Detection of micronucleus and abnormal nucleus in erythrocytes from the gill and kidney of Labeo bata cultivated in sewage-fed fish farms.

Determination of genotoxic effect in fish, micronucleus test as well as the study of the abnormal shape of nuclei is a suitable measure, in which the presence or absence of genotoxins can be detected in water. In the present study, micronuclei and abnormal nuclei frequencies were scored in the gill and kidney erythrocytes of the fish Labeo bata grown in the sewage-fed fish farms of East Calcutta wetlands. Three experimental sites were chosen, namely, Bantala, Chowbaga and Chingrihata (basically these sites have sewage-fed fishponds), which were compared with fishponds of no sewage influence as the control area. Highly significant differences (P<0.001) were noticed for micronucleus frequencies in the gill and kidney erythrocytes of experimental fishes, where kidney erythrocytes showed an increased value than gill erythrocytes without any statistical differences. The frequencies of nuclear abnormalities such as necrotic cells, apoptotic cells, notch nucleated cells and binucleated cells were also counted separately for gill and kidney erythrocytes, in which significantly (P<0.001, P<0.01, P<0.05) increased values were obtained in comparison to control populations. These genotoxicity results confirmed that the sewage-fed ponds contain genotoxic metals such as Cr, Zn, Cu, Pb, Mn, Fe through wastewater and sludge because of the direct use of sewage water without pretreatment which may lead to health risks among humans through chronic consumption of fish from these experimental fish ponds. Other vertebrates grown in sewage-fed ponds may also suffer a certain amount of genotoxic substances.     

In vivo genotoxicity of mercury chloride and lead acetate: Micronucleus test on acridine orange stained fish cells.

The genotoxic effects of mercury chloride and lead acetate were evaluated in vivo using the micronucleus (MN) assay on acridine-orange (AO) stained peripheral blood erythrocytes, gill and fin epithelial cells of Carassius auratus auratus. Fish were exposed to three different concentrations of mercury chloride (MC) (1 microg/, 5 microg/L and 10 microg/L) and lead acetate (LA) (10 microg/L, 50 microg/L and 100 microg/L) for 2, 4 and 6 days. A single dose of 5 mg/L cyclophosphamide was used as a positive control. In addition to micronuclei, nuclear buds (NBs) were assessed in the erythrocytes. The ratio of polychromatic and normochromatic erythrocytes (PCE/NCE) in peripheral blood was also evaluated to assess cytotoxicity. MN frequencies in all three tissues were elevated in fish exposed to both LA and MC. However, NBs showed different sensitivity to metal treatments. MN frequencies in both control and treated fish were highest in gill cells and generally lower in erythrocytes and fin cells. PCE/NCE rations decreased in relation to MC and LA treatments. The results of this study indicate that LA and MC have genotoxic and cytotoxic damage in fish and confirmed that AO staining is a suitable technique for in vivo MN test in fish.     

Genotoxicity assessment of carp (Cyprinus carpio L.) fingerlings by tissue DNA damage and micronucleus test, after environmental exposure to fenitrothion

The purpose of this study was to evaluate the adverse effects of sublethal doses of fenitrothion, an organophosphothionate insecticide on brain, gill, liver, and muscle tissues as a ratio of 8-OHdG to dG to indicate the DNA damage and erythrocyte micronucleus frequency for genotoxicity of carp (Cyprinus carpio L.) fingerlings. In our study, the mean weights and lengths of the fish (n?=?4-12) were 31.13?±?14.24?g and 12.53?±?1.41, respectively. Before the experiment, fish were maintained in aerated dechlorinated tap water at 21.8?±?1 °C and fed daily with commercial feed at a rate of 2% of their body weights. Experiments were conducted under static conditions in the aquaria. Technical grade (95%) fenitrothion was diluted in acetone to give a dosing solution of 10?mg/L. The increased lesions/10? DNA bases (p?相似文献   

Induction of micronuclei and nuclear abnormalities in Oreochromis niloticus following exposure to petroleum refinery and chromium processing plant effluents

The genotoxic effects of effluents from a petroleum refinery and a chromium processing plant were evaluated in Oreochromis niloticus (Pisces: Perciformes) using the micronucleus test. Fish were exposed to different concentrations (5, 10 and 20%, v/v) of the effluents for 3, 6 and 9 days. Micronucleus analyses were carried out on gill epithelial cells and peripheral blood erythrocytes. Nuclear abnormalities other than micronuclei, considered as genetic damage indicators, were also evaluated on erythrocytes. Cyclophosphamide at a single dose of 4 mg/L was used as a positive control. The results of this study showed that both effluents had genotoxic potential. On the other hand, the level of genetic damage induced by petroleum refinery effluent was considerably higher than that of chromium processing plant effluent. Our results further indicate that nuclear abnormalities other than micronuclei, such as blebbed and lobed nuclei, may also be used as indicators of genotoxic damage.     

Genotoxicity assessment of carp (Cyprinus carpio L.) fingerlings by tissue DNA damage and micronucleus test,after environmental exposure to fenitrothion

The purpose of this study was to evaluate the adverse effects of sublethal doses of fenitrothion, an organophosphothionate insecticide on brain, gill, liver, and muscle tissues as a ratio of 8-OHdG to dG to indicate the DNA damage and erythrocyte micronucleus frequency for genotoxicity of carp (Cyprinus carpio L.) fingerlings. In our study, the mean weights and lengths of the fish (n?=?4–12) were 31.13?±?14.24?g and 12.53?±?1.41, respectively. Before the experiment, fish were maintained in aerated dechlorinated tap water at 21.8?±?1°C and fed daily with commercial feed at a rate of 2% of their body weights. Experiments were conducted under static conditions in the aquaria. Technical grade (95%) fenitrothion was diluted in acetone to give a dosing solution of 10?mg/L. The increased lesions/10⁶ DNA bases (p?<?0.05) of liver tissue of exposure group (0.49?±?0.18) was observed when compared to control group (0.28?±?0.30). There was not any significant differences between brain tissues, no damage were detectable in gill and muscle tissues of control groups, and in exposure groups altered levels of damage were detected for gill (0.06?±?0.05) and muscle (0.16?±?0.21) tissues. The increased micronucleus frequencies (%) in erythrocytes of carp following the exposure to 48?h fenitrothion (6.43?±?3.89; p<0.05) was observed when compared to control group (1.29?±?1.03). The available data indicate that there is still lack of well-established dose–response relationships between occupational or environmental exposures and the induction of 8-OHdG. Such biomarkers may be used in assessing adverse/toxic effects of pesticides as environmental stressors.     

Comparative evaluation of the cytotoxicity sensitivity of six fish cell lines to four heavy metals in vitro.

To establish the potential use of cell cultures as a simple and sensitive biological tool to detect environmental pollutants, six cell lines established from several fish species including GCF (grass carp fins), CIK (Ctenopharyngodon idellus kidney), EPC (epithelioma papulosum cyprini), CCO (channel catfish ovary), BB (brown bullhead caudal trunk) and FHM (fathead minnow muscle) were tested and compared for their cytotoxic sensitivity to four heavy metals: cadmium (Cd), chromium (Cr), zinc (Zn), and copper (Cu). Following a 24-h exposure to these metal salts at selected concentrations, test cells were characterized by morphology, viability and proliferation. Our results indicate that all these metal salts are cytotoxic to these fish cell lines, but at varied levels. Calculated inhibitory concentration (IC(50)) values revealed that the cytotoxicity of Cr and Cd was significantly more pronounced than that of the other two metal salts. Comparative analysis of these fish cell lines showed that C. idellus kidney (CIK) cells are the most sensitive cell line to copper, epithelioma papulosum cyprini (EPC) cells are more sensitive than other cells to Cr and Zn, while channel catfish ovary (CCO) cell line is the most sensitive one to Cd. In conclusion, CIK, EPC and CCO could potentially be sensitive bio-indicators for the initial monitoring and assessment of acute cytotoxicity of heavy metals in the aquatic environment.     

Mutagenic and genotoxic assessment of atrazine-based herbicide to freshwater fish Channa punctatus (Bloch) using micronucleus test and single cell gel electrophoresis

The mutagenic and genotoxic effect of 'Rasayanzine', an atrazine herbicide, was carried out in fish Channa punctatus using micronucleus and single cell gel electrophoresis (SCGE) assays. Three sub-lethal concentrations of the test material viz., SL-I (1/5th LC(50)=～8.48 mgL(-1)), SL-II (1/8th LC(50)=～5.30 mgL(-1)) and SL-III (1/10th LC(50)=～4.24 mgL(-1)) were calculated using LC(50) value and the fish specimens were exposed to these concentrations. Erythrocyte and gill cells were sampled on day 0, 1, 3, 5, 7, 14, 21, 28 and 35 of exposure for assessment of micronuclei induction in blood erythrocytes and DNA damage using SCGE assay in both blood erythrocytes and gill cells. Significant effects (p<0.01) for both concentration and time of exposure were observed in treated fish. Micronuclei induction in erythrocytes was highest (8.4 mgL(-1)) on day 7 of exposure. The highest level of DNA damage in the SCGE was observed in both tissues on day 5 at all concentrations followed by gradual non-linear decline. This study further confirmed that the micronucleus and SCGE assays are useful in determining potential genotoxicity of water pollutants and might be appropriate as part of monitoring program.     

Differential metallothionein induction patterns in three freshwater fish during sublethal copper exposure

We assessed whether fish that tolerate higher levels of Cu exposure have a higher capacity to induce metallothionein (MT) synthesis than other, more sensitive, fish species. Furthermore, we examined if a correlation could be found between tissue Cu accumulation and MT levels. Cu accumulation and MT concentrations in gill, liver, kidney and muscle of rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gibel carp (Carassius auratus gibelio) were measured during a 1 week exposure to a sublethal Cu (1 microM). Different patterns were observed for the three species regarding Cu accumulation as well as MT induction. Virtually no Cu accumulation was seen in rainbow trout gill, while in both cyprinid species gill Cu levels increased three- to four-fold. Cu accumulated fast in common carp (within the first day), but slow in gibel carp (1 week). Gill MT induction was obvious in gibel carp only, with an increase of 156% after 1 week of exposure. Liver accumulated most Cu in rainbow trout (235% increase) and common carp (144% increase), with Cu levels in liver being significantly higher in rainbow trout compared to the carp species from the start. MT induction was pronounced in common carp liver only (138% increase). In gibel carp liver, there was no clear Cu accumulation or MT induction. In contrast, gibel carp was the only species to show Cu accumulation in kidney after 3 days of exposure (83% increase), after which levels returned to normal. Concomitantly, gibel carp kidney was also the only kidney tissue to show MT induction (192-195% increase after 3 and 7 days). In common carp, a significant decrease of kidney MT levels was observed from day 1 onwards. In muscle, Cu accumulation was clear for the two cyprinid species (three- to four-fold increase) but not for rainbow trout. Of the species studied, gibel carp is the most resistant to copper polluted environments, and showed a positive significant relationship between tissue copper concentrations and MT levels in gill, liver and muscle tissues. Common carp showed an intermediate response, with significant correlations in liver and muscle tissue. In contrast, we found low MT induction in rainbow trout, the most sensitive species, and no correlation at all between MT concentrations and tissue copper contents. Possibly, the regulatory capacity for copper homeostasis was exceeded in rainbow trout, and MT synthesis inhibited.     

Genotoxicity and cytotoxicity response to environmentally relevant complex metal mixture (Zn,Cu, Ni,Cr, Pb,Cd) accumulated in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>). Part I: importance of exposure time and tissue dependence

Health impact of metal mixture at environment realistic concentrations are difficult to predict especially for long-term effects where cause-and-effect relationships may not be directly obvious. This study was aimed to evaluate metal mixture (Zn—0.1, Cu—0.01, Ni—0.01, Cr—0.01, Pb—0.005 and Cd—0.005?mg/L, respectively for 1, 2, 4, 7, 14 and 28 days at concentrations accepted for the inland waters in EU) genotoxicity (micronuclei, nuclear buds, nuclear buds on filament), cytotoxicity (8-shaped nuclei, fragmented-apoptotic erythrocytes), bioaccumulation, steady-state and the reference level of geno-cytotoxicity in hatchery-reared Atlantic salmon tissues. Metals accumulated mostly in gills and kidneys, to the lesser extent in the muscle. Uptake of metals from an entire mixture in the fish for 14 days is sufficient to reach steady-state Cr, Pb concentrations in all tissues; Zn, Cu—in kidneys and muscle, Ni—in liver, kidneys, muscle and Cd—in muscle. Treatment with metal mixture significantly increased summed genotoxicity levels at 7 days of exposure in peripheral blood and liver erythrocytes, at 14 days of exposure in gills and kidney erythrocytes. Significant elevation of cytotoxicity was detected after 2 and 14 days of exposure in gills erythrocytes and after 28 days—in peripheral blood erythrocytes. The amount of Cu, Cr, Pb and Cd accumulated in tissues was dependent upon duration of exposure; nuclear buds, 8-shaped nuclei frequencies also were dependent upon duration of exposure. This study indicates that metals at low levels when existing in mixture causes significant geno-cytotoxicity responses and metals bioaccumulation in salmon.     

Evaluation of genotoxicity from Nilufer Stream (Bursa/Turkey) water using piscine micronucleus test

In the present study, the in vivo micronucleus (MN) test in fish erythrocytes was used to evaluate the genotoxic potentials of water samples collected from four different sites along the Nilufer Stream which empties into the Marmara Sea on the north-west of Turkey. Nilufer Stream receives discharges of industrial and domestic wastes resulting from industrialization and urbanization activities in Bursa city. Nile tilapias (Oreochromis niloticus) were exposed to collected water samples under laboratory conditions for 3 and 6 days. Micronuclei analyses were carried out in peripheral blood erythrocytes. In addition to micronuclei, other nuclear abnormalities (NAs) such as bi-nucleated cells and binuclei with nucleoplasmic bridge and cells with blebbed, notched and lobbed nuclei, were assessed in the erythrocytes. Chemical analyses were also carried out in the water samples to assess the presence of major pollutants. MN and NA frequencies were significantly elevated in fishes exposed to water from polluted areas compared to those exposed to clean water sample. The results of this study indicate that Nilufer Stream is contaminated with potential genotoxic chemicals and the genotoxicity is possibly related with the industrial, agricultural and domestic activities.     

In vivo genotoxicity evaluation of atrazine and atrazine-based herbicide on fish Carassius auratus using the micronucleus test and the comet assay

Atrazine is a selective triazine herbicide used to control broadleaf and grassy weeds mainly in corn, sorghum, sugarcane, pineapple, and other crops, and in conifer reforestation planting fields. It has been showed that atrazine is one of the most frequently detected pesticides in agricultural streams and rivers, over the past two decades. Although the toxic properties of atrazine are well known, the data on the genotoxic effects of atrazine on aquatic organisms are rather scarce. Thus, in the present study we aimed to evaluate the genotoxic effects of atrazine and an atrazine-based herbicide (Gesaprim®) on a model fish species Carassius auratus L., 1758, (Pisces: Cyprinidae) using the micronucleus test and the comet assay in peripheral blood erythrocytes. Fish were exposed to 5, 10 and 15 μg/L atrazine and to its commercial formulation for 2, 4 and 6 days. Ethyl methane sulfonate (EMS) at a single dose of 5 mg/L was used as positive control. Our results revealed significant increases in the frequencies of micronuclei and DNA strand breaks in erythrocytes of C. auratus, following exposure to commercial formulation of atrazine and thus demonstrated the genotoxic potential of this pesticide on fish.     

In vivo genotoxicity testing of the amnesic shellfish poison (domoic acid) in piscine erythrocytes using the micronucleus test and the comet assay

Domoic acid (DA) is a neurotoxic amino acid naturally produced in the marine environment by some diatom species belonging to the genus Pseudo-nitzschia. Although the neurotoxic properties of DA have been demonstrated, very little is known about in vivo genotoxicity of DA on aquatic organisms. In the present paper, an in vivo study on the genotoxic effects of domoic acid was carried out on a fish, Oreochromis niloticus, using the micronucleus test and the comet assay. The fish were exposed to three doses of domoic acid (1, 5 and 10 microg/g body weight) by intracoelomic injections. Ethyl methane sulphonate at a single dose of 5mg/l was used as positive control. Analysis of micronuclei, nuclear abnormalities and DNA damage were carried out on peripheral erythrocytes sampled 24, 48 and 72 h post-treatment. Our results revealed significant increases in the frequencies of micronuclei, nuclear abnormalities as well as DNA strand breaks and thus demonstrated the genotoxic potential of DA on fish.     

Metal concentrations in tissues of the Black Sea fish Mugil auratus from Sinop-Icliman,Turkey

Levels of five heavy metals [copper (Cu), lead (Pb), cadmium (Cd), chromium (Cr) and nickel (Ni)] were evaluated in liver and muscle tissues of fish (Mugil auratus) collected from the Black Sea at Sinop-Icliman, Turkey. Sampling and analysis methods are described. Variations of heavy metal concentrations with seasons are discussed. Cr and Ni concentrations were below the limits of detection (< 0.05 and 0.1 microg/g dry weight) in all tissues and seasons. Cu, Pb and Cd were detected within these limits, as microg/kg dry weight, in liver tissue: 0.49-1.30, 0.60-1.21 and 0.15-0.50, and in muscle tissue: 0.30-1.00, 0.57-1.12 and 0.10-0.40, respectively. Cu, Pb and Cd concentrations in these tissues were elevated and the highest heavy metal concentrations were found in the liver. While Cu, Pb and Cd concentrations were highest in fish tissues collected in August 2000, the lowest concentrations of these metals were observed in fish tissues collected in May 2000. Pb had the highest level observed in fish tissues. According to the Turkish Food Codex Regulation's residue limits, the cadmium level determined in fish tissues was high (0.1 microg/g) and the lead level, especially in liver tissue, was high in August (1 microg/g), while other metals (Cu, Cr and Ni) were within the maximum residue limits.     

A novel response of wild-type duckweed (Lemna paucicostata Hegelm.) to heavy metals

The responses of wild-type Lemna paucicostata, used as the experimental stock, to seven heavy metals were investigated with a hydroponic culture in a culture facility. The results showed that six of the metals could induce L. paucicostata to release daughter fronds from the mother frond before maturity, resulting in breakup of the colony. This phenomenon is novel and has not been reported before. Six of the heavy metals investigated-copper, cadmium, nickel, zinc, mercury, and chromium-could induce this response, but sensitivity and response of the plant varied. When Cu(2+) at a concentration of 0.2 micromol/L was added, the response was significantly different from that of the control after a 2-h exposure. The response to cadmium (0.8 micromol/L) was significantly different after a 6-h exposure; to zinc (10 micromol/L) and nickel (5 micromol/L), after 5 and 7 h, respectively. But for lead, even at a concentration of 80 micromol/L for 96 h, no colony breakup was induced. The response to heavy metals occurred within 24 h; increasing the exposure by another 24 h had nearly no additional effect on the colony breakup. When the response was over, the number of colonies with fronds varied with the inducement capacity of the metal. Nutrients could affect the sensitivity of L. paucicostata to some heavy metals, such as mercury, chromium, zinc, and nickel, but not to copper and cadmium. Sensitivity was also changed when CuCl(2) was used as the copper source instead of CuSO(4). These results indicate that some heavy metals could serve as a toxicant for inducing L. paucicostata to release daughter fronds in advance. This change might involve a stress response such as ethylene production. Also discussed is the possibility of this response as a biomarker in a bioassay.     

Assessment of genotoxic and mutagenic potential of hexavalent chromium in the freshwater fish Labeo rohita (Hamilton, 1822)

The present study was undertaken to investigate the genotoxicity and mutagenicity of sublethal concentrations of hexavalent chromium (potassium dichromate) in the Indian major carp, Labeo rohita. The 96?h LC₅₀ value of potassium dichromate estimated was 118?mg?L^?1 by probit analysis using SPSS (version 16.0) software. Based on 96?h LC₅₀ value, three sublethal test concentrations of potassium dichromate (29.5, 59.0 and 88.5?mg?L^?1) were selected and specimens were exposed in vivo to these test concentrations for 96?h. The mutagenic and genotoxic effects of potassium dichromate were evaluated in gill and blood cells using micronucleus (MN) test and comet assay. In general, significant (p?相似文献   

Acute toxicity impacts of hexavalent chromium on behavior and histopathology of gill, kidney and liver of the freshwater fish, Channa punctatus (Bloch)

Alteration in behavioral patterns and histopathology of gill, kidney and liver were studied in Channa punctatus, after acute exposure to hexavalent chromium. The 96h LC(50) of chromium salt, potassium dichromate was determined to be 41.75mg/L. The exposed fish displayed erratic swimming and became lethargic. The changes in gills were characterized by epithelial hyperplasia, lamellar fusion, oedema, epithelial lifting, epithelial necrosis and desquamation, aneurism as well as curling of secondary lamellae. Hypertrophy of epithelial cells of renal tubules with reduced lumens, atrophy of the renal tubules, glomeruli contraction in the Bowman's capsules and necrosis of haematopoietic tissues were observed in the trunk kidney of exposed fish. The interrenal cells of the head kidney exhibited distinct hypertrophy and vacuolization. The liver hepatocytes showed cytoplasmic vacuolization with the lateral nuclei arrangement. Hepatocytes atrophy and increase in sinusoidal space were also observed. The result showed that acute chromium toxicity severely affects the vital organs and normal behavior which may be deleterious for fish populations.     

Sublethal toxicity of esbiothrin relationship with total antioxidant status and in vivo genotoxicity assessment in fish (Cyprinus carpio L., 1758) using the micronucleus test and comet assay

Esbiothrin, synthetic pyrethroid with quick activity against insects, is widely used against household pests and in public health. Despite widespread use, data on ecotoxicity and genotoxic effects are extremely scarce. The aim of the present study is to evaluate the genotoxic potential of esbiothrin on a model fish species Cyprinus carpio L., 1758 (Pisces: Cyprinidae, koi) using the micronucleus test and comet assay in peripheral blood erythrocytes. Effects of two sublethal exposure concentrations on plasma total antioxidant status (TAS mmol/L), and Hct values were examined. On the basis of the 96 h LC₅₀ data from U.S. EPA ecotox database (32 μg/L) two sublethal exposure concentrations (5 and 10 μg/L) were used together with ethyl methanesulfonate (EMS) (5 mg/L) as positive control. Five fish were used for each dose/duration group (24, 48, and 72 h) under controlled laboratory conditions. The fish showed behavioral changes at the higher dose. Plasma TAS (mmol/L) levels decreased in 24 h; an increase was observed slightly for 48 and obviously for 72 h in both exposure doses. Similarly, hematocrit (Hct) values differed between exposure duration but no significant differences in mean values were found between groups of the same exposure time. The general trend was a rise after 48 h, which decreased afterwards. Our results revealed significant increases in the frequencies of micronuclei and levels of DNA strand breaks and thus demonstrated the genotoxic potential of this pesticide on fish, a nontarget organism of the aquatic ecosystem. To our knowledge this is the first study to report observable genotoxic effects of esbiothrin on fish. © 2011 Wiley Periodicals, Inc. Environ Toxicol 28:644–651, 2013.     

Inhibition of ATPase activity in the freshwater fish Labeo rohita (Hamilton) exposed to sodium cyanide

Present study concerns the effect of sodium cyanide on the Indian major carp, Labeo rohita. Fishes were exposed to lethal (0.32?mg/L) and sublethal (0.064?mg/L) concentrations of sodium cyanide. The effect of intoxication was studied on Na(+)K(+)ATPase, Mg(+2)ATPase and Ca(2+)ATPase in various physiological tissues (gill, liver, and muscle) at the end of 1, 2, 3 and 4 days of lethal and 5, 10 and 15 days of sublethal exposure periods. Sodium cyanide induced significant inhibitory effects on the ATPase activity of the fish. Inhibition of the ATPase blocked the active transport system of the gill epithelial as well as chloride cells, and thus altered the osmo-regulatory mechanism of the fish. The value of the measured responses as an indicator of stress caused by water contamination discussed. The results confirm that ATPase levels significantly decreased in treated fish, indicating that ATPases could be used as sensitive and useful biomarkers for cyanide pollution.     

Influence of urbanization of the western coast of the United Arab Emirates on trace metal content in muscle and liver of wild Red-spot emperor (Lethrinus lentjan).

We hypothesized that increased ambient concentrations of metals, as a consequence of escalating urbanization and industrialization of the Gulf region will respond in increased contamination of edible fish species. In this study, we report concentrations of chromium, manganese, cobalt, copper, zinc, arsenic, cadmium, mercury and lead in meat and liver of wild Red-spot emperor (Lethrinus lentjan) from three sampling points at the UAE coast. Analysis was performed by the ICP-MS/microwave digestion. Our study has shown that meat and liver metal content was significantly higher in areas with higher industrial activity, although metal values did not exceed permitted levels of fish for human consumption.