ethoxyresorufin-O-deethylase Research Articles

Olive leaf (Olea europaea L. folium) extract influences liver microsomal detoxifying enzymes in rats orally exposed to 2-amino-l-methyI-6-phenyI-imidazo pyridine (PhIP).

Olive tree (Olea europaea, Oleaceae) leaf extract (OLE) exerts many biological activities. One of the most common polycyclic aromatic hydrocarbons (PAHs) that pollute the environment is 2-amino-l-methyI-6-phenyI-imidazo pyridine (PhIP). It is a food-derived carcinogen that is present in fish and meat that has been cooked at high temperatures. Due to the generation of reactive electrophilic species, phase I enzymes have the potential to cause oxidative damage. In order to safely remove these reactive species from the body, phase II detoxification (conjugation) enzymes are necessary. It is not known whether OLE could influence their activities and hence reduce the carcinogenic effects of PhIP. This study evaluated whether OLE could modulate phase I detoxifying enzymes as well as phase II enzymes that metabolize PhIP in rat liver microsomes. Four groups of rats were used: group I: no treatment; group II: OLE (10mg/kg bw orally); group III: PhIP (0.1mg/kg bw orally); and group IV: PhIP followed by OLE. After 4weeks, the activities of phase I enzymes such as CYP1A1 (ethoxyresorufin O-deethylase), CYP2E1 (p-nitrophenol hydroxylase), CYP1A2 (methoxyresorufin O-demethylase), UDP-glucuronyl transferase, sulphotransferase, and glutathione-S transferase were evaluated in rat liver microsomes. Analysis of OLE by gas chromatography-mass spectrometry (GC/MS) showed various active ingredients in OLE, including 3,5-Heptadienal (C10H14O), 3,4-dimethoxy benzoic acid (C8H10O3), 4-hydroxy-3-methoxy (C8H8O4), 1,3,5-Benzenetriol (C6H6O3), hexadecanoic acid (C16H32O2), and hexadecanoic acid ethyl ester (C18H36O2). Our results showed that rats given PhIP were found to have a statistically significant (p < 0.001) reduction in the activities of CYP1A1, CYP1A2, and CYP2E1 in comparison with the control group. However, treatment with OLE enhanced their activities but not to a normal level compared with untreated groups. Administration of PhIP decreased the activities of phase II enzymes (glutathione S-transferase, UDP-glucuronyltransferase, or sulphotransferase) (p < 0.01) in comparison with the control group. Histological examination of rat livers was consistent with the biochemical changes. The administration of OLE improved the phase II enzyme activities in animals injected with PhIP. We conclude that OLE influences phase I and phase II detoxification enzymes exposed to PhIP, which may represent a new approach to attenuating carcinogenesis induced by it.

Chronic Effects of Carbamazepine, Progesterone and Their Mixtures at Environmentally Relevant Concentrations on Biochemical Markers of Zebrafish (Danio rerio).

The impact of pharmaceuticals on non-target organisms in the environment is of increasing concern and study. Pharmaceuticals and other pollutants are often present as mixtures in an environmental compartment. Studies on the toxicological implications of these drugs on fish, particularly as mixtures at environmentally relevant concentrations, are very limited. Thus, this study aimed to evaluate the chronic effects of the anticonvulsant drug carbamazepine (CBZ) and progesterone (P4) at environmentally relevant concentrations, individually and in binary mixtures, applying a suite of biomarkers at the molecular level in zebrafish (Danio rerio). The effects on biotransformation enzymes 7-ethoxyresorufin O-deethylase (EROD) and glutathione-S-transferase (GST), antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidases (GPxSe and GPxTOT), and glutathione reductase (GR), and markers of damage, such as DNA strand breaks (DNAsb), lactate dehydrogenase (LDH), lipid peroxidation (LPO), and vitellogenin-like proteins (VTG), were evaluated. Analyses of the biochemical markers indicated that a synergistic dose-ratio-dependent effect of CBZ and P4 in zebrafish occurs after chronic exposure regarding VTG, biotransformation enzymes (EROD, GST), and oxidative stress marker (DNAsb). The results suggest a synergistic effect regarding VTG, thus indicating a high risk to the reproductive success of fish if these pharmaceuticals co-occur.

Bioaccumulation, Detoxification, and Biological Macromolecular Damage of Benzo[a]pyrene in Exposure in Tissues and Subcellular Fractions of Scallop Chlamys farreri.

Because of the persistence and high toxicity of benzo[a]pyrene (B[a]P), the bioaccumulation and detoxification mechanisms of B[a]P have been studied extensively at the tissue level; but the data at the subcellular level in bivalves have not been reported. The present study was conducted to investigate the effects of B[a]P exposure on bioaccumulation, detoxification, and biomacromolecular damage in gills, digestive glands, and their subcellular fractions of the scallop Chlamys farreri. The subcellular fraction contains cytoplasm, mitochondria, microsome, nucleus, cell membrane, and overall organelle. The results demonstrated that B[a]P accumulation showed a clear time-dose effect. Based on the time-dependent accumulation of B[a]P in subcellular fractions, we speculated that the intracellular migration order of B[a]P was cell membrane, organelle, and nucleus in turn. Considering the difference of B[a]P accumulation may be related to B[a]P metabolism, we have further confirmed that the activities of B[a]P metabolizing enzymes in scallop tissues and subcellular fractions were significantly tempted by B[a]P (p < 0.05), including 7-ethoxyresorufin O-deethylase (increased), glutathione-S-transferase (GST; decreased), and superoxide dismutase (increased). First, GST was detected in bivalve cytoplasm and microsome. Second, B[a]P exposure also caused biomacromolecules damage. The results demonstrated that mitochondria and microsome were more vulnerable to lipid peroxidation than cell membrane and nucleus. Taken together, the present study fills some of the gaps in our knowledge of the bioaccumulation and detoxification mechanisms of C. farreri exposed to B[a]P in subcellular fractions and deeply explores the transportation and the main metabolic and damage sites of polycyclic aromatic hydrocarbons (PAHs) in cells, which helped us to comprehensively understand the toxic mechanism of PAHs on bivalves. Environ Toxicol Chem 2022;41:2353-2364. © 2022 SETAC.

E3 ubiquitin ligases STUB1/CHIP contributes to the Th17/Treg imbalance via the ubiquitination of aryl hydrocarbon receptor in rheumatoid arthritis.

STIP1-homologous U-Box containing protein 1 (STUB1) is involved in the development of immune pathologies and the regulation of T cell. However, the potential role of STUB1 in the pathogenesis of rheumatoid arthritis (RA), especially in the regulation of T cells, remains elusive. Here we show that STUB1 promotes the imbalance of Th17/Treg cells through non-degradative ubiquitination of aryl hydrocarbon receptor (AHR). Using Western blot and flow cytometry analysis, we observe that the level of STUB1 was increased in RA patients compared with healthy controls. In particular, the expression of STUB1 protein was different in Th17 cells and Treg cells of RA patients. We also demonstrated that STUB1 facilitates Th17/Treg imbalance by up- or downregulating the expression of STUB1. In a subsequent series of in vitro experiments, we revealed that STUB1 promoted the imbalance of Th17 and Treg cells through non-degradative ubiquitination of AHR. Both knockdown of the AHR expression by siRNA and assays of CYP1A1 enzymatic activity by ethoxyresorufin-O-deethylase (EROD) supported this conclusion. Furthermore, we explored the ubiquitination sites of AHR responsible for STUB1-mediated ubiquitination and revealed that STUB1 promotes ubiquitination of AHR via K63 chains. Together, STUB1 may induce the imbalance of Th17/Treg cells via ubiquitination of AHR and serve as a potential therapeutic target for RA.

Environmental microplastics disrupt swimming activity in acute exposure in Danio rerio larvae and reduce growth and reproduction success in chronic exposure in D. rerio and Oryzias melastigma

Microplastics (MPs), widely present in aquatic ecosystems, can be ingested by numerous organisms, but their toxicity remains poorly understood. Toxicity of environmental MPs from 2 beaches located on the Guadeloupe archipelago, Marie Galante (MG) and Petit-Bourg (PB) located near the North Atlantic gyre, was evaluated. A first experiment consisted in exposing early life stages of zebrafish (Danio rerio) to MPs at 1 or 10 mg/L. The exposure of early life stages to particles in water induced no toxic effects except a decrease in larval swimming activity for both MPs exposures (MG or PB). Then, a second experiment was performed as a chronic feeding exposure over 4 months, using a freshwater fish species, zebrafish, and a marine fish species, marine medaka (Oryzias melastigma). Fish were fed with food supplemented with environmentally relevant concentrations (1% wet weight of MPs in food) of environmental MPs from both sites. Chronic feeding exposure led to growth alterations in both species exposed to either MG or PB MPs but were more pronounced in marine medaka. Ethoxyresorufin-O-deethylase (EROD) and acetylcholinesterase (AChE) activities were only altered for marine medaka. Reproductive outputs were modified following PB exposure with a 70 and 42% decrease for zebrafish and marine medaka, respectively. Offspring of both species (F1 generation) were reared to evaluate toxicity following parental exposure on unexposed larvae. For zebrafish offspring, it revealed premature mortality after parental MG exposure and parental PB exposure produced behavioural disruptions with hyperactivity of F1 unexposed larvae. This was not observed in marine medaka offspring. This study highlights the ecotoxicological consequences of short and long-term exposures to environmental microplastics relevant to coastal marine areas, which represent essential habitats for a wide range of aquatic organisms.

Biochemical and cellular responses of the freshwater mussel, Hyriopsis bialata, to the herbicide atrazine

The present study aimed to evaluate biochemical and cellular responses of the freshwater mussel, Hyriopsis bialata, to the herbicide atrazine (ATZ). The mussels were exposed to environmentally-relevant concentrations of ATZ (0, 0.02 and 0.2 mg/L) and a high concentration (2 mg/L) for 0, 7, 14, 21 and 28 days. Tissues comprising male and female gonads, digestive glands and gills were collected and assessed for ethoxyresorufin-O-deethylase (EROD) activity, glutathione S-transferase (GST) activity, multixenobiotic resistance mechanism (MXR), histopathological responses, DNA fragmentation and bioaccumulation of ATZ and its transformation derivatives, desethylatrazine (DEA) and desisopropylatrazine (DIA). Additionally, circulating estradiol levels were determined. It appeared that ATZ did not cause significant changes in activities of EROD, GST and MXR. There were no apparent ATZ-mediated histopathological effects in the tissues, with the exception of the male gonads exhibiting aberrant aggregation of germ cells in the ATZ-treated mussels. Contrarily, ATZ caused significant DNA fragmentation in all tissues of the treated animals in dose- and time-dependent manners. In general, the circulating estradiol levels were higher in the females than in the males. However, ATZ-treated animals did not show significant alterations in the hormonal levels, as compared with those of the untreated animals. Herein, we showed for the first time differentially spatiotemporal distribution patterns of bioaccumulation of ATZ, DEA and DIA, with ATZ and DEA detectable in the gonads of both sexes, DEA and DIA in the digestive glands and only DEA in the gills. The differential distribution patterns of bioaccumulation of ATZ and its derivatives among the tissues point to different pathways and tissue capacity in transforming ATZ into its transformation products. Taken together, the freshwater mussel H. bialata was resistant to ATZ likely due to their effective detoxification. However, using DNA damage as a potential biomarker, H. bialata is a promising candidate for biomonitoring aquatic toxicity.

Diluted bitumen-induced alterations in aerobic capacity, swimming performance, and post-exercise recovery in juvenile sockeye salmon (Oncorhynchus nerka).

The transportation of heavy crudes such as diluted bitumen (dilbit) sourced from Canadian oil sands through freshwater habitat requires the generation of information that will contribute to risk assessments, spill modelling, management, and remediation for the protection of aquatic organisms. Juvenile sockeye salmon (Oncorhynchus nerka) were exposed acutely (96 h) or subchronically (28 d) to the water-soluble fraction (WSFd) of Cold Lake Blend dilbit at initial total polycyclic aromatic compound (TPAC) concentrations of 0, 13.7, 34.7, and 124.5 µg/L. A significant induction (>3-fold) of hepatic liver ethoxyresorufin-O-deethylase (EROD) activity was induced by 96 h in fish exposed to [TPAC] ≥ 34.7 µg/L and at ≥13.7 µg/L for a 28 d exposure. Exposure resulted in a typical physiological stress response and disturbance of ion homeostasis; this included elevations in plasma [cortisol], [lactate], [Na+], and [Cl-], and significant reductions in muscle [glycogen]. Critical swimming speed (Ucrit) was significantly reduced (28.4%) in the acute exposure at [TPAC] 124.5 µg/L; reductions of 14.2% and 35.4% were seen in fish subchronically exposed at the two highest concentrations. Reductions in Ucrit were related to significant reductions in aerobic scope (24.3-46.6%) at [TPAC]s of 34.7 and 124.5 µg/L, respectively. Exposure did not impair the ability to mount a secondary stress response following burst exercise, however, the time required for biochemical parameters to return to baseline values was prolonged. Alterations in critical systems supporting swimming, exercise recovery and the physiological stress response could result in decreased salmonid fitness and contribute to population declines if a dilbit spill occurs.

A screening study of relationships among concentrations of algal toxins, PFAS, thiamine deficiency and biomarkers in the European flounder from the southern Baltic Sea

In the disturbed Baltic Sea ecosystem, several animal species display failing health related to exposure to toxic compounds, reduced energy metabolism and immune system disorders. In order to investigate possible associations between fish health and exposure to chemicals affecting the energy metabolism and immune defence, the levels of algal toxins (bromophenols, hydroxylated polybrominated diphenyl ethers and nodularin), perfluorinated alkyl substances (PFAS) and thiamine (vitamin B1) were determined in European flounder (Platichthys flesus). Several biomarkers indicating health status were examined in the fish, including ethoxyresorufin-O-deethylase (EROD), and activities of glutathione reductase, glutathione S-transferase and catalase, in addition to a large set of blood variables. The fish were collected from Hanöbukten in the south-western parts of the Baltic Sea in late August 2018. Regression analyses of algal toxins, PFAS and thiamine concentration displayed several significant associations with biomarkers associated with detoxification and liver function, immune system function and blood status of the fish.

Impacts of size-fractionation on toxicity of marine microplastics: Enhanced integrated biomarker assessment in the tropical mussels, Perna viridis

Accumulation of microplastics (MP) in oceanic waters is eroding the health of marine biota. We investigated how size-fractionated MP influence the toxicity risks towards a tropical keystone species, Perna viridis. Tissue-specific bioaccumulation and in vivo toxicity of polystyrene (PS) particles (0.5, 5, and 50 μm) were measured upon continuous exposure for 7 days, followed by 7 days depuration. P. viridis were exposed to equivalent mass (0.6 mg/L), corresponding to 4.0–4.6 particles/mL, 4.6–7.1 × 103 particles/mL, and 1.1–4.8 × 106 particles/mL for 50 μm, 5 μm and 0.5 μm PS particles, respectively. Onset toxicity were quantified through the enhanced integrated multi-biomarker response (EIBR) model, measured by weighting of biological organisation levels of eight biomarkers: (i) molecular (i.e., DNA damage (comet), 7-ethoxy resorufin O-deethylase (EROD), Catalase (CAT), Superoxide dismutase (SOD), Ferric Reducing Antioxidant Power (FRAP)); (ii) cellular (i.e., Neutral red retention (NRR), phagocytosis); and (iii) physiological (i.e., filtration rate). Data showed slightly elevated lysosomal instability (NRR) and antioxidant defences (FRAP, SOD, CAT, EROD) in specimens exposed to nano-PS (0.5 μm) compared to micro-PS (5 and 50 μm). Immunotoxicity (phagocytosis) and genotoxicity (comet) for haemocyte cells were significantly higher in specimens exposed to nano-PS (p < 0.05). EIBR index corroborated increasing toxicity modulated by MP sizes in descending order: 0.5 μm > 5 μm > 50 μm, with nano-PS exerted significantly higher biological effects (EIBR = 19.77 ± 5.89) than the unexposed group (EIBR = 10.97 ± 2.02; p < 0.05). Symptomatic organismal depression was manifested by the depleting filtering proficiency and weakened defence against invasive Zymosan bioparticles in the phagocytosis assay. Although impaired mussels duly recovered during depuration, individuals affected by nano-PS showed immunocompetence deficiency and gill responses that were not readily reversible, which could potentially increase their vulnerability towards further environmental stressors.

Steroid Hormones Protect against Fluoranthene Ethoxyresorufin-O-Deethylase (EROD) Activity Inhibition

The physiological conditions of an organism may influence its ability to cope with environmental stressors, such as contaminants. Biotransformation and the endocrine system interact with each other to promote animal’s fitness. However, little is known regarding the interaction between hormones and response to pollutants such as polycyclic aromatic hydrocarbons (PAHs). In this in vitro study, we aimed to increase the knowledge regarding the effects of steroid hormones on ethoxyresorufin-O-deethylase (EROD) activity inhibited by contaminants. The effects on in vivo induced EROD activity of Anguilla anguilla were assessed by conducting single and combined exposures to fluoranthene (FL) and to physiological levels of two major steroid hormones (cortisol and 17ß-estradiol). Hepatic microsome exposure to the lowest concentrations of FL (0.1 and 0.3 µM), as well as to cortisol and 17ß-estradiol (E2), led to significant EROD activity induction. However, the highest tested concentrations of FL (0.9 and 2.7 µM) significantly inhibited this enzymatic activity. When microsomes were simultaneously exposed to 0.9 µM FL and one of the hormones, both cortisol and E2 were able to decrease the inhibitory effects, with the former completely reverting EROD activity inhibition. These findings support the idea that cortisol and E2 can help prevent the inhibitory effects of PAHs over biotransformation enzymes, highlighting the physiological relevance of these hormones.

Quantification of differential tissue biomarker responses to microplastic ingestion and plasticizer bioaccumulation in aquaculture reared sea bream Sparus aurata

Marine aquaculture is considered a potential source of microplastics (MPs). MPs can induce oxidative stress and damage in marine species. In this study we evaluated the impact of MPs intake in the commercial fish, Sparus aurata, from aquaculture facilities and the antioxidant response associated to this MPs ingestion in caged specimens for 120 days. Sampling was carried out at the beginning of the study (T0), at 60 days (T60) and at 120 days (T120). At each sampling stage, gastrointestinal tract, blood, plasma, liver and muscle samples were obtained to analyse MPs intake (gastrointestinal tract), oxidative stress markers (blood, plasma and liver) and plasticizers bioaccumulation (muscle). Fish sampled at T60 presented the highest MPs intake and plasticizers accumulated in muscle over time, but with a different pattern according to type: bisphenols and phthalates. This indicates MPs ingestion induces a differential tissue response in S. aurata. Similarly, stress biomarkers presented a differential response throughout the study, depending on the analysed tissue. In the case of oxidative damage markers, for malondialdehyde (MDA) an increase throughout the study was observed both in liver and blood cells but with a progressive decrease in plasma. In the case of phase I detoxifying enzyme activities in liver, 7-ethoxyresorufin O-deethylase (EROD), 7-benzyloxy-4-[trifluoromethyl]-coumarin-O-debenzyloxylase (BFCOD) and carboxylesterases (CE), showed a comparable decrease at T60 with a slight recovery at T120. In contrast, glutathione-S-transferase (GST) activity was significantly enhanced at T60 compared to the other sampling stages. In conclusion, MPs ingestion occurs in aquaculture reared seabream where potentially associated plasticizers accumulate in the muscle and both could be responsible for plasma and liver oxidative stress damage and alterations on detoxifying biomarkers responses.

Characteristics of cytochrome P450-dependent metabolism in the liver of the wild raccoon, Procyon lotor.

Wildlife is exposed to a wide range of xenobiotics in the natural environment. In order to appropriately assess xenobiotic-induced toxicity in wildlife, it is necessary to understand metabolic capacities. Carnivores, in general, have low metabolic abilities, making them vulnerable to a variety of chemicals. Raccoons (Procyon lotor) in the wild have been found to have high levels of xenobiotics. However, little is known about the metabolic capacity of the cytochrome P450 (CYP) enzymes in this species. Thus, this study used liver samples to investigate the characteristics of CYP enzymes in wild raccoons. In 22 wild raccoons, CYP concentrations in hepatic microsomes were examined. To better understand the properties of CYP-dependent metabolism, in vitro metabolic activity studies were performed using ethoxyresorufin, pentoxyresorufin and testosterone as substrates. In addition, three raccoons were fed commercial dog food in the laboratory for one week, and the effects on CYP-dependent metabolism were investigated. In comparison to other mammalian species, raccoons had very low concentrations of CYP in their livers. In an in vitro enzymatic analysis, raccoons' ethoxyresorufin O-deethylase (EROD) and pentoxyresorufin O-depentylase (PROD) metabolic capacities were less than one-fifth and one-tenth of rats', respectively. These results indicate the possible high risk in raccoons if exposed to high levels of environmental xenobiotics because of their poor CYP activity. In this study, the features of CYP-dependent metabolism in wild raccoons are described for the first time.

Synthesis, antioxidant activity, molecular docking and ADME studies of novel pyrrole-benzimidazole derivatives.

Several 5-(alkylsulfonyl)-1-substituted-2-(1H-pyrrol-2-yl)-1H-benzo[d]imidazole derivatives were synthesized and their antioxidant activities were investigated using lipid peroxidation (LPO) and 7-ethoxyresorufin O-deethylase (EROD) assays. Docking analysis with Human NAD[P]H-Quinone oxidoreductase 1 (NQO1) was also performed to gather thorough information about these compounds that have antioxidant activities. Moreover, their molecular descriptors and ADME properties were calculated using the SwissADME online program. As a result, most of our compounds possessed better affinity and created ample interactions with NQO1. The most potent compound 5j had LP inhibition value of 3.73 nmol/mg/min. Other compounds exhibited moderate activity on LP levels comparing to standard butylated hydroxy toluene (BHT). However, the inhibitory effect on EROD activity was not significant.

Characterization of marine-derived halogenated indoles as ligands of the aryl hydrocarbon receptor

The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor thought to mediate a number of physiological roles in the body, is becoming a target of interest for the development of new therapeutics. However, previous research has demonstrated that the downstream effects of AhR ligands cannot be predicted based simply on whether a ligand acts as an agonist or antagonist and the persistence of AhR signaling is thought to be a key determining feature. The current study investigated the AhR activity of four halogenated indoles isolated from the New Zealand red alga, Rhodophyllis membranacea: 4,7-dibromo-2,3-dichloroindole (4DBDCI), 7-bromo-2,3-dichloro-6-iodoindole (BDCII), 6,7-dibromo-2,3-dichloroindole (6DBDCI) and 2,6,7-tribromo-3-chloroindole (TBCI). Their ability to activate AhR signaling, measured as CYP1A1 activity via the ethoxyresorufin O-deethylase (EROD) assay, was determined in human HepG2, mouse Hepa1c1c7 and rat H4IIE liver cancer cells. All four compounds induced CYP1A1 activity in HepG2 cells, suggesting they all acted as AhR agonizts. 4DBDCI was particularly efficacious, inducing an 11-fold increase. Hepa1c1c7 and H4IIE cells, however, were generally less responsive to the halogenated indoles. All four compounds were persistent AhR agonizts, inducing peak CYP1A1 activity after 72 h. Moreover, the 2,3,6,7-substituted BDCII, 6DBDCI and TBCI, but not 4DBDCI, competed with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for AhR binding as observed by the inhibition of TCDD-induced CYP1A1 activity. Overall, the current study has characterized four previously untested AhR ligands, highlighting differences in species sensitivity and persistence of signaling to provide a framework for their potential future use.

Synthesis and Antioxidant Properties of New Benzimidazole Derivatives

In this study, new benzimidazole derivatives were synthesized. After purity checks were made with Thin Layer Chromatography (TLC) and melting points, their structures were proved by mass spectroscopy, 1H, 13C NMR and elemental analysis technique. Antioxidant activities of the obtained materials were searched in terms of their effect on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, lipid peroxidation inhibition and ethoxyresorufin O-deethylase (EROD) enzyme activity. It was observed the compounds were generally moderately active in terms of DPPH radical scavenging activity and lipid peroxidation inhibition compared to the standard compound and had a good effect on EROD enzyme activity.

Correlating seasonal changes of naturally produced brominated compounds to biomarkers in perch from the Baltic Sea.

Hydroxylated polybrominated diphenyl ethers (OH-PBDEs), naturally produced by algae and cyanobacteria in the Baltic Sea, are potent disrupters of energy metabolism as well as endocrine disruptors and neurotoxins. In this study, European perch (Perca fluviatilis) from the Baltic Sea were sampled from May until October. OH-PBDEs and ten biomarkers were measured in each individual (n=84 over 18 sampling time points) to study potential correlations between exposure to OH-PBDEs and changes in biomarkers. Several biomarkers showed significant non-linear seasonal variation. In the perch, ethoxyresorufin-O-deethylase (EROD) activity, plasma lactate concentration, and plasma glucose concentration showed a significant positive log-linear correlation with OH-PBDEs, whereas lipid percentage and liver somatic index showed a significant negative log-linear correlation with OH-PBDEs. These results strengthen the concern that OH-PBDEs could cause negative health effects for fish in the Baltic Sea.

A meta-analytic review of fish antioxidant defense and biotransformation systems following pesticide exposure

Pesticides reach aquatic ecosystems and interact with various targets in cells of fish and other living organisms. Toxicity originates during the metabolization process, which may produce toxic metabolites or reactive oxygen species (ROS). Ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) activities, and levels of reduced glutathione (GSH) indicate toxicants interacted with drug-metabolizing and antioxidant systems, i.e., they are biomarkers of biotransformation and oxidative stress. We meta-analytically quantified the impact of pesticides on the mean response and variability of these biomarkers. Our goals were to verify (i) the overall effect of pesticides on oxidative stress and biotransformation, and how each biomarker respond to exposure; (ii) how the life stage of fish (juvenile and adult) influence biomarkers variability and mean activity; (iii) to what extent fish sex (male, female or mixed-sex groups) modify pesticides toxicity; (iv) how different classes of pesticides, and the combination of their concentration and time of exposure, affect each biomarker. Overall, pesticides induced oxidative stress and the biotransformation system. Regardless of life stage, EROD mean activity increased significantly. In exposed juveniles, CAT and GST variability decreased and increased, respectively. CAT mean activity was higher in females, while EROD and GST activities increased in males after pesticide exposure. Organophosphorus (OPs) and organochlorine insecticides, along with imidazole and triazole fungicides, affected biomarkers the most, however the combined effect of concentration and time of exposure of OPs was not detected. Notably, imidazoles and triazoles classes increased EROD by more than 100%. Additionally, we identified research gaps, such as the lack of effect estimates of relevant pesticides on EROD (e.g., pyrethroids and neonicotinoids) and the small number of studies evaluating GSH on female fish. Future researchers may use these gaps as a guide towards enhanced experimental designs and, consequently, a better understanding of pesticide toxic effects on fish.

Tris (2,3-Dibromopropyl) Isocyanurate (TDBP-TAZTO or TBC) Shows Different Toxicity Depending on the Degree of Differentiation of the Human Neuroblastoma (SH-SY5Y) Cell Line

Tris (2,3-dibromopropyl) isocyanurate (TDBP-TAZTO or TBC) is a heterocyclic hexabromated flame retardant. It is widely used during the production of many synthetic compounds. High concentrations of TDBP-TAZTO were found in river water, surface sediments, soil, earthworms, and carp tissues. Moreover, it has been shown that this compound can cross the blood–brain barrier and accumulate in the gut and brain of carp. The aryl hydrocarbon receptor (AhR) has been characterized as a multifunctional intracellular sensor and receptor. AhR is an activator of cytochrome P450 1A1 and 1A2, which metabolize various toxic compounds. The aim of the study was to explain how/whether TDBP-TAZTO increases the expression and/or activity of the CYP1A1 enzyme and the AhR and TUBB3 expression during SH-SY5Y cell differentiation. SH-SY5Y cells were differentiated for 7 and 14 days using retinoic acid. Cell viability, ethoxyresorufin-O-deethylase (EROD) activity, and mRNA expression of CYP1A1, AhR, and TUBB3 were assessed. Our experiment showed that, during the differentiation process, the ability of TDBP-TAZTO to induce EROD activity in SH-SY5Y cells subsequently decreased, which may have been an effect of cell differentiation into neurons. Moreover, the results suggest that TDBP-TAZTO can affect the differentiation process. Since no CYP2B6 mRNA expression was detected, the CAR receptor may not be involved in the TDBP-TAZTO mechanism of action. However, more research is needed in this field to elucidate this mechanism precisely.

An integrated approach using chemical ecological risk assessment and multi-integrated biomarker indexes approach to assess pollution: A case study of Ruditapes philippinarum in four bays on the Shandong Peninsula in China

Considering the ecological risks of polycyclic aromatic hydrocarbons (PAHs) to the marine environment, it is urgent to find scientific and effective monitoring methods. In this study, an integrated approach combining chemical ecological risk assessment and multi-integrated biomarker indexes approach was used to assess the marine environment. Samples included seawater, sediments, and clam Ruditapes philippinarum were collected from four bays on the Shandong Peninsula, China in the four seasons of 2019. The concentrations, composition, potential sources, and ecological risk of PAHs were investigated in seawater and sediments. Risk quotient (RQ) and sediment quality guidelines (SQGs) were calculated to assess the ecological risks of PAHs in seawater and sediment, respectively. And then, clam Ruditapes philippinarum's multi-level biological response, including its ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), superoxide dismutase (SOD), lipid peroxidation (LPO), and acetylcholinesterase (AChE) were investigated in-depth, by which multi-integrated biomarker indexes approach were calculated to evaluate marine environmental quality. Taken together, the results showed that the concentration of PAHs was in good agreement with the response of biomarkers, and the usefulness of the combined use of chemical ecological risk assessment and integrated biomarker indexes to assess PAHs pollution was verified.

Suspended solids-associated toxicity of hydraulic fracturing flowback and produced water on early life stages of zebrafish (Danio rerio)

Hydraulic fracturing flowback and produced water (HF-FPW), which contains polyaromatic hydrocarbons (PAHs) and numerous other potential contaminants, is a complex wastewater produced during the recovery of tight hydrocarbon resources. Previous studies on HF-FPW have demonstrated various toxicological responses of aquatic organisms as consequences of combined exposure to high salinity, dissolved organic compounds and particle/suspended solids-bound pollutants. Noteworthy is the lack of studies illustrating the potentially toxic effects of the FPW suspended solids (FPW-SS). In this study, we investigated the acute and sublethal toxicity of suspended solids filtered from six authentic FPW sample collected from two fracturing wells, using a sediment contact assay based on early-life stages of zebrafish (Danio rerio). PAHs profiles and acute toxicity tests provided initial information on the toxic potency of the six samples. Upon exposure to sediment mixture at two selected doses (1.6 and 3.1 mg/mL), results showed adverse effects in larval zebrafish, as revealed by increased Ethoxyresorufin-O-deethylase (EROD) activity. Transcriptional alterations were also observed in xenobiotic biotransformation (ahr, pxr, cyp1a, cyp1b1, cyp1c1, cyp1c2, cyp3a65, udpgt1a1, udpgt5g1), antioxidant response (sod1, sod2, gpx1a, gpx1b) and hormone receptor signaling (esr1, esr2a, cyp19a1a, vtg1) genes. The results demonstrated that even separated from the complex aqueous FPW mixture, FPW-SS can induce toxicological responses in aquatic organisms' early life stages. Since FPW-SS could sediment to the bottom of natural wetland acting as a continuous source of contaminants, the current findings imply the likelihood of long-term environmental risks of polluted sediments on aquatic ecosystems due to FPW spills.