Biotransformation of Organophosphate Diesters Characterized via In Vitro Metabolism and In Vivo Screening.

Organophosphate diesters (di-OPEs), as additives in industrial applications and/or transformation products of emerging environmental pollutants, such as organophosphate triesters (tri-OPEs), have been found in the environment and biological matrices. The metabolic fate of di-OPEs in biological media is of great significance for tracing the inherent and precursor toxicity variations. This is the first study to investigate the metabolism of a suite of di-OPEs by liver microsomes and to identify any metabolite of metabolizable di-OPEs in in vitro and in vivo samples. Of the 14 di-OPEs, 5 are significantly metabolizable, and their abundant metabolites with hydroxyl, carboxyl, dealkylated, carbonyl, and/or epoxide groups are tentatively identified. More than half of the di-OPEs are detectable in human serum and/or wild fish tissues, and dibenzyl phosphate (DBzP), bis(2,3-dibromopropyl) phosphate (BDBPP), and isopropyl diphenyl phosphate (ip-DPHP) are first reported at a detectable level in humans and wildlife. Using an in vitro assay and a known biotransformation rule-based integrated screening strategy, 2 and 10 suspected metabolite peaks of DEHP are found in human serum and wild fish samples, respectively, and are then identified as phase I and phase II metabolites of DEHP. This study provides a novel insight into fate and persistence of di-OPE and confirms the presence of di-OPE metabolites in humans and wildlife.

Aerobic Microbial Transformation of Fluorinated Liquid Crystal Monomer: New Pathways and Mechanism.

Fluorinated liquid crystal monomers (FLCMs) have been suggested as emerging contaminants, raising global concern due to their frequent occurrence, potential toxic effects, and endurance capacity in the environment. However, the environmental fate of the FLCMs remains unknown. To fill this knowledge gap, we investigated the aerobic microbial transformation mechanisms of an important FLCM, 4-[difluoro(3,4,5-trifluorophenoxy)methyl]-3, 5-difluoro-4'-propylbiphenyl (DTMDPB), using an enrichment culture termed as BG1. Our findings revealed that 67.5 ± 2.1% of the initially added DTMDPB was transformed in 10 days under optimal conditions. A total of 14 microbial transformation products obtained due to a series of reactions (e.g., reductive defluorination, ether bond cleavage, demethylation, oxidative hydroxylation and aromatic ring opening, sulfonation, glucuronidation, O-methylation, and thiolation) were identified. Consortium BG1 harbored essential genes that could transform DTMDPB, such as dehalogenation-related genes [e.g., glutathione S-transferase gene (GST), 2-haloacid dehalogenase gene (2-HAD), nrdB, nuoC, and nuoD]; hydroxylating-related genes hcaC, ubiH, and COQ7; aromatic ring opening-related genes ligB and catE; and methyltransferase genes ubiE and ubiG. Two DTMDPB-degrading strains were isolated, which are affiliated with the genus Sphingopyxis and Agromyces. This study provides a novel insight into the microbial transformation of FLCMs. The findings of this study have important implications for the development of bioremediation strategies aimed at addressing sites contaminated with FLCMs.

Liquid Crystal Monomer: A Potential PPARγ Antagonist.

Liquid crystal monomers (LCMs) are a large family of artificial ingredients that have been widely used in global liquid crystal display (LCD) industries. As a major constituent in LCDs as well as the end products of e-waste dismantling, LCMs are of growing research interest with regard to their environmental occurrences and biochemical consequences. Many studies have analyzed LCMs in multiple environmental matrices, yet limited research has investigated the toxic effects upon exposure to them. In this study, we combined in silico simulation and in vitro assay validation along with omics integration analysis to achieve a comprehensive toxicity elucidation as well as a systematic mechanism interpretation of LCMs for the first time. Briefly, the high-throughput virtual screen and reporter gene assay revealed that peroxisome proliferator-activated receptor gamma (PPARγ) was significantly antagonized by certain LCMs. Besides, LCMs induced global metabolome and transcriptome dysregulation in HK2 cells. Notably, fatty acid ß-oxidation was conspicuously dysregulated, which might be mediated through multiple pathways (IL-17, TNF, and NF-kB), whereas the activation of AMPK and ligand-dependent PPARγ antagonism may play particularly important parts. This study illustrated LCMs as a potential PPARγ antagonist and explored their toxicological mode of action on the trans-omics level, which provided an insightful overview in future chemical risk assessment.

High-Resolution Mass Spectrometry Screening of Emerging Organophosphate Esters (OPEs) in Wild Fish: Occurrence, Species-Specific Difference, and Tissue-Specific Distribution.

There is a dearth of information regarding the pollution status of emerging organophosphate esters (OPEs) in wild fish. Here, we optimized and validated a quick, easy, cheap, effective, rugged, and safe (QuEChERS) pretreatment method, which was further applied for target, suspect, and nontarget screening of OPEs in n = 48 samples of wild fishes from Taihu Lake (eastern China). This integrated technique allows us to fully identify 20 OPEs, and 9 out of them are emerging OPEs detected in wild fish for the first time. Importantly, some of the emerging OPEs, i.e., tris(2,4-di-tert-butylphenyl) phosphate (TDtBPP), 4-tert-butylphenyl diphenyl phosphate (BPDP), and 2-isopropylphenyl diphenyl phosphate (IPDP), exhibited greater or at least comparable contamination levels as compared to traditional ones. There were no statistically significant interspecies (n = 6) differences regarding OPE concentrations. However, we observed significant differences on OPE concentrations among different tissues of silver carp (Hypophthalmichthys molitrix), for which the intestine has the highest OPE mean concentration (46.5 ng/g wet weight (ww)), followed by the liver (20.1 ng/g ww) ≈ brain (20.0 ng/g ww) > gill (14.8 ng/g ww) > muscle (11.4 ng/g ww). An interesting exception is IPDP, which presents an unexpectedly high concentration in the brain (0.510 ng/g ww). Collectively, this study expands our understanding of OPE contamination in wild fish and clearly shows that emerging TDtBPP, IPDP, and BPDP could play an equally important role as traditional OPEs in contribution of OPE pollution in wild fish samples.

Suspect Screening of Liquid Crystal Monomers (LCMs) in Sediment Using an Established Database Covering 1173 LCMs.

Recent studies have suggested that liquid crystal monomers (LCMs) are emerging contaminants in the environment, and knowledge of this class of substances is very rare. Here, we reviewed existing LCM-related documents, i.e., publications and patents, and established a database involving 1173 LCMs. These 1173 LCMs were further calculated for their physicochemical properties, i.e., persistence (P), bioaccumulation (B), long-range transport potential (LRTP), and Arctic contamination and bioaccumulation potential (ACBAP). We found that 476 out of them were P&B chemicals (99% of them were halogenated), and 320 of them could have ACBAP properties (67% of them were halogenated). This LCM database was further applied for suspect screening of LCMs in n = 33 sediment samples by use of gas chromatography coupled to quadrupole time-of-flight mass spectrometry (GC-QTOF/MS). We tentatively identified 26 LCM formulas, which could have 43 chemical structures. Two out of these 43 suspect LCM candidates, 1-butoxy-2,3-difluoro-4-(4-propylcyclohexyl) benzene (3cH4OdFP) and 1-ethoxy-2,3-difluoro-4-(4-pentyl cyclohexyl) benzene (5cH2OdFP), were fully confirmed by a comparison of unique GC and MS characteristics with their authentic standards. Overall, our present study expanded the previous LCM database from 362 to 1173, and 1173 LCMs in this database were calculated for their physicochemical properties. Meanwhile, taking n = 33 sediment samples as an exercise, we successfully developed a suspect screening strategy tailored for LCMs, and this strategy could have promising potential to be extended to other environmental matrices.

Electronic-Waste-Driven Pollution of Liquid Crystal Monomers: Environmental Occurrence and Human Exposure in Recycling Industrial Parks.

Liquid crystal monomers (LCMs) in liquid crystal displays (LCDs) may be released into the environment, especially in electronic waste (e-waste) recycling industrial parks with a high pollution risk. However, little has been known about the environmental release and human exposure to LCMs until now. Herein, a total of 45 LCMs were detected in LCDs of commonly used smartphones and computers by high-resolution mass spectrometry with suspect screening analysis. Fluorinated biphenyls and their analogs were the dominant LCMs. Based on available standards of the screening results and previous studies, 55 LCMs were quantified in samples from an e-waste recycling industrial park in Central China. The LCMs were frequently detected in outdoor dust (n = 43), workshop #1 indoor dust (n = 53), and hand (n = 43) and forehead wipes (n = 43), with median concentrations of 6950 ng/g, 67,400 ng/g, 46,100 ng/m2, and 62,100 ng/m2, respectively. The median estimated daily intake values of the LCMs via dust ingestion and dermal absorption were 48.3 and 16.5 ng/kg body weight/day, respectively, indicating a high occupational exposure risk of these compounds. In addition, 16 LCMs were detected in the serum of eight elderly people (≥60 years old) with over 5 years of experience in e-waste dismantling operations, resulting in a total concentration range of 3.9-26.3 ng/mL.

Persistent, bioaccumulative, and toxic properties of liquid crystal monomers and their detection in indoor residential dust.

Liquid crystal monomers (LCMs) are used widely in liquid crystal displays (LCDs), which are dramatically changing the world due to the provision of convenient communication. However, there are essentially no published reports on the fate and/or effects of LCMs in the environment. Of 362 currently produced LCMs, 87 were identified as persistent and bioaccumulative (P&B) chemicals, which indicated that these chemicals would exhibit resistance to degradation and exhibit mobility after entering the environment. Following exposure to mixtures of LCM collected from 6 LCD devices, significant modulation of 5 genes, CYP1A4, PDK4, FGF19, LBFABP, and THRSP, was observed in vitro. Modulation of expressions of mRNAs coding for these genes has frequently been reported for toxic (T) persistent organic pollutants (POPs). In LCM mixtures, 33 individual LCMs were identified by use of mass spectrometry and screened for in 53 samples of dust from indoor environments. LCMs were detectable in 47% of analyzed samples, and 17 of the 33 LCMs were detectable in at least 1 sample of dust. Based on chemical properties, including P&B&T of LCMs and their ubiquitous detection in dust samples, the initial screening information suggests a need for studies to determine status and trends in concentrations of LCMs in various environmental matrices as well as tissues of humans and wildlife. There is also a need for more comprehensive in vivo studies to determine toxic effects and potencies of LCMs during chronic, sublethal exposures.

Industrial Production of Organophosphate Flame Retardants (OPFRs): Big Knowledge Gaps Need to Be Filled?

Since the phase-out of traditional halogenated flame retardants (HFRs), interests of research are gradually being shifted to organophosphate flame retardants (OPFRs), and this can be reflected by the increasing number of publications on OPFRs year by year. Here, an extensive survey is conducted in an attempt to generate a list of OPFRs that are being produced in factories, and to investigate the annual production volume (APV). This survey suggests that at least n = 56 OPFR monomers and n = 62 OPFR mixtures are being currently produced in 367 factories around the world, and 201 out of them are in Mainland China. APV of OPFRs was estimated as 598,422 metric tons, and this number could be underestimated due to the limitation of available information. We also notice that current researches are confined to a limited number of OPFRs, especially for OP esters (OPEs), and other OPFRs with different structures from OPEs has been rarely studied. Based on all the collected datasets, we provide five recommendations for how to proceed with future research to more comprehensively understand the currently-produced OPFRs in the environment.

Identifying Citric Acid Esters, a Class of Phthalate Substitute Plasticizers, in Indoor Dust via an Integrated Target, Suspect, and Characteristic Fragment-Dependent Screening Strategy.

Citrate acid esters (CAEs) have been proposed as a class of phthalate substitute plasticizers; however, information on their occurrence in indoor environments is rare. By using liquid chromatography coupled with a quadrupole-Orbitrap mass spectrometer, we developed an integrated strategy that can be applied for target, suspect, and characteristic fragment-dependent screening of CAEs. In n = 50 indoor dust samples collected from Nanjing City (China), three CAEs, namely, acetyl tributyl citrate (ATBC; mean: 412,000 ng/g), tributyl citrate (TBC, 11,600 ng/g), and triethyl citrate (TEC, 10,900 ng/g), exhibited the greatest contamination levels. Total concentrations of CAEs (∑8CAEs) were statistically significantly (p < 0.01) greater than those of common organophosphate triesters (OPTEs), a class of ubiquitous contaminants in dust. Suspect and characteristic fragment-dependent screening (m/z 111.0078 ([C5H3O3]+) and m/z 129.0181 ([C5H5O4]+)) of CAEs were further conducted for the same batch of samples. We tentatively identified six novel CAEs, and the most frequent and abundant CAE was fully identified as tributyl aconitate (TBA). Statistically significant correlation relationships were observed on dust levels between TBA vs ATBC (r = 0.650; p < 0.01) and TBA vs TBC (r = 0.384; p < 0.01), suggesting their similar sources in dust samples.

Establishment of a Target, Suspect, and Functional Group-Dependent Screening Strategy for Organophosphate Esters (OPEs): "Into the Unknown" of OPEs in the Sediment of Taihu Lake, China.

Current environmental monitoring studies are generally confined to several target organophosphate esters (OPEs), and there is a lack of strategies for comprehensively screening all potential OPEs in environmental samples. Here, an effective and accurate strategy was developed for the target, suspect, and functional group-dependent screening of OPEs by the use of ultrahigh-performance liquid chromatography-Q Exactive hybrid quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS), and this strategy was applied for the analysis of n = 74 sediment samples (including 23 surface sediment samples and 51 sediment core samples) collected from Taihu Lake (eastern China) in 2019. In these analyzed samples, we successfully identified n = 35 OPEs, and 23 of them were reported in this region for the first time. In addition, this strategy also presented other interesting findings, i.e., (1) OPE concentrations decreased with increasing distance from the coast of the lake; (2) the newly identified 3-hydroxyphenyl diphenyl phosphate (meta-OH-TPHP) was not statistically significantly correlated with triphenyl phosphate (TPHP; r = 0.02494, p = 0.9101) but with resorcinol bis(diphenyl phosphate) (RDP) (r = 0.9271, p < 0.0001) and three other OPEs; and (3) the summed concentrations of aryl OPEs (∑arylOPEs) in sediment core samples exhibited significantly increasing trends as the depth decreased. Collectively, this study provided an effective strategy that was successfully applied for comprehensive screening of OPEs in the sediments of Taihu Lake, and this strategy could have promising potential to be extended to other environmental matrices or samples.

Liquid Crystal Monomers (LCMs) in Sediments: Method Validation and Detection in Sediment Samples from Three Typical Areas.

Liquid crystal monomers (LCMs) have been proposed to be persistent, bioaccumulative, and toxic (PBT) substances; however, there is a dearth of information regarding their occurrence in sediment samples. Here, an analytical method was developed for the quantitative determination of LCMs in sediment samples, and n = 76 sediment samples were collected and analyzed to determine accurate concentrations of LCMs. Our results indicated that the developed pretreatment procedure was applicable for the determination of LCM concentrations in sediments. We observed that LCMs were detected in 75 out of the 76 sediment samples, and 23, 18, and 14 out of the 39 target LCMs were quantified in at least one of the analyzed sediments from rivers around LCM or liquid crystal device (LCD) manufacturers, Taihu Lake, and rivers around e-waste recycling sites, respectively. The LCMs in the samples from rivers around LCM/LCD manufacturers exhibited the greatest mean concentrations of 26.1 ng/g dry weight (dw), followed by those from e-waste recycling site areas (1.15 ng/g dw) and Taihu Lake (0.076 ng/g dw). Collectively, this study provided the first analytical method that was able to quantify the concentrations of LCMs in sediment samples and provided the first evidence for the occurrence of LCMs in sediment samples.

Life Cycle Exposure to Environmentally Relevant Concentrations of Diphenyl Phosphate (DPhP) Inhibits Growth and Energy Metabolism of Zebrafish in a Sex-Specific Manner.

Due to commercial uses and environmental degradation of aryl phosphate esters, diphenyl phosphate (DPhP) is frequently detected in environmental matrices and is thus of growing concern worldwide. However, information on potential adverse effects of chronic exposure to DPhP at environmentally realistic concentrations was lacking. Here, we investigated the effects of life cycle exposure to DPhP on zebrafish at environmentally relevant concentrations of 0.8, 3.9, or 35.6 µg/L and employed a dual-omics approach (metabolomics and transcriptomics) to characterize potential modes of action. Exposure to DPhP at 35.6 µg/L for 120 days resulted in significant reductions in body mass and length of male zebrafish, but did not cause those same effects to females. Predominant toxicological mechanisms, including inhibition of oxidative phosphorylation, down-regulation of fatty acid oxidation, and up-regulation of phosphatidylcholine degradation, were revealed by integrated dual-omics analysis and successfully linked to adverse outcomes. Activity of succinate dehydrogenase and protein content of carnitine O-palmitoyltransferase 1 were significantly decreased in livers of male fish exposed to DPhP, which further confirmed the proposed toxicological mechanisms. This study is the first to demonstrate that chronic, low-level exposure to DPhP can retard growth via inhibiting energy output in male zebrafish.

Functional Group-Dependent Screening of Organophosphate Esters (OPEs) and Discovery of an Abundant OPE Bis-(2-ethylhexyl)-phenyl Phosphate in Indoor Dust.

There is increasing scientific interest in environmental pollution and the effect on public health caused by organophosphate esters (OPEs). Using liquid chromatography coupled to a hybrid quadrupole Orbitrap high-resolution mass spectrometer, a novel, robust, and untargeted screening strategy for the identification of novel OPEs in indoor dust samples was presently developed based on the characteristic molecular fragmentation pathways, and 12 previously reported OPEs and six previously unrecognized OPEs were detected in the combined extracts of indoor dust samples, collected in Nanjing, eastern China. One of the six detected OPEs, bis-(2-ethylhexyl)-phenyl phosphate (BEHPP), was identified by comparison of unique LC and MS characteristics with a synthesized pure standard. Accurate concentrations of BEHPP were determined in n = 50 individual indoor dust samples with 100% detection frequency with a median concentration range of 50-1530 ng/g dry weight, which were generally greater or at least comparable to traditional OPEs, that is, triphenyl phosphate and 2-ethylhexyl diphenyl phosphate (EHDPP), in the same dust samples. Statistically significant, positive correlations were found for log-transformed concentrations of BEHPP versus EHDPP (r2 = 0.7884, p < 0.0001), and BEHPP versus tris(2-ethylhexyl)phosphate (r2 = 0.4054, p < 0.0001), suggesting their similar commercial applications and sources in the environment.

Organophosphate Ester, 2-Ethylhexyl Diphenyl Phosphate (EHDPP), Elicits Cytotoxic and Transcriptomic Effects in Chicken Embryonic Hepatocytes and Its Biotransformation Profile Compared to Humans.

The effects of 2-ethylhexyl diphenyl phosphate (EHDPP) on cytotoxicity and mRNA expression, as well as its metabolism, were investigated using a chicken embryonic hepatocyte (CEH) assay. After incubation for 36 h, the lethal concentration 50 (LC50) was 50 ± 11 µM, suggesting that EHDPP is one of a small cohort of highly toxic organophosphate esters (OPEs). By use of a ToxChip polymerase chain reaction (PCR) array, we report modulation of 6, 11, or 16/43 genes in CEH following exposure to 0.1, 1, or 10 µM EHDPP, respectively. The altered genes were from all nine biological pathways represented on the ToxChip including bile acids/cholesterol regulation, glucose metabolism, lipid homeostasis, and the thyroid hormone pathway. After incubation for 36 h, 92.5% of EHDPP was transformed, and one of its presumed metabolites, diphenyl phosphate (DPHP), only accounted for 12% of the original EHDPP concentration. Further screening by use of high-resolution mass spectrometry revealed a novel EHDPP metabolite, hydroxylated 2-ethylhexyl monophenyl phosphate (OH-EHMPP), which was also detected in a human blood pool. Additional EHDPP metabolites detected in the human blood pool included EHMPP and DPHP. Overall, this study provided novel information regarding the toxicity of EHDPP and identified a potential EHDPP metabolite, OH-EHMPP, in both avian species and humans.

Multigenerational Effects and Demographic Responses of Zebrafish ( Danio rerio) Exposed to Organo-Bromine Compounds.

Long-term exposure to toxic chemicals often has deleterious effects on aquatic organisms. In order to support appropriate environmental management of chemicals, a mathematical model was developed to characterize the effects of chemicals on multigenerational population dynamics in aquatic animals. To parametrize the model, we conducted a multigenerational laboratory toxicity test in zebrafish ( Danio rerio) exposed to 2-bromo-4,6-dinitroaniline (BDNA). Long-term exposure to BDNA considerably reduced the fecundity of adult zebrafish (F0 and F1) and caused deformities in the offspring (F2). Life history data, including changes in fecundity and population growth, were then integrated into the model to predict population dynamics of zebrafish exposed to two novel brominated flame retardants, bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH) and 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (TBB). The model predicted that the fecundity of adult zebrafish would be significantly impaired after exposure to 90.36 µM TBPH and 99.16 µM TBB. Thus, prolonged exposure to such levels over multiple generations could result in population extinction within 20 years. Our results provide an intensive temporal perspective to investigate a keystone that connects with individual response to chemicals, population dynamics, and ultimately ecosystem influences.

Isomer-Specific Hexabromocyclododecane (HBCDD) Levels in Top Predator Fish from Across Canada and 36-Year Temporal Trends in Lake Ontario.

Hexabromocyclododecane (HBCDD) is a high concern environmental pollutant due to its persistent, bioaccumulative, and toxic properties. The spatial distribution of HBCDD was investigated in top predator fish (lake trout, walleye, or brook trout) collected in 2013 ( n = 165) from 19 sampling sites and in 2015 ( n = 145) from 20 sites across Canada. HBCDD was measurable in at least one sample at each sampling site regardless of sampling year with the exception of walleye from the south basin of Lake Winnipeg (2013). Sampling sites in or near the Laurentian Great Lakes had greater ΣHBCDD concentrations compared to locations to the west or east. The greatest mean ΣHBCDD concentration was 72.6 ng/g lw in fish from Lake Huron-Goderich (2015). Regardless of the sampling sites, α-HBCDD was the dominant congener followed by γ-HBCDD, whereas ß-HBCDD was barely detectable. In fish from the same waterbody there were comparable α/γ isomer concentration ratios. The greatest ratio was 20.8 in fish from Lake Ontario, whereas the lowest ratio was 6.3 for fish from Lac Memphrémagog (Québec) likely related to more recent emissions of a technical HBCDD mixture. Temporal trends of HBCDD in lake trout from Lake Ontario showed a significant decreasing trend for γ-HBCDD with a half-life estimate of 10 years over a 36-year period (1979-2015), and for α-HBCDD with a half-life of 11 years over the years of 2008 to 2015. The proportion of α-HBCDD to ΣHBCDD increased significantly during 1979 to 2015. The present study provided novel information on the isomer-specific HBCDDs in Canada freshwater fish.

Halogenated Flame Retardants in Predator and Prey Fish From the Laurentian Great Lakes: Age-Dependent Accumulation and Trophic Transfer.

The identification, persistence, accumulation and trophic transfer of 25 polybrominated diphenyl ether (PBDE) congeners, 23 non-PBDE halogenated flame retardants (NPHFRs), 4 polybrominated-diphenoxybenzenes (PB-DiPhOBzs) and 6 methoxylated (MeO-) PB-DiPhOBzs were investigated in predator and prey fish collected in 2010 from sites in the North American Great Lakes of Ontario (n = 26) and Erie (n = 39). Regardless of locations or species, 20 PBDEs and 12 NPHFRs were quantifiable in at least one of the 65 analyzed samples, and polybrominated-1,4-diphenoxybenzenes (PB-DiPhOBzs) and MeO-PB-DiPhOBzs were not detectable in any of analyzed samples. Among the FRs, the greatest concentrations were the ∑PBDE, ranging from 1.06 (Rainbow Smelt, Lake Erie) to 162 (Lake Trout, Lake Ontario) ng/g wet weight (ww), which was followed by mean HBCDD concentrations ranging ND to 17.3 (Lake Trout, Lake Ontario) ng/g ww. The remaining FRs were generally not detectable or at sub-ppb levels. In most of cases, FR concentrations in samples from Lake Ontario were greater than those from Lake Erie. Strong and significant positive linear relationships occurred between log-normalized FR concentrations (ww or lipid weight (lw)) and ages of the top predator Lake Trout (n = 16, from Lake Ontario), and the estimated FR doubling ages (T2) were 2.9-6.4 years. For Walleye from Lake Erie, significantly positive linear relationships were also observed for some FRs, but the linear relationships generally became negative after FR concentrations were normalized with lipid weight. This study provides novel information on FR accumulation in aquatic organisms, and for the first time, significant positive linear relationships are reported between log-normalized FR concentrations (lw or ww) and ages of Lake Trout from the Great Lakes.

Whole-Life-Stage Characterization in the Basic Biology of Daphnia magna and Effects of TDCIPP on Growth, Reproduction, Survival, and Transcription of Genes.

Toxicity tests of chemicals have mainly focused on the partial life-cycle evaluation of model animals. Limited information is available for the evaluation of effects of chemicals from a whole-life-stage exposure perspective. The objective of this study was to perform a whole-life-stage characterization in the basic biology of Daphnia magna (D. magna) and evaluate the effects of a known organophosphate ester (OPE) contaminant, tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), on growth, reproduction, survival, and transcription of genes. The whole-life-stage characterization in growth, reproduction, and survival of D. magna was conducted, and representative sampling time points for the three developmental stages were identified (day 6, day 32, and day 62). Transcriptomic profiles for these three stages were compared, and stage-specific PCR arrays of D. magna were developed. The whole-life-stage exposure to environmentally relevant or greater concentrations of TDCIPP significantly inhibited growth and reproduction of D. magna and decreased survival at the later stage of the exposure experiment (≥32 days). Such adverse effects were not observed in the early stage of the exposure (<32 days), suggesting that short-term toxicity tests, such as the standard 21-day test, might underestimate the environmental risk of TDCIPP. Furthermore, expressions of genes selected at day 6, day 32, and day 62 were significantly changed after TDCIPP exposure, and the changes in the expressions of partial genes were correlated to the inhibitory effects on growth, reproduction, and survival.

Environmentally relevant organophosphate triesters in herring gulls: In vitro biotransformation and kinetics and diester metabolite formation using a hepatic microsomal assay.

The in vitro biotransformation and kinetics of six organophosphate triester (OPE) flame retardants were investigated in herring gulls (Larus argentatus) from the Great Lakes using a hepatic microsomal metabolism assay. Administration of each individual OPE (tri-n-butyl phosphate (TNBP), tris(2-butoxyethyl) phosphate (TBOEP), triphenyl phosphate (TPHP), triethyl phosphate (TEP), tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) and tris(2-chloroisopropyl) phosphate (TCIPP)) to the in vitro assay (concentration range 0.01 to 10µM) resulted in rapid depletion with the exception of TEP. Following the Michaelis-Menten enzyme kinetics model, a preliminary 2-minute incubation period was used to estimate the Vmax (±SE) values (i.e., the maximal rate of reaction for a saturated enzyme system), which ranged from 5.0±0.4 (TPHP) to 29±18pmol/min/mg protein (TBOEP), as well as the KM (±SE) values (i.e., the OPE concentration corresponding to one half of the Vmax), which ranged from 9.8±1 (TPHP) to 189±135nM (TBOEP). Biotransformation assays over a 100-minute incubation period revealed that TNBP was metabolized most rapidly (with a depletion rate of 73±4pmol/min/mg protein), followed by TBOEP (53±8pmol/min/mg), TCIPP (27±1pmol/min/mg), TPHP (22±2pmol/min/mg) and TDCIPP (8±1pmol/min/mg). In vitro biotransformation of OP triesters was clearly structure-dependent where non-halogenated alkyl OP triesters were metabolized more rapidly than halogenated alkyl triesters. Halogenated OP triesters were transformed to their respective diesters more efficiently relative to non-halogenated OP triesters. To our knowledge, this is the first study to investigate OP triester metabolism and OP diester formation in an avian or wildlife model system, which is important to understand the fate and biological activity of OPEs in an exposed organism.

In Vitro Metabolism of Photolytic Breakdown Products of Tetradecabromo-1,4-diphenoxybenzene Flame Retardant in Herring Gull and Rat Liver Microsomal Assays.

Tetradecabromo-1,4-diphenoxybenzene (TeDB-DiPhOBz) is used as a flame retardant chemical and has been hypothesized to be the precursor of methoxylated polybrominated diphenoxybenzene (MeO-PB-DiPhOBz) contaminants reported in herring gulls from sites across the Laurentian Great Lakes. Here, by irradiating the parent TeDB-DiPhOBz (solution 1) with natural sunlight or UV, we prepared three solutions where solution 2 was dominated by the Br8-11-PB-DiPhOBzs, along with Br5-8-PB-DiPhOBzs (solution 3) and Br4-6-PB-DiPhOBzs (solution 4). The in vitro metabolism of TeDB-DiPhOBz and PB-DiPhOBzs was investigated using harvested wild herring gull (Larus argentatus) and adult male Wister-Han rat liver microsomal assays. After a 90 min incubation period of solution 1 in gull or rat microsomal assays, there was no significant (p > 0.05) depletion of TeDB-DiPhOBz. OH-PB-DiPhOBz metabolites were detectable after gull and rat microsomal assay incubation with solutions 3 or 4, and showed clear species-specific differences. Also detected were two polybrominated hydroxylated metabolites having polybenzofuran structures. Overall, this study suggested that TeDB-DiPhOBz is slowly metabolized in vitro, and also indicated that if wild herring gulls are exposed (e.g., via the diet) to photolytic products of TeDB-DiPhOBz, OH-PB-DiPhOBz and other metabolites could be formed. OH-PH-DiPhOBz are likely precursors to MeO-PB-DiPhOBz contaminants that we reported previously in eggs of wild Great Lakes herring gulls.