Evaluation of genotoxic and non-genotoxic effects of organic air pollution using in vitro bioassays

In this study, organic extracts of total suspended particles (TSP) and the particulate matter (PM) with the size below 2.5 microm (PM(2.5)) combined with organic extracts of the gas phase (GP) collected at two urban and two background localities were analyzed with a bacterial genotoxicity test, SOS chromotest, and an in vitro test for the dioxin toxicity determination, using a modified cell-line of rat hepatoma H4IIE.luc. In addition, the samples of TSP and GP were analyzed for PAHs contents. The PAHs concentrations and both of the toxic activities at the urban localities were much higher than ones at the background localities. Predominantly, traffic was a source of the urban air pollution there which was also confirmed by the evaluation of portions of certain PAHs (BaP/BPE, PYR/BaP) at the localities. On the other hand, the background localities were apparently affected by a long-distance transport of the pollutants from urban and industrial centers. The results of the bioassays indicated potential health risks for the population exposed to the organic air pollutants, especially at the urban localities. Based on the collected samples, distribution of the organic pollutants with the toxic effects in the air was evaluated. The significant portion of the direct genotoxins was bound to the particles larger than 2.5 microm. On the contrary, the indirect genotoxins were bound predominantly to the particles with the size below 2.5 microm. However, in the urban air they may be also bound to the larger particles, as well. While the direct genotoxicity may be related with the presence of PAH-derivatives as well as some polar organic pollutants, the indirect genotoxicity is related with the detected carcinogenic PAHs. But besides the above specified pollutants it is also necessary to consider the presence of other toxic components of the complex organic air pollution mixture that may also show potential health risks. This study demonstrates application of the combination of the screening bioassays for the evaluation of organic air pollution and identification of its health risks.     

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.     

Assessing noxious effects of dietary exposure to methylmercury,PCBs and Se coexisting in environmentally contaminated rice in male mice

Polychlorinated biphenyls and methylmercury are two of the most ubiquitous environmental contaminants in Guizhou province. Rice is eaten with almost every meal and provides more calories than any single food in Guizhou province. The estimated tolerable daily intake of total mercury, MeHg, Se and PCBs from Guizhou contaminated rice by Chinese people showed that MeHg and/or PCBs exceeded the corresponding limits. The aim of this study was to characterize the effects of exposure to environmental contaminated rice on neurobehavioral development and neurobiological disruptions in mice. Animals were treated from postnatal day (PND) 22 to 91. At PND 26–91 days of age, mice were tested for neurobehavioural development and neurochemical level changes. We showed that dietary exposure to environmentally contaminated rice gave rise to different changes in antioxidants. Reduced superoxide dismutase (SOD) activity and excess increased nitric oxide (NO) indicated aggravation of oxidative status after long-term dietary intake of Hg and PCBs. Neurobehavioral derangement in the central nervous system and significant delay in the Morris water maze test response on PND 91 are correlated with the increased of c-fos/c-jun expression levels in the cerebral cortex. These results suggest that MeHg neurotoxicity might be a greater hazard than that associated with PCB, but PCB may augment the neurobehavioral deficits caused by increased levels of mercury exposure. The simultaneous intake of selenium might have a protective effect on Hg accumulation in the body, and vitamin C might protect mice against the toxic effects of PCBs. However, the protective role of Se and vitamin C is very limited for multiple-agent pollution. Immediately early genes in the brain response to contaminated rice might be dependent on interaction among NO, NO synthase (NOS), SOD and reduced glutathione (GSH). We should be alert to mental health problems in human beings when any kind of Hg- and PCB-polluted food is consumed.     

Deleterious effects in mice of fish-associated methylmercury contained in a diet mimicking the Western populations' average fish consumption

Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutant through fish consumption. Only a few contradictory epidemiological studies are currently available examining the impact of fish consumption on human populations. In the present study, we wanted to address whether a diet mimicking the fish consumption of Western populations could result in observable adverse effects in mice, and whether beneficial nutriments from fish were able to counterbalance the deleterious effects of MeHg, if any. In Europe and the United States, fish consumption varies widely between countries, from 11 to 100 g fish/day. A mid-range value of 25 g fish/day corresponds to a fish contribution to the total diet of 1.25% on a dry weight basis. We decided to supplement a vegetarian-based mouse diet with 1.25% of lyophilized salmon flesh (SAL diet), or 1.25% of a blend of lyophilized cod, tuna, and swordfish (CTS diet). Total mercury contents were 1.15±0.15, 2.3±0.1 and 35.75±0.15 ng Hg/g of food pellets for the control, SAL and CTS diets, respectively. After two months feeding, the CTS diet resulted in significant observable effects as compared to the control and SAL diets, encompassing decreased body growth, altered behavioral performance and increased anxiety level, modification of mitochondrial respiratory protein subunit concentrations in kidney and brain structures, modified gene expression patterns in kidneys, liver and muscles, and a decrease of dopamine concentrations in the hypothalamus and striatum. Our findings have health implications, firstly because 1.25% of CTS flesh in the diet corresponds to an average exposure to MeHg below the WHO provisory tolerable weekly intake (PTWI) (1.6 μg MeHg/kg of body weight/week), and secondly because many people in Western populations, among them women of child-bearing age, are exceeding the PTWI value (for instance, 35% of the French population inhabiting the Atlantic and Mediterranean coasts).     

Toxic and genotoxic impact of fibrates and their photoproducts on non-target organisms

Lipid regulators have been detected in effluents from sewage treatment plants and surface waters from humans via excretion. This study was designed to assess the ecotoxicity of fibrates, lipid regulating agents. The following compounds were investigated: Bezafibrate, Fenofibrate and Gemfibrozil and their derivatives obtained by solar simulator irradiation. Bioassays were performed on bacteria, algae, rotifers and microcrustaceans to assess acute and chronic toxicity, while SOS Chromotest and Ames test were utilized to detect the genotoxic potential of the investigated compounds. The photoproducts were identified by their physical features and for the first risk evaluation, the environmental impact of parental compounds was calculated by Measured Environmental Concentrations (MEC) using the available data from the literature regarding drug occurrence in the aquatic environment and the Predicted No Effect Concentrations (PNEC) based on our toxicity data. The results showed that acute toxicity was in the order of dozens of mg/L for all the trophic levels utilized in bioassays (bacteria, rotifers, crustaceans). Chronic exposure to these compounds caused inhibition of growth population on rotifers and crustaceans while the algae seemed to be slightly affected by this class of pharmaceuticals. Genotoxic and mutagenic effects were especially found for the Gemfibrozil photoproduct suggesting that also byproducts have to be considered in the environmental risk of drugs.     

The genotoxic contribution of wood smoke to indoor respirable suspended particles

The effect of wood burning stoves on the genotoxicity of indoor respirable organic matter was investigated for four homes during the winter and spring of 1986. Paired samples, one collected when the stove was not used and one when wood was burned, were extracted with dichloromethane and acetone. Aliquots of the dichloromethane extracts were analyzed with and without metabolic activation using the Microscreen bioassay. The Microscreen is a rapid, sensitive bioassay which measures a broad genotoxic endpoint, λ-prophage induction. Per nanogram of organic material, wood smoke proved to be a major source of indirect (observed with metabolic activation) but not direct genotoxins in homes. The increase in indirect genotoxicity for extracts from aerosol containing wood smoke is probably due to higher concentrations of polycyclic aromatic hydrocarbons in the wood smoke aerosol as well as other unidentified classes. The direct genotoxicity observed for extracts of aerosol not containing wood smoke decreased with metabolic activation. This direct genotoxicity may be related to cooking activities in the homes. The trends in genotoxicity observed per nanogram of organic material are more pronounced when expressed per m³ of air due to the higher percentage of extractable material in aerosol containing wood smoke.     

Rice consumption contributes to low level methylmercury exposure in southern China

Fish consumption is considered as the primary pathway of human methylmercury (MeHg) exposure. However, recent studies highlighted that, rice, rather than fish, is the main route of human MeHg exposure in Guizhou, inland China. China is considered as the largest anthropogenic source of mercury (Hg) emission in the world, which has led to serious environmental Hg pollution. But there are no comprehensive studies regarding this environmental health problem to evaluate human Hg exposure and associated health effects. This study aimed to estimate daily MeHg intake and health risk in 7 provinces in southern China, and to assess the relative contribution from rice and fish consumption. The average levels of total mercury (THg) and MeHg in rice samples were generally low at 10.1 ng·g^− 1 and 2.47 ng·g^− 1, respectively. But a total of 36 rice samples (12.7%) had THg concentration exceeding the national limit (20 ng·g^− 1). Generally, rural population had significantly higher Probable Daily Intakes (PDIs) of MeHg than urban population from rice consumption and its relative contribution to MeHg exposure increased significantly from coastal to inland area. The averages of PDIs of MeHg were 0.020 μg·kg^− 1·d^− 1 and 0.028 μg·kg^− 1·d^− 1 for urban and rural population in southern China, respectively. Despite the serious environmental Hg pollutions in China, the general population in southern China had low risk of MeHg exposure. But rice is an important route of human MeHg exposure in southern China, especially for the rural population in inland area. The findings indicate that rice consumption should be considered when evaluating MeHg exposure in rice eating population in southern China.     

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.     

Prenatal methylmercury exposure and language delay at three years of age in the Norwegian Mother and Child Cohort Study

Prenatal methylmercury (MeHg) exposure and its possible neurodevelopmental effects in susceptible children are of concern. Studies of MeHg exposure and negative health outcomes have shown conflicting results and it has been suggested that co-exposure to other contaminants and/or nutrients in fish may confound the effect of MeHg. Our objective was to examine the association between prenatal exposure to MeHg and language and communication development at three years, adjusting for intake of fish, n − 3 long chain polyunsaturated fatty acids (n − 3 LCPUFAs) and co-exposure to dioxins and dioxin like polychlorinated biphenyls (dl-PCBs).We used data from the Norwegian Mother and Child Cohort Study (MoBa) collected between 2002 and 2008. The study sample consisted of 46,750 mother-child pairs. MeHg exposure was calculated from reported fish intake during pregnancy by a FFQ in mid-pregnancy. Children's language and communication skills were measured by maternal report on the Dale and Bishop grammar rating and the Ages and Stages communication scale (ASQ). We estimated odds ratios (OR) and 95% confidence intervals (CI) using logistic regressions.Median MeHg exposure was 1.3 μg/day, corresponding to 0.14 μg/kg bw/week. An exposure level above the 90th percentile (> 2.6 μg/day, > 0.29 μg/kg bw/week) was defined as the high MeHg exposure. Results indicated an association between high MeHg exposure and unintelligible speech with an adjusted OR 2.22 (1.31, 3.72). High MeHg exposure was also associated with weaker communication skills adjusted OR 1.33 (1.03, 1.70). Additional adjustment for fish intake strengthened the associations, while adjusting for PCBs and n − 3 LCPUFA from diet or from supplements had minor impact. In conclusion, significant associations were found between prenatal MeHg exposure above the 90th percentile and delayed language and communication skills in a generally low exposed population.     

Fetal methylmercury exposure as measured by cord blood mercury concentrations in a mother-infant cohort in Hong Kong

This study was designed to examine newborn infants in Hong Kong prenatally exposed to levels of methylmercury considered to increase risk of neurotoxic effects and to examine subject characteristics that modify the degree of prenatal mercury exposure. Mercury concentrations in 1057 sets of maternal and cord blood samples and 96 randomly selected maternal hair samples were measured. Subject characteristics were measured or collected by questionnaire. Of the 1057 cord blood samples collected only 21.6% had mercury concentrations less than 29 nmol/L (5.8 micro g/L). Median maternal hair mercury concentration was 1.7 ppm. The geometric mean cord to maternal blood mercury ratio was 1.79 to 1. Increasing maternal fish consumption and maternal age were found to be associated with increased cord blood mercury concentrations. Marine fish consumption increased cord blood mercury concentrations more than freshwater fish (5.09%/kg vs 2.86%/kg). Female babies, maternal alcohol consumption and increasing maternal height were associated with decreased cord blood mercury concentrations. Pregnant women in Hong Kong consume large amounts of fish and as a result, most of their offspring have been prenatally exposed to moderately high levels of mercury. In this population, pregnant women should choose freshwater over marine fish and limit fish consumption.     

Drinking water quality: An in vitro approach for the assessment of cytotoxic and genotoxic load in water sampled along distribution system

An in vitro approach was performed to assess the quality of drinking water collected at two treatment/distribution networks located near the source (Plant #1) and the mouth of River Po (Plant #2). The water was sampled at different points of each distribution network, before (raw water) and after the chlorine dioxide disinfection, and in two points of the pipeline system to evaluate the influence of the distribution system on the amount and quality of the disinfection by-product. Cytotoxicity and genotoxicity of water extracts were evaluated in human peripheral lymphocytes and Hep-G2 cells by the use of the micronucleus (MN) test and Comet assay. Raw water samples of both plants induced cytotoxic effects, but not the increases of MN frequency in Hep-G2 cells and in human lymphocytes. Increases of DNA damage in human leukocytes was detected by Comet assay for raw water of Plant #2 at concentration ≥ 0.25 Leq/mL. The disinfection process generally has reduced the toxicity of water samples, even if potential direct DNA-damaging compounds have been detectable in drinking water samples. The proposal approach, if currently used together with chemical analysis, can contribute to improve the monitoring drinking water.     

Polymorphisms in ATP-binding cassette transporters associated with maternal methylmercury disposition and infant neurodevelopment in mother-infant pairs in the Seychelles Child Development Study

BackgroundATP-binding cassette (ABC) transporters have been associated with methylmercury (MeHg) toxicity in experimental animal models.AimsTo evaluate the association of single nucleotide polymorphisms (SNPs) in maternal ABC transporter genes with 1) maternal hair MeHg concentrations during pregnancy and 2) child neurodevelopmental outcomes.Materials and methodsNutrition Cohort 2 (NC2) is an observational mother-child cohort recruited in the Republic of Seychelles from 2008–2011. Total mercury (Hg) was measured in maternal hair growing during pregnancy as a biomarker for prenatal MeHg exposure (N = 1313) (mean 3.9 ppm). Infants completed developmental assessments by Bayley Scales of Infant Development II (BSID-II) at 20 months of age (N = 1331). Genotyping for fifteen SNPs in ABCC1, ABCC2 and ABCB1 was performed for the mothers.ResultsSeven of fifteen ABC SNPs (ABCC1 rs11075290, rs212093, and rs215088; ABCC2 rs717620; ABCB1 rs10276499, rs1202169, and rs2032582) were associated with concentrations of maternal hair Hg (p < 0.001 to 0.013). One SNP (ABCC1 rs11075290) was also significantly associated with neurodevelopment; children born to mothers with rs11075290 CC genotype (mean hair Hg 3.6 ppm) scored on average 2 points lower on the Mental Development Index (MDI) and 3 points lower on the Psychomotor Development Index (PDI) than children born to mothers with TT genotype (mean hair Hg 4.7 ppm) while children with the CT genotype (mean hair Hg 4.0 ppm) had intermediate BSID scores.DiscussionGenetic variation in ABC transporter genes was associated with maternal hair Hg concentrations. The implications for MeHg dose in the developing child and neurodevelopmental outcomes need to be further investigated.     

Health effects of indoor air pollution: Synergistic effects of nitrogen dioxide and a respirable aerosol

Comparative studies of the effects of various air pollutants on lung collagen biosynthesis have been performed. A hitherto unexpected synergism between the oxidant air pollutants ozone or nitrogen dioxide and a respirable-sized aerosol of ammonium sulfate was observed during controlled exposures of rats to these substances. In an assay system, measuring collagen biosynthesis by lung minces prepared from rats exposed for 1 week to either filtered air or to these pollutants gases, dose-response curves to either O₃ or NO₂ are altered in the presence of 5 mg/m³ of (NH₄)₂SO₄ aerosol. These observations may have broad implications for the appropriate evaluation of laboratory data in the setting of ambient air quality standards and/or the setting of threshold limit values for maintenance of occupational health and safety.     

Theoretically exploring direct and indirect chemical effects across ecological and exposure scenarios using mechanistic fate and effects modelling

Predicting ecosystem response to chemicals is a complex problem in ecotoxicology and a challenge for risk assessors. The variables potentially influencing chemical fate and exposure define the exposure scenario while the variables determining effects at the ecosystem level define the ecological scenario. In absence of any empirical data, the objective of this paper is to present simulations by a fugacity-based fate model and a differential equation-based ecosystem model to theoretically explore how direct and indirect effects on invertebrate shallow pond communities vary with changing ecological and exposure scenarios. These simulations suggest that direct and indirect effects are larger in mesotrophic systems than in oligotrophic systems. In both trophic states, interaction strength (quantified using grazing rates) was suggested a more important driver for the size and recovery from direct and indirect effects than immigration rate. In general, weak interactions led to smaller direct and indirect effects. For chemicals targeting mesozooplankton only, indirect effects were common in (simple) food-chains but rare in (complex) food-webs. For chemicals directly affecting microzooplankton, the dominant zooplankton group in the modelled community, indirect effects occurred both in food-chains and food-webs. We conclude that the choice of the ecological and exposure scenarios in ecotoxicological modelling efforts needs to be justified because of its influence on the prevalence and magnitude of the predicted effects. Overall, more work needs to be done to empirically test the theoretical expectations formulated here.     

Health effects of oxidants

Oxidants of significance to human health include ozone, nitrogen dioxide, and peroxyacetylnitrate. All of these compounds are involved in complex photochemical reactions which makes quantification and prediction of their individual health effects difficult. Ozone causes trauma to lung tissues and interferes with enzyme systems in the lungs and other tissues causing a broad range of symptoms. Measurable health impacts can occur at concentrations as low as 390 μg/m³. Acute effects of ozone exposure are reversible at normal urban concentrations (80–120 μg/m³). A special problem of concern, however, is increased susceptibility to infectious diseases contracted through the lungs. Nitrogen dioxide also causes trauma to lung tissues and interferes with enzyme systems. Measurable impacts can occur at concentrations as low as 100 μg/m³, but recovery is rapid and it is not known whether repeated exposures at this level have cumulative effects or predispose the lungs to permanent damage. Chronic exposure of laboratory animals to higher nitrogen dioxide levels can cause emphysema-like conditions and reduction in resistance to respiratory infection. Epidemiological studies of children in houses with gas stoves confirm the finding of reduced resistance to respiratory infection. The U.S. EPA estimates that health effects may occur in young children exposed to concentrations in excess of 280 to 560 μg/m³ one-hour average. These concentrations occur routinely in houses having gas stoves. Peroxyacetylnitrate is a powerful eye irritant in photochemical smog. Other health effects are similar to those of ozone, but less important because of the relatively low concentrations of this pollutant compared to other oxidants.     

Ecological effects of particulate matter

Atmospheric particulate matter (PM) is a heterogeneous material. Though regulated as un-speciated mass, it exerts most effects on vegetation and ecosystems by virtue of the mass loading of its chemical constituents. As this varies temporally and spatially, prediction of regional impacts remains difficult. Deposition of PM to vegetated surfaces depends on the size distribution of the particles and, to a lesser extent, on the chemistry. However, chemical loading of an ecosystem may be determined by the size distribution as different constituents dominate different size fractions. Coating with dust may cause abrasion and radiative heating, and may reduce the photosynthetically active photon flux reaching the photosynthetic tissues. Acidic and alkaline materials may cause leaf surface injury while other materials may be taken up across the cuticle. A more likely route for metabolic uptake and impact on vegetation and ecosystems is through the rhizosphere. PM deposited directly to the soil can influence nutrient cycling, especially that of nitrogen, through its effects on the rhizosphere bacteria and fungi. Alkaline cation and aluminum availability are dependent upon the pH of the soil that may be altered dramatically by deposition of various classes of PM. A regional effect of PM on ecosystems is linked to climate change. Increased PM may reduce radiation interception by plant canopies and may reduce precipitation through a variety of physical effects. At the present time, evidence does not support large regional threats due to un-speciated PM, though site-specific and constituent-specific effects can be readily identified. Interactions of PM with other pollutants and with components of climate change remain important areas of research in assessment of challenges to ecosystem stability.     

Fate and effects of amphoteric surfactants in the aquatic environment

Amphoteric surfactants form part of specialty surfactants available for formulators to improve or design new formulations in response to environmental, toxicity, safety and performance demands. Nevertheless, limited information on the ecological properties of amphoterics is available. In the present work, the aerobic and anaerobic biodegradability and the aquatic toxicity of different types of amphoteric surfactants (three alkyl betaines, one alkylamido betaine and three alkyl imidazoline derivatives) were studied. The amphoteric surfactants tested were readily biodegradable under aerobic conditions (CO(2) headspace test) and alkylamido betaines and alkyl imidazoline derivatives were also easily biodegradable under anaerobic conditions (test based on the ECETOC method). Toxicity to Photobacterium phosphoreum and Daphnia magna increased with the fatty chain length of the surfactant. The EC(50) toxicity values of the amphoterics tested were higher than 5 mg/L, and alkyl imidazoline derivatives, with EC(50) values from 20 to >200 mg/L, showed the lowest aquatic toxicity.     

The development and purpose of the FREDERICA radiation effects database

Any system for assessing the impact of a contaminant on the environment requires an analysis of the possible effects on the organisms and ecosystems concerned. To facilitate this, the FREDERICA radiation effects database has been developed to provide an online search of the known effects of ionising radiation on non-human species, taken from papers in the scientific peer reviewed literature. The FREDERICA radiation effects database has been produced by merging the work done on radiation effects under two European funded projects (FASSET and EPIC) and making the database available online. This paper highlights applications for the database, gaps in the available data and explains the use of quality scores to help users of the database determine which papers may benefit their research in terms of techniques and reproducibility.