Tamara Grummt

Genotoxic effects in swimmers exposed to disinfection by-products in indoor swimming pools.

Background Exposure to disinfection by-products (DBPs) in drinking water has been associated with cancer risk. A recent study (Villanueva et al. 2007; Am J Epidemiol 165:148–156) found an increased bladder cancer risk among subjects attending swimming pools relative to those not attending. Objectives We evaluated adults who swam in chlorinated pools to determine whether exposure to DBPs in pool water is associated with biomarkers of genotoxicity. Methods We collected blood, urine, and exhaled air samples from 49 nonsmoking adult volunteers before and after they swam for 40 min in an indoor chlorinated pool. We estimated associations between the concentrations of four trihalomethanes (THMs) in exhaled breath and changes in micronuclei (MN) and DNA damage (comet assay) in peripheral blood lymphocytes before and 1 hr after swimming; urine mutagenicity (Ames assay) before and 2 hr after swimming; and MN in exfoliated urothelial cells before and 2 weeks after swimming. We also estimated associations and interactions with polymorphisms in genes related to DNA repair or to DBP metabolism. Results After swimming, the total concentration of the four THMs in exhaled breath was seven times higher than before swimming. The change in the frequency of micronucleated lymphocytes after swimming increased in association with higher exhaled concentrations of the brominated THMs (p = 0.03 for bromodichloromethane, p = 0.05 for chlorodibromomethane, p = 0.01 for bromoform) but not chloroform. Swimming was not associated with DNA damage detectable by the comet assay. Urine mutagenicity increased significantly after swimming, in association with the higher concentration of exhaled bromoform (p = 0.004). We found no significant associations with changes in micronucleated urothelial cells. Conclusions Our findings support potential genotoxic effects of exposure to DBPs from swimming pools. The positive health effects gained by swimming could be increased by reducing the potential health risks of pool water.

Emerging risks from ballast water treatment: The run-up to the International Ballast Water Management Convention

Uptake and discharge of ballast water by ocean-going ships contribute to the worldwide spread of aquatic invasive species, with negative impacts on the environment, economies, and public health. The International Ballast Water Management Convention aims at a global answer. The agreed standards for ballast water discharge will require ballast water treatment. Systems based on various physical and/or chemical methods were developed for on-board installation and approved by the International Maritime Organization. Most common are combinations of high-performance filters with oxidizing chemicals or UV radiation. A well-known problem of oxidative water treatment is the formation of disinfection by-products, many of which show genotoxicity, carcinogenicity, or other long-term toxicity. In natural biota, genetic damages can affect reproductive success and ultimately impact biodiversity. The future exposure towards chemicals from ballast water treatment can only be estimated, based on land-based testing of treatment systems, mathematical models, and exposure scenarios. Systematic studies on the chemistry of oxidants in seawater are lacking, as are data about the background levels of disinfection by-products in the oceans and strategies for monitoring future developments. The international approval procedure of ballast water treatment systems compares the estimated exposure levels of individual substances with their experimental toxicity. While well established in many substance regulations, this approach is also criticised for its simplification, which may disregard critical aspects such as multiple exposures and long-term sub-lethal effects. Moreover, a truly holistic sustainability assessment would need to take into account factors beyond chemical hazards, e.g. energy consumption, air pollution or waste generation.

Anthropogenic organic micro-pollutants and pathogens in the urban water cycle: assessment, barriers and risk communication (ASKURIS)

In urban areas, water often flows along a partially closed water cycle in which treated municipal wastewater is discharged into surface waters which are one source of raw waters used for drinking water supply. A number of organic micro-pollutants (OMP) can be found in different water compartments. In the near future, climatic and demographic changes will probably contribute to an increase of OMP and antibiotic-resistant pathogens in aquatic ecosystems. The occurrence of OMP, possible adverse effects on aquatic organisms and human health and the public perception must be carefully assessed to properly manage and communicate potentially associated risks and to implement appropriate advanced treatment options at the optimum location within the water cycle. Therefore, the interdisciplinary research project ASKURIS focuses on identification and quantification, toxicological assessment and removal of organic micro-pollutants and antibiotic-resistant pathogens in the Berlin water cycle, life cycle-based economic and environmental assessment, public perception and management of potential risks.

Impact of ozonation on the genotoxic activity of tertiary treated municipal wastewater

Ozonation is an emerging technology for the removal of micropollutants from treated wastewater. Aim of the present study was to investigate the impact of ozone treatment on genotoxic and acute toxic effects of tertiary treated municipal wastewater. It is known that DNA-damaging chemicals cause adverse effects in the environment and that exposure to humans leads to cancer and other diseases. Toxicity was tested in organisms from three trophic levels namely in bacteria (Salmonella/microsome assays) which enable the detection of gene mutations, in a plant bioassay (micronucleus assay with root tip cells of Allium cepa) which reflects clastogenic and aneugenic effects and in single cell gel electrophoresis (SCGE) tests with mammalian cells which detect DNA migration caused by single-, double strand breaks and alkali labile sites. In the bacterial tests negative results were obtained with untreated samples but after concentration with C(18) cartridges a positive result was found in strains TA1537 and TA98 which are sensitive to frameshift mutagens while no mutations were induced in other tester strains (TA100, TA102 and YG1024). Ozone treatment led to a decrease of the mutagenic activity of the samples. In the SCGE experiments, DNA migration was detected with the unconcentrated effluent of the treatment plant and ozonation led to a substantial decrease of this effect. In the plant bioassays, negative results were obtained with the effluent and ozone treatment did not cause an alteration of the micronucleus frequencies. Also acute toxic effects were monitored in the different indicator organisms under all experimental conditions. The bacteriocidal/bacteriostatic effects which were seen with the concentrated samples were reduced by ozonation. In the experiments with the eukaryotic (plant and animal) cells no acute toxicity was seen with the effluents and ozonation had no impact on their viability. In conclusion findings of this study indicate that ozonation of tertiary effluents of a municipal treatment plant reduces the adverse effects caused by release of mutagens in aquatic ecosystems and does not decrease the viability of bacteria and eukaryotic cells. However, future research is required to find out if, and to which extent these findings can be generalized and which mechanisms account for the detoxification of the wastewater.

Toxicity studies of nonylphenol and octylphenol: hormonal, hematological and biochemical effects in Clarias gariepinus

Among the numerous chemicals discharged into aquatic ecosystems, nonylphenol (NP) and octylphenol (OP) have been shown to have a potent effect on the endocrine system of fish; this issue has been clearly dealt with in several studies. The objective of this study was to assess and compare the general toxicity of these estrogenic chemicals individually on Clarias gariepinus. Fish were exposed to different concentrations of both NP and OP (250, 500, 750 and 1000 μg l−1) under semi‐static conditions for a period of 7 days. The adverse effect was evaluated with use of blood cell counting, hemoglobin (Hb), hematocrit (HCT), hematimetric indices, bilirubin, protein, glucose, serum transaminases, serum phosphatases, lactate dehydrogenase and cortisol. The results showed a clear indication of anemia, increases in leukocyte count and bilirubin content and a reduction in plasma protein levels with higher concentrations of both the toxicants compared with controls. Furthermore, with all the concentrations the inevitable increase in serum cortisol and plasma glucose showed primary and secondary stress responses. Moreover, probable tissue damage gave rise to a series of fluctuations of enzyme levels at lower concentrations, but a decrease with higher concentrations showed the severity of the effect. Depending on the parameters examined, OP had a relatively greater effect than NP. Overall, these two chemicals seemingly affected hematology and the activity of some enzymes, leading to serious impairment of the metabolism and physiology of C. gariepinus. Copyright

Genotoxic effects of wastewater from an oncological ward.

Aim of this study was the evaluation of the genotoxic activities of hospital wastewaters. Samples from an oncological ward of the general hospital of Vienna, Austria, were tested in the Salmonella/microsome assay in strains TA98, TA100 and TA1535 with or without metabolic activation, and in the single-cell gel electrophoresis (SCGE) assay with primary rat hepatocytes. In the bacterial tests, consistently negative results were obtained while in the experiments with liver cells a significant and dose-dependent induction of DNA damage (up to two-fold over the background) was found. Membrane filtration resulted in a substantial (62-77%) reduction of these effects, while additional treatments (activated carbon filtration and UV-irradiation) did not lead to a further decrease of the genotoxic activity of the samples. SCGE experiments with cisplatin, carboplatin and 5-fluorouracil, which were detected in the water samples, showed that these cytostatics cause a significant induction of DNA damage only at concentrations that are substantially higher than those in the native waters. These findings indicate that other chemicals, possibly quaternary ammonium compounds, account for the effects of the hospital wastewaters.

Tox-Box: securing drops of life - an enhanced health-related approach for risk assessment of drinking water in Germany

This article introduces ‘Tox-Box’, a joint research project designed to develop a holistic approach towards a harmonized testing strategy for exposure- and hazard-based risk management of anthropogenic trace substances in drinking water to secure a long-term drinking water supply. The main task of the Tox-Box consortium is to enhance the existing health-related indicator value concept (German: GOW-Konzept - Gesundheitlicher Orientierungswert) through development and prioritization of additional end point-related testing strategies for genotoxicity, neurotoxicity, germ cell damage, and endocrine effects. In this context, substance-specific modes of action will be identified and characterized. Toxicological data collected by the 12 Tox-Box subprojects will be evaluated and weighted to structure a hierarchical testing strategy for an improved risk assessment. A technical guidance document for exposure and hazard-based risk management of anthropogenic trace substances in drinking water will eventually be prepared.ZusammenfassungDieser Artikel stellt das Verbundprojekt “Tox-Box” vor, das einen ganzheitlichen Ansatz für eine harmonisierte Teststrategie eines Expositions-bezogenen und Gefahren-basierten Risikomanagements von anthropogenen Spurenstoffen in Trinkwasser entwickeln und somit einen Beitrag zur langfristigen Sicherung der Trinkwasserversorgung leisten soll. Die Hauptaufgabe des Tox-Box-Konsortiums ist die Weiterentwicklung des bestehenden GOW-Konzeptes (Gesundheitlicher Orientierungswert) durch Erforschung und Priorisierung zusätzlicher Endpunkt-bezogener Teststrategien für Gentoxizität, Neurotoxizität, Keimzellschädigung und endokrine Effekte. In diesem Kontext werden zudem Substanz-spezifische Wirkmechanismen identifiziert und charakterisiert. Im Anschluss werden die toxikologischen Daten aus den 12 Teilprojekten evaluiert und gewichtet um eine hierarchische Teststrategie für eine verbesserte Risikobewertung zu erstellen. Zum Abschluss des Projektes wird eine technische Richtlinie für ein Expositions-bezogenes und Gefahren-basiertes Risikomanagement von anthropogenen Spurenstoffen im Trinkwasser erstellt.

Cytotoxicity of water-soluble fraction from biodiesel and its diesel blends to human cell lines.

The designation of biodiesel as a green fuel has increased its commercialization and use, making its fate in the environment a matter of concern. Fuel spills constitute a major source of aquatic pollution and, like diesel spills, biodiesel can produce adverse effects on aquatic environments, animals and humans. The present study assessed cytotoxic effects of water systems contaminated with neat biodiesel and its diesel blends by means of different procedures on human T cell leukemia (Jurkat) and human hepatocellular carcinoma (HepG2) cells [detection of changes in mitochondrial membrane potential (ΔΨ(m)) using tetramethylrhodamine ethyl ester (TMRE), apoptosis recognition by Annexin V and impedance real-time cell analyzer (xCELLigence™ system)]. The data obtained showed concordance across the different bioassays, with cytotoxic effects observed as a dose-dependent response only for waters contaminated with pure diesel (D100) and B5 blend, which is characterized by a mixture of 95% diesel and 5% biodiesel. The data can also lead us to hypothesize that diesel accounts for the harmful effects observed, and that biodiesel does not worsen the impacts caused by diesel pollution.

Application of an optimized system for the well-defined exposure of human lung cells to trichloramine and indoor pool air

In this study an in vitro exposure test to investigate toxicological effects of the volatile disinfection by-product trichloramine and of real indoor pool air was established. For this purpose a set-up to generate a well-defined, clean gas stream of trichloramine was combined with biotests. Human alveolar epithelial lung cells of the cell line A-549 were exposed in a CULTEX(®) device with trichloramine concentrations between 0.1 and 40 mg/m(3) for 1 h. As toxicological endpoints the cell viability and the inflammatory response by the cytokines IL-6 and IL-8 were investigated. A decreasing cell viability could be observed with increasing trichloramine concentration. An increase of IL-8 release could be determined at trichloramine concentrations higher than 10 mg/m(3) and an increase of IL-6 release at concentrations of 20 mg/m(3). Investigations of indoor swimming pool air showed similar inflammatory effects to the lung cells although the air concentrations of trichloramine of 0.17 and 0.19 mg/m(3) were much lower compared with the laboratory experiments with trichloramine as the only contaminant. Therefore it is assumed that a mixture of trichloramine and other disinfection by-products in the air of indoor pool settings contribute to that effect.

Genotoxicity assessment of water soluble fractions of biodiesel and its diesel blends using the Salmonella assay and the in vitro MicroFlow® kit (Litron) assay

The designation of biodiesel as an environmental-friendly alternative to diesel oil has improved its commercialization and use. However, most biodiesel environmental safety studies refer to air pollution and so far there have been very few literature data about its impacts upon other biotic systems, e.g. water, and exposed organisms. Spill simulations in water were carried out with neat diesel and biodiesel and their blends aiming at assessing their genotoxic potentials should there be contaminations of water systems. The water soluble fractions (WSF) from the spill simulations were submitted to solid phase extraction with C-18 cartridge and the extracts obtained were evaluated carrying out genotoxic and mutagenic bioassays [the Salmonella assay and the in vitro MicroFlow® kit (Litron) assay]. Mutagenic and genotoxic effects were observed, respectively, in the Salmonella/microsome preincubation assay and the in vitro MN test carried out with the biodiesel WSF. This interesting result may be related to the presence of pollutants in biodiesel derived from the raw material source used in its production chain. The data showed that care while using biodiesel should be taken to avoid harmful effects on living organisms in cases of water pollution.