nuclear abnormalities (ENA) frequency. The effects on endocrine function were evaluated as plasma cortisol and glucose. Cortisol was not affected by xeno/estrogens tested, either in single exposure or mixed with BNF. Nevertheless, the intermediary metabolism was affected since glucose concentration increased after 4 h exposure to E(2), and after all BNF+NP exposure lengths. Moreover, a synergism between BNF and NP was thoroughly demonstrated, whereas a sporadic antagonistic interaction was found at 4 h BNF + E(2) exposure. Liver EROD and GST activities were not significantly altered by single E(2) or NP exposure. However, both compounds were able to induce EROD activity in the presence of BNF. NP single exposure was able to significantly increase liver P450 content, while its mixture with BNF displayed an antagonistic interference. Considering the xeno/estrogens single exposures, only NP induced an ENA increase; however, both mixtures (BNF + E(2) and BNF + NP) displayed genotoxic effects. Fish responses to mixtures of xenobiotics are complex and the type of interaction (synergism/potentiation or antagonism) in a particular mixture can vary with the evaluated biological response.

Animals↗

PubMed2004

Responses of European eel (Anguilla anguilla L.) in two polluted environments: in situ experiments.

European eels (Anguilla anguilla L.) were caged for 8 and 48 h in two polluted areas to assess the contamination cleanup process in the Vouga River 2 years after the official closing of a bleached kraft pulp mill effluent (BKPME; experiment 1), and to monitor the effects induced by contaminated offward fishing harbor waters (experiment 2). Plasma cortisol, glucose, and lactate were evaluated as stress responses. In experiment 1, plasma cortisol, glucose, and lactate increased after 8 h of exposure in site 3, which is located farthest from the deactivated sewage outlet. However, A. anguilla seemed to adapt after 48 h of exposure in site 3, because all three parameters returned to control levels. Plasma glucose also significantly increased after 8 h of exposure at sites 1, 2, and 3, returning to control levels after 48 h. Plasma lactate levels increased after 8 h of exposure at site 3 and after 48 h of exposure at site 1. In experiment 2, A. anguilla exposed to contaminated harbor waters increased their plasma lactate after 8 and 48 h of exposure, whereas their cortisol and glucose plasma were elevated only after 48 h of exposure. The results demonstrate that even 2 years after the official closing of the BKPME sewage outlet, the river Vouga water remains contaminated by the sediment associated chemicals. Because the fishing harbor induced in A. anguilla the same type of stress responses, it is also an area of concern. The adopted stress parameters allied to a caging strategy are recommended for future environmental monitoring assessments.

Anguilla↗

PubMed2004

Glutathione protects heavy metal-induced inhibition of hepatic microsomal ethoxyresorufin O-deethylase activity in Dicentrarchus labrax L.

The in vitro effects of chromium (Cr(VI)), copper (Cu2+), iron (Fe2+), mercury (Hg2+), and zinc (Zn2+) were assessed on liver microsomal ethoxyresorufin O-deethylase (EROD) activity from a sea bass (Dicentrarchus labrax L.) preexposed under laboratory conditions to 2.7 microM beta-naphthoflavone. The reduced glutathione (GSH) protection potential against heavy metal effects was also studied. The heavy metal concentration ranges used for this study were as follows: 10 pM-5 mM Cr(VI), 10 pM-100 microM Cu2+, 10 pM-1 mM Fe2+, 10 pM-10 microM Hg2+, and 10 pM-100 microM Zn2+. Liver microsomal EROD activity was significantly inhibited after in vitro exposure to Cr(VI) (500 microM), Cu2+ (1 microM), Fe2+ (100 microM), Hg2+ (100 pM), and Zn2+ (10 microM). Heavy metals inhibitory effect on liver EROD activity was ordered as follows: Hg2+ > Cu2+ > Zn2+ > Fe2+ > Cr(VI). Protective effects against Hg2+ (1 and 10 microM), Cu2+ (1, 10, and 100 microM), and Zn2+ (10, 50, and 100 microM) were observed in the presence of 0.5 mM GSH by a decrease in liver microsomal EROD activity inhibition. However, 0.5 mM GSH did not protect liver microsomal EROD activity from Cr(VI), and Fe2+-induced inhibition. The effect of metal mixtures (Cu(2+) + Zn(2+), Zn(2+) + Fe(2+), Zn(2+) + Cr(VI), and Cr(VI) + Fe(2+)) (100 microM) on liver microsomal EROD activity was also assessed, revealing a synergistic interaction.

Animals↗

PubMed2004

Enzymatic and nonenzymatic antioxidants as an adaptation to phagocyte-induced damage in Anguilla anguilla L. following in situ harbor water exposure.

Anguilla anguilla L. were caged for 8 and 48 h in harbor water of Aveiro Lagoon, Portugal. Respiratory burst activity (RBA) of peritoneal, head kidney, and gill phagocytes was measured. Lipid peroxidation (LPO) was estimated in gill, kidney, and liver. Liver ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (Cyt P450) content, and bile metabolites were assayed. Various antioxidant enzymes, viz., glutathione peroxidase, catalase, and glutathione S-transferase and nonenzymatic antioxidant, viz., total reduced glutathione were also studied. Harbor water xenobiotics induced a significant RBA increase in gill after 8 h; whereas in peritoneum and head kidney it increased after 48 h exposure. These responses were adversely associated with tissue-specific peroxidative damage since significant LPO increase was observed in gill (8 and 48 h), kidney (48 h), and liver (48 h). The tissue most affected was gill. Moreover, liver EROD activity, Cyt P450 content and bile metabolites remain unaltered after 8 h; in contrast, 48 h exposure showed significant EROD activity decrease and pyrene-type bile metabolites increase. Decreased EROD activity may be associated with concomitant liver damage, as increased LPO was observed after 48 h. Furthermore, the tissue-specific damage corresponded to the differences in the antioxidant potentials of the tissues, since the initial exposure period caused a significant increase in liver antioxidant activities, whereas gill and kidney showed a significant decrease, demonstrating that liver is highly adaptive to oxidative damage. However, at 48 h exposure gill, kidney, and liver showed a suppressive antioxidant effect, probably due to PAHs, since a significant induction at PAH-type bile metabolites has been seen. Our results demonstrate that phagocyte activation and associated peroxidative damage are concomitantly corroborated with enzymatic and nonenzymatic antioxidant activity differences. In addition, hepatic antioxidant induction after short-term exposure may serve as a potent biomarker for water pollutants in fish.

Anguilla↗

PubMed2004

Anguilla anguilla L. plasma cortisol, lactate and glucose responses to abietic acid, dehydroabietic acid and retene.

Anguilla anguilla L. were exposed to 0, 0.1, 0.3, 0.9 and 2.7 microM abietic (AA), dehydroabietic (DHAA) acids and retene (Re) during 8, 16, 24 and 72 h. The eels plasma cortisol, glucose and lactate were measured. A significant decrease in plasma cortisol was observed at 72 h exposure to 0.9 and 2.7 microM Re. DHAA (0.1 microM) significantly decreased plasma cortisol in eels after 8 and 24 h exposure. However, a significant plasma cortisol increase was found after 16 h, 2.7 microM AA exposure and after 24 h exposure to 0.1 microM and 2.7 microM AA. Furthermore, 72 h exposure to 0.9 microM AA also induced a plasma cortisol increase. A general rise in plasma glucose was detected after all exposure periods to Re. The plasma lactate also increased after 72 h exposure to 2.7 microM AA and after 8 h exposure to 0.1 microM DHAA.

Abietanes↗

PubMed2004

Anguilla anguilla L. antioxidants responses to in situ bleached kraft pulp mill effluent outlet exposure.

This study assesses the antioxidant enzymes activities viz., catalase, glutathione peroxidase, glutathione S-transferase and nonenzymatic antioxidant molecule such as glutathione in Anguilla anguilla L. gill, kidney and liver in response to 8- and 48-h exposure to bleached kraft pulp mill effluent (BKPME). A. anguilla were caged and plunged at three different sites-50 (Site 1), 100 (Site 2) and 2000 m (Site 3) away from the closed BKPME outlet. A significant gill (8 and 48 h) and kidney (48 h) catalase activity decrease was observed at site 2 exposure whereas liver showed a significant increase in catalase activity after 8 and 48 h to site 1 exposure. Glutathione peroxidase (GPX) activity was significantly decreased in gill after 8-h exposure to site 1 and 48-h exposures to sites 1 and 2, respectively. Concerning gill, kidney and liver glutathione S-transferase (GST) activity, a significant gill GST activity decrease after 8 h at site 2 and 48 h at sites 1 and 2 was observed; in kidney, a significant decrease in its activity was observed after 48 h at sites 1 and 2, respectively, whereas in liver, the decrease was significant only at site 2 after 48-h exposure. The in situ BKPME exposure caused a significant total gill and kidney reduced glutathione (GSH) decrease after 8 h at site 2 exposure and after 48 h at site 1 and 2 exposures, respectively. However, a biphasic response was observed in liver, i.e. initial significant increase after 8 h at site 2 followed by a significant decrease after 48 h to the same site exposure. The enzymatic and nonenzymatic antioxidants pattern in gill and kidney, as observed in this study, was different than liver, demonstrating that the liver was more resistant to oxidative damage than gill and kidney. In addition, A. anguilla gill, kidney and liver antioxidants adaptation potentials may serve as a surrogate biomarker to BKPME exposure.

Adaptation, Physiological↗

PubMed2003

Naphthalene-induced differential tissue damage association with circulating fish phagocyte induction.

The effect of naphthalene on phagocytes and associated abnormal cellular activities was studied in Anguilla anguilla L. Fish were exposed to sublethal concentrations (0.1, 0.3, 0.9, 2.7 microM) of naphthalene for 8, 16, 24, 48, and 72 h. Gill, head kidney, and peritoneum phagocyte respiratory burst activity (RBA) was measured by nitroblue tetrazolium reduction assay, whereas lipid peroxidation in fish gill, kidney, and liver was measured by thiobarbituric acid reaction substance. A significant increase (P < 0.05-P < 0.001) in total cell count (TCC) of phagocytes (TCCPH) isolated from gill, head kidney, and peritoneum was observed after 8 h of naphthalene exposure and persisting at 16 h exposure. However, long-term exposures (24, 48, and 72 h) induced a significant (P < 0.05-P < 0.001) decrease in TCC at all the concentration levels. Naphthalene exposure caused a significant (P < 0.05-P < 0.001) RBA induction after 8 and 16 h in gill, head kidney, and peritoneal phagocytes, which consequently resulted in a significant (P < 0.01 and P < 0.001) peroxidative tissue damage increase measured as lipid peroxidation (LPO) in gill, kidney, and liver at the same time intervals. Considering TCCPH, RBA, and LPO, the most affected tissues were gill and kidney after 8 and 16 h exposure to all naphthalene concentrations. RBA was significantly decreased after 24, 48, and 72 h of exposure (P < 0.05-P < 0.001), whereas peroxidative damage increased significantly (P < 0.05-P < 0.001) and persisted in long-term exposures (72 h) at all concentration levels in gill and kidney. Liver short-term exposure (8 and 16 h) to all naphthalene concentrations did not alter LPO activity. Long-term exposures (48 and 72 h) caused a significant (P < 0.01 and P < 0.001) LPO increase, which was more pronounced at 72 h. The results demonstrate that the activation pattern of RBA was corroborated by the extent of phagocyte-induced peroxidative damage in the tissues, as demonstrated by a significant increase of circulating phagocytes. However, the route of exposure and mode of entry of a pollutant may affect the activation pattern of circulating fish phagocytes. It is proposed that measurement of phagocyte-induced reactive oxygen species and their association with peroxidative damage in fish tissues may prove to be useful in biomonitoring fish exposure to aquatic pollutants.

Anguilla↗