Glutathione S-tranferases and cytochrome P450 activities in Mytilus galloprovincialis from the South coast of Portugal: effect of abiotic factors

To assess the health of aquatic organisms, biomarkers that measure changes at the biochemical level have been used as effective early warning tools in ecological risk assessment. In order to develop an integrated risk assessment strategy for the south coast of Portugal, mussels Mytilus galloprovincialis were collected from six sites along the coast with different organic contaminant characteristics. Additionally, an active biomonitoring approach was followed by transplanting indigenous mussels from site 4 to 6 and vice versa (from site 6 to 4) for 28 days. PAHs and PCBs contents were measured and the associated responses of phase I and phase II detoxification mechanisms evaluated by measuring cytochrome P450 and GST activities. GST activity was also determined on different tissues (gills, digestive gland, foot, mantle and gonads) of M. galloprovincialis and the impact of abiotic parameters (temperature, salinity, pH, conductivity and dissolved oxygen) on the GST activity assessed. Results indicate that CYP 450 follow the same pattern of PAHs and a direct relationship exist between CYP 450 and PAH levels from the different sites. Although there is a decrease between GST and PAHs concentrations it was not significant. The majority of the GST activity was in the gills and the digestive gland (around of 75% of the activity measured in all tissues) followed in decreasing order by the mantle, gonads. An inverse relationship between GST activity and salinity was detected along with temperature although not significant. These two biomarkers respond to changes of these two groups of compounds and to salinity especially for GST. In conclusion CYP 450 in mussels gives a reliable response as biomarker for organic contaminants in risk assessment in the South Coast of Portugal.     

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.     

The changes of T-PAH levels and health status of mussels in Izmit bay (Turkey) after Marmara earthquake and subsequent refinery fire

As is well known, a powerful earthquake along the North Anatolian Fault struck the eastern part of the Marmara region on August 17, 1999. Izmit Bay, which is known as one of the most polluted sites of Turkey, was also affected by the quake and the subsequent refinery fire. The measurements performed just before and after the earthquake showed that T-PAH levels increased significantly after the event [Okay OS, Tolun L, Telli-Karako? F, Tüfek?i V, Tüfek?i H, Morko? E. Izmit Bay (Turkey) ecosystem after Marmara earthquake and subsequent refinery fire: the long-term data. Marine Pollution Bulletin 2001;42:361-9]. In the framework of ecotoxicological studies, the Bay ecosystem was continuously monitored for T-PAH levels in seawater, sediments and mussels (Mytilus galloprovincialis) to find out whether change occurred during the 2-year period following the earthquake. For that purpose, after the earthquake, the samples were collected six times between the period of September 1999 and March 2001 at coastal stations of the Bay situated away from the mouth of main discharges. The responses of the mussels were also measured by means of the lysosomal stability of the blood cells and feeding rate biomarker techniques at two different sites of the bay. Although the T-PAH levels in all matrices generally showed a decreasing trend, they were found to be still high especially at stations near the refinery. Both biomarker results showed that the health status of the mussels is very poor in the Bay ecosystem, based on the results obtained from the two sites monitored.     

Molecular biomarkers of energy metabolism in mussels under anthropogenic pollution of Peter the Great Bay, the Sea of Japan

The functional state of the indicator species, the Gray mussel Crenomytilus grayanus (Bivalvia), has been analyzed in five areas of Peter the Great Bay exposed to anthropogenic pollution. The following indices of the state of mussels have been used: molecular biomarkers of energy metabolism—Na⁺,K⁺-ATPase, Mg²⁺-ATPase, and total ATPase activity—as well as the level of lipid peroxidation (LPO) and glutathione concentration in the hepatopancreas, gills, and gonads of mussels. The activity of ATPases, LPO level, and glutathione concentration significantly change in mussels from polluted areas relative to those in mussels from a conventionally unpolluted area (a bay in the Far Eastern State Marine Reserve). The molecular biomarkers used in the study provide reliable information on animal metabolism in impact areas. With consideration of the data obtained, it is concluded that the state of mussels in polluted areas is impaired.     

A biological and geochemical integrated approach to assess the environmental quality of a coastal lagoon (Ravenna, Italy)

The assessment of the environmental quality of coastal lagoons is rather difficult as their intrinsic high variability limits the power of analytical chemistry and ecotoxicological approaches. Aim of the present work was to integrate geochemical and biological data which were simultaneously collected from the Pialassa Baiona lagoon (Ravenna, Italy), and evaluate the usefulness of a biomonitoring strategy based on sentinel organisms and biomarkers. Geochemical analysis on suspended, surface, and subsurface sediments were performed at five different sites of the lagoon; moreover the heavy metal bioaccumulation and a battery of biomarkers was evaluated in Mytilus galloprovincialis transplanted in the same sites. Heavy metals appeared to be diffused throughout the lagoon reaching higher levels in the southernmost sites, and mainly concentrated in suspended sediments. Lysosome membrane stability, neutral lipid and lipofuscin accumulation, and metallothionein levels in mussels indicated the development of a stress syndrome in animals exposed to the polluted sites.     

Induction of EROD activity in European eel (Anguilla anguilla) experimentally exposed to benzo[a]pyrene and beta-naphthoflavone

The induction of liver ethoxyresorufin O-deethylase (EROD) activity was investigated in the European eel, Anguilla anguilla, collected from a Mediterranean brackish environment and experimentally exposed to benzo[a]pyrene (B[a]P) and beta-naphthoflavone (BNF). Eels were injected intraperitoneally at increasing doses (0.1, 1, 10, and 50 mg/kg wet body weight) using corn oil as a carrier and sacrificed after 7 days. The main objectives of the present study are: (1). to assess of the sensitivity of EROD induction as a biomarker to polycyclic aromatic hydrocarbon (PAH) exposure; (2). to determine an EROD dose-response relationship of the contaminants used; and (3). to compare the efficiency of B[a]P and BNF as inducers of EROD activity. Results showed that both chemicals resulted in a dose-dependent EROD induction, but increases were not linear. EROD activity seemed to reach a plateau at the exposure of 10 mg/kg in both treatment groups; B[a]P was a more potent inducer than BNF was at the higher doses (10 and 50 mg/kg), while the opposite result was observed at the lower ones (0.1 and 1 mg/kg). The greatest induction occurred in eels treated with 10 mg/kg B[a]P, in which a 261-fold increase in EROD activity was observed. Results showed that EROD activity in A. anguilla is significantly induced by B[a]P and BNF exposure, responding to a wide range of concentrations of these contaminants. We infer that this tool may be suited as a diagnostic biomarker for biomonitoring PAHs pollution in Mediterranean brackish environments and further field research is suggested.     

Effects of pyrene on mussels in different experimental conditions

The toxicity of the polyaromatic hydrocarbon pyrene to Mytilus edulis and Mytilus galloprovincialis was investigated in experiments conducted in the United Kingdom for M. edulis and in Turkey for M. galloprovincialis. Experimental conditions of temperature and salinity were chosen to be appropriate to the ambient conditions in which the mussels typically live. The effect of different feeding regimes on pyrene bioaccumulation and toxicity was also investigated. Feeding rate and neutral red retention biomarker techniques were used for toxicity assessment. An experiment with M. edulis demonstrated that mussels exposed to pyrene accumulated increasing amounts of this compound throughout a 15-day exposure period and that accumulation increased in relation to exposure concentration and with increasing concentration of unicellular algal food material. However, in these experiments, which were protected from UV light, there was no clear relationship between pyrene concentration in tissues and feeding rate. A clear concentration-response relationship was observed between tissue concentration and neutral red retention for days 1 and 7 of the experiment, but this relationship was lost by day 15, with evidence of the cells recovering. A similar experiment was conducted with M. galloprovincialis at a single (high) pyrene concentration in darkness. The feeding rate of the exposed mussels was always lower than the feeding rate of the control mussels, although the difference was insignificant at 7-day exposure. A decreasing trend in health status of the mussels was indicated by the neutral red retention assay results after 7 and 15 days of exposure to pyrene. In a third experiment (with M. galloprovincialis), an illuminated algal chemostat system containing pyrene was inserted in the exposure system as the food source for the mussels. Both biomarker results showed conclusively that toxic effects correlated with pyrene bioaccumulation, though there was no clear evidence for light-enhanced toxicity. These experiments showed that experimental conditions such as salinity, temperature and feeding regime and perhaps the species of mussel used may influence whether pyrene is perceived as being a toxic molecule.     

Genotoxic and biochemical responses in caged eel (Anguilla anguilla L.) after short-term exposure to harbour waters

European eel (Anguilla anguilla L.) were caged and exposed in situ for 8 and 48 h to the Aveiro offward fishing harbour water (HW) and to clean seawater under laboratory conditions (Control). Eel liver biotransformation (Phase I) was measured as ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (P450) and glutathione S-transferase (GST) activity (Phase II). Genotoxic responses were determined as blood, liver and kidney DNA strand breaks as well as erythrocytic nuclear abnormalities (ENAs). HW failed to significant increase liver EROD, GST activities and ENA frequency. Nevertheless, P450 content was significantly increased after 8 and 48 h exposure. Genotoxicity measured as DNA integrity decrease was found in blood after 8 and 48 h exposure to HW, whereas in liver and kidney, it was observed after 48 h exposure to HW. Blood, kidney and liver genotoxicity may be due to the presence of polycyclic aromatic hydrocarbons (PAHs) which are genotoxic compounds and the main HW organic contaminants.     

Cellular alterations in Mytilus galloprovincialis (LMK) and Tapes philippinarum (Adams and Reeve, 1850) as biomarkers of environmental stress: field studies in the Lagoon of Venice (Italy)

A long-term biomonitoring study was carried out in the Lagoon of Venice (North-East Italy) with the aim of evaluating variations in biological responses to environmental stress in estuarine bivalves. Two different species, the mussel Mytilus galloprovincialis and the clam Tapes philippinarum, both widespread in the Lagoon, were studied in several sites. Two cellular biomarkers: lysosomal membrane stability in digestive cells and thickness of digestive epithelia, were evaluated in native organisms (on a seasonal basis), and in organisms which have been transferred from a reference site to several differently influenced ones. Results indicate that, to some extent, both test and organisms were able to highlight site-specific differences, but the effects of pollution were generally more easily detected by reduction in lysosomal stability than by reduction in digestive tubule epithelium. Further findings show that the inherent variability of a number of natural parameters, particularly in the reference sites, produced less effective results when biological responses in the reference organisms were compared with the polluted ones. The assessment of the two conditions was most valuable when they referred to the 25-75% range of values comprised within the seasonal medians, recorded respectively in control and polluted sites. Impaired from steady states were most effectively distinguished when the control values were medians from two reference locations. Lately, the overall results indicate that both biomarkers are more suitably deployed through the translocation approach, revealing it to be more sensitive than traditional biomonitoring, at least in the sense that it may overcome problems related to the adaptation of native organisms to sub lethal chronic pollution levels.     

Pollution biomarkers in two estuarine invertebrates,Nereis diversicolor and Scrobicularia plana,from a Marsh ecosystem in SW Spain

The polychaete worm Nereis diversicolor and the clam Scrobicularia plana were collected from several sites, affected by different types of contamination, in a littoral enclosure in the SW Spain (Caño Sancti-Petri and Rio San Pedro). N. diversicolor was present in 6 sampling sites whereas S. plana in 4 of them. The aim of our study was to relate several pollution biomarkers to chemical sources (metals and organic pollutants e.g. PCB, PAH) in these species, thereby confirming their adequacy as sentinels for this habitat. The biomarkers surveyed in the two invertebrates were the activities of the antioxidant enzyme catalase (CAT), the phase II detoxifying enzyme glutathione S-transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE). Metallothionein (MT) levels were measured as a biomarker of exposure to metals. The results suggested a different response in the two sediment-dwelling organisms, the sediment-eating polychaete and the water-filtering clam, probably as a consequence of different contamination exposures. The results also suggested that samples from the “Caño Sancti-Petri” were exposed to biologically active compounds that altered some of their biochemical responses. Of all the biomarkers tested, AChE was the most sensitive one and N. diversicolor the potentially most robust sentinel in this ecosystem. In this low to moderately polluted environment, the biochemical approach better reflected temporal trends than site-related differences although it was also able to detect punctual chemical insults.     

Targeted analytical toxicology: Simultaneous determination of 17α-ethynylestradiol and the estrogen-induced vitellogenin biomarker

In most toxicological studies, chemical concentrations and its biological/molecular response (biomarker) levels are often quantified separately. We report a method, named as Targeted Analytical Toxicology (TAT), for the simultaneous determination of levels of chemicals and protein biomarkers in organisms by UPLC/ESI–MS/MS on the basis of targeted proteomics. The TAT method is successfully applied to detect the concentrations of 17α-ethynylestradiol (EE₂) and EE₂-induced vitellogenin (a biomarker for estrogens) in male zebrafish exposed to EE₂. It is expected for TAT to have broad application in toxicology, especially in environmental toxicology, due to its high sensitivity, accuracy, reproducibility, high throughput, and the savings of time, cost and labor.     

Benthic community structure and biomarker responses of the clam Scrobicularia plana in a shallow tidal creek affected by fish farm effluents (Rio San Pedro,SW Spain)

The effects of solid organic wastes from a marine fish farm on sediments were tested using benthic community as ecological indicators and biomarkers in native clam (Scrobicularia plana) as biochemical indicators. The benthic fauna and clam samples were collected in the intertidal sediment in October 2010 from five sites of the Rio San Pedro (RSP) creek, following a gradient of contamination from the aquaculture effluent to the control site. Numbers of species, abundance, richness and Shannon diversity were the biodiversity indicators measured in benthic fauna. Morphological and reproduction status of clams were assessed using the condition factor and gonado-somatic index, respectively. Phase I and Phase II detoxification enzymatic activities (ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST)), antioxidant enzymatic activities (glutathione peroxidase (GPX), glutathione reductase (GR)) and oxidative stress parameters (Lipid Peroxidation (LPO) and DNA strand breaks) were measured in clams' digestive gland tissues. In parallel, temperature and salinity in the adjacent water, redox potential, pH and organic matter in sediment, and dissolved oxygen in the interstitial water were measured. The results suggested that RSP showed a spatial gradient characterised by hypoxia/anoxia, reduced potential, acidic conditions and high organic enrichment in sediments at the most contaminated sites. Significant (p < 0.05) decrease of biodiversity indicators were observed in the areas impacted by the aquaculture discharges. Biomarkers did not show a clear pattern and of all biochemical responses tested, GPX, DNA damage and LPO were the most sensitive ones and showed significant (p < 0.05) increase in the polluted sites. Benthic biodiversity indicators were significantly (p < 0.05) positively correlated with pH, redox potential and dissolved oxygen and negatively correlated with organic matter. On the contrary, antioxidant enzymatic responses (GPX) and oxidative stress parameters were significantly (p < 0.05) negatively correlated with those physico-chemical parameters. It has been demonstrated that effluents from fish aquaculture activities in Río San Pedro creek may produce an alteration of physico-chemical characteristics of seabed and induce oxidative stress and DNA damage in soft-sediment species which may lead to changes of the benthic population structure and health status of the exposed organisms.     

Lysosomal responses and metallothionein induction in the blue mussel Mytilus edulis from the south-west coast of Iceland

It has recently been emphasized that high levels of inorganic and organic micropollutants (particularly organometals, POPs and PAHs) may be present in coastal waters at high latitudes, stressing the need to evaluate the effects of contaminants on marine organisms from sub-arctic zones. With this aim, specimens of the blue mussel Mytilus edulis were sampled in polluted and reference areas along the south-west coast of Iceland in July 2004. Samples were collected from the intertidal zone at three sites in Reykjavik harbour which are differently exposed to contaminants, and at three reference coastal sites, two located along the Reykjanes Peninsula and the third one on the northern part of Hvalfjordur fiord. Lipofuscin content, neutral lipid accumulation and lysosomal enlargement were evaluated in digestive cells from cryostat sections of the mussel hepatopancreas, and quantified by automated image analysis. Metallothionein induction was also determined in the same tissue. Results indicate that mussels from the inner part of Reykjavik harbour, which is the most sheltered and most influenced by extensive shipping traffic, were the worst affected, with the highest values in neutral lipids, lipofuscin and lysosomal swelling. At the other two harbour sites, mussels exhibited lower values, similar to those observed in organisms collected in Hvalfjordur fiord and in bay of Osar. Mussels from Kuagerdi had the lowest values.     

Monitoring of the impact of Prestige oil spill on Mytilus galloprovincialis from Galician coast

In November 2002, the Prestige oil tanker was wrecked in front of Galician coast (NW of Spain), spilling near 63,000 tons of heavy oil until February 2003. Contamination produced was very extensive (70% of Galician beaches were reached by the oil) but heterogeneous, alternating intensely affected zones with neighbour locations where the repercussion was minimal. The objective of this study was to monitor sea environment contamination caused by Prestige oil spill during an 11-month period (August 2003-June 2004, nine samplings) in two locations of Galician coast with different geographical properties (Lira and Ancoradoiro beaches), by means of chemical determination of total polycyclic aromatic hydrocarbons (TPAH) in seawater, and using as exposure biomarker TPAH content in mussel (Mytilus galloprovincialis) tissues, and as effect biomarker DNA damage in mussel gills, evaluated by the comet assay. In addition, recovery ability of the mussels was determined after a 7-day stay in the laboratory. TPAH contents in seawater were very high in the earliest samplings, but then they maintained below 200 ng L(-)(1), similar to reference seawater. However, TPAH levels in mussel tissues were more variable: they increased again from January 2004, probably due to the adverse meteorological conditions that turned over the sea bottom and dispersed the oil accumulated in sediments. In most samplings, these levels decreased during the recovery stage. DNA damage in oil-exposed mussels was significantly higher than in reference mussels, both before and after the recovery phase, but they did not differ to one another. Comet tail length was slightly reduced during the recovery stage, indicative of a certain DNA repair in exposed mussels. This study showed up the importance of monitoring sea contamination events during an extended time, not only in evaluating the presence of the contaminants in the environment but also in determining their bioaccumulation and their effect on the exposed organisms.     

Polycyclic aromatic hydrocarbons and effects on marine organisms in the Istanbul Strait

Surficial sediments and mussels (Mytilus galloprovincialis) from the Istanbul Strait and Marmara Sea were analysed for sixteen parent polycyclic aromatic hydrocarbon (PAH) contents by gas chromatography-mass spectrometry (GC-MS) employing isotope dilution technique. Microalgae toxicity testing was applied to sediment elutriates and biological responses in terms of filtration rate and lysosomal stability were measured in mussels. Total PAH concentrations ranged from 2.1 to 3152 ng g^? 1 dry wt in sediments and from 43–601 ng g^? 1 wet weight in mussels. Molecular indices of phenanthrene/ anthracene, fluoranthene/pyrene and benzo(a)anthracene/chrysene were used to differentiate between pyrolytic and petroleum origin. Results showed that most of the contamination originates from high temperature pyrolytic inputs with some slight contribution of petrogenic PAH. PAH in sediments were frequently lower than the National Oceanic and Atmospheric Administration (NOAA)–ERM (Effects Range Medium) index. Results of sediment elutriate toxicity testing and biomarkers indicate that the cause of negative effects in sediments may result from different classes of pollutants and does not only relate with PAH contamination. Mussels from most of the stations showed both reduced lysosomal membrane stability and filtration rate indicating disturbed health although the two biomarker results did not always complement each other. The effect studies showed that the pollutants in the strait ecosystem have more pronounced effects in the middle parts than those at the Black Sea entrance.     

A study of radium bioaccumulation in freshwater mussels, Velesunio angasi, in the Magela Creek catchment, Northern Territory, Australia

Freshwater mussels, Velesunio angasi, along Magela Creek in Australia’s Northern Territory were examined to study radionuclide activities in mussel flesh and to investigate whether the Ranger Uranium mine is contributing to the radium loads in mussels downstream of the mine. Radium loads in mussels of the same age were highest in Bowerbird Billabong, located 20 km upstream of the mine site. Variations in the ratio of [Ra]:[Ca] in filtered water at the sampling sites accounted for the variations found in mussel radium loads with natural increases in calcium (Ca) in surface waters in a downstream gradient along the Magela Creek catchment gradually reducing radium uptake in mussels. At Mudginberri Billabong, 12 km downstream of the mine, concentration factors for radium have not significantly changed over the past 25 years since the mine commenced operations and this, coupled with a gradual decrease of the ²²⁸Ra/²²⁶Ra activity ratios observed along the catchment, indicates that the ²²⁶Ra accumulated in mussels is of natural rather than mine origin. The ²²⁸Th/²²⁸Ra ratio has been used to model radium uptake and a radium biological half-life in mussels of approximately 13 years has been determined. The long biological half-life and the low Ca concentrations in the water account for the high radium concentration factor of 30,000-60,000 measured in mussels from the Magela Creek catchment.     

Water and sediment toxicity assessment by use of behavioural responses of medicinal leeches

Behaviour has been shown to be a sensitive indicator of chemically induced stress and pathology in aquatic organisms. However, ecotoxicological investigations on medicinal leech are restricted because of scarce leech resources in natural waters. We used artificially bred medicinal leech (Hirudo medicinalis L.) of two different ages: young (1-2 weeks old) and adult leeches (1 year old). Animals were exposed to: (1) waters of Lake Drukshiai-the cooler of Ignalina Nuclear Power Plant, (2) sediments of the river Nemunas and (3) solution of heavy metal model mixture (HMMM). The following behavioural responses of leech were investigated: mobility (number of moving individuals within certain periods of time), avoidance response (number of individuals escaping the tested water or sediments) changes in body shape (contractions of some muscles, abnormal position of suckers) and feeding activity (longevity of attachment process, interruptions of feeding bouts, size of blood meal). Mobility of young leeches was increased in the tested waters of Lake Drukshiai, whereas adult leeches showed no changes of this index. Avoidance response as well as impaired feeding activity (prolonged attachment process or completely suppressed attachment reflex, decreased size of blood meal) was observed in young leeches exposed to the waters from Lake Drukshiai and in adult animals exposed to all three kind of trials mentioned above. Avoidance response and changes in mobility were recorded during the first hour of exposure to tested samples; therefore, these responses can be used as an express method for water and sediment pollution assessment. Impaired feeding activity was recorded after 1-3-week exposure: this response may be used in assessing a chronic toxicity of pollution. Medicinal leech, due to its sensitivity, simplicity of measured indices and ease of laboratory maintenance, can be used as a test organism in ecotoxicity studies.     

Biomonitoring Human Exposure to Household Air Pollution and Association with Self-reported Health Symptoms – A Stove Intervention Study in Peru

BackgroundHousehold air pollution (HAP) from indoor biomass stoves contains harmful pollutants, such as polycyclic aromatic hydrocarbons (PAHs), and is a leading risk factor for global disease burden. We used biomonitoring to assess HAP exposure and association with self-reported symptoms in 334 non-smoking Peruvian women to evaluate the efficacy of a stove intervention program.MethodsWe conducted a cross-sectional study within the framework of a community randomized control trial. Using urinary PAH metabolites (OH-PAHs) as the exposure biomarkers, we investigated whether the intervention group (n = 155, with new chimney-equipped stoves) were less exposed to HAP compared to the control group (n = 179, with mostly open-fire stoves). We also estimated associations between the exposure biomarkers, risk factors, and self-reported health symptoms, such as recent eye conditions, respiratory conditions, and headache.ResultsWe observed reduced headache and ocular symptoms in the intervention group than the control group. Urinary 2-naphthol, a suggested biomarker for inhalation PAH exposure, was significantly lower in the intervention group (GM with 95% CI: 13.4 [12.3, 14.6] μg/g creatinine) compared to control group (16.5 [15.0, 18.0] μg/g creatinine). Stove type and/or 2-naphthol was associated with a number of self-reported symptoms, such as red eye (adjusted OR with 95% CI: 3.80 [1.32, 10.9]) in the past 48 h.ConclusionsEven with the improved stoves, the biomarker concentrations in this study far exceeded those of the general populations and were higher than a no-observed-genotoxic-effect-level, indicating high exposure and a potential for increased cancer risk in the population.     

Interpretation of fish biomarker data for identification,classification, risk assessment and testing of endocrine disrupting chemicals

Chemical induced changes in fish biomarkers vitellogenin (VTG), secondary sex characteristics (SSC), and sex ratio indicate modes/mechanisms of action (MOAs) of EAS (estrogen, androgen and steroidogenesis) pathways. These biomarkers could be used for defining MOAs and the causal link between MOAs and adverse effects in fish for the identification of endocrine disrupting chemicals (EDCs). This paper compiled data sets of 150 chemicals for VTG, 57 chemicals for SSC and 38 chemicals for sex ratio in fathead minnow, medaka and zebrafish. It showed 1) changes in fish biomarkers can indicate the MOAs as anticipated; 2) in addition to EAS pathways, chemicals with non-EAS pathways induced changes in fish biomarkers; 3) responses of fish biomarkers did not always follow the anticipated patterns of EAS pathways. These responses may result from the interaction of chemical-induced multiple MOAs and confounding factors like fish diet, infection, culture conditions, general toxicity and stress response. The complex response of fish biomarkers to a chemical of interest requires EDC testing at multiple biological levels. Interpretation of fish biomarker data should be combined with relevant information at different biological levels, which is critical for defining chemical specific MOAs. The utility of fish biomarker data for identification, classification, PBT assessment, risk assessment, and testing of EDCs in the regulatory context was discussed. This paper emphasizes the importance of fish biomarker data in the regulatory context, a weight of evidence approach for the interpretation of fish biomarker data and the need for defining levels of evidence for the identification of EDCs.     

Marine water quality assessment using transplanted oyster larvae

Active bio-monitoring in terms of biomarkers was attempted using Crassostrea gigas larvae produced in the laboratory and transplanted using special containers to two sites at the entrance (A) and inner part (P) of the harbour of Arcachon (French Atlantic Coast). The larvae were kept in the medium for 48 h. Their physiological status and their biomarker levels : acetylcholinesterase AChE, catalase CAT and glutathione S-transferase GST activities were determined together with metallothionein MT and Thiobarbituric Acid Reactive Substances TBARS concentrations. Copper and PAH (polycyclic aromatic hydrocarbon) concentrations were determined in the exposed larvae and in the sediments collected under the containers. Cadmium, lead and zinc could be also analyzed in the sediments. Toxicity tests demonstrate that the larvae are in better physiological conditions in A compared to P. Larvae transplanted in the inner harbour (P) present relatively high GST activity (869.1+/-39.3 nmol min(-1)mg protein(-1)), TBARS (2.74+/-0.19 nmol mg protein(-1)), compared to those exposed at the harbour entrance (A). Copper measured in the sediments (65+/-1 mg kg(-1) d.w.) collected under the cages at P is higher than at A. Larvae placed in A present higher total PAH concentrations compared to the inner part. The data tend to reveal a lower copper and higher PAH contamination in A than in P. Therefore larvae, developing in the natural medium, show different responses according to their immersion sites. These responses, obtained within 48 h, may be related to the chemical contamination of the environment and may be used for seawater quality assessment in future studies.