Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.)

The aim of this study was to characterize biomarker responses in three-spined sticklebacks exposed to prochloraz (Pcz). For this purpose, adult sticklebacks were exposed for 2 weeks to prochloraz at 0, 10, 50, 100 and 500 microg/L prior to one week of depuration in clean water. At days 7, 14 and 21, several hepatic biomarkers were measured including 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), total glutathione (GSH) content and thiobarbituric acid reactive substances (TBARS). Pcz induced a transient increase of antioxidant enzymes and a depletion of glutathione content during the first 7 days of exposure. This study showed that EROD activity and antioxidants were disrupted in a transient manner. GST was rapidly induced in a dose-dependent manner and this induction was persistent and observed also after depuration. GST appeared as a valuable biomarker to assess the exposure to Pcz.     

Field validation of a battery of biomarkers to assess sediment quality in Spanish ports

Two marine invertebrates, the crab Carcinus maenas and the clam Ruditapes philippinarum, were used as bioindicator species to assess contamination when exposed in situ to sediment from different sites from four Spanish ports Cadiz (SW Spain), Huelva (SW Spain), Bilbao (NE Spain) and Pasajes (NE Spain). In an attempt to determine sediments toxicity, a combination of exposure biomarkers was analyzed in both species: metallothionein-like-proteins (MTLPs), ethoxyresorufin O-deethylase (EROD), glutathione S-transferase activity (GST), glutathione peroxidase (GPX) and glutathione reductase (GR). In parallel, physical and chemical characterization of the different sediments was performed and biological responses related to the contaminants. Significant induction of MTLPs was observed when organisms were exposed to metal contaminated sediments (port of Huelva), and EROD and GPX activities after exposure to sediments containing organic compounds (port of Bilbao and Pasajes). No significant interspecies differences were observed in biomarker responses except for the GST and GR.     

Effects of chitosan on oxidative stress and metallothioneins in aquatic worm Tubifex tubifex (Oligochaeta, Tubificidae)

Chitosan is a natural polymer which has the property to elicit the natural defenses mechanism in plant and which can be an interesting biopesticides. It is then necessary to investigate the potential toxicity of chitosan for aquatic animal health. Metallothioneins (MTs) are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MTs contents had been considered as a specific biomarker of metal exposure. However recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth and anti-oxidative defenses. Therefore, the induction of MTs has been investigated in the aquatic worms Tubifex tubifex exposed to chitosan. MTs levels in exposed worm increased significantly (p > 0.05) after 2, 4, and 7 days of exposure to different concentrations of chitosan (maximum + 158.19 +/- 10.2% after 2 days of exposure to 125 mgl(-1) of chitosan). Several antioxidant parameters including glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR), and catalase (CAT) were quantified in T. tubifex after 2, 4, and 7 days of exposure to chitosan. Exposure to chitosan had a negative effect on T. tubifex growth (maximum effect -6.11 +/- 1.6% after 7 days with 125 mgl(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein contents. The activity of catalase (CAT), glutathione-S-transferase (GST), and glutathione reductase (GR) increased in response to the chitosan demonstrating an oxidative stress in the worms.     

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.     

Comparision of the waterborne and dietary routes of exposure on the effects of Benzo(a)pyrene on biotransformation pathways in Nile tilapia (Oreochromis niloticus)

BaP is one of the most studied PAH, due to its ubiquitous presence in aquatic environments and toxicity to aquatic organisms. The main goal of this study was to assess BaP effects in Nile Tilapia after waterborne and dietary exposures, through the evaluation of EROD and GST activities in liver, gills and intestine, and BaP metabolites in bile; and also to evaluate the usefulness of these commonly used biomarkers after two different routes of exposure. Waterborne exposure to BaP led to a significant induction of EROD in all tissues analyzed (644%, 1640% and 2880% in relation to solvent in liver, gill and intestine respectively) while in dietary exposures EROD was induced only in intestine (3143%) after exposure to high BaP concentrations. GST activities with CDNB were slightly induced in liver (40%) and in gill (66%) after water exposure to BaP, and in intestine after dietary exposure to low BaP concentrations (182%). BaP metabolites in bile increased after both exposure routes, and were highly correlated with EROD activity after water exposure. In summary, this work has shown that the effects of BaP on biotransformation pathways depend on the route of exposure. Moreover, barrier tissues like gills and intestine also have an important role in the first-pass metabolism of BaP, reducing the amount of parent compound that reaches the liver to be metabolized. For that reason, EROD activity as a biomarker of exposure should also be applied in extrahepatic organs, like gills and intestine, in monitoring studies. Biliary BaP type metabolites are good reflectors of contamination levels under both exposure routes, while GST activity with CDNB as substrate, as a phase II enzyme, does not seem a reliable biomarker of exposure to BaP regardless the route of exposure.     

Biochemical responses in Salmo salar muscle following exposure to ethynylestradiol and tributyltin

This study investigated the effects of nominal concentrations of two endocrine-disrupting chemicals (EDCs) on fish physiology. The effects of the synthetic pharmaceutical estrogen ethynylestradiol (EE(2)) and the antifoulant tributyltin (TBT) were investigated in exposure studies with immature Atlantic salmon, Salmo salar. Fish were exposed for 7 days to waterborne EE(2), TBT, or a combination of both. The activities of lactate dehydrogenase (LDH), acetylcholinesterase (AchE) and glutathione S-transferase (GST), and lactate and glycogen content were determined in samples of fish muscle. Fish exposed to EE(2) for 3 days responded rapidly with increasing AchE and GST activities and increasing lactate content. These responses were limited to the first 3 days of exposure and had disappeared by day 7, indicating that the fish had adapted to EE(2) exposure. Compared to the controls, TBT increased AchE and LDH activity, inhibited GST activity and had no effect on lactate content. When mixed, the highest concentration of EE(2) increased the effect of TBT on lactate content. However, fish exposed to a lower concentration of EE(2) in combination with TBT had the lowest lactate content. Effects on AchE and LDH activities were smaller when TBT was combined with EE(2) compared to TBT alone. This suggests that TBT and EE(2) influence biochemical processes in fish muscle, acting on different organizational levels, by antagonistic and synergistic mechanisms.     

Accumulation and depuration of cyanobacterial toxin nodularin and biomarker responses in the mussel Mytilus edulis

Blue mussels (Mytilus edulis) were exposed to an extract made of natural cyanobacterial mixture containing toxic cyanobacterium Nodularia spumigena (70-110 microg nodularin l(-1), 24-h exposure followed by 144-h depuration period in clean water). Toxin concentration increased from initial 400 to 1100 mg kg(-1) after 24-h exposure, measured by liquid chromatography/mass spectrometry (LC/MS). Acetylcholinesterase activity (AChE), a biomarker of direct neurotoxic effects, showed inhibition after 12 and 24h exposure but returned to control level during the depuration period. Catalase (CAT) activity, an indicator of oxidative stress, showed significantly elevated levels in exposed mussels but only 72 h after the end of the exposure. No change in the activity of glutathione-S-transferase (GST) involved in conjugation reactions could be observed. A gradual yet incomplete elimination of nodularin (from 1100 to 600 mg kg(-1)) was observed during the depuration period, and the tissue levels were 30% lower in clean water after 24 h. The observed increase in oxidative stress indicated by elevated CAT activity is likely connected to detoxification reactions leading to the production of reactive oxygen species, including an apparent time lag in this specific enzymatic defence response. That no change in GST activity was observed suggests that this enzyme is not significantly involved in the detoxification process of nodularin-containing cyanobacterial extract in M. edulis.     

Evaluation of impact of PAH on a tropical fish, Oreochromis mossambicus using multiple biomarkers

Treatment of the widely occurring tropical cichlid, Oreochromis mossambicus with a pure polycyclic aromatic hydrocarbon (PAH), phenanthrene, induced a concentration-dependant formation of the enzyme, 7-ethoxyresorufin-O-deethylase (EROD). Concomittant increase (65-669%) in the activity of sorbitol dehydrogenase in the serum (SSDH) occurring with EROD induction denoted liver cell damage, which was more severe in fish exposed to lower concentrations (0.4-4 microg g-1) of the chemical. In O. mossambicus exposed to 25% refinery effluent liver damage associated with cell death was indicated by the twin analyses of SSDH and liver somatic index. Cell injury appeared to have occurred at low PAH concentrations due to inadequately induced phase II-related detoxification of metabolites. This was indicated by the nearly 33% higher activity of hepatic glutathione S-transferase (GST) in fish exposed to higher PAH concentrations as compared to low-exposure animals. Tilapia such as O. mossambicus were found to be eminently suited for biomonitoring in tropical coastal waters. A combination of EROD and serum sorbitol dehydrogenase activity measurements serves as an excellent tool for biomonitoring sublethal effects of PAH pollution in fish.     

Anguilla anguilla L. oxidative stress biomarkers: an in situ study of freshwater wetland ecosystem (Pateira de Fermentelos, Portugal)

Pateira de Fermentelos (PF) is a natural freshwater wetland in the central region of Portugal. In the last decade, the introduction of agricultural chemicals, heavy metals, domestic wastes, as well as eutrophication and incorrect utility of resources resulted in an increased water pollution. The present research work was carried out to check the various oxidative stress biomarker responses in European eel (Anguilla anguilla L.) gill, kidney and liver due to this complex water pollution. Eels were caged and plunged at five different PF sites (A-E) for 48h. A reference site (R) was also selected at the river spring where no industrial contamination should be detected. Lipid peroxidation (LPO), catalase (CAT), glutathione peroxidase (GPX), glutathione S-transferase (GST) and reduced glutathione (GSH) were the oxidative stress biomarkers studied. In gill, site A exposure induced a significant GST activity increase and site B exposure induced CAT activity increase when compared to R. Site C exposure showed a significant CAT and GPX activity increase. Data concerning site D exposure were not determined due to cage disappearance. Site E exposure displayed a significant CAT and GST activity increase. In kidney, site A exposure induced a significant CAT and GPX decrease as well as a GST increase. Site B exposure showed a significant decrease in GPX activity and GSH content. However, site C exposure demonstrated a significant increase in CAT and a decrease in GPX. Site E exposure showed a significant decrease in GPX and increase in GST. In liver, site A exposure showed a significant GST activity decrease as well as GSH content increase. Site B exposure showed a significant CAT, GST and LPO decrease. Site C exposure showed only GST activity decrease, while site E exposure induced a significant increase in GPX. These investigation findings provide a rational use of oxidative stress biomarkers in freshwater ecosystem pollution biomonitoring using caged fish, and the first attempt reported in Portugal as a study of this particular watercourse under the previous conditions. The presence of pollutants in the PF water was denunciated even without a clear relation to the main pollution source distance. The organ specificity was evident for each parameter but without a clear pattern.     

Atrazine promotes biochemical changes and DNA damage in a Neotropical fish species

The effects of Atrazine, an herbicide used worldwide and considered as a potential contaminant in aquatic environments, were assessed on the Neotropical fish Prochilodus lineatus acutely (24 and 48 h) exposed to 2 or 10 μg L⁻¹ of atrazine by using a set of biochemical and genetic biomarkers. The following parameters were measured in the liver: activity of the biotransformation enzymes ethoxyresorufin-O-deethylase (EROD) and glutathione S transferase (GST), antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), content of reduced glutathione (GSH), generation of reactive oxygen species (ROS) and occurrence of lipid peroxidation (LPO); in brain and muscle the activity of acetylcholinesterase (AChE) and DNA damage (comet assay) on erythrocytes, gills and liver cells. A general decreasing trend on the biotransformation and antioxidant enzymes was observed in the liver of P. lineatus exposed to atrazine; except for GR, all the other antioxidant enzymes (SOD, CAT and GPx) and biotransformation enzymes (EROD and GST) showed inhibited activity. Changes in muscle or brain AChE were not detected. DNA damage was observed in the different cell types of fish exposed to the herbicide, and it was probably not from oxidative origin, since no increase in ROS generation and LPO was detected in the liver. These results show that atrazine behaves as enzyme inhibitor, impairing hepatic metabolism, and produces genotoxic damage to different cell types of P. lineatus.     

Sublethal responses in caged organisms exposed to sediments affected by oil spills

This study was performed to determine sublethal responses of two invertebrate species by using field deployments in areas affected by oil spills, which are acute in the Galician Coast (NNW, Spain) and chronic in the Bay of Algeciras (SSW, Spain). The organisms employed were the crab Carcinus maenas and the clam Ruditapes philippinarum, and during 28 days the animals were exposed to contaminated sediments in cages under field conditions. Different biomarkers of exposure were determined after a 28-day period exposure: ethoxyresorufin O-deethylase (EROD), phase I detoxification enzyme, glutathione-S-transferase (GST) phase II detoxification enzyme but also implicated in oxidative stress events, glutathione peroxidase (GPX) and glutathione reductase (GR), both antioxidant enzymes. In addition, histopathological effects in target tissues of the deployed organisms were evaluated. Biomarker measurements were linked with the concentration of chemicals in the sediments in order to elucidate the type, source and bioavailability of contaminants that produce adverse effects in the bioindicator species. Results obtained in this study have shown how the application of the selected battery of biomarkers under field bioassays allows for the identification of alternative sources of stress that are not observable in laboratory experiments.     

Effects of hexachlorobenzene on antioxidant status of liver and brain of common carp (Cyprinus carpio)

Hexachlorobenzene (HCB)-induced oxidative damages have been published in rats while the effects have not yet been reported in fishes. Juvenile common carps (Cyprinus carpio) were exposed to waterborne HCB from 2 to 200 microg l-1 for 5, 10 or 20 days. Liver and brain were analyzed for various parameters of oxidative stress. There were no significant changes of glutathione (GSH) content and superoxide dismutase (SOD) activity in liver after 5 or 10 days exposure, whereas obvious drops were observed at higher concentrations after 20 days exposure. Significant decreases of GSH content and SOD activity in brain were found during all the exposure days. In brain, HCB also significantly elevated the contents of reactive oxygen species (ROS), thiobarbituric acid- reactive substances (TBARS, as an indicator of lipid peroxidation products), glutathione disulfide (GSSG), and activities of nitric oxide synthase (NOS), glutathione peroxidase (GPx), and glutathione reductase (GR), and inhibited activities of acetylcholinesterase (AchE) and glutathione S-transferase (GST). The results clearly demonstrated that environmentally possible level of HCB could result in oxidative stress in fish and brain was a sensitive target organ of HCB toxicity.     

Effects of prochloraz and malathion on the red-legged partridge: a semi-natural field study

A semi-natural field study was carried out to assess the likelihood of a potentiation of toxicity between the ergosterol biosynthesis inhibiting (EBI) fungicide, prochloraz, and the organophosphorus (OP) insecticide, malathion, in the red-legged partridge (Alectoris rufa). Groups of partridges kept in four large grassland enclosures were exposed to either prochloraz-treated or control wheat for 7 days after which two of the enclosures were sprayed with malathion whilst the remaining two were sham-sprayed. Cytochrome P-450, aldrin epoxidase and 7-ethoxyresorufin-O-deethylase (EROD) activities were found to be significantly higher in the group exposed to prochloraz alone compared to controls, suggesting that induction of the hepatic microsomal monooxygenase system had occurred by ingestion of prochloraz-treated wheat. However, the level of induction produced was not sufficient to cause a potentiation of malathion toxicity. There was evidence for induction of several forms of P-450 recognised by antibodies raised against 1A1, 2C6 and 4A1 in the prochloraz-exposed partridges.     

Methyl parathion toxicity to and removal efficiency by Typha latifolia in water and artificial sediments

Methyl parathion (MeP) is a very hazardous pesticide freely used in agriculture in Mexico. This pesticide and others, arriving through different processes, exert significant effects on water quality with serious consequences for environmental and human health. This study evaluates the removal efficiency of common cattail Typha latifolia L. on MeP in water and artificial sediments. The effects of the pesticide on this macrophyte after 10 days of exposure were determined using a concentration range of 0-200 mg l(-1), 198.1+/-1.79 g average biomass, pH 7.0, 18-22 degrees C temperature and natural daylight/darkness periods, using chlorophyll production as a biomarker. Removal kinetics were conducted under similar conditions on days 0, 3, 7, 9, 11 and 14 of exposure, using 6 mg l(-1) in each system. Pesticide concentration, chlorophyll content and glutathione S-transferase (GST) activity were quantified. Results show a high removal efficiency of cattails on MeP in water and sediments relative to controls. An increase in GST activity and a decline in chlorophyll content in the test systems were not significantly different relative to controls. Cattails may thus be a good candidate for development of a phytoremediation system for MeP-contaminated water and artificial sediments.     

Oxidative stress and hepatotoxicity in the frog,Rana chensinensis,when exposed to low doses of trichlorfon

Trichlorfon is an organophosphate insecticide that is widely used in aquaculture and agriculture against parasitic infestations and has caused aquatic toxicity to non-target organisms. To evaluate the effects of low doses of trichlorfon on the oxidative stress and hepatotoxicity in amphibians, Chinese brown frogs (Rana chensinensis) were exposed to trichlorfon at concentrations of 0, 0.01, 0.1, and 1.0 mg/L for 2 and 4 weeks. Then, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and the content of malondialdehyde (MDA) in hepatic tissue were examined to evaluate the effects of oxidative stress and lipid peroxidation. The histopathological alternations to the liver were observed through light and transmission electron microscopy (TEM). The results showed that SOD and CAT activities were increased in the livers of frogs exposed to various concentrations of trichlorfon. The GST activity showed no significant changes at any concentration after 2 weeks of exposure, whereas there was an initial increase after exposure to 0.1 mg/L of trichlorfon at 4 weeks. The content of MDA revealed a significant decrease after exposure. Histopathological and ultrastructural studies showed that trichlorfon induced hyalinization, vacuolation, nucleus necrosis, and cellular swelling in hepatocytes. These results suggest that low doses of trichlorfon could induce oxidative stress, lipid peroxidation, and hepatic lesions in frogs, which shows that even lower, non-lethal doses of trichlorfon are potentially toxic to amphibians.     

Biomarker responsiveness in different tissues of caged Ruditapes philippinarum and its use within an integrated sediment quality assessment

Biomarkers comprising activities of biotransformation enzymes (ethoxyresorufin-O-deethylase -EROD-, dibenzylfluorescein dealkylase -DBF-, glutathione S-transferase -GST), antioxidant enzymes (glutathione reductase -GR- and glutathione peroxidase -GPX), lipid peroxidation -LPO- and DNA strand breaks were analyzed in the clam Ruditapes philippinarum caged at Cádiz Bay, Santander Bay and Las Palmas de Gran Canaria (LPGC) Port (Spain). Sediments were characterized. Digestive gland was the most sensitive tissue to sediment contamination. In Cádiz Bay, changes in LPO regarding day 0 were related with metals. In LPGC Port, DBF, EROD, and GST activity responses suggested the presence of undetermined contaminants which might have led to DNA damage. In Santander Bay, PAHs were related with EROD activity, organic and metal contamination was found to be associated with GR and GST activities and DNA damage presented significant (p < 0.05) induction. R. philippinarum was sensitive to sediment contamination at biochemical level. Biomarkers allowed chemical exposure and sediment quality assessment.     

Effects of long-term alachlor exposure on hepatic antioxidant defense and detoxifying enzyme activities in crucian carp (Carassius auratus)

Alachlor has been widely used in agriculture all over the world. It is suggested that it may be a carcinogen and also an environmental estrogen. In this paper, the physiological and biochemical perturbations of crucian carp (Carassius auratus) exposed to alachlor at different concentrations over 60 days were investigated. The gonadosomatic index (GSI) and hepatosomatic index (HSI) were measured. The activity of hepatic antioxidant defense and detoxifying enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) and the content of glutathione (GSH) were determined and compared with the control group. The result showed that GSI and HSI decreased significantly (P<0.05) in almost all treatments. The activities of SOD, CAT and GST were induced continuously (P<0.05), while the content of reduced glutathione (GSH) was inhibited on the whole. These changes reflect that the antioxidant systems of the tested fishes were affected. The possible defense mechanistic implications about the changes were thus discussed. Furthermore, hepatic SOD and GST were sensitive to alachlor at low concentration, indicating that they might be potential biomarkers in early detection of alachlor contamination in aquatic ecosystems.     

Accumulation pattern and biotransformation enzyme induction in rainbow trout embryos exposed to sublethal aqueous concentrations of 3,3',4,4'-tetrachlorobiphenyl

Accumulation pattern of 3,3',4,4'-tetrachlorobiphenyl (PCB 77) and the exposure time needed to activate the monooxygenase (EROD) and conjugation (GST) enzyme systems of fish at the advanced embryonic stage were studied. Eyed stage embryos of rainbow trout (Oncorhynchus mykiss) were exposed to sublethal doses (0, 1, 10, and 100 micrograms/l) of PCB 77. Results indicated direct accumulation of the chemical into the eggs, but the exposure time was not long enough for PCB 77 to reach constant steady state. However, at the two lowest test concentrations (1 microgram/l and 10 micrograms/l) a temporary plateau at chemical accumulation was reached at the third exposure day (185 and 1221 ng PCB/g egg w.w). At the highest concentration (100 micrograms/l) the decrease in the accumulation rate was already evident after the first day (2182 ng PCB/g egg w.w). The chemical uptake increased again at day 7 in all the exposure groups. That event could have been caused by the increased metabolic rate of the embryos in preparation for the upcoming hatching event. The microsomal CYP1A monooxygenase system (EROD) was shown to be a sensitive indicator of embryonic exposure, being induced at the low (1 microgram/l, 182 ng/g egg) PCB concentration and after a short (3 day) exposure time. The conjugation enzyme system (GST) was shown to be functioning already at the advanced embryonic stage, although no response to the studied chemical stress was detected.     

Microsomal monooxygenase activity in Tilapia (Oreochromis mossambicus) exposed to a bleached kraft mill effluent using different exposure systems.

Bleached kraft pulp and paper mill effluents (BKMEs) are known to have adverse effects on aquatic organisms. One of the effects of BKMEs is its ability to induce cytochrome P4501A activity in exposed fish. 7-Ethoxyresorufin O-deethylase (EROD) activity is the most common biomarker used to measure the mixed-function monooxygenase activity. In this study, Tilapia were exposed to BKMEs using different exposure systems and their hepatic EROD activity, as well as liver/somatic index (LSI), were determined. In the Phase I study, Tilapia treated with betaNF and a whole (100%) BKME using a static, non-renewal system exhibited statistically significant EROD induction, but LSI values were not altered. In the Phase II study, fish were either caged in the mill's fishpond with the whole effluent passing through or cultured in tanks receiving 100% of the BKME continuously using a flow-through system in the laboratory. Their EROD activities were then compared with the non-exposed fish (control). The EROD activities in both groups of fish were elevated significantly with the greatest induction being observed in the field-exposed group. The LSI values in all of the field-exposed fish were significantly greater than the control Tilapia. The EROD assay was sensitive in detecting biological changes in fish exposed to the BKME. Further studies are warranted to better understand the impacts of BKMEs on aquatic organisms in Taiwan.     

Response of multixenobiotic defence mechanism in Dreissena polymorpha exposed to environmental stress

Multixenobiotic defence mechanism (MXDM) has been recently described as a first line defence system in many aquatic organisms and it has raised attention for its potential as an early biomarker of exposure to environmental stress. In order to investigate the relevance of this biomarker in the freshwater zebra mussel Dreissena polymorpha, we examined its response to exposure and to cessation of exposure. For this purpose, depurated zebra mussels were transplanted to natural rivers or exposed to medium experimentally contaminated with fuel oil 2 in the laboratory. After the exposure period, mussels were submitted to depuration in clean water in the laboratory. The level of MXDM activity was assessed by the efflux method that allows a simple and fast measure of the rate of rhodamine B efflux. MXDM activity was induced following exposure and induction appears earlier after laboratory exposure (3 days) than after in the field exposure (3-14 days). The level of MXDM response could not be directly correlated to exposure dose in the laboratory and to isolated physicochemical parameters or AOX levels in the field. The behaviour of MXDM activity seems to reflect the occurrence of exposure to a global stress rather than to specific compounds. Our results highlighted the possibility to monitor MXDM response in the freshwater zebra mussel. In spite of the difficulty to interpret responses resulting from the lack of specificity of response, multixenobiotic defence mechanism displays qualities to represent a biomarker of general stress in freshwater ecosystems.