Lipid Metabolic Disorders Induced by Organophosphate Esters in Silver Carp from the Middle Reaches of the Yangtze River.

The Yangtze River fishery resources have declined strongly over the past few decades. One suspected reason for the decline in fishery productivity, including silver carp (Hypophthalmichthys molitrix), has been linked to organophosphate esters (OPEs) contaminant exposure. In this study, the adverse effect of OPEs on lipid metabolism in silver carp captured from the Yangtze River was examined, and our results indicated that muscle concentrations of the OPEs were positively associated with serum cholesterol and total lipid levels. In vivo laboratory results revealed that exposure to environmental concentrations of OPEs significantly increased the concentrations of triglyceride, cholesterol, and total lipid levels. Lipidome analysis further confirmed the lipid metabolism dysfunction induced by OPEs, and glycerophospholipids and sphingolipids were the most affected lipids. Hepatic transcriptomic analysis found that OPEs caused significant alterations in the transcription of genes involved in lipid metabolism. Pathways associated with lipid homeostasis, including the peroxisome proliferator-activated receptor (PPAR) signal pathway, cholesterol metabolism, fatty acid biosynthesis, and steroid biosynthesis, were significantly changed. Furthermore, the affinities of OPEs were different, but the 11 OPEs tested could bind with PPARγ, suggesting that OPEs could disrupt lipid metabolism by interacting with PPARγ. Overall, this study highlighted the harmful effects of OPEs on wild fish and provided mechanistic insights into OPE-induced metabolic disorders.

Exposure of Peregrine Falcons to Halogenated Flame Retardants: A 30 Year Retrospective Biomonitoring Study across North America.

Compared to aquatic ecosystem, terrestrial systems have been subjected to fewer investigations on the exposure to halogenated flame retardants (HFRs). Our study utilized peregrine falcon eggs collected from multiple habitats across North America to retrospectively explore both spatial distribution and temporal changes in legacy (e.g., polybrominated diphenyl ethers) and alternative HFRs over a 30 year period (1984-2016). The results reveal intensive HFR exposure in terrestrial ecosystems and chemical-specific spatiotemporal distribution patterns. The correlations between egg levels of the selected HFRs and human population density clearly illustrated a significant urban influence on the exposure of this wildlife species to these HFRs and subsequent maternal transfer to their eggs. Temporal analyses suggest that, unlike aquatic systems, terrestrial ecosystems may undergo continual exposure to consistently high levels of legacy HFRs for a long period of time. Our findings collectively highlight the effectiveness of using peregrine eggs to monitor terrestrial exposure to HFRs and other bioaccumulative chemicals and the need for continuous monitoring of HFRs in terrestrial ecosystems.

Developing Methods for Assessing Trophic Magnification of Perfluoroalkyl Substances within an Urban Terrestrial Avian Food Web.

We investigated the trophic magnification potential of perfluoroalkyl substances (PFAS) in a terrestrial food web by using a chemical activity-based approach, which involved normalizing concentrations of PFAS in biota to their relative biochemical composition in order to provide a thermodynamically accurate basis for comparing concentrations of PFAS in biota. Samples of hawk eggs, songbird tissues, and invertebrates were collected and analyzed for concentrations of 18 perfluoroalkyl acids (PFAAs) and for polar lipid, neutral lipid, total protein, albumin, and water content. Estimated mass fractions of PFCA C8-C11 and PFSA C4-C8 predominantly occurred in albumin within biota samples from the food web with smaller estimated fractions in polar lipids > structural proteins > neutral lipids and insignificant amounts in water. Estimated mass fractions of longer-chained PFAS (i.e., C12-C16) mainly occurred in polar lipids with smaller estimated fractions in albumin > structural proteins > neutral lipids > and water. Chemical activity-based TMFs indicated that PFNA, PFDA, PFUdA, PFDoA, PFTrDA, PFTeDA, PFOS, and PFDS biomagnified in the food web; PFOA, PFHxDA, and PFHxS did not appear to biomagnify; and PFBS biodiluted. Chemical activity-based TMFs for PFCA C8-C11 and PFSA C4-C8 were in good agreement with corresponding TMFs derived with concentrations normalized to only total protein in biota, suggesting that concentrations normalized to total protein may be appropriate proxies of chemical activity-based TMFs for PFAS, which predominantly partition to albumin. Similarly, TMFs derived with concentrations normalized to albumin may be suitable proxies of chemical activity-based TMFs for longer-chained PFAS, which predominantly partition to polar lipids.

Characterizing the Movement of Per- and Polyfluoroalkyl Substances in an Avian Aquatic-Terrestrial Food Web.

The movement of per- and polyfluoroalkyl substances (PFAS) through linked aquatic-terrestrial food webs is not well understood. Tree swallows (Tachycineta bicolor) in such systems may be exposed to PFAS from multiple abiotic and/or biotic compartments. We show from fatty acid signatures and carbon stable isotopes that tree swallow nestlings in southwestern Ontario fed on both terrestrial and aquatic macroinvertebrates. The PFAS profiles of air, terrestrial invertebrates, and swallows were dominated by perfluorooctanesulfonic acid (PFOS). Short-chain perfluoroalkyl acids (PFAAs) were largely restricted to air, surface water, and sediment, and long-chain PFAAs were mainly found in aquatic invertebrates and tree swallows. PFOS, multiple long-chain perfluorocarboxylic acids [perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorotridecanoic acid (PFTrDA)] and perfluorooctane sulfonamide precursors were estimated to bioaccumulate from air to tree swallows. PFOS bioaccumulated from air to terrestrial invertebrates, and PFOS, PFDA, and perfluorooctane sulfonamidoacetic acids (FOSAAs) bioaccumulated from water to aquatic invertebrates. PFOS showed biomagnification from both terrestrial and aquatic invertebrates to tree swallows, and PFDA and FOSAAs were also biomagnified from aquatic invertebrates to tree swallows. The movement of PFAS through aquatic-terrestrial food webs appears congener- and compartment-specific, challenging the understanding of PFAS exposure routes for multiple species involved in these food webs.

Varying Diet Composition Causes Striking Differences in Legacy and Emerging Contaminant Concentrations in Killer Whales across the North Atlantic.

Lipophilic persistent organic pollutants (POPs) tend to biomagnify in food chains, resulting in higher concentrations in species such as killer whales (Orcinus orca) feeding on marine mammals compared to those consuming fish. Advancements in dietary studies include the use of quantitative fatty acid signature analysis (QFASA) and differentiation of feeding habits within and between populations of North Atlantic (NA) killer whales. This comprehensive study assessed the concentrations of legacy and emerging POPs in 162 killer whales from across the NA. We report significantly higher mean levels of polychlorinated biphenyls (PCBs), organochlorine pesticides, and flame retardants in Western NA killer whales compared to those of Eastern NA conspecifics. Mean ∑PCBs ranged from ∼100 mg/kg lipid weight (lw) in the Western NA (Canadian Arctic, Eastern Canada) to ∼50 mg/kg lw in the mid-NA (Greenland, Iceland) to ∼10 mg/kg lw in the Eastern NA (Norway, Faroe Islands). The observed variations in contaminant levels were strongly correlated with diet composition across locations (inferred from QFASA), emphasizing that diet and not environmental variation in contaminant concentrations among locations is crucial in assessing contaminant-associated health risks in killer whales. These findings highlight the urgency for implementing enhanced measures to safely dispose of POP-contaminated waste, prevent further environmental contamination, and mitigate the release of newer and potentially harmful contaminants.

Bottom-up proteomics analysis for adduction of the broad-spectrum herbicide atrazine to histone.

Histones are the major proteinaceous components of chromatin in eukaryotic cells and an important part of the epigenome. The broad-spectrum herbicide atrazine (2-chloro-4-[ethylamino]-6-[isopropylamino]-1, 3, 5-triazine) and its metabolites are known to form protein adducts, but the formation of atrazine-histone adducts has not been studied. In this study, a bottom-up proteomics analysis method was optimized and applied to identify histone adduction by atrazine in vitro. Whole histones of calf thymus or human histone H3.3 were incubated with atrazine. After solvent-based protein precipitation, the protein was digested by trypsin/Glu-C and the resulting peptides were analyzed by high-resolution mass spectrometry using an ultra-high-performance liquid chromatograph interfaced with a quadrupole Exactive-Orbitrap mass spectrometer. The resulting tryptic/Glu-C peptide of DTNLCAIHAK from calf thymus histone H3.1 or human histone H3.3 was identified with an accurate mass shift of +179.117 Da in atrazine incubated samples. It is deduced that a chemical group with an elemental composition of C8H13N5 (179.1171 Da) from atrazine adducted with calf thymus histone H3.1 or human histone H3.3. It was confirmed by MS/MS analysis that the adduction position was at its cysteine 110 residue. Time- and concentration-dependent assays also confirmed the non-enzymatic covalent modification of histone H3.3 by atrazine in vitro. Thus, the potential exists that atrazine adduction may lead to the alteration of histones that subsequently disturbs their normal function.

Temporal change and the influence of climate and weather factors on mercury concentrations in Hudson Bay polar bears, caribou, and seabird eggs.

Temporal trends of mercury in Arctic wildlife are inconsistent within and between species and are often insignificant, which limits data interpretation. Recent multivariate analyses have shown that weather and climate factors (e.g. temperatures, sea ice conditions) are related to total Hg (THg) concentrations in wildlife tissues, though relatively few studies have explored these relationships. The present study compared time series of THg concentrations in liver of polar bear (Ursus maritimus, 2007/08-2015/16), eggs of thick-billed murres (Uria lomvia, 1993-2015) and kidney of caribou (Rangifer tarandus groenlandicus, 2006-2015) from the Hudson Bay region of Canada and statistically modelled THg over time with available climate and weather data. Significant temporal trends of THg concentrations were not detected in any species. However, in multivariate models that included time-lagged sea ice freeze up dates, THg concentrations increased 4.4% yr-1 in Qamanirjuaq caribou. Sea ice conditions were also related to THg levels in polar bear liver but not those in eggs of murres, though year was not a signifcant factor. Greater precipitation levels one to two years prior to sampling were associated with greater THg concentrations in polar bears and caribou, likely due to greater deposition, flooding and discharge from nearby wetlands and rivers. Time-lagged Arctic and/or North Atlantic Oscillation (AO/NAO) indices also generated significant, inverse models for all three species, agreeing with relationships in other time series of similar length. The magnitude and direction of many relationships were affected by season, duration of time-lags, and the length of the time series. Our findings support recent observations suggesting that temporal studies monitoring Hg in Arctic wildlife should consider including key climatic or weather factors to help identify consistent variables of influence and to improve temporal analyses of THg time series.

Mercury, legacy and emerging POPs, and endocrine-behavioural linkages: Implications of Arctic change in a diving seabird.

Arctic species encounter multiple stressors including climate change and environmental contaminants. Some contaminants may disrupt hormones that govern the behavioural responses of wildlife to climatic variation, and thus the capacity of species to respond to climate change. We investigated correlative interactions between legacy and emerging persistent organic pollutants (POPs), mercury (Hg), hormones and behaviours, in thick-billed murres (Uria lomvia) (N = 163) breeding in northern Hudson Bay (2016-2018). The blood profile of the murres was dominated by methylmercury (MeHg), followed by much lower levels of sum (∑) 35 polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB) and p,p'-dichlorodiphenyltrichloroethylene (DDE), polybrominated diphenyl ethers (PBDEs) BDE-47, -99 and BDE-100; all other measured organochlorine pesticides and replacement brominated flame retardants had low concentrations if detected. Inter-annual variations occurred in MeHg, circulating triiodothyronine (T3), thyroxine (T4), and the foraging behaviours of the murres, identified using GPS-accelerometers. Compared to the 50-year mean date (1971-2021) for 50% of sea-ice coverage in Hudson Bay, sea-ice breakup was 1-2 weeks earlier (2016, 2017) or comparable (2018). Indeed, 2017 was the earliest year on record. Consistent with relationships identified individually between MeHg and total T3, and T3 and foraging behaviour, a direct interaction between these three parameters was evident when all possible interactions among measured chemical pollutants, hormones, and behaviours of the murres were considered collectively (path analysis). When murres were likely already stressed due to early sea-ice breakup (2016, 2017), blood MeHg influenced circulating T3 that in turn reduced foraging time underwater. We conclude that when sea-ice breaks up early in the breeding season, Hg may interfere with the ability of murres to adjust their foraging behaviour via T3 in relation to variation in sea-ice.

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.

Tris(1,3-dichloro-2-propyl)phosphate Reduces Growth Hormone Expression via Binding to Growth Hormone Releasing Hormone Receptors and Inhibits the Growth of Crucian Carp.

Tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) has commonly been used as an additive flame retardant and frequently detected in the aquatic environment and in biological samples worldwide. Recently, it was found that exposure to TDCIPP inhibited the growth of zebrafish, but the relevant molecular mechanisms remained unclear. In this study, 5 day-old crucian carp (Carassius auratus) larvae were treated with 0.5, 5, or 50 µg/L TDCIPP for 90 days; the effect on growth was evaluated; and related molecular mechanisms were explored. Results demonstrated that 5 or 50 µg/L TDCIPP treatment significantly inhibited the growth of crucian carp and downregulated the expression of growth hormones (ghs), growth hormone receptor (ghr), and insulin-like growth factor 1 (igf1). Molecular docking, dual-luciferase reporter gene assay, and in vitro experiments demonstrated that TDCIPP could bind to the growth hormone releasing hormone receptor protein of crucian carp and disturb the stimulation of growth hormone releasing hormone to the expression of ghs, resulting in the decrease of the mRNA level of gh1 and gh2 in pituitary cells. Our findings provide new perceptions into the molecular mechanisms of developmental toxicity of TDCIPP in fish.

Individual Prey Specialization Drives PCBs in Icelandic Killer Whales.

Interindividual variation in prey specialization is an essential yet overlooked aspect of wildlife feeding ecology, especially as it relates to intrapopulation variation in exposure to toxic contaminants. Here, we assessed blubber concentrations of an extensive suite of persistent organic pollutants in Icelandic killer whales (Orcinus orca). Polychlorinated biphenyl (PCB) concentrations in blubber were >300-fold higher in the most contaminated individual relative to the least contaminated, ranging from 1.3 to 428.6 mg·kg-1 lw. Mean PCB concentrations were 6-to-9-fold greater in individuals with a mixed diet including marine mammals than in fish specialist individuals, whereas males showed PCB concentrations 4-fold higher than females. Given PCBs have been identified as potentially impacting killer whale population growth, and levels in mixed feeders specifically exceeded known thresholds, the ecology of individuals must be recognized to accurately forecast how contaminants may threaten the long-term persistence of the world's ultimate marine predator.

A review of 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane in the environment and assessment of its persistence, bioaccumulation and toxicity.

Following the ban of many historically-used flame retardants (FRs), numerous replacement chemicals have been produced and used in products, with some being identified as environmental contaminants. One of these replacement flame retardants is 1,2-dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH; formerly abbreviated as TBECH), which to date has not been identified for risk assessment and potential regulation. DBE-DBCH technical mixtures consist largely of α- and ß-diastereomers with trace amounts of γ- and δ-DBE-DBCH. The α- and ß-isomers are known contaminants in various environmental media. While current global use and production volumes of DBE-DBCH are unknown, recent studies identified that DBE-DBCH concentrations were among the highest of the measured bromine-based FRs in indoor and urban air in Europe. Yet our mass balance fugacity model and modeling of the physical-chemical properties of DBE-DBCH estimated only 1% partitioning to air with a half-life of 2.2 d atmospherically. In contrast, our modeling characterized DBE-DBCH adsorbing strongly to suspended particulates in the water column (~12%), settling onto sediment (2.5%) with minimal volatilization, but with most partitioning and adsorbing strongly to soil (~85%) with negligible volatilization and slow biodegradation. Our modeling further predicted that organisms would be exposed to DBE-DBCH through partitioning from the dissolved aquatic phase, soil, and by diet, and given its estimated logKow (5.24) and a half-life of 1.7 d in fish, DBE-DBCH is expected to bioaccumulate into lipophilic tissues. Low concentrations of DBE-DBCH are commonly measured in biota and humans, possibly because evidence suggests rapid metabolism. Yet toxicological effects are evident at low exposure concentrations: DBE-DBCH is a proven endocrine disruptor of sex and thyroid hormone pathways, with in vivo toxic effects on reproductive, metabolic, and other endpoints. The objectives of this review are to identify the current state of knowledge concerning DBE-DBCH through an evaluation of its persistence, potential for bioaccumulation, and characterization of its toxicity, while identifying areas for future research.

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.

Perfluoroalkyl acids and sulfonamides and dietary, biological and ecological associations in peregrine falcons from the Laurentian Great Lakes Basin, Canada.

Perfluoroalkyl substances (PFAS) are a large, diverse group of chemicals and several perfluoroalkyl acids (PFAAs) are known environmental contaminants. Wildlife exposure to PFAAs and precursors has been shown, but less is known regarding replacements such as shorter-chain PFAS. In the present study, exposure to a suite of PFAAs and associations with dietary, biological and ecological factors were investigated in populations of a sentinel apex species - the peregrine falcon (Falco peregrinus). Nestling blood (n = 57) and sibling eggs (n = 9) were sampled in 2016 and 2018 from nests in rural and urban regions across the Laurentian Great Lakes Basin, Canada. PFSAs (perfluorinated sulfonic acids) including PFHxS, PFOS, and PFDS were detected in most egg and plasma samples, whereas 11 PFCAs (perfluorinated carboxylic acids; C5-C14, C16) compared to eight PFCAs (C8-C14, C16) were detected in most eggs and plasma, respectively. Shorter-chain C8-C10 PFCAs were more dominant in plasma and longer-chain C12-C14 PFCAs in eggs, but profiles were similar for PFOS, PFDS, PFUdA and PFHxDA. The exposure to PFAAs in peregrine falcons is likely mediated by dietary factors such as foraging location (δ13C and δ34S) and trophic position (δ15N) given the associations observed in eggs and nestling plasma, respectively. Moreover, significant relationships were observed for circulating ΣPFCAs and region (rural/urban), and nestling body condition after adjusting for sampling year and dietary tracers, suggesting that compared to rural nestlings, urban nestlings may be more exposed to ΣPFCAs and prone to their potential physiological impacts. Our findings highlight the importance of integrating dietary, biological and ecological factors when studying PFAS exposure in birds.

Promotion effect of liver tumor progression in male kras transgenic zebrafish induced by tris (1, 3-dichloro-2-propyl) phosphate.

A previous study reported that exposure to tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) could promote the progression of hepatocellular carcinoma (HCC) in female HCC model zebrafish. Due to the existence of gender disparity in the development of HCC between females and males, whether the promotion effect of TDCIPP still exists in male HCC model zebrafish remains unclear. In this study, Tg(fabp10:rtTA2s-M2; TRE2:EGFP-krasG12V), referred as kras transgenic zebrafish which was shown to be an inducible liver tumor model, was applied as experimental model to assess the promotion potential of TDCIPP for HCC in males. In brief, kras males were exposed to 20 mg/L doxycycline (DOX), 0.3 mg/L TDCIPP and a binary mixture of 20 mg/L DOX with 0.3 mg/L TDCIPP, and after exposure liver size, histopathology and transcriptional profiles of liver from these treatments were examined. With the involvement of TDCIPP, the liver size was significantly increased and the lesion of hepatocyte became more aggressive. Furthermore, expressions of genes involved in DNA replication and inflammatory response were simultaneously up-regulated in the treatment of TDCIPP compared with the solvent control and in the treatment of the binary mixture of the two chemicals compared to the single DOX treatment. Overall, our results suggested that TDCIPP had promotion effect on the progression of liver tumor in kras males.

Tetrabromobisphenol-A-Bis(dibromopropyl ether) Flame Retardant in Eggs, Regurgitates, and Feces of Herring Gulls from Multiple North American Great Lakes Locations.

The occurrence of tetrabromobisphenol-A-bis(2,3,-dibromopropyl ether) (TBBPA-BDBPE) flame retardant is generally unknown in wildlife. A highly sensitive, gas chromatography-mass spectrometry-based method was developed for TBBPA-BDBPE with optimized parameters for large volume injection. We report on TBBPA-BDBPE and temporal and spatial trends in herring gull egg pools and individuals from 14 colony sites across the Laurentian Great Lakes of North America. TBBPA-BDBPE identification was confirmed using liquid chromatography time-of-flight mass spectrometry and quantification with liquid chromatography tandem mass spectrometry analysis. TBBPA-BDBPE was quantifiable in 95% of egg pools from all colonies sampled in 2013-2017, and retrospective analysis of archived eggs (2001-2017) at 3 of the 14 colonies indicated that TBBPA-BDBPE concentrations were greater in pools from eggs collected in more recent years (

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.

In ovo exposure to brominated flame retardants Part I: Assessment of effects of TBBPA-BDBPE on survival, morphometric and physiological endpoints in zebra finches.

Tetrabromobisphenol A bis(2,3-dibromopropyl) ether (TBBPA-BDBPE) is an additive flame retardant used in polyolefins and polymers. It has been detected in biota, including in avian eggs, yet little is known of its effects. We assessed the pattern of TBBPA-BDBPE concentrations in songbird eggs over the incubation period, and the effects of embryonic exposure to TBBPA-BDBPE in a model songbird species, the zebra finch (Taeniopygia guttata). To assess concentrations during embryo development, eggs were injected on the day they were laid with the vehicle control (safflower oil) or 100 ng TBBPA-BDBPE/g egg, and whole egg contents were collected throughout embryonic development on day 0 (unincubated), 5, 10 and 13. To evaluate effects of embryonic exposure to TBBPA-BDBPE, eggs were injected at Hamburger-Hamilton stage 18 (∼80 h after initiation of incubation) with safflower oil only, 10, 50 or 100 ng TBBPA-BDBPE/g egg (albumin injection volume 1 µl/g). Eggs were monitored for hatching success, and nestlings were monitored for growth and survival. At 15 days post-hatch, tissues were collected to assess physiological effects. TBBPA-BDBPE was incorporated into the egg as the embryo developed, and concentrations started declining in late incubation, suggesting biotransformation by the embryo. There were no effects on hatching success, nestling survival, growth, organ somatic indices, or thyroid hormone homeostasis; however, there was evidence that body condition declined in a dose-dependent manner towards the end of the rapid nestling growth phase. This decreased body condition could be a delayed effect of early developmental exposure, or it may be the result of increased exposure to biotransformation products of TBBPA-BDBPE produced over the nestling period, which are predicted to be more bioaccumulative and toxic than the parent compound.

In ovo exposure to brominated flame retardants Part II: Assessment of effects of TBBPA-BDBPE and BTBPE on hatching success, morphometric and physiological endpoints in American kestrels.

Tetrabromobisphenol A bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) and 1,2-bis(2,4,6-tribromophenoxy)ethane (BTPBE) are both brominated flame retardants (BFRs) that have been detected in birds; however, their potential biological effects are largely unknown. We assessed the effects of embryonic exposure to TBBPA-BDBPE and BTBPE in a model avian predator, the American kestrel (Falco sparverius). Fertile eggs from a captive population of kestrels were injected on embryonic day 5 (ED5) with a vehicle control or one of three doses within the range of concentrations that have been detected in biota (nominal concentrations of 0, 10, 50 or 100 ng/g egg; measured concentrations 0, 3.0, 13.7 or 33.5 ng TBBPA-BDBPE/g egg and 0, 5.3, 26.8 or 58.1 ng BTBPE/g egg). Eggs were artificially incubated until hatching (ED28), at which point blood and tissues were collected to measure morphological and physiological endpoints, including organ somatic indices, circulating and glandular thyroid hormone concentrations, thyroid gland histology, hepatic deiodinase activity, and markers of oxidative stress. Neither compound had any effects on embryo survival through 90% of the incubation period or on hatching success, body mass, organ size, or oxidative stress of hatchlings. There was evidence of sex-specific effects in the thyroid system responses to the BTBPE exposures, with type 2 deiodinase (D2) activity decreasing at higher doses in female, but not in male hatchlings, suggesting that females may be more sensitive to BTBPE. However, there were no effects of TBBPA-BDBPE on the thyroid system in kestrels. For the BTPBE study, a subset of high-dose eggs was collected throughout the incubation period to measure changes in BTBPE concentrations. There was no decrease in BTBPE over the incubation period, suggesting that BTBPE is slowly metabolized by kestrel embryos throughout their ∼28-d development. These two compounds, therefore, do not appear to be particularly toxic to embryos of the American kestrel.

Polychlorinated Diphenylsulfides Activate Aryl Hydrocarbon Receptor 2 in Zebrafish Embryos: Potential Mechanism of Developmental Toxicity.

It is hypothesized that polychlorinated diphenyl sulfides (PCDPSs) induce lethal toxicity in zebrafish which is mediated by aryl hydrocarbon receptor 2 (Ahr2) activation. In this study an assay was developed based on in vivo exposure of wild-type and Tg(cyp1a:gfp) transgenic zebrafish embryos/larvae to PCDPS congeners (i.e., six dichloro- to heptachloro-diphenyl sulfides) coupled with a zebrafish Ahr2-luciferase reporter gene (LRG) expression. Waterborne PCDPSs were found to be accumulated in zebrafish larvae, and exposure to PCDPSs led to a significant increase in mortality and cyp1s mRNA expression. Furthermore, treatment with PCDPSs caused a significant induction of Ahr2-LRG activity in COS-7 cells, and extremely significant correlations were observed between the in vivo median lethal concentrations and the levels of cyp1s mRNA expression and Ahr2 activation. Molecular dynamics simulations indicated the interaction between dioxins/dioxin-like compounds (DLCs) and six key amino acid residues in the ligand-binding domain of Ahr2 probably determined the susceptibility to dioxins/DLCs in zebrafish. These results strongly support the hypothesis that early life-stage mortality of zebrafish is initiated and mediated by Ahr2 activation.