Use of biomarkers in Nile tilapia (Oreochromis niloticus) to assess the impacts of pollution in Bolgoda Lake, an urban water body in Sri Lanka

The present study reports the first analysis of water pollutants in Sri Lankan waters using a suite of biomarkers in Nile tilapia (Oreochromis niloticus) residing in Bolgoda Lake which receives urban, industrial and domestic wastes from multiple sources. The fish were collected from the lake in the dry period (April 2005) and wet periods (September 2005, October 2006) and the levels of biomarkers viz. hepatic ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), metallothioneins, biliary fluorescent aromatic compounds, brain and muscle cholinesterases (ChE) were compared with those of the laboratory reared control fish and the fish obtained from a less polluted water body, Bathalagoda reservoir (reference site). The results revealed that biomarker levels of the fish collected from the reference site were not significantly different from the controls. Hepatic EROD and GST activities in fish from Bolgoda Lake were induced 4.2-16.6 folds and 1.4-3.3 folds respectively compared with the control fish. Analysis of bile in the lake fish revealed recent uptake of naphthalene, pyrene and benzo(a)pyrene type polycyclic aromatic hydrocarbons (PAHs). The induction of EROD activities in feral fish reflects the exposure of fish to aryl hydrocarbon receptor agonists including PAHs present as pollutants in the Bolgoda Lake. Cholinesterase activity in the fish inhabiting one sampling site of Bolgoda Lake was lower (22-40% inhibition) than the activity measured in the control fish indicating the presence of anticholinesterase pollutants in the area. Hepatic metallothionein levels in the lake fish were higher (1.9-3.2 folds) in comparison to the controls indicating metal exposure. The results support the potential use of these biomarkers in Nile tilapia in assessing pollution in tropical water bodies.     

Influence of feeding procedure on biomarkers in caged rainbow trout (Oncorhynchus mykiss) used in environmental monitoring

Biomarkers in feral fish can be a useful tool for environmental monitoring of aquatic ecosystems. Drawbacks, however, are that suitable fish species are not always available and that natural variations can bias the results. An alternative strategy is to use farmed fish placed in cages. There is, however, still a risk that factors other than pollution level could have an impact on the biomarkers and the observed responses in the fish. The present study evaluates the effects of feeding procedure on biomarkers in caged fish. Two feeding rations (2% and 8% weekly feeding) have been examined for a large number of biomarkers in caged rainbow trout (Oncorhynchus mykiss). Significant effects of feeding rations were found on hepatic ethoxyresorufin-O-deethylase (EROD) and catalase activity, PAH metabolites in bile, plasma ion concentrations and the presence of immature red blood cells. The influence on EROD activity and PAH metabolites seems to be caused by elevated uptake of pollutants when feeding ratios are higher. The effects on other biochemical and physiological variables are more likely caused by stress due to insufficient feeding. According to these results, valid comparison of fish groups in environmental monitoring requires standardized feeding levels.     

Concentration of four heavy metals (cadmium, lead, mercury, and arsenic) in organs of two cyprinid fish (Cyprinus carpio and Capoeta sp.) from the Kor River (Iran)

Concentration of heavy metals in aquatic animals mainly occurs due to industrial contamination. In this study, the concentrations of four heavy metals (cadmium, lead, mercury, and arsenic) in organs of two cyprinid fish and in water collected from three sections of the Kor River, Iran were determined using the inductively coupled plasma method. Pathological and hormonal changes due to metal contamination were also measured. The concentrations of heavy metals in tissue of fish from the middle sampling zone were significantly higher (p?<?0.05) than those from the other two sampling zones, whereas no significant differences (p?>?0.05) were detected between the two sexes and species. High levels of metals were found in the ovaries and testes; estradiol in females and progesterone and testosterone in males from the middle study site were significantly (p?<?0.05) lower than values from the other two sites. Pathological changes in blood cells, liver, and kidneys of fishes were significantly higher in highly polluted areas (middle sampling zone). These results show that industrial activities have polluted the river and that the maximum concentrations of Cd, Pb, and Hg were higher than the permissible levels for human consumption.     

Monitoring Pollution in River MureŞ, Romania, Part III: biochemical effect markers in fish and integrative reflection

Along a downstream stretch of River Mureş, Romania, adult males of two feral fish species, European chub (Leuciscus cephalus) and sneep (Chondrostoma nasus) were sampled at four sites with different levels of contamination. Fish were analysed for the biochemical markers hsp70 (in liver and gills) and hepatic EROD activity, as well as several biometrical parameters (age, length, wet weight, condition factor). None of the biochemical markers correlated with any biometrical parameter, thus biomarker reactions were related to site-specific criteria. While the hepatic hsp70 level did not differ among the sites, significant elevation of the hsp70 level in the gills revealed proteotoxic damage in chub at the most upstream site, where we recorded the highest heavy metal contamination of the investigated stretch, and in both chub and sneep at the site right downstream of the city of Arad. In both species, significantly elevated hepatic EROD activity downstream of Arad indicated that fish from these sites are also exposed to organic chemicals. The results were indicative of impaired fish health at least at three of the four investigated sites. The approach to relate biomarker responses to analytical data on pollution was shown to fit well the recent EU demands on further enhanced efforts in the monitoring of Romanian water quality.     

Evaluating the power of surface attendance counts to detect long-term trends in populations of crevice-nesting auklets

This study assessed metals in water and different tissues of Labeo rohita and the impact of these metals on DNA and proteins as biomarkers of gills and muscles of these fish from three different polluted sites (reference or low = KW, medium = CH and high = SK) of the Indus River, Pakistan. The Mn, Pb, Cu, Zn, Hg, and Cr levels in water, gills, liver, muscles, and skin of these fish were compared with the international permissible levels. All metals except Pb and Hg in water were within the acceptable limits of drinking water. In contrast, the Mn, Hg, and Cr levels in the fish tissues were higher than their permissible limits for fish as a human food. Here, the gills contained higher metals than the other tissues. Different patterns of biomarkers were found in fish from these sites. While the gills did not show four protein bands (55, 30, 18.4, and 16.4 kDa), the muscles showed four new protein bands (100, 85, 45, and 20 kDa) for fish from the medium and high polluted sites as compared to the reference or low polluted site. The fish from the CH and SK sites of the Indus River contained low molecular weight DNA in their gills but high molecular weight DNA in their muscles when compared with the KW site. This study suggests that the proteins and DNA profiles of L. rohita could be used as biomarkers to assess the impact of potential environmental stressors such as metals on the freshwater systems.     

Kinetic of Biomarker Responses in Juveniles of the Fish Sparus aurata Exposed to Contaminated Sediments

Sediments in the National Park of the Atlantic Islands (Galicia, Spain) were affected by the spill of the tanker Prestige (November, 2002) and still present high levels of Polycyclic aromatic hydrocarbons. The adverse effects associated with the contaminants in sediments were tested using a chronic bioassay, exposing juveniles of the fish Sparus aurata (seabream). A toxicokinetic approach is proposed to evaluate sediment quality by linking chemical and ecotoxicological data along the time. Sediment samples were physicochemically characterized and the concentration of contaminants (Polycyclic aromatic hydrocarbons – PAHs – and metals) was measured. Fishes were exposed to contaminated sediments, and samples from different tissues were collected every 15 days throughout the 60 days that lasted the experiment. A biomarker of exposure (ethoxyresorufin O-deethylase activity – EROD activity) and a biomarker of effect (histopathology) were analyzed during the exposure period. Results show a relationship between the biomarkers and the concentrations in sediments of polycyclic aromatic hydrocarbons—PAHs. Besides, the toxicokinetic approach links biomarkers response providing information about the relationship between the detoxification process and the damages observed in the different tissues. The frequency of the histological damage is highest when the EROD activity slightly decreases in accordance with the mechanism of detoxification of this enzymatic system against PAHs and other organic contaminants.     

Biomarkers of environmental contaminants in the coastal waters of Estonia (Baltic Sea): effects on eelpouts (Zoarces viviparus)

The eastern Baltic Sea near the Estonian coast is heavily navigated by numerous cargo ships and oil tankers. Hundreds of accidents and oil spills happen yearly in this area. Yet, there is a lack of data concerning the distribution and effects of the environmental contaminants, especially polycyclic aromatic hydrocarbons (PAHs). Different parts of the Baltic Sea have different levels of contamination; therefore a wide range of monitoring stations in coastal areas in the Gulf of Finland and Gulf of Riga were chosen. The aim of the present research was to document the responses of chosen biomarkers of environmental contaminants in different unstudied areas of the Estonian coastal sea. During 2009 and 2010 we measured PAH metabolites, EROD activities, geno- and cytotoxicity, histology, parasites and other biomarkers from the eelpout (Zoarces viviparus), a resident benthic fish species. The results showed that fish from the Gulf of Riga emitted lower levels of fluorescence in fixed wavelength analyses (representing equivalents of PAH metabolites in bile and urine), and consistently, showed less geno- and cytotoxicity and parasite infection, higher liver somatic index (LSI) and a higher condition factor (CF) than fish inhabiting areas close to the Baltic proper and in the Gulf of Finland. The results point to the effect of long-range contaminant transportation, whether atmospheric or hydrodynamic, and also to the intensive shipping activity in international routes. This study fills the gap of knowledge in this area that has persisted until now. Nevertheless, more studies in this area on the different groups of contaminants are necessary, to specify the factors that are responsible for observed biological effects.     

Determination of 7-ethoxyresorufin-o-deethylase (EROD) induction in leaping mullet (Liza saliens) from the highly contaminated Aliaga Bay, Turkey

Pollution of the aquatic environment is a global concern owing to the devastating effects of contaminants whose levels are growing at an alarming rate, and it has become a major threat for marine organisms, as well as to humans as consumers. This study has been carried out on leaping mullet obtained from Aliaga Bay, which is located on the west coast of the Aegean Sea near Izmir and hosts the world’s fifth largest shipyard, plus a broad range of industrial activities, including an oil refinery and a paper factory. The waste from these industries, combined with municipal sewer discharges, is the main cause of pollution in this region. There is no national documentation or research on the determination of pollution resulting from the industrial activities in this area. In the present study, the degree of induction of CYP4501A-associated 7-ethoxyresorufin O-deethylase (EROD) activity and immunochemical detection of CYP1A1 in the liver of leaping mullet (Lisa saliens) were used as biomarkers for the assessment of polycyclic aromatic hydrocarbon (PAH)-type organic pollutants in Aliaga Bay. Mullet caught from different locations of the bay had approximately 52 times more EROD activity than the feral fish sampled from a clean reference site near Foca, Izmir. The results of this study indicate that Aliaga Bay is highly contaminated with PAH-type organic pollutants.     

Genotoxic effects of water pollution on two fish species living in Karasu River,Erzurum, Turkey

Karasu River, which is the only river in the Erzurum plain, is the source of the Euphrates River (Eastern Anatolia of Turkey). The river is in a serious environmental situation as a result of pollution by agricultural and industrial sewage and domestic discharges. The present study aims to evaluate genotoxic effects of toxic metals in chub, Leuciscus cephalus, and transcaucasian barb, Capoeta capoeta, collected from contaminated site of the Karasu River, in comparison with fish from an unpolluted reference site. Heavy metal concentrations in surface water of the river were determined. The condition factor (CF) was taken as a general biomarker of the health of the fish, and genotoxicity assays such as micronucleus (MN) and other nuclear abnormalities (NA) were carried out on the fish species studied. MN and NA such as kidney-shaped nucleus, notched nucleus, binucleated, lobed nucleus, and blebbed nucleus were assessed in peripheral blood erythrocytes, gill epithelial cells, and liver cells of the fish. A significant decrease in CF values associated with a significant elevation in MN and NA frequencies was observed in fish collected from the polluted sites compared with those from the reference site. Results of the current study show the significance of integrating a set of biomarkers to identify the effects of anthropogenic pollution. High concentrations of heavy metals have a potential genotoxic effects, and the toxicity is possibly related to industrial, agricultural, and domestic activities.     

Multivariate analysis of mixed contaminants (PAHs and heavy metals) at manufactured gas plant site soils

Principal component analysis (PCA) was used to provide an overview of the distribution pattern of polycyclic aromatic hydrocarbons (PAHs) and heavy metals in former manufactured gas plant (MGP) site soils. PCA is the powerful multivariate method to identify the patterns in data and expressing their similarities and differences. Ten PAHs (naphthalene, acenapthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo[a]pyrene) and four toxic heavy metals — lead (Pb), cadmium (Cd), chromium (Cr) and zinc (Zn) — were detected in the site soils. PAH contamination was contributed equally by both low and high molecular weight PAHs. PCA was performed using the varimax rotation method in SPSS, 17.0. Two principal components accounting for 91.7% of the total variance was retained using scree test. Principle component 1 (PC1) substantially explained the dominance of PAH contamination in the MGP site soils. All PAHs, except anthracene, were positively correlated in PC1. There was a common thread in high molecular weight PAHs loadings, where the loadings were inversely proportional to the hydrophobicity and molecular weight of individual PAHs. Anthracene, which was less correlated with other individual PAHs, deviated well from the origin which can be ascribed to its lower toxicity and different origin than its isomer phenanthrene. Among the four major heavy metals studied in MGP sites, Pb, Cd and Cr were negatively correlated in PC1 but showed strong positive correlation in principle component 2 (PC2). Although metals may not have originated directly from gaswork processes, the correlation between PAHs and metals suggests that the materials used in these sites may have contributed to high concentrations of Pb, Cd, Cr and Zn. Thus, multivariate analysis helped to identify the sources of PAHs, heavy metals and their association in MGP site, and thereby better characterise the site risk, which would not be possible if one uses chemical analysis alone.     

Use of the land snail Helix aspersa for monitoring heavy metal soil contamination in Northeast Algeria

The objective of this study was to investigate the impact of anthropogenic activities on soil quality using the land snail Helix aspersa as a bioindicator. Soil samples and snails were collected from several sites in Northeast Algeria during the summer and winter of 2010. All of the sites were chosen due to their proximity to industrial factories—a potential source of soil pollution via heavy metal contamination. The concentration of heavy metals (Pb, Cd, Mn, and Fe) in soil samples was analyzed using atomic absorption spectrophotometry. Activity levels of glutathione S-transferase (GST) and acetylcholinesterase (AChE), indicators of oxidative stress and neurotoxicity, respectively, were measured in snails collected from each site. GST and AChE activity were found to vary between sites and by season. The highest levels of GST activity were registered during the summer at sites closest to potential sources of pollution. AChE activity levels also peaked during the summer with the highest values recorded at the site in El Hadjar. These increased levels of bioindicative stress response correlated with increasing metal concentration in soil samples collected at each site.     

Multimarker approach analysis in common carp Cyprinus carpio sampled from three freshwater sites

The aim of this study is to assess the response of a multimarker approach in common carp Cyprinus carpio sampled from three Tunisian dam lakes selected according to different environmental and ecological characteristics. Glutathione-S-transferase (GST) activity was analyzed in carp liver and used as a phase II detoxification enzyme, hepatic metallothionein content (MTs) was used as a metallic stress indicator, and cholinesterase activities were analyzed in muscle and brain and used as neurotoxicity biomarker. Micronucleus frequency test (MN) as a genotoxicity marker. GST and MT levels showed an increase in fish from the Bir Mcherga site and a decrease in Sidi Saâd site with respect to fish from Nebhana site. Results showed a strong inhibition of cholinesterase activities in fish from Bir Mcherga and Sidi Saâd sites compared to Nebhana site. Relatively high level of MN is reported specially in fish blood from the Bir Mcherga site.     

Oxidative stress in Nile tilapia (Oreochromis niloticus) and armored catfish (Pterygoplichthys anisitsi) exposed to diesel oil

Considering that diesel oil is one of the most common aquatic contaminants, we compare the oxidative stress between two species of fish with different habitats (Pterygoplichthys anisitsi, benthic and Oreochromis niloticus, nektonic) exposed to diesel oil. Malondialdehyde concentrations (MDA) and the activities of ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase were all analyzed in the fishes' livers and gills after 2 and 7 days of exposure to different concentrations of diesel (0.1 and 0.5 mL/L). In the tilapia, MDA levels and the activities of EROD and GST activity in the liver, as well as MDA levels and the activities of GST and SOD in the gill had statistically significant differences between the treatments and between the times of exposure. For the catfish, the same occurred in the case of MDA, EROD, and SOD in the liver and in CAT and SOD in the gills. There were significant differences in the enzyme activity and lipid peroxidation between the species. Although the activity of most enzymes seemed to be more expressive and responsive to diesel in O. niloticus, diesel oil also caused significant effects on oxidative stress parameters in P. anisitsi, even though this species is benthic and thus has less access to insoluble fractions of diesel oil. Therefore, both species can be used as sentinel organisms in environmental biomonitoring of diesel contamination.     

Heavy metals in Patella caerulea (Mollusca, Gastropoda) in polluted and non-polluted areas from the Iskenderun Gulf (Mediterranean Turkey)

The concentrations of Cd, Cu, Zn, Fe, Pb, Ni, and Co were measured in gastropod mollusks Patella caerulea in the Mediterranean area. The organisms were collected at two coastal sites in Iskenderun Gulf during winter, spring, summer, and autumn 2008. Samples of the digestive gland, gill, and muscle were analyzed for heavy metals. The aim of study is to determine heavy metal levels in tissues of P. caerulea in different locations. Tissues of P. caerulea from the polluted site showed metal concentrations appreciably higher than unpolluted organisms. The highest metal levels were registered in the digestive gland of P. caerulea. Generally, digestive gland and gills showed higher metal concentrations than muscle. The average concentrations of heavy metals analyzed exhibited the following decreasing order: Fe > Zn > Cu > Ni > Cd > Pb > Co for both stations 1 and 2. Seasonal changes in metal concentrations were observed in the tissues of P. caerulea from a polluted and an unpolluted population.     

Biotransformation and Genotoxic Biomarkers in Mullet Species (LIZA SP.) From a Contaminated Coastal Lagoon (Ria De Aveiro, Portugal)

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

Depurated fish as an alternative reference for field-based biomarker monitoring

The whole of the Swan-Canning Estuary, in the south-west of Australia, is impacted by human activity, and the selection of a local reference site to assess the impact of environmental contamination on the health of biota is not possible. To determine whether fish depurated under laboratory conditions could be used as an alternative to a reference site; adult black bream (Acanthopagrus butcheri) were collected from the estuary and maintained in clean water (S24) for 3 months. A suite of biomarkers of fish health were assessed, and the results were compared with field-captured black bream from three sites within the estuary (Ascot, Claisebrook, and Riverton). Comparisons of a subset of biomarkers were also made between hatchery-bred juvenile fish and the depurated fish. Biomarker levels were up to 3.8 times higher in field captured fish compared with depurated fish, while DNA integrity was lower. EROD activity was comparable in the hatchery-bred black bream to the depurated fish while s-SDH levels were two times higher in the hatchery fish. From the results obtained, field-captured black bream depurated for 3 months are suitable to determine reference/baseline levels for biomarker of health studies in estuarine environments.     

Spatial distribution and source identification of persistent pollutants in marine sediments of Hong Kong

A data matrix, obtained during a 3-year monitoring period (2007–2009) from 45 sampling sites in Hong Kong marine, was subjected to determine the spatial characterization and identify the sources of main pollutants. Indicator analyses indicated that polycyclic aromatic hydrocarbons (PAHs), nickel, manganese, and arsenic (As) were at safe levels. Five heavy metals (zinc, lead, cupper, cadmium, chromium (Cr)) were moderate to severe enrichment at some sites. Inner Deep Bay and Victoria Harbor were considered as hot spots for PAHs and the heavy metals, while Tolo Harbor was highly polluted by the heavy metals. Cluster analysis classified the 45 sampling sites into three groups, representing different pollution levels. Principal component analysis/factor analysis identified four principal components (PCs) and explained 84.9 % of the total variances, standing for persistent pollution, N factor, P and Cr factor, and As factor, respectively. Group A was highly polluted by persistent pollution, group B was the less polluted group, and subgroup B1 was less affected by PC3 and PC4 than subgroup B2. Group C, considered as the moderately polluted group, was greatly affected by N factor or persistent pollution, while subgroup C2 received more N pollution than subgroup C1.     

The polycyclic aromatic hydrocarbon and geochemical biomarker composition of sediments from sea lochs on the west coast of Scotland

Sediments from twelve sea lochs on the west coast of Scotland were analysed for parent and branched 2- to 6-ring polycyclic aromatic hydrocarbons (PAHs), n-alkanes and geochemical biomarkers (triterpanes). Where possible at least fourteen sediment samples were collected at random from each sea loch. All sea lochs were remote, most had limited industrial and urban inputs, although all had fish farms. Four lochs had moderate total PAH concentrations and eight lochs had high total PAH concentrations. Total PAH concentration was related to organic carbon content and particle size distribution, with sandier sediments having lower PAH concentrations. The highest total PAH concentrations, normalised for organic carbon, were in Loch Linnhe and Ballachulish Bay (Loch Leven), close to an aluminium smelter. PAH concentration ratios showed that pyrolysis was the main source of PAHs in most lochs. Only sediments from Loch Clash showed evidence of petrogenic input based on their geochemical biomarker (triterpane and sterane) and n-alkane profiles. PAH profiles were similar across lochs apart from Loch Linnhe and Ballachulish Bay, which had a greater proportion of heavy parent PAHs. West coast sediments had a smaller proportion of heavy PAHs than sediments collected from voes in Shetland and a smaller proportion of alkylated PAHs relative to sediments collected from coastal waters around Orkney.     

Effects of Acid Rock Drainage on Stocked Rainbow Trout (Oncorhynchus mykiss): An In-Situ, Caged Fish Experiment

In-situ caged rainbow trout (Oncorhynchus mykiss) studies reveal significant fish toxicity and fish stress in a river impacted by headwater acid rock drainage (ARD). Stocked trout survival and aqueous water chemistry were monitored for 10 days at 3 study sites in the Snake River watershed, Colorado, U.S.A. Trout mortality was positively correlated with concentrations of metals calculated to be approaching or exceeding conservative toxicity thresholds (Zn, Mn, Cu, Cd). Significant metal accumulation on the gills of fish stocked at ARD impacted study sites support an association between elevated metals and fish mortality. Observations of feeding behavior and significant differences in fish relative weights between study site and feeding treatment indicate feeding and metals-related fish stress. Together, these results demonstrate the utility of in-situ exposure studies for stream stakeholders in quantifying the relative role of aqueous contaminant exposures in limiting stocked fish survival.