Polybrominated Flame Retardants Research Articles

Exposure to persistent organic pollutants as potential risk factors for developing diabetes

Persistent organic pollutants (POPs), such as dioxins, polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs), which are synthetic chemicals or by-products with an intrinsic resistance to natural degradation processes, are released into the environment, resulting in the widespread dispersal and accumulation in the environment, as well as in human and ecological food chains. Due to their ubiquity in the environment and lipophilic properties, there is emerging concern over the potential risks of human exposure to POPs. Extensive growing evidence indicated that exposure to POPs might be strongly associated with increased risk of a worldwide epidemic of diabetes, especially type 2 diabetes, suggesting that POPs might play a key role in their pathogenesis. Based on summary of the related studies, this paper reviews the epidemiologic and experimental data that addresses the association between increased risk of diabetes and POP exposure, including dioxins, PCBs, OCPs, polybrominated flame retardants (PBFRs), and some environmental estrogens. The potential mechanisms whereby POPs cause diabetes were discussed, such as alterations in lipid metabolism, in glucose transport, in insulin signaling pathway, in steroid metabolism, and disruption of endocrine system, induction of tumor necrosis factor-α (TNF-α). However, with respect to diabetes, some of the evidence on POPs linked to risk of diabetes was suggestive of a direct or indirect association but was limited or inconclusive. Future research is urgently needed for determining the relative contribution of POPs to diabetes and elucidating the exact mechanisms.

Prenatal Exposure to Organohalogens, Including Brominated Flame Retardants, Influences Motor, Cognitive, and Behavioral Performance at School Age

BackgroundOrganohalogen compounds (OHCs) are known to have neurotoxic effects on the developing brain.ObjectiveWe investigated the influence of prenatal exposure to OHCs, including brominated flame retardants, on motor, cognitive, and behavioral outcome in healthy children of school age.MethodsThis study was part of the prospective Groningen infant COMPARE (Comparison of Exposure-Effect Pathways to Improve the Assessment of Human Health Risks of Complex Environmental Mixtures of Organohalogens) study. It included 62 children in whose mothers the following compounds had been determined in the 35th week of pregnancy: 2,2′-bis-(4 chlorophenyl)-1,1′-dichloroethene, pentachlorophenol (PCP), polychlorinated biphenyl congener 153 (PCB-153), 4-hydroxy-2,3,3′,4′,5-pentachlorobiphenyl (4OH-CB-107), 4OH-CB-146, 4OH-CB-187, 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47), BDE-99, BDE-100, BDE-153, BDE-154, and hexabromocyclododecane. Thyroid hormones were determined in umbilical cord blood. When the children were 5–6 years of age, we assessed their neuropsychological functioning: motor performance (coordination, fine motor skills), cognition (intelligence, visual perception, visuomotor integration, inhibitory control, verbal memory, and attention), and behavior.ResultsBrominated flame retardants correlated with worse fine manipulative abilities, worse attention, better coordination, better visual perception, and better behavior. Chlorinated OHCs correlated with less choreiform dyskinesia. Hydroxylated polychlorinated biphenyls correlated with worse fine manipulative abilities, better attention, and better visual perception. The wood protective agent (PCP) correlated with worse coordination, less sensory integrity, worse attention, and worse visuomotor integration.ConclusionsOur results demonstrate for the first time that transplacental transfer of polybrominated flame retardants is associated with the development of children at school age. Because of the widespread use of these compounds, especially in the United States, where concentrations in the environment are four times higher than in Europe, these results cause serious concern.

Determination of Halogenated Natural Products in Passive Samplers Deployed along the Great Barrier Reef, Queensland/Australia

Halogenated natural products (HNPs) have been increasingly reported to occur in marine wild life from all oceans. Several HNPs, such as 2,3,3',4,4',5,5'-heptachloro-1'-methyl-1,2'-bipyrrole (1) and 4,6-dibromo-2-(2',4'-dibromo)phenoxyanisole (2'-MeO-BDE 68 or BC-2), were detected at particularly high concentrations in dolphins from Queensland/Australia. About half of the coastline of Queensland (approximately 2500 km) is covered by the Great Barrier Reef, a rich ecosystem hosting a huge variety of species, many of which are known to produce natural compounds. In this study, semipermeable membrane devices (SPMDs) were deployed as passive samplers for about 30 days at 12 marine and 2 nonmarine sites (i.e., rivers) along the Great Barrier Reef as part of a routine monitoring program during November 2007 and May 2008. Q1 and 2'-MeO-BDE 68 were detected at the marine sites with frequencies of about 65% but not in any sample from the two rivers. Further HNPs (2,4,6-tribromophenol, TBP; 2,4,6-tribromoanisole, TBA; 2,2'-dimethoxy-3,3'5,5'-tetrabromobiphenyl, 2,2'-diMeO-BB 80 or BC-1; 3,5-dibromo-2-(2',4'-dibromo)phenoxyanisole, 6-MeO-BDE 47 or BC-3; and 3,5-dibromo-2-(3',5'-dibromo,2'-methoxy)phenoxyanisole, 2',6-diMeO-BDE 68 or BC-11) were detected as well with frequencies of 18-97% in the marine samples, but no polybrominated flame retardants were detected. The highest amount of a single HNP, 2.3 microg/SPMD, was determined for TBP, which had a frequency of detection of only 46%. The maximum (average) amount in the SPMDs from marine sites was 44 ng (12 ng) for (1 and 115 ng (20 ng) for 2'-MeO-BDE 68. A first order kinetic model was used to estimate concentrations of the HNPs in the water phase. Based on the depuration of performance reference compounds obtained at one of the sites, we assumed a sampling rate of 16 L/day. We used this sampling rate to estimate that the highest and average available concentrations of Q1 in the water during the deployment of the SPMD were 97 and 25 pg/L, respectively. The estimated maximum water concentrations of 2'-MeO-BDE 68, 2,2'-diMeO-BB 80, 6-MeO-BDE 47, and 2',6-diMeO-BDE 68 were on average 2-5.5 fold higher than that of Q1. The results confirm that the HNPs are produced throughout the Great Barrier Reef, which appears to be a significant source of these compounds.

Behavioral changes in aging but not young mice after neonatal exposure to the polybrominated flame retardant decaBDE.

BackgroundAfter several decades of commercial use, the flame-retardant chemicals polybrominated diphenyl ethers (PBDEs) and their metabolites are pervasive environmental contaminants and are detected in the human body. Decabrominated diphenyl ether (decaBDE) is currently the only PBDE in production in the United States.ObjectivesLittle is known about the health effects of decaBDE. In the present study we examined the effects of neonatal decaBDE exposure on behavior in mice at two ages.MethodsNeonatal male and female C57BL6/J mice were exposed to a daily oral dose of 0, 6, or 20 mg/kg decaBDE from postnatal days 2 through 15. Two age groups were examined: a cohort that began training during young adulthood and an aging cohort of littermates that began training at 16 months of age. Both cohorts were tested on a series of operant procedures that included a fixed-ratio 1 schedule of reinforcement, a fixed-interval (FI) 2-min schedule, and a light–dark visual discrimination.ResultsWe observed minimal effects on the light–dark discrimination in the young cohort, with no effects on the other tasks. The performance of the aging cohort was significantly affected by decaBDE. On the FI schedule, decaBDE exposure increased the overall response rate. On the light–dark discrimination, older treated mice learned the task more slowly, made fewer errors on the first-response choice of a trial but more perseverative errors after an initial error, and had lower latencies to respond compared with controls. Effects were observed in both dose groups and sexes on various measures.ConclusionsThese findings suggest that neonatal decaBDE exposure produces effects on behavioral tasks in older but not younger animals. The behavioral mechanisms responsible for the pattern of observed effects may include increased impulsivity, although further research is required.

Development of the First Certified Reference Materials for Several Brominated Flame Retardants in Polymers

The first reference materials certified for several polybrominated flame retardants in polymers were developed. Commercially available polyethylene and polypropylene were fortified with technical mixtures of Pentabrominated diphenylether (Penta-BDE), Octa-BDE, Deca-BDE, and Decabrominated biphenyl (BB) (where the capitalized forms refer to the technical mixtures). Homogeneity was tested on 20 units of each material, and between-unit variation was confirmed to be below 4% for all congeners. Stability was assessed after storage of samples for 1 year at 4, 18, and 60 degrees C. Uncertainty of degradation during transport was found negligible for all congeners, whereas uncertainty of degradation for storage of 24 months at 4 degrees C was estimated between 2% and 11%. A characterization intercomparison involving 16 laboratories was organized. After exclusion of technically doubtful results, between-laboratory standard deviations ranged from 3% to 12%, making this intercomparison the best for this field of analysis so far. Statistical analysis revealed that the use of isotopically labeled internal standards did not improve analytical precision in this study. The good comparability, together with the independent confirmation of the assigned mass fractions via the total bromine content as well as by using non-GC/MS-based methods, allowed for the first time the certification of polymer materials for several brominated flame retardants.

Optimization of microwave-assisted extraction with saponification (MAES) for the determination of polybrominated flame retardants in aquaculture samples

The efficiency of microwave-assisted extraction with saponification (MAES) for the determination of seven polybrominated flame retardants (polybrominated biphenyls, PBBs; and polybrominated diphenyl ethers, PBDEs) in aquaculture samples is described and compared with microwave-assisted extraction (MAE). Chemometric techniques based on experimental designs and desirability functions were used for simultaneous optimization of the operational parameters used in both MAES and MAE processes. Application of MAES to this group of contaminants in aquaculture samples, which had not been previously applied to this type of analytes, was shown to be superior to MAE in terms of extraction efficiency, extraction time and lipid content extracted from complex matrices (0.7% as against 18.0% for MAE extracts). PBBs and PBDEs were determined by gas chromatography with micro-electron capture detection (GC-μECD). The quantification limits for the analytes were 40–750 pg g−1 (except for BB-15, which was 1.43 ng g−1). Precision for MAES-GC-μECD (%RSD < 11%) was significantly better than for MAE-GC-μECD (%RSD < 20%). The accuracy of both optimized methods was satisfactorily demonstrated by analysis of appropriate certified reference material (CRM), WMF-01.

Perfluoroalkane Acids and Fetal Growth

In the November issue of Environmental Health Perspectives, Apelberg et al. (2007) reported an inverse relationship between umbilical cord blood concentrations of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) and ponderal index and head circumference in children delivered vaginally in Baltimore, Maryland. In the same issue, Fei et al. (2007) reported an inverse relationship between first trimester maternal blood PFOA (but not PFOS) concentration and birth weight in Danish infants born to normal-weight women. Although these studies do not necessarily support one another (Fei et al. also collected cord blood but did not report these results), they raise the important question of whether low-level exposure to perfluoroalkane acids might affect fetal growth. In both articles, the authors called attention to the inconsistency between these findings and those in experimental animal studies, in which fetal growth effects occur only at blood concentrations several orders of magnitude higher than were measured in human umbilical cord or maternal blood. The question was reasonably posed by both groups whether a confounder could be responsible for the observed associations. The Baltimore group (Apelberg et al. 2007) identified two candidate confounders that may explain their findings: diet and plasma volume. Perfluoroalkanesulfonamides, which may be metabolized to PFOS, have been used in grease- and water-repellant packaging for foods, particularly pizza, french fries, and other fried foods. The Canadian Total Diet Study (Tittlemier et al. 2006) detected perfluoroalkanesulfonamides in all foods tested, but the highest concentrations were found in pizza, microwave popcorn, egg breakfast sandwiches, french fries, chicken nuggets, and fish burgers. Fluorotelomer alcohols, which can be converted to the corresponding alkane acids, have been used in coatings for paper, including microwave popcorn bags. 8-2 Fluorotelomer alcohol can be converted atmospherically and metabolically to PFOA, and gavage treatment of pregnant mice with 8-2 fluorotelomer alcohol results in the appearance of PFOA in fetuses (Henderson and Smith 2007). Both 8-2 fluorotelomer alcohol and PFOA have been found in popcorn bags and in the vapor produced after cooking microwave popcorn (Begley et al. 2005; Sinclair et al. 2007). The pregnancies studied by Fei et al. (2007) occurred in 1996–2002, a period during which perfluorinated compounds were commonly used in fast-food packaging. The use of perfluorinated compounds in food packaging decreased some years before 2004–2005, the study period of Apelberg et al. (2007); however, PFOS and PFOA have long half-lives and may still have been present as markers of a high intake of fast-food. A high intake of fast food may in turn be a marker of poor nutrition. The Danish National Birth Cohort (Fei et al. 2007) included a food frequency questionnaire. It would be interesting to know if a relationship between nutrition and maternal blood perfluoroalkane acid concentration was detected. PFOA and PFOS repel fat and are distributed in body water, particularly plasma. Women with a reduced plasma or body water volumes would distribute the same body burden of perfluoroalkane acids in a smaller space, producing higher perfluoroalkane acid concentrations. Fat-free body mass and total body water volumes are important predictors of birth weight (Butte et al. 2003; Lederman et al. 1999; Mardones-Santander et al. 1998; Sanin Aguirre et al. 2004), giving rise to the possibility that higher maternal blood (and therefore fetal blood) concentrations of PFOS and PFOA are markers of reduced plasma or total body water volumes, producing an apparent inverse association between the perfluoroalkane acid concentrations and fetal growth. A reasonable next step in addressing the question of whether perfluoroalkane acids (at current human blood concentrations) play a role in fetal growth will be studies in which maternal nutrition and body composition, as opposed to body weight, are considered as possible confounders.

Extraction of components of composite materials: ionic liquids in the extraction of flame retardants from plastics

AbstractBACKGROUND: The presence of certain polybrominated flame retardants in electrical and electronic equipment plastics can make recycling of the raw materials difficult. We have investigated the use of ionic liquids to selectively remove these harmful components and a series of imidazolium‐ and pyridinium‐based ionic liquids have been prepared in a microwave reactor. Microwave synthesis is rapid and the products obtained are identical to those obtained by conventional synthetic methods.RESULTS: The solubilities of the flame retardants decabromodiphenyl ether (deca‐BDE), antimony trioxide and high‐impact polystyrene (HIPS) in the ionic liquids, identified 1‐hexylpyridinium bromide as the most suitable for solid–liquid and liquid–liquid extraction, for the selective recovery of materials from the plastic composite. Optimum extraction of deca‐BDE and antimony trioxide was achieved by liquid–liquid extraction from an 8% solution of HIPS in ethyl acetate, with a reaction time of 10 min at 77 °C. The percentages of deca‐BDE and Sb2O3 extracted are 92.7% and 99.9%, respectively. The chain length of the polymer is not degraded by the extraction process, giving a residue that can be recycled.CONCLUSIONS The results show that ionic liquids can be designed to achieve selective material recovery from composites. Copyright © 2008 Society of Chemical Industry

Xeno-estrogenic compounds in precipitation

The exposure to some chemicals can lead to hormone disrupting effects. Presently, much attention is focused on so-called xeno-estrogens, synthetic compounds that interact with hormone receptors causing a number of reactions that eventually lead to effects related to reproduction and development. The current study was initiated to investigate the presence of a number of such compounds in precipitation as a follow-up on a previous study in which pesticide concentrations in air and precipitation were determined. Rainwater samples were collected at about 50 locations in The Netherlands in a four week period. The samples were analysed for bisphenol-A, alkylphenols and alkylphenol ethoxylates, phthalates, flame retardants and synthetic musk compounds. The results clearly indicated the presence of these compounds in precipitation. The concentrations ranged from the low ng l(-1) range for flame retardants to several thousands of ng l(-1) for the phthalates. Bisphenol-A was found in 30% of the samples in concentrations up to 130 ng l(-1), while alkylphenols and alkylphenol ethoxylates were found in virtually all locations in concentrations up to 920 ng l(-1) for the individual compounds. Phthalates were by far the most abundant xeno-estrogens in the precipitation samples and were found in every sample. Di-isodecyl phthalate was found in a surprisingly high concentration of almost 100 000 ng l(-1). Polybrominated flame retardants were found in the low ng l(-1) range and generally in less than 20% of the samples. Noticeable was the finding of hexabromocyclododecane, a replacement for the polybrominted diphenyl ethers at one location in a concentration of almost 2000 ng l(-1). Finally, as expected, synthetic musk compounds were detected in almost all samples. This is especially true for the polycyclic musks HHCB and AHTN. Nitro musks were found, but only on a few locations. Kriging techniques were used to calculate precipitation concentrations in between actual sampling locations to produce contour plots for a number of compounds. These plots clearly show located emission sources for a number of compounds such as bisphenol-A, nonylphenol ethoxylate, phthalates and AHTN. On the contrary, the results for HHCB and some phthalates indicated diffuse emission patterns, probably as the result of the use of consumer products containing these compounds.

Comparison of toxicity of congener-153 of PCB, PBB, and PBDE to Daphnia magna

Acute and reproductive toxicity tests were performed to assess the effects of two polybrominated flame retardants, polybrominated biphenyl (PBB)-153, polybrominated diphenyl ether (PBDE)-153, and polychlorinated biphenyl (PCB)-153 on Daphnia magna. According to the 24-h acute toxicity test, up to concentration of 210 μg/L, these chemicals were not toxic to Daphnia. In the reproductive tests, we used concentrations of 12.5, 25, 50, and 100 μg/L. At the concentrations of 50 and 100 μg/L, the Daphnia started to die after the second test day, and there were no Daphnia alive on the fourth test day. The testing was continued using the concentrations of 12.5 and 25 μg/L. The reproductive tests were then repeated using only concentrations of 12.5 and 25 μg/L. The results, based on the weight per volume (w/v) concentrations, showed that of the three compounds we tested PCB-153 had the most severe effects on the Daphnia reproduction. PBDE-153 was the least toxic, although in this group, too, offspring production was significantly lower and mortality significantly higher than those in the control group. Also, it was observed that PCB-153 accumulated into the parent Daphnia nearly three times more actively than the two other chemicals we tested. These chemicals, although having very similar structures, seem to affect differently the reproduction of Daphnia. They also seem to accumulate differently into Daphnia. Thus, further research data are needed to understand the mechanisms responsible for the effects caused by these kinds of chemicals and to assess their risks accurately.

Results of an interlaboratory comparison on the determination of polybrominated flame retardants in poly(ethyleneterephthalate)

An intercomparison to establish the performance of routine laboratories in the determination of polybrominated flame retardants in polymers was organised. Commercial poly(ethyleneterephthalate) was fortified with technical pentabromodiphenyl ether, octabromodiphenyl ether and decabromodiphenyl ether mixtures and with a decabromobiphenyl technical mixture at 0.4-0.8 g/kg. Homogeneity and stability of the total Br content in the material was confirmed. Thirty-seven laboratories from Europe, Asia and the Americas submitted results. Relative repeatability standard deviations for individual congeners ranged from 7 to 17%. Relative between-laboratory standard deviations ranged from 22 to 61%. No significant influence of a common standard, application of a standard method or method parameters could be identified. The quality and uncertainty of the results of this study are significantly worse than those reported in the environmental field and indicate a clear need for a learning process among the laboratories involved. [figure: see text] Mandel's h (between labs): critical level: 1.91.

Guest Editorial: Biomarkers of Perfluorinated Chemicals and Birth Weight

Guest Editorial: Biomarkers of Perfluorinated Chemicals and Birth Weight

Presence of polybrominated flame retardants in sewage sludges and sediments in the Czech Republic

Presence of polybrominated flame retardants in sewage sludges and sediments in the Czech Republic

Cytotoxicity of TBBPA and effects on proliferation, cell cycle and MAPK pathways in mammalian cells

Poly-brominated flame retardants are ecotoxicologically relevant chemicals that can show high persistency in environmental samples and bioaccumulation in marine and fresh water animals. One of the most widely used compound is tetrabromobisphenol A (TBBPA). Until today, the toxicological data are rather fragmentary. Our studies on acute and sub-acute toxic effects with established cell lines demonstrate that TBBPA interferes with cellular signaling pathways. Cell viability is significantly reduced in a time- and concentration-dependent manner. The observed EC50 for rat kidney cells (NRK) was 52 microM (27 mg/l), 168 microM (90 mg/l) for A549 human lung cells, and 200 microM (108 mg/l) for Cal-62 human thyroid cells, respectively. The comparison of TBBPA with the non-brominated substance bisphenol A (BPA) clearly demonstrates that only the brominated compound exerts these effects on proliferation and cell viability. Cell cycle regulation was influenced considerably in Cal-62 cells, showing an explicit G2/M arrest in the cell cycle at TBBPA concentrations higher than 75 microM. Cellular signaling pathways directly connected to these affected parameters, e.g. the mitogen activated protein kinase (MAPK) cascades, are partly influenced in a cell specific and dose dependent manner. The extracellular-signal regulated kinase (ERK) is deactivated in NRK and A549 cells and activated in Cal-62 cells with increasing TBBPA concentrations.

Measuring emissions of organophosphate flame retardants using a passive flux sampler

Flame retardants are used in polymers to reduce the flammability of building materials, electric appliances, fabric and papers. In recent years, organophosphate flame retardants have been used as substitutes for polybrominated flame retardants (BFRs). In Japan, the amount of organophosphate flame retardants used in 2001 was about five times more than in 2000. Recently, several studies have shown the health concerns for some organophosphate flame retardants. Little research has been performed on the emission of organophosphate flame retardants, especially the relationship between content and emissions. In this study, a new type of passive sampler was developed to measure emissions of organophosphate flame retardants from plastic materials. With this sampler, emissions from polyvinyl chloride wallpaper samples with different content of tris(2-chloroisopropyl)phosphate (TCPP) at different temperatures were examined. The observed maximum emissions of TCPP from 1, 3, 5, 10 and 20 w/w% content wallpaper materials were 262.3, 452.6, 644.8, 1119.1 and 2166.8 μg m −2 h −1, respectively. Emissions from 5% TCPP content materials at 40 and 60 °C were 1135.7 and 2841.2 μg m −2 h −1, respectively. A significantly positive correlation between the flux of TCPP and the TCPP content of the wallpaper samples was observed. A linear relationship was found between the inverse of temperature and the logarithm of TCPP emission. The results imply that the use of materials with a high organophosphate flame retardant content can lead to high emission rates in high-temperature indoor environments.

Indoor organophosphate and polybrominated flame retardants in Tokyo

In Japan, organophosphate and polybrominated flame retardants are used in building materials and electric appliances to protect them from fire hazards. In this study, to identify the emission sources of these flame retardants to indoor air, the migration rates (flux) of organophosphate and polybrominated flame retardants from building materials and electrical appliances to solid extraction disks that were placed in contact with the interior surfaces were measured. In addition to the migration test, indoor air and outdoor air concentrations of these flame retardants were investigated. With regard to building materials in a newly built house, triethylphosphate (TEP) and tributylphosphate (TBP) were detected in the wall and ceiling coverings, and tris(2-butoxyethyl)phosphate (TBEP) was detected in the wooden flooring cleaned with a floor polish agent. With regard to electrical appliances, triphenylphosphate (TPHP) was predominantly detected in computer monitors and tris(2-chloroethyl) phosphate (TCEP) in television (TV) sets, with the highest median levels. Among the polybrominated compounds, only 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) was detected from a few old TV sets manufactured before 1995. In an indoor and outdoor air survey, nine organophosphates and nine polybrominated flame retardants were detected from indoor air. In outdoor air, only four organophosphate flame retardants were detected. The maximum level of indoor organophosphate compounds was 1260 ng/m(3) with tris(2-chloro-1-methylethyl) phosphate (TCPP), and that of polybrominated compounds was 29.5 ng/m(3) with hexabromocyclododecane (HBCD). Tetrabromobisphenol A (TBBPA) was not detected in this study, although it has the largest demand among flame retardants in Japan. The results of the migration test and the indoor air survey revealed that in indoor air, organophosphate compounds were more predominant than polybrominated compounds in Tokyo. Polybrominated biphenyls (PBB) and polybrominated diphenyl ethers (PBDE) are commonly used as flame retardants in plastics. The use of these two compounds in electric appliances will be banned in 2007 by the EU Directives on waste electrical and electronic equipment (WEEE) and on the restriction of the use of certain hazardous substances (RoHS) in electrical and electronic equipment. In Japan, the use of PBB was banned and that of PBDE diminished in the early 1990s by the self-imposed controls of the Japanese Flame Retardants Conference (Akutu and Hori, 2004). In Japan, the predominantly used organic flame retardants were tetrabromobisphenol A and organophosphate compounds. Tetrabromobisphenol A has been reported to disrupt endocrine systems (Kitamura et al., 2005), and some organophosphate flame retardants were recently reported to have neurochemical hazardous effects. Furthermore, organophosphate compounds were suspected to cause endocrine-disrupting effects (Fang et al., 2003; Ohyama et al., 2005) or attention deficit hyperactivity disorder (ADHD) (Winrow et al., 2003). In this study, organophosphate and polybrominated flame retardants were surveyed in indoor environments in Tokyo.

Identification of Proteomic Signatures of Exposure to Marine Pollutants in Mussels (Mytilus edulis)

Bivalves and especially mussels are very good indicators of marine and estuarine pollution, and so they have been widely used in biomonitoring programs all around the world. However, traditional single parameter biomarkers face the problem of high sensitivity to biotic and abiotic factors. In our study, digestive gland peroxisome-enriched fractions of Mytilus edulis (L., 1758) were analyzed by DIGE and MS. We identified several proteomic signatures associated with the exposure to several marine pollutants (diallyl phthalate, PBDE-47, and bisphenol-A). Animals collected from North Atlantic Sea were exposed to the contaminants independently under controlled laboratory conditions. One hundred and eleven spots showed a significant increase or decrease in protein abundance in the two-dimensional electrophoresis maps from the groups exposed to pollutants. We obtained a unique protein expression signature of exposure to each of those chemical compounds. Moreover a set of proteins composed a proteomic signature in common to the three independent exposures. It is remarkable that the principal component analysis of these spots showed a discernible separation between groups, and so did the hierarchical clustering into four classes. The 14 proteins identified by MS participate in alpha- and beta-oxidation pathways, xenobiotic and amino acid metabolism, cell signaling, oxyradical metabolism, peroxisomal assembly, respiration, and the cytoskeleton. Our results suggest that proteomic signatures could become a valuable tool to monitor the presence of pollutants in field experiments where a mixture of pollutants is often present. Further studies on the identified proteins could provide crucial information to understand possible mechanisms of toxicity of single xenobiotics or mixtures of them in marine ecosystems.

Specificity of the peroxisome proliferation response in mussels exposed to environmental pollutants

Peroxisome proliferation has been proposed as novel biomarker of exposure to organic pollutants in aquatic organisms. Peroxisome proliferator compounds comprise a heterogeneous group of substances known for their ability to cause massive proliferation of peroxisomes and liver carcinogenesis in sensitive species such as rodents. Recently, several marine organisms (mussels and fish) have been shown as target species of peroxisome proliferators. In the present work, we aimed to investigate the specificity of the peroxisome proliferation response in mussels. For this purpose, mussels ( Mytilus edulis) were exposed for three weeks to North Sea crude oil (NSO), a mixture of NSO, alkylphenols and extra PAHs (MIX), diallylphthalate (DAP), bisphenol-A (BPA) and tetrabromodiphenylether (TBDE), or transplanted for three weeks to four stations showing different copper concentrations in a copper mine. Peroxisome proliferation was assessed by measuring the activity of the peroxisomal β-oxidation enzyme acyl-CoA oxidase (AOX) and the volume density occupied by peroxisomes ( V VP) in the digestive gland. Mussels exposed to NSO and MIX showed significantly increased AOX activities and V VP compared to control animals. Significantly higher V VP was also found in DAP and TBDE exposed mussels. V VP did not vary in mussels transplanted into a copper concentration gradient. Our results confirm the usefulness and specificity of peroxisome proliferation as a suitable biomarker of exposure to organic contaminants such as oil derived hydrocarbons, phthalate plasticizers and polybrominated flame retardants in mussels.

Effects of Developmental Exposure to 2,2′,4,4′,5-Pentabromodiphenyl Ether (PBDE-99) on Sex Steroids, Sexual Development, and Sexually Dimorphic Behavior in Rats

Increasing concentrations of polybrominated flame retardants, including polybrominated diphenyl ethers (PBDEs), in breast milk cause concern about possible developmental effects in nursed babies. Because previous studies in rats have indicated effects on sex steroids and sexually dimorphic behavior after maternal exposure to polychlorinated biphenyls (PCBs), our goal in the present study was to determine if developmental exposure to 2,2′,4,4′,5-pentabromodiphenyl ether (PBDE-99) induces similar endocrine-mediated effects. Pregnant rats were exposed to vehicle or PBDE-99 (1 or 10 mg/kg body weight, daily during gestational days 10–18). For comparison, we also included a group exposed to the technical PCB mixture Aroclor 1254 (30 mg/kg body weight, daily). PBDE exposure resulted in pronounced decreases in circulating sex steroids in male offspring at weaning and in adulthood. Female offspring were less affected. Anogenital distance was reduced in male offspring. Puberty onset was delayed in female offspring at the higher dose level, whereas a slight acceleration was detected in low-dose males. The number of primordial/primary ovarian follicles was reduced in females at the lower dose, whereas decline of secondary follicles was more pronounced at the higher dose. Sweet preference was dose-dependently increased in PBDE-exposed adult males, indicating a feminization of this sexually dimorphic behavior. Aroclor 1254 did not alter sweet preference and numbers of primordial/primary and secondary follicles but it did affect steroid concentrations in males and sexual development in both sexes. PBDE concentrations in tissues of dams and offspring were highest on gestational day 19. These results support the hypothesis that PBDEs are endocrine-active compounds and interfere with sexual development and sexually dimorphic behavior.

In vitro estrogenicity of polybrominated flame retardants

Estrogenicity of five brominated flame retardants (BFRs), namely BDE-47, BDE-99, BDE-205, PBB-153 and technical Firemaster BP-6™, were assessed by in vitro assays developed to detect chemicals with estrogenic properties. Recombinant yeast cells containing a human estrogen receptor gene failed to give any response to the chemicals tested. However, the positive control compound, estradiol-17β, showed that the yeast cell assays had worked properly. The freshly separated fish hepatocyte assay based on the synthesis and secretion of vitellogenin from the isolated liver cells produced a clear dose–response curve in the presence of all tested flame retardants except Firemaster BP-6™. The toxicity of the BFRs was detected by determining the cell ethoxyresorufin-O-deethylase activity (EROD). The BFRs tested induced hepatic EROD activity at low test concentrations, but started to inhibit activity at higher concentrations. The decreased detoxification capacity of the hepatocytes resulted in a decrease in the vitellogenin production of the cells. The capability of in vitro assays to detect estrogenic properties of chemicals seems to vary. Thus, further work is needed to understand the mechanisms responsible for these reactions.