Ethoxyresorufin O-deethylase Activity Research Articles

Differential effects of oxytetracycline on detoxification and antioxidant defense in the hepatopancreas and intestine of Chinese mitten crab under cadmium stress

This study aims to evaluate the effects of oxytetracycline (OTC) on detoxification and oxidative defense in the hepatopancreas and intestine of Chinese mitten crab (Eriocheir sinensis) under cadmium (Cd) stress. The crab was exposed to 0.6 μM Cd, 0.6 μM OTC, and 0.6 μM Cd plus 0.6 μM OTC for 42 days. Our results showed that in the intestine, OTC alone enhanced protein carboxylation (PC) and malondialdehyde (MDA) contents, which was associated with the increased OTC accumulation. Compared to Cd alone, Cd plus OTC increased Cd and OTC contents, and reduced detoxification (i.e., glutathione (GSH) content, gene expressions of cytochrome P450 (CYP) isoforms, 7-ethoxyresorufin O-deethylase (EROD) activity, mRNA levels and activities of glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST)), and antioxidant defense (i.e., gene expressions and activities of catalase (CAT) and superoxide dismutase (SOD)) in the intestine, leading to the increased in PC and MDA contents, suggesting that OTC had a synergistic effect on Cd-induced oxidative damage. In the hepatopancreas, although OTC alone increased OTC accumulation, it did not affect PC and MDA contents. Compared to Cd alone, Cd plus OTC reduced MDA content, which was closely related to the improvement of detoxification (i.e., GSH content, mRNA levels of CYP isoforms, EROD activity, gene expressions and activities of GPx, GR and GST), and antioxidant defense (gene expressions and activities of CAT and SOD, metallothionein content). Aryl hydrocarbon receptor (AhR) and nuclear factor E2-related factor 2 (Nrf2) transcriptional expressions were positively correlated with most detoxification- and antioxidant-related gene expressions, respectively, indicating that AhR and Nrf2 were involved in the regulation of these gene expressions. Our results unambiguously demonstrated that OTC had tissue-specific effects on Cd-induced toxicological effect in E. sinensis, which contributed to accurately evaluating Cd toxicity modulated by TCs in crab.

Metabolism of β-naphthoflavone by cytochrome P450 isoforms in three air breathing fish, Channa punctatus, Heteropneustes fossilis and Clarias batrachus

The crucial phase I xenobiotic metabolising enzyme cytochrome P450 (CYP 450) has a variety of isoforms and a wide range of substrate specificities. While the roles of mammalian CYP 450 are well established, those of non-mammalian species are much less defined. These roles are frequently extrapolated from mammalian data and assumed to be similar across vertebrate species. In the present study, three air breathing fish species, Channa punctatus, Heteropneustes fossilis and Clarias batrachus were injected intraperitoneally (IP) with 50 mg/kg β-naphthoflavone (β-NF) and its effects on hepatic CYP 450 system were evaluated. β-NF treatment resulted in significant increase (p<0.01) in CYP 450 content and CYP1A mediated EROD (Ethoxyresorufin O-deethylase) activity in all three treated fish groups compared to control. CYP2B mediated N,N-dimethylaniline demethylase activity and CYP2E1 mediated aniline hydroxylase activity showed no response suggesting insensitivity to β-NF treatment. All three fish species showed an increase in CYP3A4 mediated erythromycin N-demethylase activity, but only H. fossilis displayed a significant difference (p<0.05). Of all the activities investigated, EROD activity mediated by CYP1A responded towards β-NF treatment the best and could serve as a sensitive biomarker to gauge the effects of chemical pollutants in the aquatic environment.

Assessment of pollution variability across the central part of the Büyük Menderes River (Turkey) using water physicochemical parameters and biomarker responses in the non-biting midge Chironomus riparius (Diptera: Chironomidae)

ABSTRACT This study aimed to examine the pollution level in the central part of the Büyük Menderes River (in both the main tributary and Çürüksu stream) in Denizli (Turkey) using water physicochemical parameters and biomarker responses in Chironomus riparius (Diptera: Chironomidae). In physicochemical parameter evaluations, Sığma, Korucuk, and Güzelköy stations were determined to have higher mean temperature, pH, salinity, conductivity, and total dissolved solids (TDS) values and lower dissolved oxygen (dO2) and saturated oxygen (sO2) values than Bekilli, Mahmutlu, and Sarayköy stations. In biomarker response evaluations, C. riparius larvae from Sarayköy, Sığma, Korucuk, and Güzelköy stations were found to exhibit inhibited acetylcholinesterase (AChE) activities (55%−68%) and elevated 7-ethoxyresorufin O-deethylase (EROD) activities (191%−281%), glutathione S-transferase (GST) activities (138%−187%), and metallothionein (MT) contents (155%−231%) when compared with the control values. This study proves that different stations are exposed to different pollution sources, the pollution level of a station varies depending on the season, and the stations on Çürüksu stream, among which Güzelköy appears to be the most contaminated, are exposed to more pollutants than those on the main tributary of the river.

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.

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.

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.

Early Toxic Effects in a Central American Native Fish (Parachromis dovii) Exposed to Chlorpyrifos and Difenoconazole.

In Costa Rica, agriculture is one of the most important economic activities. Chlorpyrifos and difenoconazole have been identified as agrochemicals widely used in banana and pineapple crops in the Caribbean area of the country and are constantly recorded in aquatic ecosystems. The toxicity of these pesticides in Parachromis dovii was studied. Median lethal concentrations (LC50s) for each substance were obtained from 96-h acute tests. Then, fish were exposed to sublethal concentrations of both substances (10% of LC50), individually and in mixture, to evaluate biomarker responses. Ethoxyresorufin-O-deethylase (EROD), catalase, and glutathione S-transferase activities as well as lipid peroxidation were measured in liver and gill tissues as markers of biotransformation and oxidative stress processes. Cholinesterase activity in brain and muscle tissue was also quantified as a biomarker of toxicity. The LC50s were 55.34 μg/L (95% confidence interval [CI] 51.06-59.98) for chlorpyrifos and 3250 μg/L (95% CI 2770-3810) for difenoconazole. Regarding the biomarkers, a significant inhibition of brain and muscle cholinesterase activity was recorded in fish exposed to 5.50 μg/L of chlorpyrifos. This activity was not affected when fish were exposed to the mixture of chlorpyrifos with difenoconazole. Significant changes in lactate dehydrogenase activity were observed in fish exposed to 325 μg/L of difenoconazole, whereas fish exposed to the mixture showed a significant increase in EROD activity in the liver. These results suggest harmful effects of chlorpyrifos insecticide at environmentally relevant concentrations. There is also evidence for an interaction of the 2 substances that affects the biotransformation metabolism at sublethal levels of exposure. Environ Toxicol Chem 2021;40:1940-1949. © 2021 SETAC.

Toxicity and sublethal effects of methylparaben on zebrafish (Danio rerio) larvae and adults.

Among the parabens, methylparaben (MeP) is the most commonly found in personal care products. Due to the continuous use of MeP and low removal efficiency by wastewater treatment plants (WWTPs), it reaches aquatic environments, where it is incorporated by organisms inhabiting these waters. The present study aimed to evaluate the effects of MeP on the zebrafish Danio rerio larvae and adults through toxicity tests and physiological and biochemical biomarkers in order to assess possible harmful effects of MeP. For biomarker measurements, fish were exposed to the environmental concentration of 30 μg/L of MeP and the non-effect concentration (NOEC) estimated for larvae (60 mg/L) and adult (50 mg/L) in toxicity tests. The median lethal concentration (LC50) of MeP was 105.09 mg/L for adults and 211.12 mg/L for larvae. These values unexpectedly indicated that adults were more sensitive to MeP compared to larvae. In adult fish, exposure to 50 mg/L MeP induced a significant decrease in phase 1 biotransformation (ethoxyresorufin O-deethylase activity) and an increase in lipoperoxidation (LPO) in gills, as well as an increase in frequency of micronuclei in erythrocytes of these fish. Biomarkers results were integrated (integrated biomarker response [IBR] index), and it observed lower IBR scores in tissues of fish exposed to MeP, suggesting a suppression of biological responses. In addition, LPO contributed mostly to the IBR score estimated for gills of fish exposed to 50 mg/L MeP. Based on LPO quantity, sublethal exposure of MeP (30 μg/L and 60 mg/L) did not cause toxicity to larvae. Hence, we investigated whether the difference in sensitivity between adults and larvae could be associated with the antimicrobial action of MeP that could affect the intestinal microbiota of adults. We only found an increase in the number of carbon sources consumed by them without effects on diversity and abundance. This outcome can be considered an adaptation to environmental stress, but not a negative effect. However, the LPO and genotoxicity caused by MeP to zebrafish adults call direct attention to the importance of regulating the presence of this compound in the environment and improve cleaning processes adopted by WWTP.

Effects of Benzo[a]pyrene, Cortisol, and 17ß-Estradiol on Liver Microsomal EROD Activity of Anguilla anguilla: An In Vitro Approach

Fish liver ethoxyresorufin-O-deethylase (EROD) activity is widely used as biomarker of exposure to chemicals such as polycyclic aromatic hydrocarbons (PAHs). It is known that endocrine system plays a major role in fish stress mechanism. Despite the considerable scientific information about steroid hormone’s response, namely cortisol and 17ß-estradiol (E2), to stress situations, little is known about the influence of these hormones on enzymes involved on the biotransformation process. Thus, this study aimed to assess the in vitro effects of environmentally relevant concentrations of benzo[a]pyrene (B[a]P) (0.1, 0.3, 0.9, and 2.7 µM) and of two steroid hormones (cortisol and 17ß-estradiol) in a physiologically relevant concentration (5.997 ng/mL), alone or in combination, on Anguilla anguilla liver microsomal EROD activity, previously induced by 4 mg/kg β-naphthoflavone intraperitoneal injection. Hepatic microsomes in vitro exposure to the tested B[a]P concentrations induced a dose response inhibition of EROD activity, whereas exposure to cortisol significantly induced the activity of this enzyme. The steroid hormones were able to decrease the inhibitory effects of B[a]P on microsomal EROD activity, thus revealing a protective effect of these hormones over enzyme activity inhibited by contaminants.

The sensitivity of multiple ecotoxicological assays for evaluating Microcystis aeruginosa cellular algal organic matter and contribution of cyanotoxins to the toxicity

Secondary metabolites of cyanobacteria and algae released during algal blooms often exhibit toxic effects, but only a small number of the metabolites are the subject of routine analytical screenings. Alternatively, ecotoxicological assays offer a better representation of the overall negative effects. The aim of this work was to compare multiple assays in their sensitivity towards cellular algal organic matter (COM) of the toxin-producing cyanobacterium Microcystis aeruginosa. Multiple endpoints were investigated: mortality, growth inhibition, bioluminescence inhibition, genotoxicity, endocrine-disrupting effects, oxidative stress, and the induction of ethoxyresorufin-O-deethylase (EROD). Three rainbow trout (Oncorhynchus mykiss) cell lines as well as representatives of bacteria, yeasts, algae, vascular plants, and crustaceans were employed, and the results were expressed per mg of dissolved organic carbon (DOC) in the COM. M. aeruginosa COM was toxic to the RTgill-W1, RTG-2, and RTL-W1 cell lines (EC50 values ranging from 0.48 ± 0.02 to 1.9 ± 0.1 mgDOC/L), to the crustacean Thamnocephalus platyurus (LC50 = 20 ± 1 mgDOC/L), and to Lepidium sativum (IC50 = 241 ± 13 mgDOC/L). In contrast, no effect was observed for bacteria and yeasts, and the growth of the alga Desmodesmus subspicatus was even stimulated. No genotoxicity, endocrine-disrupting effects or increase in oxidative stress or EROD activity was detected. The content of six microcystins (MC-LR, MC-RR, MC-YR, MC-LY, MC-LW, and MC-LF), anatoxin-a, cylindrospermopsin, and nodularin in the M. aeruginosa COM was determined by liquid chromatography–tandem mass spectrometry. An artificially prepared mixture of the detected cyanotoxins in the corresponding concentrations did not induce response in the O. mykiss cell lines and T. platyurus, suggesting that other cyanobacterial metabolites are responsible for the toxicity of M. aeruginosa.

Marine Fish Primary Hepatocyte Isolation and Culture: New Insights to Enzymatic Dissociation Pancreatin Digestion.

Primary cell cultures from wild organisms have been gaining relevance in ecotoxicology as they are considered more sensitive than immortalized cell lines and retain the biochemical pathways found in vivo. In this study, the efficacy of two methods for primary hepatocyte cell isolation was compared using liver from two marine fish (Sparus aurata and Psetta maxima): (i) two-step collagenase perfusion and (ii) pancreatin digestion with modifications. Cell cultures were incubated in L-15 medium at 17 ± 1 °C and monitored for up to six days for cell viability and function using the trypan blue exclusion test, MTT test, lactate dehydrogenase (LDH) activity, and ethoxyresorufin O-deethylase (EROD) activity after Benzo[a]Pyrene exposure. The results showed significant differences between the number of viable cells (p < 0.05), the highest number being obtained for the pancreatin digestion method (average = 4.5 ± 1.9 × 107 cells). Moreover, the hepatocytes showed solid adherence to the culture plate and the rounded shape, changing into a triangular/polygonal shape. The cell viability and function obtained by pancreatin digestion were maintained for five days, and the EROD induction after exposure to the B[a]P showed that cells were metabolically active. This study shows that the optimized pancreatin digestion method is a valid, cost-effective, and simple alternative to the standard perfusion method for the isolation of primary hepatocytes from fish and is suitable for ecotoxicological studies involving marine pollutants, such as PAHs.

Evaluation of the Antioxidant Activity of Some Imines Containing 1H-Benzimidazoles.

The in vitro antioxidant properties of some 2-(2-phenyl)-1H-benzo(d)imidazol-1-yl)-N'-(arylmethylene) acetohydrazide derivatives (1-12) were investigated in this study. The in vitro antioxidant activity of compounds 1-12 was explored by determination of rat liver microsomal nicotinamideadenine dinucleotide phosphate dependent inhibition on lipid peroxidation (LPO) levels and microsomal ethoxyresorufin O-deethylase (EROD) activity. All synthesised compounds had LPO inhibitory activity (15-57%) except compound 6, which contains a thiophene ring. Almost all the compounds displayed slightly inhibitory activity (2-20%) on EROD. The most active compound, 3 bearing a p-bromophenyl substituent at the second position of the benzimidazole ring, caused 57% inhibition of LPO level, while butylated hydroxytoluene showed 65% inhibition. None of the synthesised compounds had a marked inhibitory effect on EROD activity.

Mugil liza (Valenciennes, 1836) biomarker responses in a hypersaline lagoon in southeastern Brazil before and after a microalgal bloom

Araruama lagoon is one of the largest permanent hypersaline lagoons in the world and has been impacted for twenty years by sewage dumping. In early 2005, a Prasinophyceae (Pseudoscourfieldia sp.) phytoplankton bloom occurred, leading to fish mortality during subsequent summers (2006 and 2007). In this context, the aim of the present study was to accompany this historical period through the use of biochemical biomarkers in mullet (Mugil liza) during and after the algal bloom. Mullet were collected from Araruama Lagoon in June and July 2005 and August 2007, as well as from a reference lagoon with no algal blooms, Saquarema Lagoon. Several enzymatic activities in liver were analyzed, namely glutathione peroxidase (GPX), catalase (CAT), total glutathione S-transferase (GSTt), specific GSTπ (pi) and GSTμ (mi) classes and ethoxyresorufin-O-deethylase (EROD), as well as muscle acetylcholinesterase activity (AChE). In 2005, only EROD activity was significantly increased at Araruama Lagoon, indicating induction of the biotransformation enzyme system (CYP 1A1). GSTπ, on the other hand, was lower in the Araruama Lagoon, especially in 2005, when compared to mullet from Saquarema lagoon (reference area). These results highlight the importance of developing actions that go beyond the physical-chemical monitoring of these environments, since the study of ecological relationships and the physiology of organisms affected by algal blooms also include biochemical parameters to evaluate new approaches to icthyotoxicity.

Induction of Cytochrome P4501 Genes by Various Inducers in Rainbow Trout (Oncorhynchus mykiss)

Gao, K.; Tian, J.; Wu, Y.; Mei, P.; Zhang, Z.; Jonsson, M., and Brandt, I., 2021. Induction of cytochrome P4501 genes by various inducers in rainbow trout (Oncorhynchus mykiss). Journal of Coastal Research, 37(1), 143–148. Coconut Creek (Florida), ISSN 0749-0208.Juvenile rainbow trout is widely found in various aquatic systems, such as freshwater, coastal, and marine habitats all over the world. Expression patterns of the cytochrome P4501 (CYP1) genes exposed to various stressors in such fish could be informative and useful in biomonitoring. In the present study, the induction of CYP1 in rainbow trout by indigo, humic acid (HA), and benzo(a)pyrene (BaP) was studied through a 12-hour exposure experiment by using rainbow trout as an experimental object. First, 7-ethoxyresorufin O-deethylase (EROD) activity and CYP1 expression levels in fish exposed to different concentrations of indigo were analyzed. The EROD activities in fish gills were greatly affected by the concentration of indigo (1, 18, 50, and 80 nM). The expression level of CYP1A1 and CYP1A3 in gills was higher compared with CYP1B1 and CYP1Cs. Second, the gill EROD activity and CYP1 mRNA expressions levels were compared when induced by indigo, HA, and BaP at the same concentration (1 nM). Results indicated that rainbow trout were sensitive to all three inducers. Among them, the HA affected CYP1A1 and CYP1A3 most obviously. CYP1A, as an important biomarker, was more sensitive than CYP1B and CYP1C to those inducers. The observed effects of the different CYP1 inducers could provide a useful tool for monitoring of AhR-active pollutants in the aquatic environment.

Occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants and e-waste soil-derived extracts.

We investigated the occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants (BFRs) and e-waste soil-derived extracts. Analysis of the e-waste site samples using AAS and GC-MS techniques revealed the presence of a range of toxic metals as well as persistent and toxic organic pollutants, respectively, in the vicinity of the e-waste sites. As expected, the occurrence (%) of all the detected chemical pollutants in experimental soils significantly (P < 0.05) differs from occurrence (%) in control soil. The calculated LC50 values on RBL-2H3 cells of the detected tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) were 3.75 μM and 4.2 μM, respectively. Tribromophenol (TBP), dibromobiphenyl (DBB), and decabromodiphenyl ether (DBDE) were remarkably less toxic on RBL-2H3 cells compared with TBBPA and HBCD as they did not reduce RBL-2H3 cell viability below 50% in the tested concentration range (0-20 μM). The study revealed that TBBPA and HBCD could induce significant RBL-2H3 cell death through caspase-dependent apoptosis. The study further shows that the cytotoxicity of some of these BFRs could increase synergistically when in mixtures and potentially activate inflammation through the stimulation of mast cell degranulation. The e-waste soil-derived extracts induced a concentration-dependent increase in EROD activity in the exposed RTG-W1 cells. Ultimately, nonpolar extracts had higher EROD-inducing potency compared with polar extracts and hence suggesting the presence in higher amounts of AhR agonists in nonpolar e-waste soil-derived extracts than polar extracts. Overall, there is urgent need for actions in order to improve the environmental quality of the e-waste sites.

Biomonitoring the effects of urban-stream waters on the health status of pale chub (Zacco platypus): A comparative analysis of biological indexes and biomarker levels

The objective of this study was to biomonitor the effects of potential environmental pollutants in urban-stream waters, on fish health. Pale chub (Zacco platypus), a dominant species in the Korea urban stream waters, was chosen and biomonitoring indicators for the different spatial characteristics were tailored in an urban watershed. Biological responses including biotic-somatic index as well as gonadal development phase and plasma steroids levels, and the biochemical responses, ethoxyresorufin-o-deethylase (EROD) and acetylcholinesterase (AChE) activities, were measured. No significant difference was observed in the length–weight relationship between the up-stream waters and the down-stream waters. However, changes in the gonad-somatic index (GSI) levels, plasma 17β-estradiol (E2) levels, and mature oocyte frequencies in the female fish collected during the spawning season were observed in the down-stream waters at each monitoring site. Moreover, intersex condition (testis-ova) in the male fish in down-stream waters was recorded, even if it was just one fish. Although no significant difference was observed in the EROD and AChE activities between the up-stream waters and the down-stream waters, changes in the reproductive biomarker levels, including the GSI levels, plasma E2 levels, and gonadal maturation, lead to variable biomonitoring endpoints between the spatial different sites. These results imply that exposure to the down-stream waters can cause reproductive impairment in wild Z. platypus, individual variability in the biological responses further indicate the reproductive health was affected more by the down-stream waters than the up-stream waters. The finding from this study can provide the biomonitoring endpoint on the wild fish health in urban watershed that is crucial to the early risk assessment of its biological impacts. More multi-biomarkers studies reflecting the variation in the biological organization of wild fish and, therefore, the effects of urban-stream waters in the fish health are warranted.

Biomonitoring of polycyclic aromatic hydrocarbons (PAHs) from Manila clam Ruditapes philippinarum in Laizhou, Rushan and Jiaozhou, bays of China, and investigation of its relationship with human carcinogenic risk

This study examined the marine environment and seafood safety using chemical monitoring and multiple biomarkers. Samples were collected from three bays on the Shandong Peninsula in China, Laizhou, Rushan and Jiaozhou, in March, May, August, and October of 2018 and 2019. The polycyclic aromatic hydrocarbon (PAH) concentrations in sediments and tissue samples from the clam Ruditapes philippinarum and multiple biomarkers were measured. All the sampling sites were found to be medium-PAH-contaminated areas (100–1000 ng/g d.w.). According to the correlation analysis, ethoxyresorufin-o-deethylase (EROD) and superoxide dismutase (SOD) activity in the clam's digestive gland were sensitive to PAHs (p < .05), but the incremental lifetime cancer risk (ILCR) was lower than the priority risk level (10−4) at most sampling sites. EROD, SOD and acetylcholinesterase activity exhibited significant correlations with the ILCR values (p < .01), suggesting that they may serve as good indicators for assessing safe seafood consumption levels for human beings.

Monitoring of pollution in the western Black Sea coast of Turkey by striped red mullet (Mullus surmuletus).

The striped red mullet (Mullus surmuletus) is an economically important demersal fish species. In this study, our aim was to monitor the pollution in the western Black Sea coast of Turkey using striped red mullet as a bioindicator species. Fish samples were caught from four different locations in the western Black Sea coast of Turkey in 2006, 2009-2011, and 2016. Highly elevated cytochrome P4501A (CYP1A)-related 7-ethoxyresorufin O-deethylase (EROD) activities were measured in striped red mullet caught from Zonguldak Harbor in all of the sampling years. The lowest EROD activities were measured in fish samples caught from Kefken. In addition to the EROD activity measurements, glutathione S-transferase (GST), glutathione reductase, and catalase activities were also measured in the striped red mullet samples. Higher GST and catalase activities were measured in the striped red mullet samples caught from Zonguldak Harbor than from Kefken in 2016. These results indicate that the striped red mullet is responsive to CYP1A inducer pollutants. This study covers intermittent measurements of the biomonitoring data from the striped red mullet caught around the western Black Sea coast of Turkey, over a 10-year period.

Biological uptake, depuration and biochemical effects of diclofenac and carbamazepine in Carassius carassius

The uptake and depuration kinetics of diclofenac and carbamazepine alone at an environmentally relevant nominal concentration of 2 μg/L and in combination at a concentration ratio of 1:1 with total concentration of 4 μg/L were evaluated in Carassius carassius after 7 d uptake and depuration. Also, the biochemical effects of both drugs alone at nominal concentrations of 2 and 10 μg/L as well as in combination with total concentrations of 4 and 20 μg/L were investigated in Carassius carassius after 7 d exposure followed by 10 d recovery. In the single treatments, steady-state BCFs measured after the 7 d exposure were 73.05, 49.71, 38.01 and 24.93 L/kg for diclofenac and 9.25, 8.99, 5.29 and 4.11 L/kg for carbamazepine in the liver, brain, gill and muscle of Carassius carassius, respectively. Comparatively lower BCFs were measured in the tissues of Carassius carassius for both drugs in the combined treatments. Acetylcholinesterase activity in the brain was significantly induced by diclofenac while carbamazepine and the mixtures significantly inhibited it during all the exposure days as well as after the 10 d recovery in all treatments. This indicates that Carassius carassius could not recover from the neurotoxic effects caused by carbamazepine unlike the inductive effect caused by diclofenac which was recoverable after 10 days. A significant increase in the activities of 7-ethoxyresorufin O-deethylase and glutathione s-transferase for individual and mixed pharmaceuticals suggest that metabolism and detoxification of both drugs took place in the liver of Carassius carassius. Also, a significant increase in the activities of superoxide dismutase, catalase, glutathione reductase and malondialdehyde contents in the individual and mixture treatments mean that the antioxidant defence system of Carassius carassius was triggered to fight against oxidative stress but lipid peroxidation still occurred. However, Carassius carassius recovered from all these increases (superoxide dismutase, catalase, glutathione reductase and malondialdehyde) after the 10 d recovery, suggesting that oxidative damage is reversible. Our results indicate that both drugs at environmentally relevant concentrations might cause adverse effects in Carassius carassius and other fish species.