Concentration Addition Model Research Articles

Additive and dose-dependent mixture effects of Flumite 200 (flufenzin, acaricide) and Quadris (azoxystrobin, fungicide) on the reproduction and survival of Folsomia candida (Collembola)

Chemical plant protection is still a dominant agricultural practice in the world, and usually fields are treated with several pesticides many times per year. This means that not only the single substances affect the environment and the non-target organisms, but their mixtures. Our model organism was Folsomia candida (Collembola). We aimed to gain information on the toxicity of Quadris (azoxystrobin) and Flumite 200 (flufenzine aka. diflovidazine) on survival and reproduction and whether the animals can mitigate the toxicity with soil and/or food avoidance behaviour. Also, we aimed to test the effect of the mixture of these two pesticides. We used the OECD 232 reproduction test, a soil avoidance test, and a food choice test for both single pesticides and their mixture. We prepared the mixtures based on the concentration addition model, so the 50% effective concentrations (EC50) of the single materials were used as one toxic unit with a constant ratio of the two materials in the mixture. In the end, the measured mixture EC and LC (lethal concentration) values were compared to the estimate of the concentration addition model. Both materials were toxic to the Collembola in much higher concentrations than the recommended field concentrations (Flumite 200 EC50: 1.096, LC50: 1.561, Quadris EC50: 65.568, LC50: 386.165 mg kg−1). The springtails did not consistently avoid the polluted soils, this only happened in higher concentrations. The mixtures seemed to have additive effects on the reproduction and we found dose-dependent interaction with the survival (EC50: 1.022 Toxic Unit, 0.560 Flumite 200 and 33.505 Quadris; LC50: 1.509 Toxic Unit, 0.827 Flumite 200 and 49.471 mg kg−1 Quadris). The deviation from the concentration addition model suggests that the curve starts with a synergy. but above EC50 it becomes antagonistic. We conclude that both Quadris and Flumite 200 are safe for springtails until the recommended field concentration is respected. However, if higher concentrations are used the animals cannot avoid Flumite 200 and the toxic effects can fully manifest. Consequently, the dose-dependent deviation from the concentration addition model is a reason for caution as the low concentrations were synergistic for survival. That means the field concentrations can possibly cause synergistic effects. However, to clarify that further tests are necessary.

Study on the characterization of pesticide modes of action similarity and the multi-endpoint combined toxicity of pesticide mixtures to Caenorhabditis elegans

With the widespread use of pesticides, the coexistence of multiple low-residue pesticides in environmental media has increased significantly, and the “cocktail” effect caused by this phenomenon has garnered increasing attention. However, owing to the scarcity of information regarding the modes of action (MOAs) of chemicals, the application of concentration addition (CA) models for evaluating and predicting the toxicity of mixture with similar MOAs is limited. Additionally, the joint toxicity laws of complex mixture systems to different toxicity endpoints in organisms remain unclear, and effective methods to test the mixture toxicity on lifespan and reproductive inhibition are lacking. Therefore, in this study, the similarity of pesticide MOAs was characterized using molecular electronegativity-distance vector (MEDV-13) descriptors based on eight pesticides (aldicarb, methomyl, imidacloprid, thiamethoxam, dichlorvos, dimethoate, methamidophos and triazophos). Additionally, the methods of lifespan and reproduction inhibition microplate toxicity analysis of elegans (EL-MTA and ER-MTA) were established to test the lifespan and reproduction inhibition toxicity of Caenorhabditis elegans. Finally, a unified scale synergistic-antagonistic heatmap (SAHscale) method was proposed to explore the combined toxicity of the mixtures on the lifespan, reproduction, and mortality of nematodes. The results showed that the MEDV-13 descriptors could effectively characterize the similarity in MOAs. The lifespan and reproductive ability of Caenorhabditis elegans were significantly inhibited when the pesticide exposure concentration was one order of magnitude lower than the lethal dose. The sensitivity of lifespan and reproductive endpoints to mixtures was dependent on the concentration ratio. The same rays in the mixture had consistent toxicity interactions on the lifespan and reproductive endpoints of Caenorhabditis elegans. In conclusion, we demonstrated the feasibility of MEDV-13 in characterizing the similarity of MOAs, and provided a theoretical basis for exploring the mechanism of chemical mixtures by studying their apparent toxicity of mixtures on nematode lifespan and reproduction endpoints.

Reproductive toxicity of combined effects of endocrine disruptors on human reproduction.

Confluence of environmental, genetic, and lifestyle variables is responsible for deterioration of human fecundity. Endocrine disruptors or endocrine disrupting chemicals (EDCs) may be found in a variety of foods, water, air, beverages, and tobacco smoke. It has been demonstrated in experimental investigations that a wide range of endocrine disrupting chemicals have negative effects on human reproductive function. However, evidence on the reproductive consequences of human exposure to endocrine disrupting chemicals is sparse and/or conflicting in the scientific literature. The combined toxicological assessment is a practical method for assessing the hazards of cocktails of chemicals, co-existing in the environment. The current review provides a comprehensive overview of studies emphasizing the combined toxicity of endocrine disrupting chemicals on human reproduction. Endocrine disrupting chemicals interact with each other to disrupt the different endocrine axes, resulting in severe gonadal dysfunctions. Transgenerational epigenetic effects have also been induced in germ cells, mostly through DNA methylation and epimutations. Similarly, after acute or chronic exposure to endocrine disrupting chemicals combinations, increased oxidative stress (OS), elevated antioxidant enzymatic activity, disrupted reproductive cycle, and reduced steroidogenesis are often reported consequences. The article also discusses the concentration addition (CA) and independent action (IA) prediction models, which reveal the importance of various synergistic actions of endocrine disrupting chemicals mixtures. More crucially, this evidence-based study addresses the research limitations and information gaps, as well as particularly presents the future research views on combined endocrine disrupting chemicals toxicity on human reproduction.

Combined toxicity of erythromycin and roxithromycin and their removal by Chlorella pyrenoidosa

The ecological effects of antibiotics in surface water have attracted increasing research attention. In this study, we investigated the combined ecotoxicity of erythromycin (ERY) and roxithromycin (ROX) on the microalgae, Chlorella pyrenoidosa, and the removal of ERY and ROX during the exposure. The calculated 96-h median effect concentration (EC50) values of ERY, ROX, and their mixture (2:1 w/w) were 7.37, 3.54, and 7.91 mg∙L-1, respectively. However, the predicted EC50 values of ERY+ROX mixture were 5.42 and 1.51 mg∙L-1, based on the concentration addition and independent action models, respectively. This demonstrated the combined toxicity of ERY+ ROX mixture showed an antagonistic effect on Chlorella pyrenoidosa. During the 14-d culture, low-concentration (EC10) treatments with ERY, ROX, and their mixture caused the growth inhibition rate to decrease during the first 12 d and increase slightly at 14 d. In contrast, high-concentration (EC50) treatments significantly inhibited microalgae growth (p < 0.05). Changes in the total chlorophyll contents, SOD and CAT activities, and MDA contents of microalgae suggested that individual treatments with ERY and ROX induced higher oxidative stress than combined treatments. After the 14-d culture time, residual Ery in low and high concentration Ery treatments were 17.75% and 74.43%, and the residual Rox were 76.54% and 87.99%, but the residuals were 8.03% and 73.53% in ERY+ ROX combined treatment. These indicated that antibiotic removal efficiency was higher in combined treatments than that in individual treatments, especially at low concentrations (EC10). Correlation analysis suggested that there was a significant negative correlation between the antibiotic removal efficiency of C. pyrenoidosa and their SOD activity and MDA content, and the enhanced antibiotic removal ability of microalgae benefited from increased cell growth and chlorophyll content. Findings in this study contribute to predicting ecological risk of coexisting antibiotics in aquatic environment, and to improving biological treatment technology of antibiotics in wastewater.

Estrogenic potency of endocrine disrupting chemicals and their mixtures detected in environmental waters and wastewaters

Endocrine disrupting chemicals such as natural and synthetic steroid hormones and bisphenols are among the most important pollutants in the aquatic environment. We performed an environmental chemical analysis of five Slovenian water samples, two rivers, one groundwater, and the influent and effluent of wastewater treatment plants, with a highly sensitive analysis of twenty-five endocrine-disrupting compounds belonging to the groups of natural hormones, synthetic hormones, and bisphenols. Since these compounds are simultaneously present in the environment, it is important to study their individual effects as well as the effects of mixtures. We investigated in vitro the estrogenic potency of selected natural and synthetic steroid hormones and bisphenols detected in surface, ground and waste water in Slovenia using the OECD-validated transactivation assay on the cell line Hela9903. We predicted their mixture effects using the concentration addition model and compared them with experimentally determined values. Two mixing designs were used: a balanced design in which chemicals were combined in proportion to their individual EC50 values, and an unbalanced design with compounds in proportion to their measured concentrations in the environmental samples. The estrogenic effects of the experimental mixtures followed the concentration addition model. Real water samples exhibited weaker estrogenic effects, showing the great heterogeneity of the real water samples.

Predictive in silico models for aquatic toxicity of cosmetic and personal care additive mixtures

As emerging environmental contaminants, cosmetic and personal care additives (CPCAs) may have less oversight than other consumer products. Their continuous release and pseudopersistence could cause long-term harm to the aquatic environment. Since CPCAs generally exist in the form of mixtures in the environment, prediction and analysis of their mixture toxicity are crucial for ecological risk assessment. In this study, the acute toxicity of five typical CPCA mixtures to Daphnia magna was tested. The combined toxicity of binary mixtures was examined with the traditional concentration addition (CA) and independent action (IA) model. Overall, the synergistic effect of the five CPCAs may be caused mainly by methylparaben. In addition, reliable approaches for quantitative structure-activity relationship (QSAR) model development were explored. Specifically, 18 QSAR models were developed by three dataset partitioning techniques (Kennard-Stone's algorithm division, Euclidean distance based division, and sorted activity based division), two descriptor filtering methods (genetic algorithm and stepwise multiple linear regression) and three regression methods (multiple linear regression, partial least squares and support vector machine). Sixteen equations were applied for the calculation of the mixture descriptors to screen the functional expression of the mixture descriptors with the largest contribution to the mixture toxicity. A new comprehensive parameter that integrates internal and external validation was proposed for QSAR models evaluation. The mixture toxicity is mainly related the 3D distribution of atomic masses and the spatial distribution of the molecule electronic properties. Rigorously validated and externally predictive QSAR models were developed for predicting the toxicity of binary CPCAs mixtures with any ratio, in the applicability domain. The best possible work frame for construction and validation of QSAR models to provide reliable predictions on the mixture toxicity was proposed.

Ecotoxicity of pesticide formulations and their mixtures: the case of potato crops in Costa Rica.

Despite their environmental implications, ecotoxicological information regarding pesticide mixtures is relatively scarce. This study aimed to determine the ecotoxicity of individual pesticide formulations and their mixtures (insecticides and fungicides), which are applied during the production cycle of potato, according to agricultural practices from a Latin American region in Costa Rica. Two benchmark organisms were employed: Daphnia magna and Lactuca sativa. First, the evaluation of individual formulations (chlorothalonil, propineb, deltamethrin+imidacloprid, ziram, thiocyclam and chlorpyrifos) revealed differences between available EC50 for active ingredients (a.i.) and their respective formulations toward D. magna; on the contrary, no information could be retrieved from scientific literature for comparison in the case of L. sativa. In general, acute toxicity was higher toward D. magna than L. sativa. Moreover, interactions could not be determined on L. sativa, as the chlorothalonil formulation was not toxic at high levels and the concentration-response to propineb could not be fitted to obtain an IC50 value. The commercial formulation composed of deltamethrin+imidacloprid followed the concentration addition model (when compared with parameters retrieved from individual a.i.) and the other three mixtures evaluated (I: chlorothalonil-propineb-deltamethrin+imidacloprid; II: chlorothalonil-propineb-ziram-thiocyclam; III: chlorothalonil-propineb-chlorpyrifos) produced an antagonistic effect on D. magna, thus suggesting less acute toxicity than their individual components. Subsequent chronic studies showed that one of the most toxic mixtures (II) negatively affected D. magna reproduction at sublethal concentrations indicating that this mixture poses a risk to this species if these pesticides co-exist in freshwater systems. These findings provide useful data to better estimate the impact of real agricultural practices related to the use of agrochemicals.

Combined effects of fluoroquinolone antibiotics and organophosphate flame retardants on Microcystis aeruginosa.

As freshwater harmful algal blooms continue to rise in frequency and severity, increasing focus is made on the effects of mixed pollutants and the dominant cyanobacterial species Microcystis aeruginosa (M. aeruginosa). However, few studies have investigated whether M. aeruginosa has a synergistic relationship with two common pollutants, namely, organophosphate flame retardants (OPFRs) and fluoroquinolone antibiotics (FQs). In this paper, three FQs and three OPFRs commonly detected in freshwaters were selected to construct a ternary mixture of FQs, a ternary mixture of OPFRs, and a six-component mixture of OPFRs and FQs. The effects of single substance and mixture on the growth of M. aeruginosa were determined at 24, 48, 72, and 96h, and the toxicities of the mixture were evaluated by concentration addition model and independent action model. The results showed that the mixture of FQs and the mixture of OPFRs do not show toxicological interaction. However, partial mixtures of OPFRs and FQs showed antagonism or synergism at different concentrations and times. This indicated that combined toxicities of OPFRs and FQs on M. aeruginosa were mixture ratio dependent, concentration dependent and time dependent. This study improves our understanding of the role of OPFRs and FQs in cyanobacterial outbreaks of Microcystis.

Quantitative analysis of dose interval effect of Pb-Cd interaction on Oryza sativa L. root

Combined pollution of cadmium (Cd) and lead (Pb) occurs frequently in agriculture lands, which has received increasing research attention. However, little is known about the interaction behaviors of Cd and Pb at various concentrations in the mixture. This study evaluated the single and combined effects of Cd and Pb on rice (Oryza sativa L.) root elongation through acute exposure test. The combined pollution was analyzed with the concentration addition (CA) model, independent action (IA) model and mathematical statistical methods. The dose-response results revealed that the interaction could weaken the toxicity of both Pb and Cd, and Cd had a more significant inhibitory effect on Pb toxicity. The predicted values of CA and IA models were consistently lower than the observed values in the relative root elongation range of 0–60%. Further, combining the CA or IA model with mathematical statistical methods, the interaction of Pb and Cd at similar concentrations showed a significant antagonistic effect on rice root elongation. At low Pb concentrations (Cd > 0.0195, Pb < 0.015 mg/L), there was a synergistic effect of the mixture on rice root; at high Pb concentrations (Cd < 0.225, Pb ≥ 1.25 mg/L), Pb dominated the toxicity on rice root. This is the first report of a systematic method for assessing heavy metal interaction at different concentration levels, which may facilitate the formulation of control standards of heavy metal combined pollution in agricultural land.

Prediction of herb-drug interactions involving consumption of furanocoumarin-mixtures and cytochrome P450 1A2-mediated caffeine metabolism inhibition in humans

Herb-drug interactions (HDI) has become important due to the increasing popularity of natural health product consumption worldwide. HDI is difficult to predict as botanical drugs usually contain complex phytochemical-mixtures, which interact with drug metabolism. Currently, there is no specific pharmacological tool to predict HDI since almost all in vitro-in vivo-extrapolation (IVIVE) Drug-Drug Interaction (DDI) models deal with one inhibitor-drug and one victim-drug. The objectives were to modify-two IVIVE models for the prediction of in vivo interaction between caffeine and furanocoumarin-containing herbs, and to confirm model predictions by comparing the DDI predictive results with actual human data. The models were modified to predict in vivo herb-caffeine interaction using the same set of inhibition constants but different integrated dose/concentration of furanocoumarin mixtures in the liver. Different hepatic inlet inhibitor concentration ([I]H) surrogates were used for each furanocoumarin. In the first (hybrid) model, the [I]H was predicted using the concentration-addition model for chemical-mixtures. In the second model, the [I]H was calculated by adding individual furanocoumarins together. Once [I]H values were determined, the models predicted an area-under-curve-ratio (AUCR) value of each interaction. The results indicate that both models were able to predict the experimental AUCR of herbal products reasonably well. The DDI model approaches described in this study may be applicable to health supplements and functional foods also.

The key constituents underlying the combined toxicity of eight cosmetic contaminants towards Vibrio qinghaiensis.

Cosmetic additives (ADDs) and packaging plasticizers (PLAs) probably present potential risks and dangers to the environment and human body as emerging pollutants. To investigate their potential risks and dangers, five ADDs including methyl paraben (MET), ethyl paraben (ETH), propyl paraben (PRO), butyl-hydroxy anisole (BHA), and salicylic acid (SAL), as well as three PLAs including bisphenol A (BPA), bisphenol S (BPS) and tris(2-butoxyethyl) phosphate (TBEP) were selected as research objects, and ten mixture rays (R1-R10) composed of the eight components were designed by the uniform design ray (UD-Ray) method. The toxicities of the eight cosmetic pollutants and their eight-component mixture system towards Vibrio qinghaiensis sp.-Q67 (Q67) were systematically determined by the time-dependent microplate toxicity analysis (t-MTA) method. The three-dimensional (3D) surface of deviation from the concentration addition model (dCA) was utilized to qualitatively and quantitatively analyse the toxicity interaction of the mixtures and the correlation between toxicity interaction and the components' concentration ratios. Finally, eight individual pollutants and representative rays with significant inhibitory and interactive effects were selected to analyse DNA and soluble proteolysis as well as the microstructure and morphology of Q67 after treatment with single chemicals and their mixtures. The results showed that the eight cosmetic pollutants had conspicuous concentration-dependent toxicity and acute toxicity, and none of them, except BPS, BPA and ETH, had time-dependent toxicity. All rays had time/concentration-dependent toxicity and acute toxicity. At the same time, the toxicity interaction of these mixture rays was predominantly antagonism and the strongest antagonism appeared at high concentrations at 12 h. Nevertheless, the components' concentration ratio (pi) was the decisive factor for the type of mixture interaction. The correlation analysis revealed a significant positive linear correlation between mixture toxicity and pETH and pBPA, which indicated that ETH and BPA were the key components of the toxic effects. However, there was a significant negative linear correlation between the antagonism intensity and pBPA and pTBEP, which demonstrated that BPA and TBEP were the key components of the antagonism intensity. Pollutants and their mixtures can also damage cellular structures, and mixtures can exacerbate the dissolution of DNA and soluble proteins.

Individual and combined ecotoxic effects of water-soluble polymers.

Water-soluble polymers (WSPs) are a class of high-molecular-weight compounds which are widely used in several applications, including water treatment, food processing, and pharmaceuticals. Therefore, they pose a potential threat for water resources and aquatic ecosystems. We assessed the ecotoxicity of four WSPs-non-ionic polyacrylamide (PAM) and polyethylene glycol (PEG-200), anionic homopolymer of acrylic acid (P-AA), and cationic polyquaternium-6 (PQ-6)-as single compounds and in mixture. For this purpose in vitro and in vivo assays were used to record baseline toxicity, mutagenic potential, endocrine effects, and growth inhibition in the freshwater alga Raphidocelis subcapitata. Furthermore, the mixture toxicity of the two polymers P-AA and PQ-6 which showed effects in the algae tests was evaluated with the concentration addition (CA), independent action (IA), and generalized concentration addition (GCA) model and compared with experimental data. No toxic effects were observed among the polymers and their mixtures in the in vitro assays. On the contrary, in the growth inhibition test with R. subcapitata the cationic PQ-6 caused high inhibition while the anionic P-AA and its mixture with the cationic polymer caused low inhibition. The non-ionic polymers PEG-200 and PAM showed no effect in R. subcapitata in the tested concentration range up to 100 mg/L. The IA model represented the mixture effect of the combination experiment better than the CA and GCA models. The results indicate (1) that the toxic effects of anionic and cationic polymers are most likely due to interactions of the polymers with the surfaces of organisms or with nutrients in the water and (2) that the polymers elicit their effects through different mechanisms of action that do not interact with each other.

Chemotherapy's impact on a few blood parameters

Cyclophosphamide (CP), Ifosfamide (IFO), Paclitaxel (PTX) and Doxorubicin (DOX) are commonly used cytostatic drugs. The present study investigates the ecotoxicity and genotoxicity of CP, DOX, and PTX, their human metabolites/transformation products (TPs) cyclophosphamide (NCP) as individual compounds and as a mixture. The three-parent compounds ( Further ecotoxicity studies of metabolites. The measured toxicity of the cross was lower than the toxicity predicted by the concentration addition model indicating potentiating effects of the CPCOOH toxicity. Revealed genotoxic activity of CP and the mixture in the presence The degradation study with UV irradiation of samples containing (CP, cyclo, and DOX ) showed efficient degradation of compounds and remained toxic. They are suggesting that no stable with adverse effects was formed. This is the first study describing the ecotoxicity and genotoxicity of the commonly used cytostatics CP, cyclo, and DOX, their known metabolites, and their mixture. The results indicate the importance of toxicological evaluation and monitoring drug metabolites as they may be more hazardous to humans than parent compounds. Keywords. Paclitaxel. Doxorubicin. Cyclophosphamide. toxicity

Joint Action Toxicity of Arsenic (As) and Lead (Pb) Mixtures in Developing Zebrafish.

Arsenic (As) and lead (Pb) are environmental pollutants found in common sites and linked to similar adverse health effects. Multiple studies have investigated the toxicity of each metal individually or in complex mixtures. Studies defining the joint interaction of a binary exposure to As and Pb, especially during the earliest stages of development, are limited and lack confirmation of the predicted mixture interaction. We hypothesized that a mixture of As (iAsIII) and Pb will have a concentration addition (CA) interaction informed by common pathways of toxicity of the two metals. To test this hypothesis, developing zebrafish (1-120 h post fertilization; hpf) were first exposed to a wide range of concentrations of As or Pb separately to determine 120 hpf lethal concentrations. These data were then used in the CA and independent action (IA) models to predict the type of mixture interaction from a co-exposure to As and Pb. Three titration mixture experiments were completed to test prediction of observed As and Pb mixture interaction by keeping the Pb concentration constant and varying As concentrations in each experiment. The prediction accuracy of the two models was then calculated using the prediction deviation ratio (PDR) and Chi-square test and regression modeling applied to determine type of interaction. Individual metal exposures determined As and Pb concentrations at which 25% (39.0 ppm Pb, 40.2 ppm As), 50% (73.8 ppm Pb, 55.4 ppm As), 75% (99.9 ppm Pb, 66.6 ppm As), and 100% (121.7 ppm Pb, 77.3 ppm As) lethality was observed at 120 hpf. These data were used to graph the predicted mixture interaction using the CA and IA models. The titration experiments provided experimental observational data to assess the prediction. PDR values showed the CA model approached 1, whereas all PDR values for the IA model had large deviations from predicted data. In addition, the Chi-square test showed most observed results were significantly different from the predictions, except in the first experiment (Pb LC25 held constant) with the CA model. Regression modeling for the IA model showed primarily a synergistic response among all exposure scenarios, whereas the CA model indicated additive response at lower exposure concentrations and synergism at higher exposure concentrations. The CA model was a better predictor of the Pb and As binary mixture interaction compared to the IA model and was able to delineate types of mixture interactions among different binary exposure scenarios.

Dose-dependent joint resistance action of antibacterial mixtures in their hormetic effects on bacterial resistance based on concentration addition model

The judgment of joint resistance action is significant for evaluating the resistance risk of antibacterial mixture. Using bacterial mutation frequency (MF) and conjugative transfer frequency (CTF) to respectively characterize the bacterial endogenous and exogenous resistance, mutation unit and conjugative transfer unit have been proposed to judge the joint resistance action of antibacterial mixture at a certain dose. However, these methods could not evaluate the antibacterial mixture's joint resistance action at a larger concentration-range. In this study, the concentration addition for bacterial resistance (CA-BR) approach was used to judge the joint resistance actions between kanamycin sulfate (KAN) and some other typical antibacterial agents, including sulfonamides (SAs), sulfonamide potentiators (SAPs), and silver antibacterial compounds (SACs). Through comparing the hormetic dose-response curves of the binary mixtures on the MF (or CTF) in Escherichia coli (E. coli) and the corresponding CA-BR curves calculated from the hormetic dose-responses of the single agents, the joint resistance actions between KAN and other agents were judged to exhibit dose-dependent feature: with the increase of mixture concentration, the joint mutation actions between KAN and SAs (or SAPs) were fixed at synergism, and the joint mutation actions between KAN and SACs varied from antagonism to synergism; the joint conjugative transfer actions between KAN and other agents changed from antagonism to synergism. Mechanistic explanation suggested that the heterogeneous pattern of joint resistance action had a close relationship with the interplays among the agents' modes of action, and meanwhile was significantly influenced by their joint survival pressure on E. coli. This study reveals the dose-dependent feature for the joint resistance action of antibacterial mixture and highlights the importance of exposure concentration, which will benefit clarifying the resistance risk of antibacterial mixture in the environment.

Antibiotic Toxicity Isolated and as Binary Mixture to Freshwater Algae Raphidocelis subcapitata: Growth Inhibition, Prediction Model, and Environmental Risk Assessment.

Antibiotics in aqueous environments can have extremely adverse effects on non-targeted organisms. However, many research projects have only focused on the toxicological evaluation of individual antibiotics in various environments. In the present work, individual and binary mixture toxicity experiments have been conducted with the model organism Raphidocelis subcapitata (R. subcapitata), and a mixture concentration-response curve was established and contrasted with the estimated effects on the basis of both the concentration addition (CA) and the independent action (IA) models. In addition, different risk assessment methods were used and compared to evaluate the environmental risk of binary mixtures. The toxic ranking of the selected antibiotics to R. subcapitata was erythromycin (ERY) &gt; sulfamethoxazole (SMX) &gt; sulfamethazine (SMZ). In general, the conclusion of this study is that the adverse effects of binary mixtures are higher than the individual antibiotics. The CA model and RQSTU are more suitable for toxicity prediction and risk assessment of binary mixtures. This study reveals the potential ecological risks that antibiotics and their mixtures may pose to water ecosystems, thus providing scientific information for environmental quality regulation.

Meta-analysis addressing the characterization and risk identification of antibiotics and antibiotic resistance genes in global groundwater

Antimicrobial resistance (AMR) is one of the significant global issues to public health. Compared to other aquatic environments, research on AMR in groundwater is scarce. In the study, a meta-analysis was conducted to explore the characteristics and risks of antibiotics and antibiotic resistance genes (ARGs) in global groundwater, using a data set of antibiotic concentrations collected from publications during 2000–2021 and a large-scale metagenomes of groundwater samples (n = 330). The ecotoxicological risks of antibiotics in the global groundwater were evaluated using mixture risk quotient with concentration addition model to consider the synergistic effects of multiple antibiotics. Bioinformatic annotations identified 1413 ARGs belonging to 37 ARG types in the global groundwater, dominated by rifamycin, polyketide, and quinolone resistance genes and including some emerging ARGs such as mcr-family and carbapenem genes. Relatively, the level of ARGs in the groundwater from spring was significantly higher (ANOVA, p < 0.01) than those from the riparian zone, sand and deep aquifer. Similarly, metal resistance genes (MRGs) were prevalent in the global groundwater, and network analysis suggested the MRGs presented non-random co-occurrence with the ARGs in such environments. Taxonomic annotations showed Proteobacteria, Actinobacteria, Eukaryota, Acidobacteria and Thaumarchaeota were the dominant phylum in the groundwater, and the microbial community largely shaped profile of ARGs in the environment. Notably, the ARGs presented co-occurrence with mobile genetic elements, virulence factors and human bacterial pathogens, indicating potential dissemination risk of ARGs in the groundwater. Furthermore, an omics-based approach was used for health risk assessment of antibiotic resistome and screened out 152 risk ARGs in the global groundwater. Comparatively, spring and cold creek presented higher risk index, which deserves more attention to ensure the safety of water supply.

Seasonal variation and ecological risk assessment of Pharmaceuticals and Personal Care Products (PPCPs) in a typical semi-enclosed bay — The Bohai Bay in northern China

The Bohai Bay as a typical semi-enclosed bay in northern China with poor water exchange capacity and significant coastal urbanization, is greatly influenced by land-based inputs and human activities. As a class of pseudo-persistent organic pollutants, the spatial and temporal distribution of Pharmaceuticals and Personal Care Products (PPCPs) is particularly important to the ecological environment, and it will be imperfect to assess the ecological risk of PPCPs for the lack of systematic investigation of their distribution in different season. 14 typical PPCPs were selected to analyze the spatial and temporal distribution in the Bohai Bay by combining online solid-phase extraction (SPE) and HPLC-MS/MS techniques in this study, and their ecological risks to aquatic organisms were assessed by risk quotients (RQs) and concentration addition (CA) model. It was found that PPCPs widely presented in the Bohai Bay with significant differences of spatial and seasonal distribution. The concentrations of ∑PPCPs were higher in autumn than in summer. The distribution of individual pollutants also showed significant seasonal differences. The high values were mainly distributed in estuaries and near-shore outfalls. Mariculture activities in the northern part of the Bohai Bay made a greater contribution to the input of PPCPs. Caffeine, florfenicol, enrofloxacin and norfloxacin were the main pollutants in the Bohai Bay, with detection frequencies exceeding 80 %. The ecological risk of PPCPs to algae was significantly higher than that to invertebrates and fish. CA model indicated that the potential mixture risk of total PPCPs was not negligible, with 34 % and 88 % of stations having mixture risk in summer and autumn, respectively. The temporary stagnation of productive life caused by Covid-19 weakened the input of PPCPs to the Bohai Bay, reducing the cumulative effects of the pollutants. This study was the first full-coverage investigation of PPCPs in the Bohai Bay for different seasons, providing an important basis for the ecological risk assessment and pollution prevention of PPCPs in the bay.

Contrasting toxicity of polystyrene nanoplastics to the rotifer Brachionus koreanus in the presence of zinc oxide nanoparticles and zinc ions

Emerging contaminants such as nanoplastics and nanoparticles likely experience similar environmental behaviours, fate and effects but our knowledge of their combined toxicity is scanty. This study, therefore, investigated the joint toxicity of polystyrene nanoplastics (PNPs) and zinc oxide nanoparticles (ZnO-NPs) to an ecologically important rotifer Brachionus koreanus, and compared with the joint toxicity of PNPs and Zn ions (Zn-IONs from ZnSO4·7H2O). With increasing concentration, ZnO-NPs formed significant agglomeration with PNPs for up to 1.3 times of the original hydrodynamic size of ZnO-NPs, alongside doubling in their sedimentation and thereby losing 58% of their released Zn ions. In contrast, the availability of Zn-IONs was less affected by the agglomeration and sedimentation of PNPs, with only a loss of 18% of Zn ions at the highest concentration of PNPs. Consequently, as suggested by Concentration Addition and Independent Action models and the Model Deviation Ratios, ZnO-NPs and PNPs exerted an antagonistic interaction whereas Zn-IONs and PNPs exhibited an additive effect. We also advocate the use of the Nonparametric Response Surface method, which is more useful to predict the toxicity of chemical mixtures with interacting effects. Our findings suggested a potential difference between particle-particle and particle-ion interactions, especially at higher test concentrations, which may eventually affect their toxicity. We, therefore, call for a more systematic evaluation of commonly coexisting chemical mixtures which consist of nanoplastics and manufactured nanomaterials.

Acute toxicity of pesticide mixtures to honey bees is generally additive, and well predicted by Concentration Addition

Understanding the frequency of non-additive effects of pesticides (synergism and antagonism) is important in the context of risk assessment. The goal of this study was to investigate the prevalence of non-additive effects of pesticides to honey bees (Apis mellifera). We investigated a large set of mixtures including insecticides and fungicides of different chemical modes of action and classes. The mixtures included represent a relevant sample of pesticides that are currently used globally. We investigated whether the experimental toxicity of the mixtures could be predicted based on the Concentration Addition (CA) model for acute contact and oral adult bee toxicity tests. We measured the degree of deviation from the additivity predictions of the experimental toxicity based on the well-known Mixture Deviation Ratio (MDR). Further, we investigated the appropriate MDR thresholds that should be used for the identification of non-additive effects based on acceptable rates for false positive (alpha) and true positive (beta) findings. We found that a deviation factor of MDR = 5 is a sound reference for labeling potential non-additive effects in acute adult bee experimental designs when assuming a typical Coefficient of Variation (CV%) = 100 in the determination of the LD50 of a pesticide (a factor of 2× deviation in the LD 50 resulting from inter-experimental variability). We found that only 2.4 % and 9 % of the mixtures evaluated had an MDR > 5 and MDR < 0.2, respectively. The frequency and magnitude of deviation from additivity found for bees in this study are consistent with those of other terrestrial and aquatic taxa. Our findings suggest that additivity is a good baseline for predicting the toxicity of pesticide mixtures to bees, and that the rare cases of synergy of pesticide mixtures to bees are not random but have a mechanistic basis.