Toxicity Of Mixtures Research Articles

Lethal and Sublethal Toxicity of Un-Ionized Ammonia to Early-Life Stages of Danio rerio.

Un-ionized ammonia (NH3) is a prevalent contaminant found in aquatic ecosystems, frequently associated with complex mixtures of other toxic contaminants. Early embryo-larval stages of zebrafish Danio rerio became an important model for water quality monitoring, and it is necessary to characterize its susceptibility to NH3 exposure. Fertilized eggs were exposed to NH3 concentrations ranging from 0.02 to 5.23 mg NH3 L-1 until 168 h postfertilization (hpf). The lethal concentration to 50% of exposed zebrafish during 96 h was 2.07 mg NH3 L-1, 25% above the median value reported values for early developmental stages of fishes. Sublethal toxicity endpoints indicated the lowest observed effect concentrations for slow blood circulation at 24 hpf, decreased heart ventricular contractions at 72 hpf, incomplete yolk sac absorption at 96 hpf, failure in swim bladder inflation at 96 hpf, developmental retardation at 96 hpf, decreased total length, decreased swimming speed, altered trajectories, and acetylcholinesterase inhibition at 168 hpf of 0.85, 0.06, 0.15, 0.06, 0.15, 0.61, 1.35, 0.35, and 0.85 mg NH3 L-1, respectively. Environmentally relevant NH3 concentrations can affect zebrafish's early development and larval viability, and our results help discriminate NH3 contribution to the toxicity of complex environmental mixtures when zebrafish is used in water quality monitoring.

Comprehensive Study on Environmental Behaviour and Degradation by Photolytic/Photocatalytic Oxidation Processes of Pharmaceutical Memantine

Memantine is a pharmaceutical used to treat memory loss, one of the main symptoms of dementia and Alzheimer’s disease. The use of memantine is expected to continue to grow due to the increasing proportion of the elderly population worldwide. The aim of this work was to conduct a comprehensive study on the behaviour of memantine in the environment and the possibilities of its removal from wastewater. Abiotic elimination processes (hydrolysis, photolysis and sorption) of memantine in the environment were investigated. Results showed that memantine is stable in the environment and easily leached from river sediment. Therefore, further investigation was focused on memantine removal by advanced oxidation processes that would prevent its release into the environment. For photolytic and photocatalytic degradation of memantine, ultraviolet (UV) lamps with the predominant radiation wavelengths of 365 nm (UV-A) and 254/185 nm (UV-C) were used as a source of light. TiO2 in the form of a nanostructured film deposited on the borosilicate glass wall of the reactor was used for photocatalytic experiments. Photodegradation of memantine followed pseudo-first-order kinetics. The half-life of photocatalytic degradation by UV-A light was much higher (46.3 min) than the half-life obtained by UV-C light (3.9 min). Processes degradation efficiencies and evaluation of kinetic constants were based on the results of HPLC-MS/MS analyses, which also enable the identification of memantine oxidation products. The acute toxicity of the reaction mixture during the oxidation was evaluated by monitoring the inhibition of the luminescence of Vibrio fischeri bacteria. The results showed that memantine and its oxidation products were not harmful to Vibrio fischeri.

In silico assessment of mixture toxicity mechanisms involved in the pathogenesis of thyroid diseases: The combination of toxic metal(oid)s and decabrominated diphenyl ether

The current study aimed to assess the connection between the mixture of lead (Pb), cadmium (Cd), arsenic (As), methylmercury (MeHg) and decabrominated diphenyl ether (decaBDE) and thyroid function, by using in silico toxicogenomic data-mining approach. To obtain the linkage between investigated toxic mixture and thyroid diseases (TDs), the Comparative Toxicogenomics Database (CTD) was used, while gene ontology (GO) enrichment analysis was performed by ToppGeneSuite portal. The analysis has shown 10 genes connected to all chemicals present in the mixture and TDs (CAT, GSR, IFNG, IL1B, IL4, IL6, MAPK1, SOD2, TGFB1, TNF), most of which were in co-expression (45.68%), or belonged to the same pathway (30.47%). Top 5 biological processes and molecular functions affected by the investigated mixture emphasized the role of two common mechanisms – oxidative stress and inflammation. Cytokines and inflammatory response was listed as the main molecular pathway that may be triggered by simultaneous exposure to toxic metal(oid)s and decaBDE and connected to TDs. The direct relations between Pb/decaBDE and redox status impairment in thyroid tissue was confirmed by our chemical-phenotype interaction analysis, while the strongest linkage between Pb, As and decaBDE and thyroid disorders was found. The obtained results provide better understanding of molecular mechanisms involved in the thyrotoxicity of the investigated mixture, and can be used to direct further research.

Ecotoxicological and biochemical effects of a binary mixture of pesticides on the marine diatom Thalassiosira weissflogii in a scenario of global warming

Under the current scenario of global warming, it is ecologically relevant to understand how increased temperature influences the combined toxicity of pesticides to aquatic species. Hence, this work aims to: a) determine the temperature effect (15 °C, 20 °C and 25 °C) on the toxicity of two pesticides (oxyfluorfen and Copper (Cu)), on the growth of Thalassiosira weissflogii; b) assess whether temperature affects the type of toxicity interaction between these chemicals; and c) assess the temperature effect on biochemical responses (fatty acids (FA) and sugar profiles) of the pesticides on T. weissflogii. Temperature increased the tolerance of the diatoms to the pesticides with EC50 values between 3.176 and 9.929 μg L−1 for oxyfluorfen and 42.50–230.75 μg L−1 for Cu, respectively, at 15 °C and 25 °C. The mixtures toxicity was better described by the IA model, but temperature altered the type of deviation from dose ratio (15 °C and 20 °C) to antagonism (25 °C). Temperature, as well as the pesticide concentrations, affected the FA and sugar profiles. Increased temperature increased saturated FA and decreased unsaturated FA; it also affected the sugar profiles with a pronounced minimum at 20 °C. Results highlight effects on the nutritional value of these diatoms, with potential repercussion on food webs.

Toxicity of Binary Metal Mixtures to the Tropical Ostracod Strandesia Trispinosa.

The ecotoxicity of metals is generally assessed individually, in part because current knowledge does not allow for the accurate prediction of the toxicity of metal mixtures to aquatic organisms. The objective of this study was to investigate the toxic effects of binary combinations of metal salts (copper sulphate-CuSO4, cadmium chloride-CdCl2, mercury chloride-HgCl2 and manganese sulphate-MnSO4) on the tropical ostracod Strandesia trispinosa through acute toxicity tests. To this end, ostracods were exposed to each individual metal salt as well as to their combinations by applying a full factorial design. The model that best explained the effects of the mixtures CuSO4 x CdCl2, CuSO4 x HgCl2 and CuSO4 x MnSO4 on the survival of S. trispinosa was Concentration Addition, whereas this was Independent Action for the CdCl2 x HgCl2 mixture. The observed synergistic interactions are likely to result in unacceptable risks to aquatic ecosystems under real field conditions. This is especially the case if CuSO4 predominates the metal mixture, as observed for its combination with mercury and manganese.

A critical review on the interaction of polymer particles and co-existing contaminants: Adsorption mechanism, exposure factors, effects on plankton species.

This review considers the interaction of microplastics (MPs)/nanoplastics (NPs) and co-existing contaminants, including organic contaminants, potentially toxic elements (PTEs), and metal/metal-oxide nanoparticles. Stronger adsorption between plastic particles and co-existing contaminants can either facilitate or prevent more contaminants to enter plankton. The characteristics of MPs/NPs, such as polymer type, size, functional groups, and weathering, affect combined effects. Mixture toxicity is affected by those factors simultaneously and also affected by the type of co-existing contaminants, their concentrations, exposure time, dissolved organic matter, and surfactant. For co-exposure involving organics and metal nanoparticles, marine Skeletonema costatum generally had antagonistic effects, while marine Chlorella pyrenoidosa, Platymonas subcordiformis, and Tetraselmis chuii, showed synergistic effects. For co-exposure involving organics and PTEs, both Chlorella sp. and Microcystis aeruginosa generally demonstrated antagonistic effects. Freshwater Chlorella reinhardtii and Scenedesmus obliquus had synergistic effects for co-exposure involving metal/metal oxide nanoparticles. Zooplankton shows more unpredicted sensitivity towards the complex system. Different co-existing contaminants have different metabolism pathways. Organic contaminants could be biodegraded, which may enhance or alleviate mixture toxicity. PTEs could be adsorbed and desorbed under changing environments, and further affect the combined effects. The presence of metal/metal-oxide nanoparticles is more complicated, since some may release ion metals, increasing contaminant composition.

Heavy metals in riverine/estuarine sediments from an aquaculture wetland in metropolitan areas, China: Characterization, bioavailability and probabilistic ecological risk

Aquaculture wetlands, particularly those located within urban areas, are fragile ecosystems due to urban and aquaculture impacts. However, to date, there are no reports on the combined toxicity of heavy metal mixtures in aquatic biota in sediments from aquaculture wetlands in metropolitan areas. Thus, the characterization, bioavailability, and ecological probability risk of heavy metals were studied in the riverine/estuarine sediments of the Rongjiang River in an aquaculture wetland in Chaoshan metropolis, South China. In the study area, the average total concentrations (mg/kg) were 2.38 (Cd), 113.40 (Pb), 88.27 (Cr), 148.25 (Ni), 62.08 (Cu), 125.18 (Zn), 45,636.44 (Fe), and 797.18 (Mn), with the Cd pollution being regarded as extremely serious based on the enrichment factor (EF). There are two main sources of heavy metals in the study area; Ni, Pb, Zn, Fe and Mn are mainly from domestic waste, while Cr, Cd and Cu are possibly associated with industrial production activities. The bioavailability of most heavy metals accounted for more than 20% of the total concentration. The combined toxicity of heavy metal mixtures based on probabilistic risk assessment suggests that the surface sediments of the Rongjiang River and its estuary had a 15.71% probability of toxic effects on aquatic biota.

Comprehensive micropollutant characterization of wastewater during Covid-19 crisis in 2020: Suspect screening and environmental risk prioritization strategy

Micropollutants monitoring in wastewater can serve as a picture of what is consuming society and how it can impact the aquatic environment. In this work, a suspect screening approach was used to detect the known and unknown contaminants in wastewater samples collected from two wastewater treatment plants (WWTPs) located in the Basque Country (Crispijana in Alava, and Galindo in Vizcaya) during two weekly sampling campaigns, which included the months from April to July 2020, part of the confinement period caused by COVID-19. To that aim, high-resolution mass spectrometry was used to collect full-scan data-dependent tandem mass spectra from the water samples using a suspect database containing >40,000 chemical substances. The presence of > 80 contaminants was confirmed (level 1) and quantified in both WWTP samples, while at least 47 compounds were tentatively identified (2a). Among the contaminants of concern, an increase in the occurrence of some compounds used for COVID-19 disease treatment, such as lopinavir and hydroxychloroquine, was observed during the lockdown. A prioritization strategy for environmental risk assessment was carried out considering only the compounds quantified in the effluents of Crispijana and Galindo WWTPs. The compounds were scored based on the removal efficiency, estimated persistency, bioconcentration factor, mobility, toxicity potential and frequency of detection in the samples. With this approach, 33 compounds (e.g. amantadine, clozapine or lopinavir) were found to be considered key contaminants in the analyzed samples based on their concentration, occurrence and potential toxicity. Additionally, antimicrobial (RQ-AR) and antiviral (EDRP) risk of certain compounds was evaluated, where ciprofloxacin and fluconazole represented medium risk for antibiotic resistance (1 > RQ-AR > 0.1) in the aquatic ecosystems. Regarding mixture toxicity, the computed sum of toxic unit values of the different effluents (> 1) suggest that interactions between the compounds need to be considered for future environmental risk assessments.

Coupling Environmental Whole Mixture Toxicity Screening with Unbiased RNA-Seq Reveals Site-Specific Biological Responses in Zebrafish.

Passive sampling device (PSD) extracts paired with developmental toxicity assays in Danio Rerio (zebrafish) are excellent sensors for whole mixture toxicity associated with the bioavailable non-polar organics at environmental sites. We expand this concept by incorporating RNA-Seq in 48-h post fertilization zebrafish statically exposed to PSD extracts from two Portland Harbor Superfund Site locations: river mile 6.5W (RM 6.5W) and river mile 7W (RM 7W). RM 6.5W contained higher concentrations of polycyclic aromatic hydrocarbons (PAHs), but the diagnostic ratios of both extracts indicated similar PAH sourcing and composition. Developmental screens determined RM 6.5W to be more toxic with the most sensitive endpoint being a "wavy" notochord malformation. Differential gene expression from exposure to both extracts was largely parallel, although more pronounced for RM 6.5W. When compared to the gene expression associated with individual chemical exposures, PSD extracts produced some gene signatures parallel to PAHs but were more closely matched by oxygenated-PAHs. Additionally, differential expression, reminiscent of the wavy notochord phenotype, was not accounted for by either class of chemical, indicating the potential of other contaminants driving mixture toxicity. These techniques offer a compelling method for non-targeted hazard characterization of whole mixtures in an in vivo vertebrate system without requiring complete chemical characterization.

Toxicity Characterization of Environment-Related Pollutants Using a Biospectroscopy-Bioreporter-Coupling Approach: Potential for Real-World Toxicity Determination and Source Apportionment of Multiple Pollutants.

Exposure to environmental pollutants occurs ubiquitously and poses many risks to human health and the ecosystem. Although many analytical methods have been developed to assess such jeopardies, the circumstances applying these means are restricted to linking the toxicities to compositions in the pollutant mixtures. The present study proposes a novel analytical approach, namely, biospectroscopy-bioreporter-coupling (BBC), to quantify and apportion the toxicities of metal ions and organic pollutants. Using a toxicity bioreporter ADPWH_recA and Raman spectroscopy, both bioluminescent signals and spectral alterations had similar dosage- and time-response behavior to the toxic compounds, validating the possibility of coupling these two methods from practical aspects. Raman spectral alterations successfully distinguished the biomarkers for different toxicity mechanisms of individual pollutants, such as ring breathing mode of DNA/RNA bases (1373 cm-1) by Cr, reactive oxygen species-induced peaks of proteins (1243 cm-1), collagen (813 cm-1), and lipids (1255 cm-1) by most metal ions, and indicative fingerprints of organic toxins. The support vector machine model had a satisfactory performance in distinguishing and apportioning toxicities of individual toxins from all input data, achieving a sensitivity of 88.54% and a specificity of 97.80%. This work set a preliminary database for Raman spectral alterations of whole-cell bioreporter response to multiple pollutants. It proved the state-of-the-art concept that the BBC approach is feasible to rapidly quantify and precisely apportion toxicities of numerous pollutant mixtures.

Battery of In Vitro Bioassays: A Case Study for the Cost-Effective and Effect-Based Evaluation of Wastewater Effluent Quality

Wastewater treatment plants (WWTPs) represent an important input of contaminants in the environment. Therefore, it is critical to continuously monitor the performance of WWTPs to take appropriate action and avoid an influx of contaminants in the environment. In this study, a battery of seven in vitro bioassays covering a selected spectrum of toxicity effects is proposed for quality control of wastewater effluents. The bioassays address mixture toxicity, which is the combined adverse effect of multiple contaminants and can act as an early warning system. The proposed battery was applied to samples from 11 WWTPs of representative technology from the Danube River Basin (DRB). The order of toxic effects in terms of extent of exceedance of effect-based trigger values (EBTs) was PAH (PAH activity) > PXR (xenobiotic metabolism) > ERα (estrogenic activity) > PPARγ > Nrf2 (oxidative stress) > anti-AR > GR. A mitigation plan for WWTP operators based on EBT exceedance is proposed. This study demonstrates that the proposed effect-based monitoring battery is a complementary tool to the chemical analysis approach. A regular application of such time- and cost-effective bioanalytical tools in the WWTPs of the DRB is proposed to provide a ‘safety net’ for aquatic ecosystems.

Chemical Element Mixtures and Kidney Function in Mining and Non-Mining Settings in Northern Colombia.

The exposure to chemical mixtures is a problem of concern in developing countries and it is well known that the kidney is the major target organ for toxic elements. This cross-sectional study aimed to estimate the individual and composite mixture effect of a large number of chemical elements on kidney function in gold-mining and surrounding non-mining populations in northeast Colombia. We measured concentrations of 36 chemical elements in hair as indicators of chronic exposure from 199 adult participants. We estimated the effect of exposure to mixtures of chemical elements on estimated glomerular filtration rate (eGFR) using weighted quantile sum regression (WQS). The WQS index of the mixture was associated with reduced eGFR (Coefficient -2.42; 95%CI: -4.69, -0.16) being Be, Cd, Pb, As, and Mn, the principal contributors of the toxic mixture. Mining activities and Hg concentration were not associated with decreased kidney function. Our results suggest that complex mixtures of chemical elements, mainly heavy metals, act as nephrotoxic in these populations and therefore the analysis of chemical element mixtures is a better approach to identify environmental and occupational chemical risks for kidney damage.

Profiles of novel high-molecular-weight synthetic antioxidants in urine and associated child exposure in China

The aim of this study is to investigate the exposure of novel high-molecular-weight (HMW) synthetic antioxidants (AOs), including nine synthetic phenolic antioxidants (SPAs), one low-molecular-weight (LMW) SPA, two organophosphite antioxidants (OPAs) as well as one transformation product in children's urine from eastern (n = 82) and western (n = 105) China. For the first time, all analytes were detected in children's urine such as the representative HMW SPAs pentaerythritol tetrakis(3-(3,5-di-tert-butyl-4-hydroxyphenyl) propionate) (AO1010, median = 0.447 ng/mL), octadecyl-3-(3,5-di-tert-butyl-4-hydroxyphenyl) propionate (AO1076, median = 0.0300 ng/mL), and 1,3,5-tris[(3,5-di-tert-butyl-4-hydroxyphenyl)methyl]-1,3,5-triazinane-2,4,6-trione(1,2-dioxoethylene)bis(iminoethylene) (AO3114, median = 0.0166 ng/mL) and representative OPAs bis(2,4-di-tert-butylphenyl) pentaerythritol diphosphite (AO626, median = 0.00216 ng/mL), tris(2,4-di-tert-butylphenyl) phosphite (AO168, median = 0.0296 ng/mL) as well as its transformation product tris(2,4-di-tert-butylphenyl) phosphate (AO168O, median = 1.53 ng/mL). Significant differences were observed in the concentrations of AO1010, AO3114, AO168, and AO168O between urine samples from eastern and western China (p < 0.01). The high-frequency combination of AOs from binary to a mixture of six AOs was acquired, which would provide a better investigation of the mixture toxicity. The high estimated daily intakes of AO1010 (85.4 ng/kg/day), AO1076 (10.2 ng/kg/day), AO3114 (4.50 ng/kg/day), and AO168 (1231 ng/kg/day) were less than the values of the tolerable daily intake (3,020,000, 1,500,000, 10,000,000, and 580,000 ng/kg/day for AO1010, AO1076, AO3114, and AO168, respectively), indicating low health risk to children. Our findings showed the co-occurrence of those novel AOs and transformation products in children, the overall risks associated with the mixture of transformation products and the mixture with other emerging pollutants need to be considered when assessing the risks of AOs in further studies.

Mixture toxicity of pyraclostrobine and metiram to the zebrafish (Danio rerio) and its potential mechanism.

The interplay between pesticides plays a critical role in ecotoxicology since these chemicals rarely emerge as single substances but rather in mixtures with other chemicals. In the present work, we purposed to clarify the combined toxic impacts of pyraclostrobine (PYR) and metiram (MET) on the zebrafish by using numerous indicators. Results exhibited that the 4-day LC50 value of MET to fish embryos was 0.0025mg a.i. L-1, which was lower compared with PYR (0.019mg a.i. L-1). Combinations of PYR and MET presented a synergetic impact on fish embryos. Contents of POD, CYP450, and VTG were drastically increased in the plurality of the single and joint treatments relative to the baseline value. Three genes, including vtg1, crh, and il-8, related to the endocrine and immune systems, were also surprisingly up-regulated when fish were challenged by the individual and mixture pesticides compared with the baseline value. These results afforded valuable information on the latent toxicity mechanisms of co-exposure for PYR and MET in the early growth stage of fish. Moreover, our data also revealed that frequent application of these two pesticides might exert a potentially ecotoxicological hazard on aquatic ecosystems. Collectively, the present study provided valuable guidance for the risk evaluation of chemical combinations.

Zinc homeostasis may reverse the synergistic neurotoxicity of heavy metal mixtures in Caenorhabditis elegans

Heavy metal mixtures can cause nerve damage. However, the combined effects of metal mixtures are extremely complex and rarely studied. Zinc (Zn) homeostasis plays an integral role in neural function, but the role of Zn homeostasis in the toxicity of metal mixtures is not well understood. Here, we investigated the combined effects of manganese (Mn), lead (Pb) and arsenic (As) on nerves and the effect of Zn homeostasis on metal toxicity. Caenorhabditis elegans (Maupas, 1900) were exposed to single and multiple metals for 8 days, their movement, behavior, neurons and metal concentration were detected to evaluate the combined effect of metal mixtures. After nematodes were co-treated with metal mixtures and Zn, the nerve function, Zn concentration and redox balance were detected to evaluate the effect of Zn homeostasis on metal toxicity. The results showed that Mn + Pb and Pb + As mixtures induced synergistic toxicity for nematode nerves, which damaged movement, behavior and neurons, and decreased Zn concentration. While Zn supplementation recovered Zn homeostasis and promoted redox balance on nematodes, and then improved the nerve function. Our study demonstrated the combined effects of metal mixtures and the neuroprotective effect of Zn homeostasis. Therefore, assessment of metal mixtures toxicity should consider their interaction and the impacts of essential metals homeostasis.

Mixture toxicities of tetrachlorantraniliprole and tebuconazole to honey bees (Apis mellifera L.) and the potential mechanism

The extensive use of pesticides has negative effects on the health of insect pollinators. Although pollinators in the field are seldom exposed to individual pesticides, few reports have assessed the toxic impacts of pesticide combinations on them. In this work, we purposed to reveal the combined impacts of tetrachlorantraniliprole (TET) and tebuconazole (TEB) on honey bees (Apis mellifera L.). Our data exhibited that TET had greater toxicity to A. mellifera (96-h LC50 value of 298.2 mg a.i. L-1) than TEB (96-h LC50 value of 1,841 mg a.i. L-1). The mixture of TET and TEB displayed acute synergistic toxicity to the pollinators. Meanwhile, the activities of CarE, CYP450, trypsin, and sucrase, as well as the expressions of five genes (ppo, abaecin, cat, CYP4G11, and CYP6AS14) associated with immune response, oxidative stress, and detoxification metabolism, were conspicuously altered when exposed to the mixture relative to the individual exposures. These results provided an overall comprehension of honey bees upon the challenge of sublethal toxicity between neonicotinoid insecticides and triazole fungicides and could be used to assess the intricate toxic mechanisms in honey bees when exposed to pesticide mixtures. Additionally, these results might guide pesticide regulation strategies to enhance the honey bee populations.

Combined toxicity of acetamiprid and cadmium to larval zebrafish (Danio rerio) based on metabolomic analysis

Emerging contaminants, such as neonicotinoid pesticide acetamiprid (Ace), are frequently detected in the water environment, which can interact with existing heavy metal cadmium (Cd) to produce unpredicted influence. Limited studies have evaluated the effects of multiple pollutant exposures on aquatic animals. Here, we characterized the joint toxicity of Ace and Cd exposure to zebrafish (Danio rerio). The results revealed that Cd and its combined exposure with Ace had an inhibitory effect on the growth of larval zebrafish and induced morphological defects. Combined exposure to high doses of Ace and Cd could significantly reduce the levels of TG, glucose, and pyruvate in larval zebrafish. Untargeted metabolomics revealed that Cd treatment (285) produced more differentially expressed metabolites (DEMs) than Ace treatment (115), and combined treatment produced the most DEMs (294). The KEGG pathway enrichment analysis showed that they could disrupt riboflavin metabolism, amino acid metabolism, and glycolipid metabolism in the larvae of D. rerio. ELISA showed that VB2, FMN, and FAD levels were significantly increased. In addition, gene expression analysis exhibited that the mRNA levels of essential genes related to glycolipid metabolism were substantially affected, such as PK, PEPckc, PPAR-α, and FABP6. Furthermore, targeted amino acid metabolomics confirmed that both single exposure to Cd and combined exposure to Ace and Cd altered the levels of amino acids in larvae, including ALA, ARG, MET, PRO, TYR, VAL, GLY, ORN, and PHE. Taken together, exposure to Ace and Cd, alone or in combination, exerted harmful effects on the individual development, riboflavin metabolism, glycolipid metabolism, and amino acid metabolism disorder of D. rerio. These findings highlighted that more attention should be paid to the compound toxicity of chemical mixtures to aquatic organisms.

Mixture Toxicity of Three Unconventional Gas Fracking Chemicals, Barium, O-Cresol, and Sodium Chloride, to the Freshwater Shrimp Paratya australiensis.

The 96-h acute toxicity of barium (Ba2+ ), o-cresol, and sodium chloride (NaCl) to Paratya australiensis was assessed in single, binary, and ternary combinations in addition to three biochemical assays: glutathione S-transferase, acetylcholinesterase, and sodium-potassium adenosine triphosphatase. The 96-h lethal concentrations that expressed 50% mortality (LC50) in the single-toxicant exposures were Ba2+ = 23.4 mg/L, o-cresol = 12.2 mg/L, and NaCl = 4198 mg/L. Mortality from o-cresol exposure occurred between 11 and 22 mg/L, whereas Ba2+ was more gradual across 10-105 mg/L, and most of the NaCl mortality occurred between 2050 and 4100 mg/L. Toxic units were used to assess the binary and ternary interactions of the toxicants. A more than additive effect was observed for most combinations in the binary chemical exposures, with the ternary combinations yielding highly synergistic interactions. Greater synergism was observed with the 96-h LC50 of o-cresol in combination with the three concentrations of NaCl (1025, 2050, and 3075 mg/L) compared with Ba2+ , with toxic units of 0.38, 0.48, and 0.10 (o-cresol) and 0.71, 0.67, and 0.50 (Ba2+ ). No notable enzyme activity trends were observed in the enzyme biomarker responses from both individual and mixture exposures. Although acute single-species toxicity tests tend to underestimate the effects of Ba2+ , o-cresol, and NaCl on populations, communities, and ecosystems in seminatural (e.g., mesocosms) and natural systems, there are currently no published acute toxicity data available for P. australiensis and the three toxicants used in the present study. The present study shows that chemicals with different toxicity mechanisms can potentially lead to more synergistic responses. Environ Toxicol Chem 2023;42:481-494. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Investigation on the Toxicity of Nanoparticle Mixture in Rainbow Trout Juveniles

The environmental impacts of nanoparticle mixtures in the aquatic environment is not well understood. The purpose of this study examined the sub-lethal toxicity of low concentrations (ug/L range) of selected nanoparticles alone and in mixtures in juvenile trout. Fish were exposed to to individual and two environmentally relevant mixtures of silver (nAg), copper oxide (nCuO) and cerium oxide (nCeO) nanoparticles for 96 h at 15 °C. After the exposure period, fish were depurated overnight and tissue levels in Ag, Ce, Cu and Zn were determined along with a suite of effects biomarkers such as oxidative stress/inflammation, denatured protein tagging (ubiquitin), DNA strand breaks (genotoxicity) and acetylcholinesterase (AChE) activity. The data showed that these nanoparticles behaved as suspended matter but were nevertheless bioavailable for fish with bioconcentration factors of 6, 8 and 2 for nAg, nCeO and nCuO respectively. Only nCuO alone increased malonaldehyde (lipid peroxidation) contents but all nanoparticles increased DNA damage, protein-ubiquitin labeling, and decreased AChE activity. Globally, the toxicity of nCeO and nCuO was generally stronger than nAg, and antagonist effects were found in the mixtures. The interactions involved in these antagonisms are not well understood but do not involve the liberation of free ions and labile zinc in tissues. In conclusion, the bioavailability and toxicity of these nanoparticles are influenced by mixtures of nanoparticles, which is likely to occur in contaminated environments.

Toxic effects of a mixture of pharmaceuticals in Mytilus galloprovincialis: The case of 17α-ethinylestradiol and salicylic acid

The impact of pharmaceuticals on marine invertebrates has been a topic of rising concern, with an increasing number of studies regarding the impacts on bivalves. However, very few investigated the toxicity of mixtures of pharmaceuticals. This knowledge gap was investigated in the present study, where the toxicity of 17α-ethinylestradiol (EE2) and salicylic acid (SA) mixture was evaluated. To this end, Mytilus galloprovincialis mussels were chronically subjected to both pharmaceuticals, acting alone and in combination, and the effects at the cellular level were measured. The Independent Action (IA) model was performed aiming to compare obtained with predicted responses. The integrated biomarker response (IBR) index was used to assess the overall biochemical response given by mussels. The results obtained revealed that the most stressful condition was caused by the combined effect of EE2 and SA, with the highest metabolic capacity, antioxidant (catalase activity) and biotransformation (carboxylesterases activity) activation and cellular damage in organisms exposed to the mixture of both drugs in comparison to responses observed when each drug was acting alone. Predicted responses obtained from the IA model indicate that caution should be paid as frequent deviations to observed responses were found. This study highlights the need for future studies considering the mixture of pollutants, mimicking the actual environmental conditions.