How to evaluate the ecological risk of transgenic technology is a focus of scientists because of the safety concerns raised by genetically modified (GM) organisms. Nevertheless, most studies are based on individual chemicals and always analyze the GM organism as a type of toxicant. In this study, we changed the approach and used GM organisms as the test objects with normal chemical exposure. Three types of chemicals (two substituted phenols, 4-chlorophenol and 4-nitrophenol; two ionic liquids, 1-butylpyridinium chloride and 1-butylpyridinium bromide; two pesticides, dichlorvos and glyphosate) were used to construct a six-component mixture system. The lethality to wild-type (N2) and sod-3::GFP (SOD-3) Caenorhabditis elegans was determined when they were exposed to the same mixture system after 12 and 24 h. The results showed that the pEC50 values of all of the single chemicals on SOD-3 were greater than those on N2 at 24 h. The toxicities of the single chemicals and nine mixture rays on the two strains increased with time. Notably, we discovered a significant difference between the two strains; time-dependent synergism occurred in mixtures on N2, but time-dependent antagonism occurred in mixtures on SOD-3. Finally, the strength of the synergism or antagonism turned to additive action on the two strains as the exposure time increased. These findings illustrated that the GM factor of the nematode influenced the mixture toxicological interaction at some exposure times. Compared with N2, SOD-3 were more sensitive to stress or toxic reactions. Therefore, the influence of the GM factor on mixture toxicological interactions in environmental risk assessment must be considered.

We analyzed the acute toxicity of the 48% glyphosate (GLY)-based Credit®, the 57.71% dicamba (DIC)-based Kamba®, and the 83.5% 2,4-dichlorophenoxyacetic acid (2,4-D)-based Weedar® Full, alone and as mixtures on the fish Cnesterodon decemmaculatus. Mortality revealed the LC50 96h values of 91.73 mg L−1 (range: 86.80–98.00 mg L−1), 1401.57 mg L−1 (range: 1243.78–1527.35) and 678.04 mg L−1 (range: 639.35–718.04 mg L−1) for GLY, DIC and 2,4-D, respectively. Mean values for the toxic unit (TU) that induced 50% mortality (TU50 96h) of fish exposed to equitoxic mixtures were 1.67 (range: 1.65–1.69) for Credit® and Kamba® and 1.28 (range: 1.20–1.36) for Credit® and Weedar® Full suggesting that both mixtures are antagonic. Non-equitoxic combinations demonstrated an antagonistic interaction of herbicides Credit® and Kamba®, whereas a synergistic effect was observed for Credit® and Weedar® Full formulations. GLY and DIC as a mixture demonstrated lower toxicity on non-target species compared to GLY and 2,4-D in combination, at least for C. decemmaculatus, leading to the conclusion that the former combination could be strongly recommended in further agricultural practices.

In this study, a novel biochar-supported zero valent iron (BC-nZVI) was synthesized through a green method. A high performance on the simultaneous removal of Cu²⁺ and bisphenol A (BPA) by a combination of BC-nZVI with persulfate (BC-nZVI/PS) system was successfully achieved. The simultaneous efficiencies of Cu²⁺ and BPA could reach 96 and 98% within 60 min, respectively. Both HO• and SO₄•⁻ were two major reactive species in BC-nZVI/PS system, and SO₄•⁻ was primary radical responsible for the degradation of BPA. Four kinds of Cu species, such as Cu(OH)₂, CuO, Cu₂O and Cu⁰ were generated via the adsorption and reduction of the BC-nZVI, whereas six kinds of products of BPA including p-isopropenyl phenol and 4-isopropylphenol were generated via the combined oxidation of SO₄•⁻ and HO•. The possible reaction mechanism for the simultaneous removal of Cu²⁺ and BPA by BC-nZVI/PS system contained a synergistic effect between the reduction of Cu²⁺ and the oxidation of BPA. This is the first report on the feasibility of the remediation of coexistence of heavy metal and organic compound in aquatic environment using the BC-nZVI/PS system.

Assessment of the impact of pharmaceutical residues on living organisms is a very complex subject. Apart from taking into account the toxicity of individual compounds, environmental factors should also be taken into account. In this paper, attempts were made to assess the impact of coexisting inorganic ions and changes in pH on the toxicity of ten selected pharmaceuticals. Two bioassays were used to measure the estrogenic and androgenic effects (XenoScreen YES/YAS – Saccharomyces cerevisiae) and acute toxicity (Microtox® – Vibrio fischeri).The Microtox® test gave the most definitive outputs concerning the determination of interaction type between drugs and chemical species. Synergism was proven for almost all drugs and chemical species, and only two cases of antagonism were found. Significant drug/pH interactions were rare.Regarding the XenoScreen YES/YAS bioassay, when estrogenic and androgenic agonistic effects (YES+ and YAS+, respectively) were studied, many cases of well-expressed synergism for all inorganic ions with limited number of drugs (diazepam, fluoxetine, estrone, chloramphenicol for the YES+ test and diazepam, progesterone, androstenedione, and estrone for the YAS+ test) were found. Antagonism was also proven for the YES+ test, especially for diclofenac and androstenedione interacting with cations. On the other hand, the YES- and YAS- tests (estrogenic and androgenic, respectively, antagonistic effects) did not indicate cases of synergetic interaction except for the couples Br−/diazepam and NH4+/ketoprofen. Antagonistic drug/ion interactions were detected only with diclofenac and fluoxetine. It is interesting that well-expressed (antagonism or synergism) drug/pH interactions were rare.Both tests were found utilizable in performing studies on impact of ions/pH fluctuations on drugs mixtures' toxicity confirming in most cases synergic impact of parameters studied on toxicity. The approach proposed in the paper seems to be proven as a reliable tool in assessing impact of abiotic factors on toxicity and endocrine potential of complex mixtures of pharmaceuticals' mixtures.

Pesticide contamination is more often found as a mixture of different pesticides in water bodies rather than individual compounds. However, regulatory risk evaluation is mostly based on the effects of individual pesticides. In the present study, we aimed to investigate the individual and joint toxicities of triazophos (TRI) and imidacloprid (IMI) to the zebrafish (Danio rerio) using acute indices and various sublethal endpoints. Results from 96-h semi-static test indicated that the LC₅₀ values of TRI to D. rerio at multiple life stages (embryonic, larval, juvenile and adult stages) ranged from 0.49 (0.36–0.71) to 4.99 (2.06–6.81) mg a.i. L⁻¹, which were higher than those of IMI ranging from 26.39 (19.04–38.01) to 128.9 (68.47–173.6) mg a.i. L⁻¹. Pesticide mixtures of TRI and IMI displayed synergistic response to zebrafish embryos. Activities of carboxylesterase (CarE) and catalase (CAT) were significantly changed in most of the individual and joint exposures of pesticides compared with the control group. The expressions of 26 genes related to oxidative stress, cellular apoptosis, immune system, hypothalamic-pituitary-thyroid and hypothalamic-pituitary-gonadal axis at the mRNA level revealed that zebrafish embryos were affected by the individual or joint pesticides, and greater changes in the expressions of six genes (Mn-sod, CXCL-CIC, Dio1, Dio2, tsh and vtg1) were observed when exposed to joint pesticides compared with their individual pesticides. Taken together, the synergistic effects indicated that it was highly important to incorporate joint toxicity studies, especially at low concentrations, when assessing the risk of pesticides.

Metal contaminants are rarely present in the environment individually, yet environmental quality guidelines are derived from single-metal toxicity data. Few metal mixture studies have investigated more than binary mixtures and many are at unrealistically high effect concentrations to freshwater organisms. This study investigates the toxicity of five metals (Cd, Cu, Ni, Pb, and Zn) to the Antarctic marine microalgae Phaeocystis antarctica and Cryothecomonas armigera. Two mixtures were tested: (i) an equitoxic mixture of contaminants present at their single-metal EC10 concentrations, and (ii) an environmental mixture based on the ratio metal concentrations in a contaminated Antarctic marine bay.Observed toxicity, as chronic population growth rate inhibition, was compared to Independent Action (IA) and Concentration Addition (CA) predictions parameterised to use EC10 values. This allowed for the inclusion of metals with low toxicities. The biomarkers chlorophyll a fluorescence, cell size and complexity, and intracellular lipid concentrations were assessed to investigate possible mechanisms behind metal-mixture interactions.Both microalgae had similar responses to the equitoxic mixture: non-interactive by IA and antagonistic by CA. Toxicity from the environmental mixture was antagonistic by IA to P. antarctica; however, to C. armigera it was concentration-dependent with antagonism at low toxicities and synergism at high toxicities by both IA and CA. Differences in dissolved organic carbon production and detoxification mechanisms may be responsible for these responses and warrants further investigation.This study shows that mixture toxicity interactions can be ratio, species, and concentration dependent. The responses of the microalgae to different mixture ratios highlight the need to assess toxicity at environmentally realistic metal ratios. Parameterising IA and CA reference models to use EC10s allowed for the inclusion of metals at low effect concentrations, which may otherwise be ignored. Reference mixture models are generally suitable for predicting chronic toxicity of metals to these marine microalgae at environmentally realistic ratios and concentrations.

The problem of effective assessment of risk posed by complex mixtures of toxic chemicals in the environment is a major challenge for government regulators and industry. The biological effect of the individual contaminants, where these are known, can be measured; but the problem lies in relating toxicity to the multiple constituents of contaminant cocktails. The objective of this study was to test the hypothesis that diverse contaminant mixtures may cause a greater toxicity than the sum of their individual parts, due to synergistic interactions between contaminants with different intracellular targets. Lysosomal membrane stability in hemocytes from marine mussels was used for in vitro toxicity tests; and was coupled with analysis using the isobole method and a linear additive statistical model. The findings from both methods have shown significant emergent synergistic interactions between environmentally relevant chemicals (i.e., polycyclic aromatic hydrocarbons, pesticides, biocides and a surfactant) when exposed to isolated hemocytes as a mixture of 3 & 7 constituents. The results support the complexity-based hypothesis that emergent toxicity occurs with increasing contaminant diversity, and raises questions about the validity of estimating toxicity of contaminant mixtures based on the additive toxicity of single components. Further experimentation is required to investigate the potential for interactive effects in mixtures with more constituents (e.g., 50–100) at more environmentally realistic concentrations in order to test other regions of the model, namely, very low concentrations and high diversity. Estimated toxicant diversity coupled with tests for lysosomal damage may provide a potential tool for determining the toxicity of estuarine sediments, dredge spoil or contaminated soil.

Environment pollution often occurs as an obvious combined effect involving two (or more) elements, and this effect changes with the concentrations of the different elements. The effects on barley root elongation were studied in hydroponic systems to investigate the toxicity of Cu–Ni combined at low doses and at a fixed concentration ratio. For low doses of Cu–Ni, the addition of Ni (<0.5 μM) to Cu significantly decreased Cu toxicity for barley, but the addition of Cu (<0.25 μM) had no significant effect on Ni toxicity. At a fixed concentration ratio, according to the single effective concentration (EC) (barley root elongation inhibitory concentration) values of Cu and Ni, five sets of Cu–Ni fixed ratios were used: ECn(Cu)+ECm(Ni) (n + m = 100) (ECn and ECₘ indicate toxicity unit value for n% and m% inhibition of barley root length, respectively). The calculated toxicity unit value for 50% inhibition of root length ranged from 0.44 to 0.98 (i.e., <1), indicating a synergistic effect. To consider the interactions between the metal ions, the extended concentration addition model (e-CA) was established by integrating the Cu–Ni interaction into the concentration addition model (CA), and the data of two groups (the low doses of Cu–Ni and at a fixed concentration ratio) were respectively fitted. The e-CA accurately predicted the root length of barley under the Cu–Ni combined action. The correlation coefficient (r) and the root-mean-square error (RMSE) between predicted and observed values were 0.97 and 6.6 (low-dose group) and 0.96 and 8.12 (fixed-ratio group), respectively, and e-CA significantly improved the prediction accuracy compared to the traditional CA model without consideration of the Cu–Ni competition (r = 0.89, RMSE = 14.16). The results provided a theoretical basis for evaluation and remediation of soil contaminated with heavy metal composites.

Heavy metal pollution can decrease the soil microbial biomass and significantly alter microbial community structure. In this study, a long-term field experiment (5 years) and short-term laboratory experiment (40 d) were employed to evaluate the effects of heavy metals (Cd, Cu, Zn), and their combinations at different concentrations, on the soil microbial biomass and the bacterial community. The ranges of heavy metal concentration in the long-term and short-term experiments were similar, with concentration ranges of Cd, Cu and Zn of about 0.3–1.5, 100–500, and 150–300 mg kg⁻¹, respectively. Microbial biomass decreased with increasing soil heavy metal concentrations in both the long-term and short-term experiments. The interaction between soil physicochemical factors (pH, TN, TC) and heavy metals (Cd, Cu, Zn) played a major role in change in the bacterial community in long-term polluted soil. In the laboratory experiment, although each heavy metal had an adverse effect on the microbial biomass and community structure, Cu appeared to have a greater role in the changes compared to Cd and Zn. However, the synergistic effects of the heavy metals were greater than those of the single metals and the synergistic effect between Cu and Cd was greater than that of Cu and Zn.

Obesity and exposure to particular matter (PM) have become two leading global threats to public health. However, the exact mechanisms and tissue-specificity of their health effects are largely unknown. Here we investigate whether a metabolic challenge (early nutritional obesity) synergistically interacts with an environmental challenge (PM exposure) to alter genes representing key response pathways, in a tissue-specific manner. Mice subjected to 7 weeks obesogenic nutrition were exposed every other day during the final week and a half to aqueous extracts of PM collected in the city of London (UK). The expression of 61 selected genes representing key response pathways were investigated in lung, liver, white and brown adipose tissues. Principal component analysis (PCA) revealed distinct patterns of expression changes between the 4 tissues, particularly in the lungs and the liver. Surprisingly, the lung responded to the nutrition challenge. The response of these organs to the PM challenge displayed opposite patterns for some key genes, in particular, those related to the Nrf2 pathway. While the contribution to the variance in gene expression changes in mice exposed to the combined challenge were largely similar among the tissues in PCA1, PCA2 exhibited predominant contribution of inflammatory and oxidative stress responses to the variance in the lungs, and a greater contribution of autophagy genes and MAP kinases in adipose tissues. Possible involvement of alterations in DNA methylation was demonstrated by cell-type-specific responses to a methylation inhibitor. Correspondingly, the DNA methyltransferase Dnmt3a2 increased in the lungs but decreased in the liver, demonstrating potential tissue-differential synergism between nutritional and PM exposure. The results suggest that urban PM, containing dissolved metals, interacts with obesogenic nutrition to regulate diverse response pathways including inflammation and oxidative stress, in a tissue-specific manner. Tissue-differential effects on DNA methylation may underlie tissue-specific responses to key stress-response genes such as catalase and Nrf2.