Pilar Fernandez

What's in the pool? A comprehensive identification of disinfection by-products and assessment of mutagenicity of chlorinated and brominated swimming pool water

Background Swimming pool disinfectants and disinfection by-products (DBPs) have been linked to human health effects, including asthma and bladder cancer, but no studies have provided a comprehensive identification of DBPs in the water and related that to mutagenicity. Objectives We performed a comprehensive identification of DBPs and disinfectant species in waters from public swimming pools in Barcelona, Catalonia, Spain, that disinfect with either chlorine or bromine and we determined the mutagenicity of the waters to compare with the analytical results. Methods We used gas chromatography/mass spectrometry (GC/MS) to measure trihalomethanes in water, GC with electron capture detection for air, low- and high-resolution GC/MS to comprehensively identify DBPs, photometry to measure disinfectant species (free chlorine, monochloroamine, dichloramine, and trichloramine) in the waters, and an ion chromatography method to measure trichloramine in air. We assessed mutagenicity with the Salmonella mutagenicity assay. Results We identified > 100 DBPs, including many nitrogen-containing DBPs that were likely formed from nitrogen-containing precursors from human inputs, such as urine, sweat, and skin cells. Many DBPs were new and have not been reported previously in either swimming pool or drinking waters. Bromoform levels were greater in brominated than in chlorinated pool waters, but we also identified many brominated DBPs in the chlorinated waters. The pool waters were mutagenic at levels similar to that of drinking water (~ 1,200 revertants/L-equivalents in strain TA100–S9 mix). Conclusions This study identified many new DBPs not identified previously in swimming pool or drinking water and found that swimming pool waters are as mutagenic as typical drinking waters.

Persistent organic pollutants in snow from European high mountain areas

Snow cores were collected in the catchment area of five remote mountain lakes in Europe. They were analysed for polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB), and organochlorine pesticides, namely DDTs, hexachlorobenzene (HCB) and hexachlorocyclohexanes (HCH). PAH are found in higher amounts in the Tatra and Caledonian mountains, PCB are higher in the Alps and HCH are highest in the Alps and Pyrenees. The qualitative PAH distributions are dominated by low molecular weight compounds, phenanthrene being the most abundant PAH in all but in one site. These compounds also occur predominantly in the gas phase in the atmosphere. Their high abundance in the snowpack witness the occurrence of effective transfer mechanisms from gas to snow flakes. In Starolesnianske (Tatra mountains), a higher contribution of high molecular weight compounds is found. This site exhibits the highest snow PAH and suspended particulate levels. Transformation of the concentration values of these compounds into annual deposition rates and correction for catchment/lake area indicates that in Scandinavia and the Alps a large proportion of PAH incorporation is mediated by snowfallout whereas in the Tatra mountains snow deposition only accounts for a small fraction of the compounds stored in the lake sediments. Among organochlorine compounds, only PCB and HCH have been found above method detection limit in most of the samples. The PCB congener distributions changes significantly between sites, although a predominance of the less chlorinated congeners have generally been observed.

On the Global distribution of persistent organic pollutants

The global distribution of persistent organic pollutants (POPs) has become one of the main environmental problems in the last decade. This article gives an overview of the main contributions to the knowledge of the atmospheric transport and accumulation mechanisms of POPs in remote areas, based on their analysis in selected environmental compartments from high altitude mountain regions of Europe. The studies indicated that transport and deposition of polycyclic aromatic hydrocarbons are mainly linked to atmospheric particles. Consequently, wet and dry deposition are the main removal processes of these compounds from the atmosphere, resulting in a significant influence of regional sources. In contrast, gas exchange seems to be the main input mechanism of organochlorine compounds (OCs) from the atmosphere to terrestrial and aquatic systems. For these compounds, an altitude dependence of their accumulation in fish muscle and sediments was detected, with a major retention of the less volatile compounds (vapor pressure < 10 - 2 . 5 Pa) in the locations situated at higher altitude, those of lower annual average temperature, whereas no relationship was observed for more volatile OCs. The results represent a new aspect in the Global Distillation Effect theory as proposed for semivolatile persistent pollutants, indicating that this transport mechanism not only involves transfer from low to high latitudes, but also preferential accumulation of the less volatile compounds in high altitude regions of mid-latitude areas.

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.

Mercury and Organochlorine Contamination in Brown Trout (Salmo Trutta) and Arctic Charr (Salvelinus Alpinus) from High Mountain Lakes in Europe and the Svalbard Archipelago

High concentration of Hg, less volatile PCB congeners and p,p′-DDE in Arctic charr from an arctic lake was mainly causedby biomagnification in the food chain where cannibalism was thedriving force. We suggest that low sediment fluxes of Hg, low net production of methyl mercury, and short food chains excludingpiscivory explain the low levels of Hg in the invertebrate feeding fish population in five European high mountain lakes.Concentrations of less volatile PCB congeners in insectivorous fish populations from the European high mountain lakes were mainly influenced by fish age and atmospheric deposition, indicated by the sediment inventory. Atmospheric deposition influenced by local sources may explain the higher concentrationsof pesticides (p,p′-DDT, p,p′-DDE and γ-HCH) observedin fish from the Pyrenees compared to the other sites. Theconcentrations of Hg and organochlorines did not exceedthe guidelines for fish consumption, except for Hg levelsin the oldest fish from the arctic lake.

Short-Term Changes in Respiratory Biomarkers after Swimming in a Chlorinated Pool

Background Swimming in chlorinated pools involves exposure to disinfection by-products (DBPs) and has been associated with impaired respiratory health. Objectives We evaluated short-term changes in several respiratory biomarkers to explore mechanisms of potential lung damage related to swimming pool exposure. Methods We measured lung function and biomarkers of airway inflammation [fractional exhaled nitric oxide (FeNO), eight cytokines, and vascular endothelial growth factor (VEGF) in exhaled breath condensate], oxidative stress (8-isoprostane in exhaled breath condensate), and lung permeability [surfactant protein D (SP-D) and the Clara cell secretory protein (CC16) in serum] in 48 healthy nonsmoking adults before and after they swam for 40 min in a chlorinated indoor swimming pool. We measured trihalomethanes in exhaled breath as a marker of individual exposure to DBPs. Energy expenditure during swimming, atopy, and CC16 genotype (rs3741240) were also determined. Results Median serum CC16 levels increased from 6.01 to 6.21 μg/L (average increase, 3.3%; paired Wilcoxon test p = 0.03), regardless of atopic status and CC16 genotype. This increase was explained both by energy expenditure and different markers of DBP exposure in multivariate models. FeNO was unchanged overall but tended to decrease among atopics. We found no significant changes in lung function, SP-D, 8-isoprostane, eight cytokines, or VEGF. Conclusions We detected a slight increase in serum CC16, a marker of lung epithelium permeability, in healthy adults after they swam in an indoor chlorinated pool. Exercise and DBP exposure explained this association, without involving inflammatory mechanisms. Further research is needed to confirm the results, establish the clinical relevance of short-term serum CC16 changes, and evaluate the long-term health impacts.

Bioassay-directed chemical analysis of genotoxic components in urban airborne particulate matter from Barcelona (Spain)

Organic extracts of airborne particulate matter, collected in the city of Barcelona, were subjected to three-level, bioassay-directed, chemical fractionation, including gel permeation chromatography (GPC) and normal-phase (NP) and reversed-phase (RP) liquid chromatography (LC). The chemical characterization, directed by the Salmonella microsome mutagenicity assay (TA98, TA98NR and TA98/1,8DNP6 +/- S9), was carried out by capillary GC (CGC) coupled to selective detection systems, and by GC-MS techniques. The results obtained with the nitroreductase deficient strains show the important contribution of nitroaromatic compounds. Detailed chemical analysis of the mutagenic fractions led to the identification of 82 aromatic compounds and revealed the large contribution of chemical classes that are more polar than polycyclic aromatic hydrocarbons such as aromatic ketones, quinones and aldehydes.

Differences in cytochrome P450 enzyme activities between fish and crustacea: Relationship with the bioaccumulation patterns of polychlorobiphenyls (PCBs)

Variations in cytochrome P450 enzyme (CYPs) distribution and function between animal groups could result in differential metabolism and elimination kinetics for certain contaminants. Although a number of studies have suggested that differences in polychlorobiphenyl (PCB) accumulation profiles between crustacea and fish might result from differential CYP patterns, the relationship between PCB bioaccumulation and CYP capacities has not been demonstrated in these organisms. In the present study we investigated the hepatic microsomal catalytic activities in three deep-sea fish species, Alepocephalus rostratus (Alepocephalidae), Coelorinchus mediterraneus (Macrouridae), and Lepidion lepidion (Moridae), and the decapod crustacean Aristeus antennatus (Decapoda), using six fluorescent CYP-mediated substrates, namely ER (7-ethoxyresorufin), PR (7-pentoxyresorufin), BR (7-benzyloxyresorufin), CEC (3-cyano-7-ethoxycoumarin), DBF (dibenzylfluorescein) and BFC (7-benzyloxy-4-trifluoromethylcoumarin). Furthermore, we related the metabolic activities to the accumulation patterns of 41 PCB congeners in the muscle of these organisms. The results indicated a marked difference in the presence and activities of CYP isoforms between fish and the crustacean A. antennatus. Liver microsomes of the three selected fish species were capable of metabolizing all six CYP-mediated substrates and enzymes were identified as primarily belonging to CYP1A and CYP3A subfamilies. In contrast, hepatopancreas microsomes from A. antennatus only showed activity for PR and DBF substrates, generally related to mammalian CYP2-like enzymes. Furthermore, a direct relationship between metabolic activities and PCB accumulation profiles could be established. Results revealed that A. antennatus accumulated significantly higher proportions of PCBs 28, 52, 118, 138, 158 and 169 than fish, which is in accordance with the previously observed lack of CYP1A-like biotransformation capacities. Moreover, A. antennatus exhibited lower levels of PCBs 87, 149, 153, 170, 180, 183, 194 and 206 indicating that this crustacean is able to metabolize congeners considered mammalian CYP2B inducers. Hence, the present findings highlight the role of CYP-mediated metabolism in the congener-specific accumulation of PCBs in aquatic organisms and stress the need to further investigate quantitative and qualitative differences in xenobiotic metabolism among animal groups.

Use of off-line gel permeation chromatography-normal- phase liquid chromatography for the determination of polycyclic aromatic compounds in environmental samples and standard reference materials (air particulate matter and marine sediment)

Abstract A method involving two levels of fractionation, semi-preparative gel permeation chromatography (GPC) and normal-phase liquid chromatography (NP-LC), and capillary gas chromatography with flame ionization and nitrogenphosphorus detection coupled with mass spectrometry, was developed for the determination of polar substituted aromatic compounds (PACs) in environmental matrices. A GPC clean-up procedure (BioBeads SX-12-dichloromethane) efficiently removed lipidic matter from organic extracts, yielding an enriched PAC fraction. NP-LC (μPorasil) of that fraction provided a selectivity based on chemical classes and moderate to high recoveries for standard PAC. The application of this method to environmental samples and reference materials, air particular matter (NIST 1649) and marine sediment (HS-4), demonstrated the validity of the procedure for the determination of polycyclic aromatic hydrocarbons (PAHs) from both qualitative and quantitative points of view. Further, a variety of aromatic ketones, quinones and aldehydes (oxy-PAHs) were determined in both matrices.

The C-12 and C-20 configurations of some neo-clerodane diterpenoids isolated from Teucrium species

Abstract A convenient and conclusive method for determining the C-12 stereochemistry of neo-clerodan-20,12-olide derivatives, even when only one epimer is available, is by 1 H NMR NOE measurements. The C-12 configuration of 26 neo-clerodane diterpenoids isolated from Teucrium species has been re-examined by using this type of experiment. The results indicated that all the previous assignments were correct, except for teupyreinin, where the previously assigned C-12( S ) configuration must be amended to C-12( R ). This was confirmed by chemical transformations and additional 1 H and 13 C NMR studies. Furthermore, the NOE experiments allowed the assignment of a C-20( S ) configuration for teuflavin, the structure of which had previously been reported without this feature, and indicated that the previously assigned C-20( S ) configuration for teupyreinidin must be amended to C-20( R ). The validity of using NOE experiments to establish the C-12 and C-20 configurations in these neo-clerodane derivatives has been confirmed by applying it to compounds whose structure had already firmly established by X-ray diffraction analyses.