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.     

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.     

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.     

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.     

Tissue distribution and temperature-dependence of Anguilla anguilla L. EROD activity following exposure to model inducers and relationship with plasma cortisol, lactate and glucose levels

Anguilla anguilla L. ethoxyresorufin-O-deethylase (EROD) elevation by 2.7 microM beta-naphthoflavone (BNF) 3 days water exposure, or 4 mg/kg ip exposure, was studied in four different organs--liver, kidney, gills, and intestine. The results demonstrated a significant increase in liver EROD activity for the two previous conditions, whereas kidney EROD activity only increased during the intraperitoneal exposure. A. anguilla was also exposed during 8, 16, and 24 h to water contaminated with 2.7 microM BNF or benzo[a]pyrene (BaP) at 20 degrees C and 25 degrees C. Both compounds significantly increased liver EROD activity from 8 up to 24 h. There was no significant difference in liver EROD activity elevation by both compounds, either at 20 degrees C or 25 degrees C. Liver EROD activity was demonstrated to be one of the first warning systems concerning the presence of polycyclic aromatic hydrocarbons (PAHs) in water. A. anguilla 3 h exposure to diesel oil water-soluble fraction (DWSF) significantly increased plasma cortisol and significantly decreased plasma lactate. A prolonged exposure beyond 3 h, i.e. 4 h, 2, 3, 4, and 6 days to the previous conditions demonstrated a significant liver EROD activity elevation from Day 2 up to 6, and a significant increase in erythrocytic nuclear abnormalities (ENA) at Day 6.     

Fly ash leachate induces oxidative stress in freshwater fish Channa punctata (Bloch)

Oxidative stress inducing potential of fly ash leachate (FAL) was studied in a freshwater fish, Channa punctata (Bloch). Fish were exposed to fly ash leachate for 24 h and lipid peroxidation (LPO) was studied as a marker of oxidative stress. Catalase (CAT), glutathione S-transferase (GST) activities and levels of reduced glutathione (GSH) were also estimated in the exposed fish. FAL (1 ml/l) induced LPO in all the organs and most prominent response was in the gill. It also caused induction of enzymes and glutathione. Liver showed highest level of induction of enzyme activities. The results of this study demonstrate that fly ash constituents have potential to induce oxidative stress in fish and gills are the most vulnerable organs. It is also suggested that in case of exposure to FAL, along with LPO antioxidant defense is also activated to counteract the reactive oxygen species (ROS) at least partly in the initial stages of exposure.     

Free radicals and chromosome aberrations in leaves of woody plants as a test system for the genotoxicity of the urban environment

A new method for estimating the genotoxicity of the urban environment is proposed. The method is based on the analysis of the lipid peroxidation (LPO) rate in leaves of woody plants. The mutagenicity of the urban environment was estimated using standard test systems, including plant root meristem (kidney beans germinated in water with suspended particles collected at different points along the Temernik River) and the Ames test (silt samples were tested). Sensitivities of elm, willow, and poplar leaf meristem cells to unidentified environmental mutagenes were compared. Frequencies of chromosome aberrations (CAs) in leaves of woody plants growing at different points along the Temernik River within the Rostov-on-Don city districts differed from those in the control zone. The frequency of CAs was found to correlate with the concentration of LPO products. This allows a biochemical estimation of the malonic dialdehyde (MDA) concentration in woody plants to be used as an express test for the mutagenicity of environmental factors. An advantage of the proposed test is the possibility of monitoring the level of unidentified environmental mutagens over many years, independently of its sporadic temporal variations.     

Effects of moderate pollution on toxic and trace metal levels in calves from a polluted area of northern Spain

This study evaluated the contribution of anthropogenic pollution to toxic metal residues (Cd, Pb and As) in cattle in an industrialized area of Asturias (northern Spain), and investigated possible implications of toxic metal exposure for metabolism of essential metal elements (Cu, Zn, Fe, Mn). Samples of liver, kidney, muscle and blood from animals aged 9-12 months were obtained from an industrialized area (78 calves) and from a rural area (92 calves). Samples were acid-digested, and levels of metals determined by atomic absorption spectrophotometry. Cadmium and lead contents in the liver and kidney were moderately and significantly higher in calves from the industrialized area (Cd: liver 29.6, kidney 161; Pb: liver 38.1, kidney 38.3 microg/kg wet weight) than in calves from the rural area (Cd: liver 22.9, kidney 96.4; Pb: liver 20.7, kidney 15.9 microg/kg kg wet weight). Although these toxic metals were only moderately raised, our results suggest that they may nevertheless have interfered with trace element metabolism, as reported previously for severely polluted regions. Notably, copper levels in calves from the industrialized area were generally low, and nearly half of these animals showed tissue copper levels indicating risk of copper deficiency.     

Global genomic methylation levels in the liver and gonads of the three-spine stickleback (Gasterosteus aculeatus) after exposure to hexabromocyclododecane and 17-beta oestradiol

Alteration of DNA methylation is a major epigenetic mechanism associated with the effects of nongenotoxic carcinogens. We evaluated the effects of two environmental pollutants, hexabromocyclododecane (HBCD), 17-beta oestradiol (E(2)) as well as 5-aza 2' deoxycytidine (5AdC) on global DNA methylation levels (5-methyl 2' deoxycytidine) in the liver and gonads of the three-spine stickleback (Gasterosteus aculeatus). HBCD at 30 and 300 ng/L of water did not produce statistically significant differences in global genomic methylation in liver of female stickleback. On the other hand, the methylation inhibitor, 5-aza-2'-deoxycytidine, significantly lowered hepatic global methylation levels in these fish by 14% (P<0.05). The naturally occurring oestrogen, 17-beta oestradiol (E(2)) at 100 ng/L also decreased global DNA methylation levels in female liver but this effect was not statistically significant. In contrast, both E(2) and 5AdC caused statistically significant (P<0.001 and P<0.01 respectively) global genomic hypermethylation in the gonads of male sticklebacks although the increase seen in the female gonads was not statistically significant. The male gonad effect though unexplained may potentially be an indirect response to hypomethylation in other tissues (such as the liver) and may have important implications regarding oestrogenic effects in fish. The contrasting effects of HBCD and E(2) on global DNA methylation in stickleback should contribute to the integrated risk assessment of these environmental chemicals.     

The toxicity of copper contaminated soil using a gnotobiotic Soil Multi-species Test System (SMS)

A gnotobiotic multi-species study was designed to consist of a food-web of soil-dwelling animals. The food-web was exposed to five concentrations copper (Cu) spiked soil for three exposure durations i.e. 28, 56 and 84 days. Based on multivariate analysis the food-web was significantly affected by Cu exposure at and above 300 mg Cu kg(-1) soil (lowest tested concentration). The number of animals present in the 2500 mg Cu kg(-1) (highest tested concentration) was at all sampling occasions below the starting point level. Based on analysis of the individual species the lowest 10% effect concentration (EC10) observed was 50 mg Cu kg(-1) soil, for Enchytraeus crypticus. Using the EC10 for the individual species the HC5 (Hazard Concentration at the 5% level) was estimated to be between 25 and 36 mg Cu kg(-1) soil, depending on the exposure duration. A similar experiment but using a reduced design was performed employing soil contaminated with Cu in the field more than 80 years ago. The trend in the field-contaminated soil was similar to that observed for the spiked soil.     

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.     

Internal distribution of uranium and associated genotoxic damages in the chronically exposed bivalve Corbicula fluminea

Uranium (U) internal distribution and involved effects in the bivalve Corbicula fluminea have been studied after direct chronic exposure (90 d, 10 μg.L-1). U distribution was assessed at the subcellular level (Metal Rich Granules -MRG-, pellets and cytosol fractions) in two main organs of the bivalve (gills and visceral mass). Micro-localisation was investigated by TEM-EDX analysis in the gills epithelium. DNA damage in gill and hemolymph samples was measured by the Comet assay. The 90-d exposure period led to a significant increase of U concentration in gills over time (×5) and a large U quantity in subcellular granules in gills. Finally, a significant increase (×2) in DNA damage was noted in exposed gills and haemocytes. This study shows that the accumulation levels and consequently the potential toxicity cannot be successfully predicted only on the basis of concentration in water or in tissues and subcellular fractions after chronic exposure.     

Occurrence and fate of hormone steroids in the environment

Hormone steroids are a group of endocrine disruptors, which are excreted by humans and animals. In this paper, we briefly review the current knowledge on the fate of these steroids in the environment. Natural estrogenic steroids estrone (E1), 17beta-estradiol (E2) and estriol (E3) all have a solubility of approximately 13 mg/l, whereas synthetic steroids 17alpha-ethynylestradiol (EE2) and mestranol (MeEE2) have a solubility of 4.8 and 0.3 mg/l, respectively. These steroids have a moderate binding on sediments and are reported to degrade rapidly in soil and water. Estrogenic steroids have been detected in effluents of sewage treatment plants (STPs) in different countries at concentrations ranging up to 70 ng/l for E1, 64 ng/l for E2, 18 ng/l for E3 and 42 ng/l for EE2. E2 concentrations in river waters from Japan, Germany, Italy and the Netherlands ranged up to 27 ng/l. In addition, E2 concentrations ranging from 6 to 66 ng/l have also been measured in mantled karst aquifers in northwest Arkansas. This contamination of ground water has been associated with poultry litter and cattle manure waste applied on the land. Although hormone steroids have been detected at a number of sources worldwide, currently, there is limited data on the environmental behaviour and fate of these hormone steroids in different environmental media. Consequently, the exposure and risk associated with these chemicals are not adequately understood.     

Genotoxic effects of occupational exposure to lead and influence of polymorphisms in genes involved in lead toxicokinetics and in DNA repair

Lead is still widely used in many industrial processes and is very persistent in the environment. Although toxic effects caused by occupational exposure to lead have been extensively studied, there are still conflicting results regarding its genotoxicity. In a previous pilot study we observed some genotoxic effects in a population of lead exposed workers. Thus, we extended our study analysing a larger population, increasing the number of genotoxicity endpoints, and including a set of 20 genetic polymorphisms related to lead toxicokinetics and DNA repair as susceptibility biomarkers. Our population comprised 148 workers from two Portuguese factories and 107 controls. The parameters analysed were: blood lead levels (BLL) and δ-aminolevulinic acid dehydratase (ALAD) activity as exposure biomarkers, and T-cell receptor (TCR) mutation assay, micronucleus (MN) test, comet assay and OGG1-modified comet assay as genotoxicity biomarkers. Lead exposed workers showed markedly higher BLL and lower ALAD activity than the controls, and significant increases of TCR mutation frequency (TCR-Mf), MN rate and DNA damage. Oxidative damage did not experience any significant alteration in the exposed population. Besides, significant influence was observed for VDR rs1544410 polymorphism on BLL; APE1 rs1130409 and LIG4 rs1805388 polymorphisms on TCR-Mf; MUTYH rs3219489, XRCC4 rs28360135 and LIG4 rs1805388 polymorphisms on comet assay parameter; and OGG1 rs1052133 and XRCC4 rs28360135 polymorphisms on oxidative damage. Our results showed genotoxic effects related to occupational lead exposure to levels under the Portuguese regulation limit of 70μg/dl. Moreover, a significant influence of polymorphisms in genes involved in DNA repair on genotoxicity biomarkers was observed.     

The amount and identity of [14C] residues in bluegills (Lepomis macrochirus) exposed to [14C] triclopyr

The level and identity of [¹⁴C] residues in bluegills (Lepomis macrochirus)_exposed to 2.5 mg/L [¹⁴C] triclopyr (3,5,6-trichloro-2-pyridinyloxyacetic acid) have been determined. The highest level of radioactivity observed in the flesh of a fish at any time point (0.13 mg/kg, calculated as equivalent mg triclopyr/kg fish) was less than 5% of the fish exposure level of 2.5 mg/L, while the maximum level in the remainder (head, skin, and viscera) was about 95% (2.33 mg/kg) of the fish exposure level, indicating no concentrating effect. The principal components observed in the fish tissues were triclopyr, 3,5,6-trichloro-2-pyridinol, 2-methoxy-3,5,6-trichloropyridine and a conjugate. These components accounted for greater than 75% of all the residues observed.     

Monitoring of environmental heavy metals in fish from Nasser Lake

Lake ecosystems are, in particular, vulnerable to heavy metal pollution. Tilapia nilotica is one of the aquatic organisms affected by heavy metals. Therefore, heavy metals Co, Cr, Cu, Fe, Mn, Ni, Sr and Zn were determined in different tissues of T. nilotica (ages 1, 1.5, 2, 2.5 and 3 years), which include the muscle, gill, stomach, intestine, liver, vertebral column and scales, to assess the fish pollution with heavy metals. In addition, the study was extended to determine these elements in the aquatic plant (Najas armeta), sediment and water of Khor E1-Ramel in Nasser Lake (Egypt). The study showed that of all the fish parts, fish liver accumulated the highest levels of Cu and Zn. Manganese presented in the intestine and stomach in the highest concentration. Scales exhibited the highest levels of Co, Cr, Ni and Sr, while the gill and vertebral column contains the lowest level of the studied elements. Heavy metals in different parts of T. nilotica differ with the fish growth and extraction rate of these elements from sediment, aquatic plant and lake water. Heavy metals under study in the edible parts of the investigated fish were in the safety permissible levels for human uses.     

Naphthalene and beta-naphthoflavone effects on Anguilla anguilla L. hepatic metabolism and erythrocytic nuclear abnormalities

The effects of a polycyclic aromatic hydrocarbon (PAH) such as naphthalene (NAP)--an environmental contaminant--and beta-naphthoflavone (BNF)--a model substance (PAH-like compound)--were investigated in European eel (Anguilla anguilla L.) over 3-, 6-, and 9-day exposure (0.1-2.7 microM). Both xenobiotics revealed to be strong biotransformation (phase I) inducers. After 3-day exposure, liver ethoxyresorufin O-deethylase (EROD) activity was significantly increased by all NAP and BNF tested concentrations. At 6 and 9 days, liver EROD activity was significantly induced mainly by the highest NAP and BNF concentrations. Liver cytochrome P450 content was significantly induced after 3-day exposure to 0.9 and 2.7 microM BNF and 9-day exposure to 0.1, 0.3 and 0.9 microM NAP. Liver alanine transaminase (ALT) activity was measured as an indicator of hepatic health condition, revealing a significant decrease after 6-day exposure to 0.9 microM BNF. Genotoxicity measured as erythrocytic nuclear abnormalities (ENA) was detected in all BNF treated fish on day 6, whereas on day 9, ENA frequencies returned to control levels, significantly decreasing at 0.9 microM BNF exposure. Immature erythrocytes (IE) frequency demonstrated a decreasing tendency along the BNF experiment and concomitantly with the above ENA response. The present experimental results elect EROD activity in A. anguilla as a useful short- to medium-term biomarker of exposure to both PAH and PAH-like compounds. However, some problems can emerge in the presence of high xenobiotic concentrations. Concerning genotoxicity, it is hypothesized that ENA response depends on different factors such as the exhaustion of the detoxification process, the balance erythropoiesis/erythrocytic catabolism and the DNA repairing capacity.     

Toxicity of zinc oxide nanoparticles in the earthworm, Eisenia fetida and subcellular fractionation of Zn

The extensive use of nanoparticles (NPs) in a variety of applications has raised great concerns about their environmental fate and biological effects. This study examined the impact of dissolved organic matter (DOM) and salts on ZnO NP dispersion/solubility and toxicity to the earthworm Eisenia fetida. To be able to better evaluate the toxicity of NPs, exposure in agar and on filter paper was proposed for enabling a comparison of the importance of different uptake routes. A dose-related increase in mortality was observed in earthworms exposed in agar with almost 100% mortality after 96 h exposure to the highest concentration (1000 mg ZnO/kg agar). Scanning electron microscopy (SEM) showed that the addition of salts enhanced the aggregation of ZnO NPs in agar and consequently affected the dissolution behavior and biological availability of the particles. On filter paper, mortality was the highest at the lowest exposure concentration (50 mg ZnO/L) and seemed to decrease with increasing exposure levels. TEM images of ZnO showed that the solubility and morphology of NPs were changed dramatically upon the addition of humic acids (HA). The subcellular distribution pattern of Zn in earthworms after 96 h exposure in agar and on filter paper showed that the Zn taken up via dietary ZnO particles (from agar) was mainly found in organelles and the cytosol while the Zn accumulated as soluble Zn from filter paper was mainly distributed in cell membranes and tissues. Antioxidant enzymatic activities (SOD, CAT, and GSH-px) were investigated in the worms surviving the toxicity tests. A slight increase of SOD activities was observed at the lowest exposure dose of ZnO (50mg/kg), followed by a decrease at 100mg/kg in the agar cubes. Activities of both CAT and GSH-Px enzymes were not significantly influenced in the worms exposed to agar, although a slight decrease at 500 and 1000 mg ZnO/kg agar was observed. A similar change trend of SOD activities was observed for the earthworms on filter paper, but a significant decrease began at a higher ZnO NP concentration of 500 mg ZnO/L. The use of soil extracts instead of deionized water (DW) to simulate a realistic exposure system significantly reduced the toxicity of the ZnO NPs on filter paper, which increases the predictive power of filter paper toxicity tests for the environmental risk assessment of NPs.     

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.     

Assessment of arsenic level in the hair of the nonoccupational Egyptian population: pilot study.

Arsenic level of hair samples of apparently healthy Egyptian was measured by means of hydride atomic absorption spectrophotometery. It ranged between 0.04 and 1.04 mg As/kg hair, about 55% of the analysed hair samples were within the range of allowable values (0.08-0.25 mg As/kg hair), but 45% were not. There were no considerable sex-related differences (0.303 and 0.292 mg As/kg hair for males and females, respectively). Different educational levels did not influence it either, when the effect of the age had been excluded. Children and adolescents proved to be more susceptible to arsenic as their mean levels (0.353 microg/g), and were significantly higher than those in the adults (0.233 microg/g). Smoking and some dietary habits had an important role in the elevation of arsenic levels among the nonoccupational Egyptian population: 60% of smokers and 66.7% of indoor passive smokers had arsenic levels >0.25 mg As/kg hair. Arsenic levels were also dependent on the kind of smoking, as hair arsenic of the subject smoking molasses tobacco was found to be significantly higher than that of cigarette smokers (0.459 and 0.209 mg As/kg hair, respectively). The frequency of meat and fish consumption per week was also found to be positively, significantly correlated with arsenic levels. On the other hand, the frequency of consumption of fruits, fresh and cooked vegetables, milk and milk products per week beneficially influenced the arsenic level of the hair samples examined. Arsenic content of the consumed water in Egypt was 0.001 mg/l, which is below the maximum drinking water level allowed by World Health Organisation (WHO). Therefore, the arsenic content of domestic tap water hardly contributed to the arsenic exposure of the Egyptian population in the regions of the study. It is likely that exposure routes by smoking, fish and animal protein consumption are the principal cause of arsenic accumulation in the general Egyptian population.