Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.     

Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea

The purpose of this study was to compare enzymatic biomarker activities in fish caged at two sites, Masan Bay (contaminated) and Haeguemgang (reference). In the present study, ethoxyresorufin O-deethylase (EROD), brain acetyl cholinesterase (bAChE), muscle acetyl cholinesterase (mAChE) and butyryl cholinesterase (mBChE) in caged rockfish (Sebastes schlegeli) were measured 0, 1, 3, 7, 14, 21 and 30 days after caging. The level of CYP1A mRNA and Protein expression was induced higher in Masan Bay at 1, 3, 7, 14 and 30 days after caging. EROD activity in the caged fish was significantly higher in Masan Bay than in Haeguemgang 3 and 7 days after caging, but not at 14 and 30 days after caging. bAChE activity was significantly inhibited at 7 and 14 days after caging in Masan Bay. However, mBChE activity was not significantly inhibited during the experiment. Taken together, the data suggest that the caged fish were exposed, at least transiently, to CYP1A inducers and ChE inhibitors, which is consistent with our previous observations.     

Fish biological effect monitoring of chemical stressors using a generalized linear model in South Sea,Korea

To evaluate the health status at six different study areas, we used the generalized linear model approach with selected biochemical markers in resident fish from uncontaminated and contaminated sites. We also confirmed the independence between the biochemical indices and the morphometric indices including the hepato-somatic index (HSI), gonado-somatic index (GSI), and condition factor (CF) in fish from the sampling areas. The effect of area on the presence of biotransformation markers (ethoxyresorufin-O-deethylase activity; EROD) was significantly high in Masan Bay. The area with the greatest effect on acetylcholinesterase (AChE) activity was Jindong Bay, while there was no significant effect of GSI, HSI, CF, and sex in the EROD model and HSI, CF and sex in the AChE model. These results clarify that fish from Masan, Gwangyang and Jindong Bay were affected by pollutant stress, and the analysis of sensitive biochemical responses allowed for an improved interpretation of the results.     

Hepatic 7-ethoxyresorufin O-deethylase Activity in eel (Anguilla Anguilla) from The Thames Estuary and Comparisons with other United Kingdom Estuaries

Hepatic microsomal 7-ethoxyresorufin O-deethylase (EROD) activities (indicative of exposure to polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs)) were measured in eel Anguilla anguilla from the Thames Estuary. Fish were collected from up to 13 sites during November 1997, May and August 1998 and October 1999. Throughout this period no clear seasonal variation could be identified at every site along the Thames. However, during the summer months, fish sampled from sites in the middle to the upper estuary (Woolwich, Greenhithe and West Thurrock) reported up to 3-fold higher EROD activities compared to sites either at the upper reaches (Richmond and Brentford) at the same time of the year, or fish sampled in winter, along the entire length of the estuary. A laboratory exposure experiment demonstrated a 3-fold elevation of EROD activity 2 days after injection with β-naphthoflavone (β-NF). However, higher levels of activity could be determined in fish sampled from the Weston canal near the Merseyside. The lowest levels of A. anguilla EROD activity were observed in fish sampled from the upper reaches of the River Tamar, Devon, and were comparable to activities determined in fish from the Wear and Humber estuaries. A. anguilla sampled along the Thames, Tyne and Tees estuaries reported between 2.5- and 7-fold higher EROD activities compared to fish collected from the Tamar. These results indicate that a low to moderate induction of A. anguilla CYP1A had occurred (indicative of low to moderate exposure to PAHs and planar PCBs) in fish collected from the Thames, Tyne, Wear, Tees, Humber and Tamar estuaries. However, the highest level of EROD activity was observed in fish from the Weston Canal (Merseyside).     

Hepatic 7-ethoxyresorufin O-deethylase activity in eel (Anguilla anguilla) from the Thames Estuary and comparisons with other United Kingdom Estuaries

Hepatic microsomal 7-ethoxyresorufin O-deethylase (EROD) activities (indicative of exposure to polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs)) were measured in eel Anguilla anguilla from the Thames Estuary. Fish were collected from up to 13 sites during November 1997, May and August 1998 and October 1999. Throughout this period no clear seasonal variation could be identified at every site along the Thames. However, during the summer months, fish sampled from sites in the middle to the upper estuary (Woolwich, Greenhithe and West Thurrock) reported up to 3-fold higher EROD activities compared to sites either at the upper reaches (Richmond and Brentford) at the same time of the year, or fish sampled in winter, along the entire length of the estuary. A laboratory exposure experiment demonstrated a 3-fold elevation of EROD activity 2 days after injection with β-naphthoflavone (β-NF). However, higher levels of activity could be determined in fish sampled from the Weston canal near the Merseyside. The lowest levels of A. anguilla EROD activity were observed in fish sampled from the upper reaches of the River Tamar, Devon, and were comparable to activities determined in fish from the Wear and Humber estuaries. A. anguilla sampled along the Thames, Tyne and Tees estuaries reported between 2.5- and 7-fold higher EROD activities compared to fish collected from the Tamar. These results indicate that a low to moderate induction of A. anguilla CYP1A had occurred (indicative of low to moderate exposure to PAHs and planar PCBs) in fish collected from the Thames, Tyne, Wear, Tees, Humber and Tamar estuaries. However, the highest level of EROD activity was observed in fish from the Weston Canal (Merseyside).     

Benzo[a]pyrene modulates the biotransformation,DNA damage and cortisol level of red sea bream challenged with lipopolysaccharide

In animals, biotransformation and the immune system interact with each other, however, knowledge of the toxic mechanism of benzo[a]pyrene (BaP) on these two systems is not well known. The present study investigated the toxic effects of BaP on the biotransformation system, cortisol level and DNA integrity of red sea bream (Pagrus major). The results showed that cortisol level was induced under the challenge of lipopolysaccharide (LPS). Short-term exposure (96 h) of BaP at environmental concentration significantly increased the cortisol level, hepatic EROD activity and CYP1A1 mRNA expression. When P. major was exposed to BaP for 14 d followed by LPS challenge this increased the cortisol level, EROD activity and hepatic DNA damage except CYP1A1 mRNA expression. Combined with our previous data, which showed that BaP exposure can modulate the immunologic response in P. major challenged with LPS, a hypothetical adverse outcome pathway of BaP on fish was suggested.     

The role of cytochrome P4501A activity inhibition in three- to five-ringed polycyclic aromatic hydrocarbons embryotoxicity of marine medaka (Oryzias melastigma)

The mode of action of PAHs that causes fish developmental malformations is unclear. The embryotoxicity of marine medaka (Oryzias melastigma) was investigated after individual exposure to three- to five-ring PAHs Phe, Py, and BaP or co-exposure with α-ANF for 18 days. We found that the relationships between EROD induction and developmental deformities of embryos showed a various pattern under different exposure scenarios of Phe, Py, and BaP, which suggested possibly different modes of action in determining the developmental toxicities. As for co-exposure scenarios of each PAH combined with ANF, it showed potentially synergistic effects. The inhibited CYP1A mediated enzyme activity by ANF after co-exposure did not effectively alleviate developmental toxicity of embryo. It showed potentially synergistic effects after co-exposure of marine fish embryos to CYP1A inhibitors and PAH-type CYP1A inducers. Heart deformities in the early life stages of marine medaka were recommended as a biomarker for indicating the extent of PAH pollution.     

In situ biomonitoring of caged, juvenile Chinook salmon (Oncorhynchus tshawytscha) in the Lower Duwamish Waterway

Contaminated sediments may have wide-ranging impacts on human and ecological health. A series of in situ caged exposure studies using juvenile Chinook salmon was conducted in the Lower Duwamish Waterway (LDW). Chemical analysis of sediment, water, and fish tissue were completed. Additionally, in 2004, DNA adducts in hepatic and gill tissues were measured. Gills contained significantly higher DNA adducts at stations B2 and B4, prompting further analysis of gills in 2006 and 2007. Fluorescent aromatic compounds (FACs) in bile, and CYP1A1 in hepatic tissue were also measured during 2006 and 2007, respectively. FACs in field-caged fish were comparable or significantly higher than wild-caught fish LDW fish and significantly higher than lab fish after only 8-10 days, demonstrating the equivalency of exposure to that of migrating salmon. Furthermore, selected biomarkers appear to be capable of detecting spikes in contamination between sampling years, emphasizing the need for multiple year data collection.     

Comparative study of 17 β-estradiol on endocrine disruption and biotransformation in fingerlings and juveniles of Japanese sea bass Lateolabrax japonicus

Estrogenic contaminants in the aquatic environment are associated with endocrine disruption and feminization in fish. The effects of endocrine disrupting chemicals (EDCs) on fish have been well documented. However, very few studies have focused on 17 β-estradiol (E2) and its effects on endocrine system and biotransformation in a single prolonged exposure. This study investigated changes in the levels of serotonin (5-hydroxytryptamine) and acetyl choline esterase (AchE) in brain, cortisol in plasma and Ethoxyresorufin-O-deethylase (EROD) activity in gill of two different size groups (fingerlings and juveniles) of Japanese sea bass (Lateolabrax japonicus) upon exposure to two sub-lethal concentrations (200 and 2000 ng L⁻¹) of E2 for 30 d. The results indicate that cortisol level and EROD activity significantly increased in both groups, whereas serotonin level increased in juveniles and decreased in fingerlings due to E2 exposure. The correlation analysis revealed that E2 significantly affected the endocrine and biotransformation systems in both age groups.     

Summary results from a pilot study conducted around an oil production platform on the Northwest Shelf of Australia

The Australian Institute of Marine Science (AIMS) conducted a pilot study around the Harriet A oil production platform on the Northwest Shelf of Australia. We evaluated hepatic ethoxyresorufin-O-deethylase (EROD) activity, fluorescent aromatic compounds (FACs) in bile and immunodetection of CYP1A-like proteins in two Australian tropical fish species, Gold-Spotted Trevally (Carangoides fulvoguttatus) and Bar-Cheeked Coral Trout (Plectropomus maculatus) to assess exposure to petroleum hydrocarbons associated with produced formation water (PFW). Additionally, the incidence of hydrocarbon-degrading bacteria isolated from the liver and bile of all fish captured was examined. Low EROD activity was found in both species, with EROD activity in C. fulvoguttatus showing significant site differences. FACs and CYP1A protein levels in C. fulvoguttatus showed a clear trend in hydrocarbon exposure consistent with hydrocarbon chemistry data: Harriet A>Harriet C>reference site. P. maculatus showed elevated levels of FACs at Harriet A as compared to the reference site and demonstrated detectable levels of CYP1A-like proteins at these two sites. Hydrocarbon-degrading bacteria were found in the liver and bile of both species, yet there was no correlation by sites. Our results demonstrate that C. fulvoguttatus and P. maculatus have potential as indicator species for assessing the effects from exposure to petroleum hydrocarbons. Both FACs and CYP1A are providing warning signs that there is potential for biological effects on fish populations exposed to PFW around the Harriet A production platform.     

EROD activity, serum SDH and PAH biliary metabolites in sand flathead (Platycephalus bassensis) collected in Port Phillip Bay, Australia

In order to evaluate the health status of fish inhabiting Port Phillip Bay, Australia, southern sand flathead (Platycephalus hassensis, N = 133) were collected at six stations throughout the Bay. Fish had a similar serum sorbitol dehydrogenase (SDH) activity level (p = 0.12), indicating that they were not experiencing hepatocellular injuries. Ethoxyresorufin-O-deethylase (EROD) activity was generally lower in the non-urbanized and non-industrialized southern part of the Bay. The highest EROD activity was observed in Hobson Bay, the closest station from Melbourne city. Naphthalene-type biliary metabolites were also highest in Hobson Bay with intermediate levels found in Corio Bay where refineries are present. An opposite trend was observed with the pyrene-type bile metabolites, the highest levels being observed in Corio Bay while intermediate levels were found in Hobson Bay. The ratio of naphthalene-type to benzo(a)pyrene (B(a)P)-type metabolites indicate that relatively to other sites sampled in Port Phillip Bay, Corio Bay is subjected to enriched petroleum hydrocarbons of pyrolytic origin. Temporal trends indicate that the availability of xenobiotics to fish remained unchanged over the 1990s.     

Antioxidant and biotransformation responses in Liza aurata under environmental mercury exposure - relationship with mercury accumulation and implications for public health

This study was carried out in the Laranjo basin (Ria de Aveiro, Portugal), an area impacted by mercury discharges. Liza aurata oxidative stress and biotransformation responses were assessed in the liver and related to total mercury (Hg_t) concentrations. A seasonal fish survey revealed a sporadic increase in total glutathione (GSH_t) and elevated muscle Hg_t levels, although Hg levels did not exceed the EU regulatory limit. As a complement study, fish were caged for three days both close to the bottom and on the water surface at three locations, and displayed higher Hg_t levels accompanied by increased GSH_t content and catalase activity as well as EROD activity inhibition. The bottom group displayed higher hepatic Hg_t and GSH_t contents compared with the surface group. Globally, both wild and caged fish revealed that the liver accumulates higher Hg_t concentrations than muscle and, thus, better reflects environmental contamination levels. The absence of peroxidative damage in the liver can be attributed to effective detoxification and antioxidant defense.     

Biomarker responses in pelagic and benthic fish over 1 year following the Hebei Spirit oil spill (Taean, Korea)

After the Hebei Spirit oil spill incident (7th December, 2007) in the west coast of Korea, contamination of biliary PAH metabolite and hepatic biomarkers in a pelagic and a benthic fish was monitored for 1 year. Concentrations of 16 PAHs and alkylated PAHs in fish muscle were highest (22.0 ng/g d.w. for 16 PAHs and 284 ng/g d.w. for alkylated PAHs) at 5 days after the spill and then decreased rapidly to background levels at 11 months after the spill. Fish from the oiled site had elevated biliary PAH metabolite concentrations immediately after the spill; these declined steadily in both species, but were still above reference site concentrations 2 months after the spill. Oiled-site fish showed hepatic CYP 1A induction whose trend closely followed those of biliary PAH metabolite concentrations, implying continuous exposure to PAHs. Brain acetylcholinesterase activity was not related to oil exposure.     

Assessment of the effects of a marine urban outfall discharge on caged mussels using chemical and biomarker analysis

To assess the presence of endocrine disruptors in treated marine outfall discharges and their possible effects, mussels (Mytilus galloprovincialis) were caged in the environmental mixing zone of the outfall of the Santander sanitation system and in one control area. After 30, 60 and 90 days, samples were collected to perform chemical analyses (metals, anionic surfactants, alkylphenols, bisphenol A, phthalates and estrogenic hormones), biomarkers of general stress (lysosomal membrane stability-LMS, histopathology) and biomarkers of endocrine disruption (vitellogenin-like proteins and gonad index). There were no significant differences between outfall and control sites on contaminant levels, except for 4-tert-octylphenol which was higher in the outfall site. Bacteriological counts were higher in the outfall area. No relevant differences in biomarkers were detected between treated and control mussels. A significant reduction in LMS occurred in both groups after 90 days caging, indicating a stress situation possibly related to caging or to post-spawning reproductive state.     

Cytochrome P4501A biomarker indication of the timeline of chronic exposure of Barrow's goldeneyes to residual Exxon Valdez oil

We examined hepatic EROD activity, as an indicator of CYP1A induction, in Barrow’s goldeneyes captured in areas oiled during the 1989 Exxon Valdez spill and those from nearby unoiled areas. We found that average EROD activity differed between areas during 2005, although the magnitude of the difference was reduced relative to a previous study from 1996/1997, and we found that areas did not differ by 2009. Similarly, we found that the proportion of individuals captured from oiled areas with elevated EROD activity (?2 times unoiled average) declined from 41% in winter 1996/1997 to 10% in 2005 and 15% in 2009. This work adds to a body of literature describing the timelines over which vertebrates were exposed to residual Exxon Valdez oil and indicates that, for Barrow’s goldeneyes in Prince William Sound, exposure persisted for many years with evidence of substantially reduced exposure by 2 decades after the spill.     

Validation of Arenicola marina in field toxicity bioassays using benthic cages: biomarkers as tools for assessing sediment quality

Sediment toxicity assessments using caged organisms present advantages over using laboratory and native community studies. The use of caged Arenicola marina in sediment toxicity assessments was evaluated. Lugworms were exposed in situ to sediments from coastal and port areas in Spain for seven days, and the activities of the biotransformation enzymes ethoxyresorufin O-deethylase, dibenzylfluorescein dealkylase and glutathione S-transferase, the activities of the antioxidant enzymes glutathione reductase and glutathione peroxidase and lipid peroxidation were then analyzed as biomarkers. Biomarker results and sediment physicochemical data were integrated. Cádiz Bay (SW Spain) sediments presented metal contamination that was not linked to a biochemical response. In LPGC Port (SW Spain), Pb contamination exhibited a moderate toxic potential, while PAHs, and presumably pharmaceuticals, provoked biochemical responses that efficiently prevented lipid peroxidation. In Santander Bay (N Spain), exposure to PAHs and, presumably, pharmaceuticals induced biomarker responses, but lipid peroxidation occurred nevertheless. These results indicated that caged A. marina were effective for the assessment of sediment quality and that the selected biomarkers were sufficiently sensitive to identify chemical exposure and toxicity.     

Effects of Scotian shelf natural gas condensate on the mummichog

The 96h LC₅₀ of the water soluble fraction (WSF) of natural gas condensate from the Sable Is., N.S. area to Fundulus heteroclitus was between 4 and 5 μg · ml⁻¹ naphthalene equivalents (initial concentration). Behavioural responses in the fish to sub-lethal concentrations of WSF were similar to those recorded for crude and fuel oils. Induction of the hepatic mixed function oxidase enzyme enthoxyresorufin O-de-ethylase (EROD) in sexually immature fish occurred at WSF concentrations below the LC₅₀, but during the prespawning period no induction occurred. EROD activity in unexposed fish showed seasonal variation consistent with differences in its inducibility.     

A multibiomarker approach in barramundi (Lates calcarifer) to measure exposure to contaminants in estuaries of tropical North Queensland

A suite of biomarkers were measured in barramundi (Lates calcarifer) from five North Queensland estuaries along a perceived pollution gradient. The biomarkers selected were 7-ethoxyresorufin-O-deethylase (EROD), cytochrome P450, fluorescent aromatic compounds (FACs), DNA integrity, RNA:DNA ratio, cholinesterase activity (ChE), condition factor and hepatosomatic index. The resulting database was subjected to uni- and multi-variate analyses in order to assess the most suitable biomarkers to assess pollution in North Queensland estuaries and to classify the environmental quality of the sites. Principal components analysis (PCA) on the biochemical markers revealed that EROD, EROD/P450, DNA damage and to a lesser extent ChE and FACs were found to be responsive to contaminants in the environment while cytochrome P450, condition factor and the hepatosomatic index were found to be less responsive biomarkers. This study has demonstrated the utility of applying a multibiomarker approach in conjunction with traditional analysis of contaminants in providing valuable information in environmental risk assessment.     

Impacts of cyclic hypoxia on reproductive and gene expression patterns in the grass shrimp: field versus laboratory comparison

Identification of organisms able to act as biological indicators of hypoxia exposure is important given the increasing frequency of hypoxic events worldwide. Grass shrimp, Palaemonetes pugio, are ubiquitous estuarine residents in the southeastern US, where they commonly experience cyclic hypoxia. We hypothesized that reproductive and gene expression patterns of grass shrimp in cyclic hypoxic field sites relative to normoxic sites would be similar to previous results from cyclic hypoxia laboratory experiments showing differential up- and down-regulation of hypoxia-responsive genes. There were few differences in gene expression of indigenous shrimp collected during summer from two Gulf of Mexico estuarine systems [East Bay, FL (EB) and Weeks Bay, AL (WB)], although down-regulation of the gene coding for vitellogenin-1 corresponded with a decreased relative fecundity and fewer ovigerous females at cyclic hypoxic field sites, suggesting an overall impact on reproduction. The gene expression profiles of grass shrimp caged for 7 days in field sites differed by estuary, but few hypoxia-responsive genes identified in laboratory studies were differentially expressed in field shrimp. Overall, genes coding for protein synthesis, protein degradation, carbohydrate and lipid metabolism and electron transport were mostly up-regulated in EB caged shrimp but generally down-regulated in WB caged shrimp and laboratory shrimp. Thus, caged grass shrimp from different bay systems exhibited profoundly different gene expression profiles. Such profiles may serve as sensitive bioindicators of differences in water quality, habitat quality or food resources among estuaries but are not effective as indicators of hypoxia exposure.