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.     

Effects of natural and chemical stressors on Enchytraeus albidus: Can oxidative stress parameters be used as fast screening tools for the assessment of different stress impacts in soils?

Enchytraeids are important organisms of the soil biocenosis. They improve the soil pore structure and the degradation of organic matter. These organisms are used in standardized testing, using survival and reproduction (6 weeks) as endpoints. The use of biomarkers, linked to ecologically relevant alterations at higher levels of biological organization, is a promising tool for Environmental Risk Assessment. Here, enchytraeids were exposed for different time periods (two days and three weeks) to different soils (OECD artificial soil, different compositions in its organic matter, clay or pH value, and LUFA 2.2 natural soil) and different chemicals (Phenmedipham and copper). The main question addressed in the present study was if the effects of chemicals and different soil properties are preceded by alterations at the sub-cellular level, and if these endpoints may be used reliantly as faster screening tools for the assessment of different stress conditions in soils. The parameters measured in E. albidus whole body were: lipid peroxidation (LPO), total glutathione (TG), as well as the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione S-transferase (GST). The results showed that biomarker responses in E. albidus were significantly affected by the soil type (GST, CAT, GPx, GR and LPO) and the duration of exposure in OECD artificial soil (GST, GPx, GR, CAT and LPO) but not in LUFA 2.2 natural soil. For the abiotic factors studied, after 2 days, low pH decreased significantly the TG levels and the activities of CAT and GR,and low OM also significantly decreased CAT and GR activities. After 3 weeks, differences in soil properties caused a decrease in GR and GPx activities, whereas increased GST activity was observed due to low organic matter and pH. Copper significantly increased the activities of CAT, GPx and GR, and decreased the activity of GST after 2 days as well as inscreasing LPO levels after 3 weeks. Phenmedipham increased LPO levels, associated with increased levels of TG as well as increased activities of CAT and GPx and decreased GST activity after 3 weeks exposure. This study shows that both abiotic and chemical stresses could be followed through biomarker analysis and that some of these determinations are potential endpoints in a quick soil contamination assessment procedure.     

The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination

Water contaminants have a high potential risk for the health of populations. Protection from toxic effects of environmental water pollutants primarily involves considering the mechanism of low level toxicity and likely biological effects in organisms who live in these polluted waters. The biomarkers assessment of oxidative stress and metabolic alterations to cadmium exposure were evaluated in Nile tilapia, Oreochromis niloticus. The fish were exposed to 0.35, 0.75, 1.5, and 3.0 mg/l concentrations of Cd2+ (CdCl2) in water for 60 days. Fish that survived cadmium exposure showed a metabolic shift and a compensatory development for maintenance of the body weight gain. We observed a decreased glycogen content and decreased glucose uptake in white muscle. Lactate dehydrogenase (LDH) and creatine phosphokinase (CK) activities were also decreased, indicating that the glycolytic capacity was decreased in this tissue. No alterations were observed in total protein content in white muscle due to cadmium exposure suggesting a metabolic shift of carbohydrate metabolism to maintenance of the muscle protein reserve. There was an increase in glucose uptake, CK increased activity, and a clear increase of LDH activity in red muscle of fish with cadmium exposure. Since no alterations were observed in lipoperoxide concentration, while antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were changed in the liver and the red and white muscle of fish with cadmium exposure, we can conclude that oxygen free radicals are produced as a mediator of cadmium toxicity. Resistance development is related with increased activities of antioxidant enzymes, which were important in the protection against cadmium damage, inhibiting lipoperoxide formation.     

Revegetating fly ash landfills with Prosopis juliflora L.: impact of different amendments and Rhizobium inoculation

A revegetation trial was conducted to evaluate the feasibility of growing a legume species, Prosopis juliflora L., on fly ash ameliorated with combination of various organic amendments, blue-green algal biofertilizer and Rhizobium inoculation. Significant enhancements in plant biomass, photosynthetic pigments, protein content and in vivo nitrate reductase activity were found in the plants grown on ameliorated fly ash in comparison to the plants growing in unamended fly ash or garden soil. Higher growth was obtained in fly ash amended with blue-green algae (BGA) than farmyard manure or press mud (PM), a waste from sugar-processing industry, due to the greater contribution of plant nutrients, supply of fixed nitrogen and increased availability of phosphorus. Nodulation was suppressed in different amendments of fly ash with soil in a concentration-duration-dependent manner, but not with other amendments. Plants accumulated higher amounts of Fe, Mn, Cu, Zn and Cr in various fly ash amendments than in garden soil. Further, inoculation of the plant with a fly ash tolerant Rhizobium strain conferred tolerance for the plant to grow under fly ash stress conditions with more translocation of metals to the above ground parts. The results showed the potential of P. juliflora to grow in plantations on fly ash landfills and to reduce the metal contents of fly ash by bioaccumulation in its tissues.     

Juvenile sea bass (Dicentrarchus labrax L.) DNA strand breaks and lipid peroxidation response following 17beta-estradiol two mode of exposures

Juvenile Dicentrarchus labrax L. (sea bass) were exposed to water diluted 17beta-estradiol (E2) (200 ng/L and 2000 ng/L) and treated with intraperitoneal (i.p) injection E2 (0.5 mg/kg and 5 mg/kg) during 10 days in order to study its genotoxicity and peroxidative damage, measured as gill, blood, liver and kidney DNA integrity decrease using DNA strand breaks assay as well as liver, gill and kidney lipid peroxidation (LPO) respectively. Juvenile sea bass gill DNA integrity was significantly decreased for all E2 exposure conditions. However, no differences were detected either between different exposure routes or tested concentrations. Blood DNA integrity was significantly decreased by E2 5 mg/kg as well as by both water diluted E2 exposure conditions. The highest E2 dose (5 mg/kg) also promoted liver DNA integrity decrease. Liver and gill LPO significantly increased at the highest E2 i.p treatment. An increasing trend of gill and liver LPO, though statistically insignificant, was observed in D. labrax exposed to water diluted E2 in both tested concentrations. The current results demonstrated that DNA damage in juvenile sea bass is affected by the E2 exposure conditions, such as water diluted E2 versus i.p E2 injection since i.p E2 injection promoted higher genotoxicity effect, in terms of affected organs than water diluted E2. Moreover, the organ sensitivity to E2 was different, since gill showed more susceptibility than blood, liver and kidney. Concerning kidney LPO and DNA integrity no differences were found between treated and controls juvenile sea bass groups.     

Leachability of elements from sub-bituminous coal fly ash from India

Environmental concerns regarding the potential contamination of soil, surface and ground water due to the presence of soluble metal species in the ash pond leachate is of great importance. Serial batch leaching was carried out simulating the rainwater condition of the study area to understand the behaviour of elements during leaching. The leachates were analysed for the elements Al, Ca, K, Mg, Na, P, S, Si, As, Ba, Fe, Mn, Mo, Ti, V, Pb, Zn, Co, Cr, Cu, Ni and Cd by inductively coupled plasma optical emission spectrometer (ICP-OES). It was found that Cd, Co, Cr and Ni did not leach from the ash while Cu and Pb concentrations were insignificant in the leachate regardless of liquid to solid (L/S) ratio. Most of the elements showed maximum concentrations at lower L/S ratio and then decreased with increasing L/S. The total cumulative concentrations of As, Mn and Mo were found to be higher than the World Health Organization (WHO) recommended values for drinking water while the concentrations of Fe, Mn and As exceeded the maximum allowable concentrations prescribed by the United States Environmental Protection Agency (USEPA). The pre and the post leached ash samples were analysed for morphology, specific surface area and mineralogical changes. Analysis of post-leached fly ash indicated changes in the specific surface area and morphology but no change in mineralogy.     

Multiple stressor effects of radiation and metals in salmon (Salmo salar)

These experiments were designed to look at the cellular effects in key organs in Atlantic salmon (Salmo salar) after exposure in vivo to radiation and subtoxic levels of aluminum (Al) and cadmium (Cd), alone or in combination. Salmon (25g) were exposed to a single 0.5Gy dose of gamma-irradiation in water containing Cd, Al or Cd+Al. Three fish per group were sacrificed after 1h and the liver, pronephros, fin and gill of each was dissected. Small explants of each tissue were set up. After 2 days, the culture medium was harvested and filtered then placed on a reporter cell line for determination of stress signal activity (bystander effects). Radiation in combination with Cd and/or Al, caused bystander effects in tissues harvested from in vivo exposed salmon. The effects vary between different organs and are not consistently additive or synergistic for a given treatment. Tissue type appears to be critical. Liver cultures produce a toxic factor which is lethal to reporter cells, and therefore no liver data could be obtained. It is hoped that this stress signal response will prove to be a useful indicator of environmental stress in species inhabiting aquatic ecosystems.     

An in vitro/in vivo screening assay as a sensitive tool to assess endocrine disruptive activity in surface water

Adult male fathead minnow were exposed for 14 or 28-days under flow-through conditions to undiluted filtered water samples from the rivers Meuse and Rhine in the Netherlands. The experiment included two vessels per treatment each containing 10 fish and samples of five fish were taken after 14 and 28 days. Additional groups were exposed to 17alpha-ethinylestradiol (EE2) as a reference and untreated drinking water as a negative control. Major endpoints examined included induction of vitellogenin (VTG) synthesis, VTG mRNA activity, hepato- and gonadosomatic indices (HSI and GSI) and gonadal histology. No significant difference was recorded in body weight or mean GSI values between the various treatments. Only exposure to Meuse water resulted in significantly higher HSI means after 14 days. Histological examination showed no apparent effects on gonadal tissue except for eosinophilic blood plasma in fish exposed to Meuse water or EE2. After 14 and 28 days, elevated VTG and VTG mRNA levels were measured in most livers of the fish exposed to Meuse water, but not in the fish exposed to Rhine water. This was confirmed by measuring estrogenic responses in the in vitro ER CALUX assay. Induction of VTG synthesis proved to be the most sensitive endpoint in the Non Spawning Male Fish Assay for in vivo detection of bio-available estrogenic activity supplementary to a sensitive in vitro assay. The other endpoints examined varied too much and required a higher number of fish or replicates to achieve sufficient power for statistical testing making them less animal friendly.     

Acute and sub-lethal toxicity of landfill leachate towards two aquatic macro-invertebrates: demonstrating the remediation potential of air stripping

A specific leachate that contained 1.036 mg l(-1) of 2-chlorobiphenyl was used in the study (255 mg l(-1) COD and 133 mg l(-1) BOD5). Bench scale (20 l) air stripping trials were used to simulate on a small-scale the treatment potential of this method. Air stripping effectively reduced the leachates COD concentration. Regardless of the volume of air supplied (1-5 l of air per minute) the leachates COD reached a <50 mg l(-1) equilibrium after 96-h exposure, however, increasing the volume of air accelerated the process. In untreated leachate, the LC50 for Asellus aquaticus was 57% v/v leachate in deionised water and 5% for Gammarus pulex (96-h, static LC50 tests without nutrition and oxygen depleting conditions). After being exposed to air stripping, these values rose from 90% to below the LC50 threshold for Asellus when 1-5 l of air per minute were applied and 30-90% for Gammarus. Furthermore, in sub-lethal concentrations of air stripped leachate (leachate that had been exposed to 5-l of air per minute for 96-h) the population dynamics of both test species remained unaltered.     

Biomarkers and bioindicators of the health condition of Ameca splendens and Goodea atripinnis (Pisces: Goodeaidae) in the Ameca River, Mexico

Pollution of water bodies causes stress on organisms inhabiting them. Determination of biomarkers and bioindicators on fish populations reflects whether they are subject to stress. We assessed two populations of Ameca splendens and Goodea atripinnis in a reference site (spring "El Rincon") and De La Vega reservoir, which receives wastewater of a sugar industry, on Ameca River course. We analyzed level of lipid peroxidation and enzymatic activities of gamma-glutamyl-transpeptidase, acetylcholinesterase and ethoxyresorufin-O-deethylase as biomarkers; we also studied age classes and various body indexes. Environmental factors were recorded and a water quality index was assessed. Water quality was better in the spring than in the reservoir. Organisms inhabiting the reservoir presented higher oxidative stress by the lipid peroxidation levels, and neurotoxic impacts by the acetylcholinesterase and some detoxification mechanisms were evident by the gamma-glutamyl-transpeptidase and ethoxyresorufin-O-deethylase activities. Integrated Biomarker Response demonstrate that De La Vega reservoir is a more stressing place to organisms living there, particularly for A. splendens. In both species, males were more affected than females. Condition and reproductive parameters in reservoir showed evidences of physiological changes due to xenobiotics exposure and suggest a tactic of the organisms to survive in this site. Both biomarkers and body indexes revealed that A. splendens is a more sensitive species than G. atripinnis to environmental stress.     

PCB congener specific oxidative stress response by microarray analysis using human liver cell line

In this study we have examined the effect of exposure to different congeners of PCBs and their role in oxidative stress response. A metabolically competent human liver cell line (HepG2) was exposed with two prototype congeners of PCBs: coplanar PCB-77 and non-coplanar PCB-153. After the predetermined times of exposure (0–24 h) at 70 μM concentration, the HepG2 cells showed significant apoptotic changes by fluorescent microscopy after 12 h of exposure. Gene set enrichment analysis (GSEA) identified oxidative stress as the predominant enrichment. Further, paraquat assay showed that PCB congeners lead to oxidative stress to different extents, PCB-77 being more toxic. This study, with emphasis on all recommended microarray quality control steps, showed that apoptosis was one of the most significant cellular processes as a result of oxidative stress, but each of these congeners had a unique signature gene expression, which was further validated by Taqman real time PCR and immunoblotting. The pathways involved leading to the common apoptotic effect were completely different. Further in-silico analysis showed that PCB-153 most likely acted through the TNF receptor, leading to oxidative stress involving metallothionein gene families, and causing apoptosis mainly by the Fas receptor signaling pathway. In contrast, PCB-77 acted through the aryl hydrocarbon receptor. It induced oxidative stress through the involvement of cytochrome P450 (CYP1A1) leading to apoptosis through AHR/ARNT pathway.     

Development and analysis of high-performance green concrete in the urban infrastructure

Cement production accounts for approximately 5% of total global CO₂ emissions from all human activities. In addition, the consumption of virgin aggregates for concrete infrastructure has created virgin material scarcity issues in many areas of the USA. High-performance green concrete (HPGC) with fly ash and recycled aggregates can help reduce the demand for material inputs and reduce pollution outputs associated with bulk material flow of urban concrete. Structural and durability tests showed that HPGC containing fly ash and 50% recycled aggregate (100% of the coarse aggregate fraction) performed equally or better than 100% ordinary Portland cement concrete with the same cementitious content. Durability improvements were more significant with Class F than Class C fly ash. For both Class F and Class C fly ash, greater per cent replacement of Portland cement with fly ash led to slower and lower strength gain, but still within acceptable strength criteria for Colorado Department of Transportation Class B concrete. This paper quantifies the sustainability of HPGC in urban infrastructure by addressing structural performance, environmental, economic and resource depletion impacts.     

Toxic effects of waterborne polychlorinated biphenyls and sex differences in an endangered goodeid fish (Girardinichthys viviparus)

Polychlorinated biphenyls (PCBs) elicit toxic effects in different species. PCBs undergo biotransformation by enzymes associated with the mixed functional oxidase system, such as cytochrome P450 (cyt P450), this biotransformation being sex-dependent. No other metabolic pathways are known, however, in connection with this process. Thus, the present study aims to evaluate the effects of waterborne PCBs at sublethal nominal concentrations (0.92 mg PCBs/L) on the black-fin goodeid Girardinichthys viviparus, an endangered fish native to the Valley of Mexico, as well as any sex differences related to PCB biotransformation. Eight-month-old adult fish born in the laboratory were exposed for 1, 2, 4, 8 and 16 days to half the LC(0) (calculated concentration at which no deaths occurred after 96 h) determined by an acute toxicity test. Specimens were sacrificed following exposure and the liver was used to evaluate different biomarkers: cyt P450 concentration and alcohol dehydrogenase (ADH) and ethoxyresorufin O-deethylase (EROD) activities. Results show sexual differentiation with regard to all biomarkers in both the control group and PCB-treated fish, with higher values found in males. The induction rate of cyt P450 remained constant throughout the study in males. In females, induction peaked on day 4, coinciding with maximum EROD activity, and fell significantly thereafter. EROD was lower in PCB-treated males than in the control group, but was greater in magnitude. ADH was significantly induced in both sexes from day 2 to day 16 of exposure. The highest response as compared to the control group occurred on day 8 in females. A correlation was found between ADH activity and exposure to PCBs. Three possible action mechanisms, operating either individually or concurrently, are proposed.     

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.     

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.     

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.     

Phosphorus removal by fly ash

The aim of this work was to investigate the possible use of fly ash generated from thermic power stations in the removal of phosphorus contained in aqueous solutions. A series of batch tests were conducted and the influence of temperature, phosphate concentration, and fly ash dosage on phosphate removal were investigated. The effect of adsorption dosage was not significant at any temperature applied. Phosphate removal in excess of 99% was obtained in these studies. Maximum removals were achieved at 40°C. The tests were also carried out on a continuous basis in an adsorption column. It was found that fly ash is an efficient adsorbent for phosphate removal. This was expected due to the high concentration of calcite present in the fly ash (338 g/kg). The phosphate removal decreased to 80.4% at the end of 72 h.