Are individual NOEC levels safe for mixtures? A study on mixture toxicity of brominated flame-retardants in the copepod Nitocra spinipes

In aquatic ecosystems organisms are exposed to mixtures of pollutants. Still, risk assessment focuses almost exclusively on effect characterization of individual substances. The main objective of the current study was therefore to study mixture toxicity of a common group of industrial substances, i.e., brominated flame-retardants (BFRs), in the harpacticoid copepod Nitocra spinipes. Initially, 10 BFRs with high hydrophobicity but otherwise varying chemical characteristics were selected based on multivariate chemical characterization and tested individually for effects on mortality and development using a partial life cycle test (six days) where silica gel is used as a carrier of the hydrophobic substances. Based on these findings, six of the 10 BFRs were mixed in a series of NOEC proportions (which were set to 0.008, 0.04, 0.2, 1, and five times the NOEC concentrations for each individual BFR), loaded on silica gel and tested in a full life cycle test (26 days). Significantly increased mortality was observed in N. spinipes after six and 26 days exposure at a NOEC proportion that equals the NOEC LDR value (x1) for each BFR in the mixture (p=0.0015 and p=0.0105, respectively). At the NOECx5 proportion all animals were dead. None of the other NOEC proportions caused significant negative responses related to development and reproduction. This shows that low concentrations of individual substances can cause toxicity if exposed in mixtures, which highlights the need to consider mixture toxicity to a greater extent in regulatory work.     

Maternal transfer of brominated flame retardants in zebrafish (Danio rerio)

In many species reproduction and embryonic development have been shown to be sensitive to environmental contaminants. Understanding embryonic exposure to environmental contaminants is thus highly important. In this study concentrations of brominated flame retardants (BFRs) were measured in zebrafish eggs after parental exposure for 42 days via the diet. Zebrafish were exposed to two doses of eleven structurally-diverse BFRs. Eight BFRs were detected in the female zebrafish and maternal transfer to eggs was evident for all eight compounds. The highest concentrations in eggs were observed for hexabromocyclododecane (HBCD) and 2,4,4'-tribromodiphenyl ether (BDE 28), followed by 2,2',3,4,4',5',6-heptabromodiphenyl ether (BDE 183) and tetrabromobisphenol A 2,3-dibromopropyl ether (TBBPA DBPE). Five potential BFR metabolites were tentatively identified in female fish and maternal transfer was observed also for these compounds. The lipid adjusted concentrations in eggs were significantly higher than the concentrations in female fish for several of the BFRs. Further, the results showed a generally higher transfer in the lower exposure level and also indicated a dependency on the physico-chemical properties of the compounds.     

Brominated flame retardant concentrations and trends in abiotic media

BFR burdens in abiotic media have been less studied than in biota, despite their essential value as an aide to identifying sources, temporal and geographic trends and exposure routes. Many polymer products consist of several percent by weight of BFRs. Global trade in these can result in rapid and wholesale BFR movement. Loss from in-use products of nonreactive BFRs may be important, particularly in respect to indoor exposure. In the case of BDE-209, discharges from publicly owned treatment works may be substantial. BFR burdens in air, water and sewage sludge respond rapidly to changes in environmental BFR inputs. PBDEs have been the most widely studied. In many locales PBDE burdens in these media now surpass those of PCBs. Air and water near sources and urban areas are typically enriched relative to rural locales. The more volatile PBDEs dominate in the vapor phase, while BDE-209 typically predominates on particulates. Evidence exists for long-range transport of the more volatile PBDEs. A greater diversity of BFRs (mostly PBDEs, HBCD and TBBP-A) has been detected in sewage sludges. Land application of these sludges on agricultural fields is one conduit for soil contamination. In general, environmental concentrations of BDE-209 appear to be increasing, while penta-BDE burdens in Europe may have peaked. Sediments function as longer-term integrators of environmental burdens. Concentrations of common BFRs therein may be substantial near point sources. Evidence for debromination in the environment has been limited to date. However, some laboratory and field observations suggest it is possible to a limited extent.     

Developmental exposure to decabromodiphenyl ether (PBDE 209): effects on thyroid hormone and hepatic enzyme activity in male mouse offspring

Decabrominated diphenyl ether (PBDE 209) is the second most used brominated flame retardant (BFRs). Many studies have shown that some of the BFRs act as endocrine disruptors via alterations in thyroid hormone homeostasis and affect development. Little is known about the effect of prenatal exposure to PBDE 209 on the development in male offspring. Using a CD-1 mouse model, we attempt to estimate the possible effect of in utero exposure to PBDE 209 on thyroid hormone and hepatic enzymes activities in male offspring. Pregnant mice were administered different doses of PBDE 209 (10, 500, and 1500 mg/kg/day) or corn oil for controls per gavage from gestational days 0-17. In adult male offspring whose mothers had been treated with 1500 mg/kg of PBD 209, hepatic enzyme activity of S9 7-ethoxyresorufin O-deethylase (EROD) was weak but significantly increased (54%). However, no significant changes were observed in S9 4-nitrophenol uridinediphosphate-glucuronosyltransferase (UDPGT) in any of the treatment groups. Serum triiodothyronine (T3) was found to have decreased significantly (ca. 21% both 10 mg/kg and 1500 mg/kg) in offspring, but not thyroxine (T4). Histopathological examination revealed that prenatal exposure of PBDE 209 might be related with cell swelling of hepatocytes in male offspring and there were mild changes in the thyroid glands in 1500 mg/kg group. These data demonstrate that PBDE 209 is likely an endocrine disrupter in male mice following exposure during development. Further studies using environmentally relevant doses are needed for hazard identification.     

Transformation of tetrabromobisphenol A in the presence of different solvents and metals

Solvent-based separation method is presumably an efficient and environmentally beneficial approach for elimination of brominated flame retardants (BFRs) from waste electrical and electronic equipment (WEEE). The overall goal of this study was to evaluate possible effects of organic solvent on the behavior of BFRs during solvent-based processing of WEEE. We initiated a set of batch experiments for examining the rates and possible pathways of transformation of a representative BFR (tetrabromobisphenol A, TBBPA) using acetone, toluene, and methanol as the solvents. Our results showed that toluene and methanol had no effect on the transformation of TBBPA, but approximately 20% of TBBPA (100 mg L⁻¹) was transformed by acetone within 2 h at 50 °C. Analysis of the products with GC-MS showed that two high-molecular-weight products (MW = 586) were major products of the transformation reactions. The role of acetone as a reactant in the transformation of TBBPA was further validated with dueterated acetone. In addition, the effects of co-existing metals in WEEE (i.e., Zn, Cu, and Ni) on the transformation of TBBPA in the solvent systems were investigated. These metals tested were found to greatly enhance the rates of TBBPA transformation. The metal facilitated solvent reactions with TBBPA may lower the extractability of TBBPA by formation of larger and less soluble products, hence potentially increasing the cost for separating the chemical from WEEE.     

Levels and sources of brominated flame retardants in human hair from urban, e-waste, and rural areas in South China

Human hair and indoor dust from urban, e-waste, and rural areas in south China were collected and analyzed for brominated flame retardants (BFRs). BFRs concentrations in hair from occupational e-waste recycling workers were higher than those from non-occupational exposed residents in other sampling areas. Polybrominated diphenyl ethers (PBDEs) and decabromodiphenyl ethane (DBDPE) are two major BFRs in hair samples. The PBDE congener profiles in hair from the e-waste area are different from those from urban and rural areas with relatively higher contribution of lower brominated congeners. DBDPE, instead of BDE209, has become the major BFR in non-e-waste recycling areas. Significant correlations were found between hair level and dust level for DBDPE and BTBPE but not for PBDEs. The different PBDE congener profiles between dust and hair may suggest that exogenous exposure to the PBDE adsorbed on dust is not a major source of hair PBDEs.     

Organochlorines, brominated flame retardants and mercury levels in six seabird species from the Gulf of St. Lawrence (Canada): Relationships with feeding ecology, migration and molt

Concentrations of organochlorines (OCs), brominated flame retardants (BFRs) and mercury (Hg) were measured in eggs of six seabird species breeding in the Gulf of St. Lawrence, Canada. Stable nitrogen (δ¹⁵N) and carbon (δ¹³C) isotopes were used as ecological tracers to measure trophic level and connectivity with benthos, respectively. Concentrations, patterns as well as ecological tracers varied significantly between species. The sum of polychlorinated biphenyls (ΣPCBs) was the most important group measured in all seabird species based on concentration followed generally by the sum of chlorinated pesticides (ΣCPs), the sum of brominated flame retardants (ΣBFRs) and finally total Hg (THg). ΣPCBs, ΣCPs and ΣBFRs increased with trophic level, whereas THg did not. Only ΣBFRs increased with a higher connectivity with the benthos. Seabird species resident to the Great Lakes-St. Lawrence ecosystem showed higher Hg and BFR levels than migratory species. Molt patterns were used to explain variations of contaminant levels.     

Dioxins: WHO's tolerable daily intake (TDI) revisited

In December 1990, the World Health Organization (WHO) established a tolerable daily intake (TDI) of 10 pg/kg b.w. (body weight) for TCDD, based on liver toxicity, reproductive effects and immunotoxicity in experimental animals, and making use of kinetic data in humans and experimental animals. Since then new epidemiological and toxicological data have emerged, in particular with respect to neurodevelopmental and endocrine effects of dioxin. Therefore, the European Centre for Environment and Health of the World Health Organization (WHO-ECEH) and the International Programme on Chemical Safety (IPCS) jointly organized a consultation on the "Assessment of the health risk of dioxins: re-evaluation of the TDI", May 1998, Geneva, Switzerland. The participants discussed the health risks for infants, cancer and non-cancer endpoints in humans and animals, mechanistic aspects, kinetic behaviour, modelling, exposure, and the applicability of the toxic equivalency (TEQ) concept. For the health risk assessment of dioxin-like compounds, the WHO Consultation focused on the most sensitive effects that are considered adverse (hormonal, reproductive and developmental effects) seen at low doses in animal studies (rats and monkeys). Human daily intakes corresponding with body burdens similar to those associated with adverse effects in animals could be estimated to be in the range of 14-37 pg/kg b.w./day. To arrive at a TDI expressed as TEQ, a composite uncertainty factor of 10 was recommended. By applying this uncertainty factor a TDI range of 1-4 pg TEQs/kg body weight was established. An extensive executive summary of the results of this WHO Consultation with all the underlying background documents will be published in Food Additives and Contaminants (in press).     

Analysis of Ah receptor pathway activation by brominated flame retardants

Brominated flame-retardants (BFRs) are used as additives in plastics to decrease the rate of combustion of these materials, leading to greater consumer safety. As the use of plastics has increased, the production and use of flame-retardants has also grown. Many BFRs are persistent and have been detected in environmental samples, raising concerns about the biological/toxicological risk associated with their use. Most BFRs appear to be non-toxic, however there is still some concern that these compounds, or possible contaminants in BFRs mixtures could interact with cellular receptors. In this study we have examined the interaction of decabromodiphenyl ether, Firemaster BP4A (tetrabromobisphenol A), Firemaster PHT4 (tetrabromophthalic anhydride), hexabromobenzene, pentabromotoluene, decabromobiphenyl, Firemaster BP-6 (2,2^′,4,4^′,5,5^′-hexabromobiphenyl) and possible contaminants of BFR mixtures with the Ah receptor. Receptor binding and activation was examined using the Gel Retardation Assay and increased expression of dioxin responsive genes was detected using the reporter gene based CALUX assay. The results demonstrate the ability of BFRs to activate the AhR signal transduction pathway at moderate to high concentrations as assessed using both assays. AhR-dependent activation by BFRs may be due in part to contaminants present in commercial/technical mixtures. This was suggested by our comparative analysis of Firemaster BP-6 versus its primary component 2,2^′,4,4^′,5,5^′-hexabromobiphenyl. Some technical mixtures of brominated flame-retardants contain brominated biphenyls, dioxins or dibenzofurans as contaminants. When tested in the CALUX assay these compounds were found to be equivalent to, or more active than their chlorinated analogues. Relative effective potency values were determined from dose response curves for these brominated HAHs.     

Levels and trends of HBCD and BDEs in the European and Asian environments, with some information for other BFRs

In this paper, we review recent data for brominated flame retardants (particularly BDEs, HBCD and TBBP-A) in samples from the European and Asian environments, including recent temporal trend studies. Research is active and we cite over 100 studies published during 2005-2007. Environmental compartments studied comprise the atmosphere, indoor and outdoor air, sewage sludges, soils and sediments and a variety of biological samples and food chains. Findings include that the lack of reference materials for use in analytical quality control and method validation identified earlier has been addressed, and certified concentrations are now available for a number of BDE congeners in six pre-existing and one new reference materials. BDE209 was certified in three samples of indoor dust. The analysis of BDE209 remains problematic, but suitable methods for its determination are available and need to be applied. The contamination of indoor environments with BFRs has been recognised as representing a significant uptake pathway, particularly via dust. Additional data for TBBP-A are needed from areas, where it is produced and used, primarily Asia, as the worst-case scenario. As a reactive flame retardant it is less likely to leach from finished products. Also, issues regarding the importance of e-waste recycling as a source of BFRs to the local populations and the local environment require urgent study, certainly in China and possibly elsewhere. Generally, trends show a levelling in concentrations of BDEs and increases in concentrations of HBCD wherever determined and BDE209 in Asia.     

Dilemmas facing regulatory and advisory bodies dealing with conflicting results.

Regulatory and advisory bodies have to take into account the possibility that conventional toxicological screening and testing methods may be failing to pick up relevant effects, with the implication that testing may need to go down to much lower exposure levels and use specifically designed studies. In the paper, attention is focused on the problems of understanding endocrine disrupting activity. The shape of the dose-response curve for endocrine disrupters may differ from that normally associated with toxic chemicals (the monotonic sigmoid curve). Thus, prediction of both the nature of the effects of potential endocrine disrupters and the intensity of those effects at different doses may be equally difficult. Endocrine effects are not currently amenable to analysis by a single apical assay. While existing toxicological tests can pick up many end-organ and functional effects that are relevant to endocrine disturbances, they are generally poor at exploring apparently subtle effects such as disturbances of brain endocrine homeostatic mechanisms which may be critical for normal development.     

Occurrence and distribution of PCB metabolites in blood and their potential health effects in humans: a review

In recent years, attention has been directed to chemicals with possible endocrine-disrupting properties. Polychlorinated biphenyls (PCBs) and their metabolites belong to one group of environmental contaminants that have been shown to interact with the endocrine system in mammals, including humans. Although recent developments have been made in terms of determination of PCB metabolites in blood samples, still limited number of studies have been able to elucidate their profiles and toxicological and health effects in humans. This review aims to evaluate and compare the levels of hydroxylated PCBs (OH-PCBs) and methyl sulfone PCBs (MeSO₂-PCBs) in blood and their relationship to parent compounds and also address the potential risks and adverse health effects in humans. Levels of OH-PCBs varied between 0.0002 and 1.6 ng g^?1 w/w in human serum/plasma from the selected literature, correlating well with ∑PCBs. In contrast, ∑OH-PCB/∑PCB ratio in animals did not show a significant correlation, which might suggest that the bioaccumulation plays an even more important role in the concentration of OH-PCBs compared to PCB metabolism. Highest levels of MeSO₂-PCBs were reported in marine mammals with high selectivity retention in the liver. Health effects of PCB metabolites included carcinogenicity, reproductive impairment, and developmental neurotoxicity, being more efficiently transferred to the brain and across the placenta from mother to fetus in comparison to the parent PCBs. Based on the lack of knowledge on the occurrence and distribution of lower chlorinated OH-PCBs in humans, further studies to identify and assess the risks associated to human exposure are essential.     

Leaching of brominated flame retardants from TV housing plastics in the presence of dissolved humic matter

In this study, we investigated the contents of several brominated compounds in TV molding plastics, as well as their leaching characteristics in the presence of DHM. The PBDE content was about 3% of the sample weight, and deca-BDE was the most abundant homologue, accounting for over 80% of the total amount. TBBPA, PBPs and PBBs content was 8100, 4700 and 250 ng/g, respectively. Despite no detection of most of the lower brominated DEs in distilled water, most homologues could be detected in DHM solution, and their solubility increased according to the contact time; those of highly brominated compounds increased to 10 times their maximum solubility in distilled water. Especially, contrary to the relatively faster equilibrium in distilled water, BFR solubility in DHM solution was maintained even after 20 days. In addition, a modified first-order model adequately reflected rapid desorption for each compound in the initial period, but slow desorption afterwards. From an overall perspective, it is clear that hydrophobic BFRs can leach out to a great extent in the presence of DHM, which is a matter of great concern in E&E waste as the potential contaminant source of BFRs, especially in landfills and open dump sites that provide the perfect conditions for exposure of BFRs to abundant DHM.     

The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

Soil contamination by brominated flame retardants in open waste dumping sites in Asian developing countries

In Asian developing countries, large amounts of municipal wastes are dumped into open dumping sites each day without adequate management. This practice may cause several adverse environmental consequences and increase health risks to local communities. These dumping sites are contaminated with many chemicals including brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDs).BFRs may be released into the environment through production processes and through the disposal of plastics and electronic wastes that contain them.The purpose of this study was to elucidate the status of BFR pollution in municipal waste dumping sites in Asian developing countries. Soil samples were collected from six open waste dumping sites and five reference sites in Cambodia, India, Indonesia, Malaysia, and Vietnam from 1999 to 2007. The results suggest that PBDEs are the dominant contaminants in the dumping sites in Asian developing countries, whereas HBCD contamination remains low. Concentrations of PBDEs and HBCDs ranged from ND to 180 μg/kg dry wt and ND to 1.4 μg/kg dry wt, respectively, in the reference sites and from 0.20 to 430 μg/kg dry wt and ND to 2.5 μg/kg dry wt, respectively, in the dumping sites. Contamination levels of PBDEs in Asian municipal dumping sites were comparable with those reported from electronic waste dismantling areas in Pearl River delta, China.     

中国居民及动植物的环境激素暴露研究现状和建议

环境激素污染严重威胁着人类及动植物的健康。综述了EDCs生物危害,简要介绍了美国、日本和欧盟的EDCs研究工作,分析了中国居民和动植物的EDCs暴露现状,归纳了现有的流行病学调查信息,并针对现状提出了建议。     

A systematic review on the adverse health effects of di-2-ethylhexyl phthalate

Di (ethylhexyl) phthalate (DEHP) is a global environmental pollutant. This study aims to systematically review the literature on health effects of exposure to DEHP including effects on reproductive health, carcinogenesis, pregnancy outcome, and respiratory system. The literature search was done through Scopus, ISI Web of Science, Google Scholar, PubMed, Medline, and the reference lists of previous review articles to identify relevant articles published to June 2016 in each subject area. The inclusion criteria were as follows: original research, cross-sectional studies, case–control studies, cohort studies, interventional studies, and review articles. Both human and animal studies were included. The search was limited to English language papers. Conference papers, editorials, and letters were not included. The systematic review was conducted and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Overall, 152 of the 407 papers met the inclusion criteria. We provided an up-to-date comprehensive and critical assessment of both human and animal studies undertaken to explore the effects of DEHP. It revealed that in experimental studies, exposure to DEHP mainly targeted the reproductive, neurodevelopment, and respiratory systems. Human studies reported that exposure to this contaminant had carcinogenic effects and influenced neurodevelopment in early life. This systematic review underscored the adverse health effects of DEHP for pregnant women and the pediatric age group. It summarizes different response of humans and experimental animals to DEHP exposure, and some suggested underlying mechanisms.     

机动车污染排放模型研究综述

过去几十年,为了掌握机动车污染排放的规律和特征,向决策者提供科学有效的机动车污染控制措施,研究者们致力于研究机动车污染物排放的物化原理和影响机动车污染的主要因素,并据此建立多种尺度的机动车排放模型,以模拟城市区域或者街道的污染物排放.为了分析机动车的瞬态排放特征,目前的机动车排放模型研究正逐渐从宏观向微观发展,排放测试方法注重获取逐秒的排放数据,排放模型模拟的时间尺度和空间尺度逐步趋向微观.此外,机动车模型研究正趋向与交通模型进行耦合,从而揭示机动车在实际道路交通流中的排放特征.从机动车排放的主要影响因素、机动车排放测试、机动车排放因子模型及机动车排放清单等4个方面综述了国内外机动车排放研究现状和发展动向,对比并评价各种机动车排放模型方法的优缺点和适用范围,对我国的机动车排放模型发展方向进行了展望.     

Levels and distribution of polybrominated diphenyl ethers in humans and environmental compartments: a comprehensive review of the last five years of research

Environmental Science and Pollution Research - Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants (BFRs), present in the environment, animals, and humans. Their...