Rare earth elements (REEs) are typically present as mixtures in the environment, but a quantitative understanding of mixture toxicity and interactions of REEs is still lacking. Here, we examined the toxicity to wheat (Triticum aestivum L.) of Y, La, and Ce when applied individually and in combination. Both concentration addition (CA) and independent action (IA) reference models were used for mixture toxicity analysis because the toxicity mechanisms of REEs remain obscure. Upon single exposure, the EC50s of Y, La, and Ce, expressed as dissolved concentrations, were 1.73 ± 0.24 μM, 2.59 ± 0.23 μM, and 1.50 ± 0.22 μM, respectively. The toxicity measured with relative root elongation followed La < Y ≈ Ce, irrespective of the dose descriptors. The use of CA and IA provided similar estimates of REE mixture interactions and toxicity. When expressed as dissolved metal concentrations, nearly additive effects were observed in Y-La and La-Ce mixtures, while antagonistic interactions were seen in Y-Ce mixtures. When expressed as free metal activities, antagonistic interactions were found for all three binary mixtures. This can be explained by a competitive effect of REEs ions for binding to the active sites of plant roots. The application of a more elaborate MIXTOX model in conjunction with the free ion activities, which incorporates the non-additive interactions and bioavailability-modifying factors, well predicted the mixture toxicity (with >92% of toxicity variations explained). Our results highlighted the importance of considering mixture interactions and subsequent bioavailability in assessing the joint toxicity of REEs.

Nitrate accumulation in aquatic reservoirs from agricultural pollution has often been overlooked as a water quality hazard, yet a growing body of literature suggests negative effects on human and wildlife health following nitrate exposure. This research seeks to understand differences in oxygen consumption rates between different routes of laboratory nitrate exposure, whether via immersion or injection, in zebrafish (Danio rerio) embryos. Embryos were exposed within 1 h post fertilization (hpf) to 0, 10, and 100 mg/L NO₃-N with sodium nitrate, or to counter ion control (CIC) treatments using sodium chloride. Embryos in the immersion treatments received an injection of 4 nL of appropriate treatment solution into the perivitelline space. At 24 hpf, Oxygen Consumption Rates (OCR) were measured and recorded in vivo using the Agilent Technologies XFᵉ96 Extracellular Flux Analyzer and Spheroid Microplate. Immersion exposures did not induce significant changes in OCR, yet nitrate induced significant changes when injected through the embryo chorion. Injection of 10 and 100 mg/L NO₃-N down-regulated OCR compared to the control treatment group. Injection of the 100 mg/L CIC also significantly down-regulated OCR compared to the control treatment group. Interestingly, the 100 mg/L NO₃-N treatment further down-regulated OCR compared to the 100 mg/L CIC treatment, suggesting the potential for additive effects between the counter ion and the ion of interest. These data support that elevated nitrate exposure can alter normal metabolic activity by changing OCR in 24 hpf embryos. These results highlight the need for regularly examining the counter ion of laboratory nitrate compounds while conducting research with developing zebrafish, and justify examining different routes of laboratory nitrate exposure, as the chorion may act as an effective barrier to nitrate penetration in zebrafish, which may lead to conservative estimates of significant effects in other species for which nitrate more readily penetrates the chorion.

Imposex is a disorder caused by organotins, mainly tributyltin, which results in the appearance of male sexual characteristics in females of gastropod mollusks. The main objective of this work was to make a critical analysis of the relationship between imposex and butyltin body burdens in Nucella lapillus and Nassarius reticulatus. Specifically, this study evaluates possible additive effects among butyltins, proposes scales of effects based on robust statistical criteria as alternatives to existing ones and defines the body burdens of TBT in N. lapillus and N. reticulatus corresponding to the assessment classes (ACs) of the Vas Deferens Sequence Index (VDSI) established by OSPAR. Data of organotin body burdens and biological effects was retrieved from the ICES Dataset and from scientific literature. All responses, except the percentage of females displaying Imposex (IMPF) in Nucella lapillus, showed a sigmoidal profile regarding to the body burden of mono- (MBT), di- (DBT) and tributyltin and sum of butyltins (SumBTs). TBT and the SumBTs were better indicators of the VDSI or Relative Penis Size Index/Relative Penis Length Index (RPSI/RPLI) responses than MBT or DBT in most cases. From a statistical point of view, RPSI/RPLI and VDSI were better indicators of contamination by TBT than IMPF, although both RPSI and RPLI showed lower sensitivity than VDSI. The model used for describing the joint effect of butyltins provided a statistically significant fitting to the data assuming a null effect for both MBT and DBT for N. lapillus, and a lower toxic contribution of MBT and DBT with respect to TBT for N. reticulatus. RPSI or RPLI values, equivalent to the ACs for VDSI, were proposed as alternative criteria when measuring moderate to high levels of imposex. TBT concentrations in N. reticulatus and N. lapillus tissues, corresponding to the ACs were calculated and provided valuable information for cross-species comparisons.

Contamination of fine plastic particles (FPs), including micrometer to millimeter plastics (MPs) and nanometer plastics (NPs), in the environment has caught great concerns. FPs are strong adsorbents for hydrophobic toxic pollutants and may affect their fate and toxicity in the environment; however, such information is still rare. We studied joint toxicity of FPs with phenanthrene to Daphnia magna and effects of FPs on the environmental fate and bioaccumulation of 14C-phenanthrene in fresh water. Within the five sizes particles we tested (from 50 nm to 10 μm), 50-nm NPs showed significant toxicity and physical damage to D. magna. The joint toxicity of 50-nm NPs and phenanthrene to D. magna showed an additive effect. During a 14-days incubation, the presence of NPs significantly enhanced bioaccumulation of phenanthrene-derived residues in daphnid body and inhibited the dissipation and transformation of phenanthrene in the medium, while 10-μm MPs did not show significant effects on the bioaccumulation, dissipation, and transformation of phenanthrene. The differences may be attributed to higher adsorption of phenanthrene on 50-nm NPs than 10-μm MPs. Our findings underlined the high potential ecological risks of FPs, and suggested that NPs should be given more concerns, in terms of their interaction with hydrophobic pollutants in the environment.

Exposure to trihalomethanes (THMs) and haloacetic acids (HAAs) has been individually associated with adverse male reproductive effects; however, their joint male reproductive toxicity is largely unknown. This study aimed to explore the joint effects of THMs and trichloroacetic acid (TCAA) on semen quality in a Chinese population. A total of 337 men presenting to the Reproductive Center of Tongjing Hospital, in Wuhan, China to seek semen analysis were included this study. Baseline blood THMs [chloroform (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM), and bromoform (TBM)] and urinary TCAA were analyzed and dichotomized at their median levels. The joint effects of THMs and TCAA on below-reference semen quality parameters were evaluated by calculating the relative excess risk due to interaction (RERI). After adjusting for potential confounders, we found a suggestive synergistic effect between Br-THMs (sum of BDCM, DBCM, and TBM) and TCAA for below-reference sperm count (RERI = 2.14, 95% CI: −0.37, 4.91) (P = 0.076); men with high Br-THMs and TCAA levels (above the median) had 3.31 times (95% CI: 1.21, 9.07) elevated risk of having below-reference sperm count than men with low Br-THMs and TCAA levels (below the median). No apparent joint effects were observed between THMs and TCAA for other semen quality parameters. Our results suggest that co-exposure to Br-THMs and TCAA is associated with additive effects on decreased semen quality. However, further studies in a larger sample size and mechanistic studies are needed to confirm the findings.

Molting is a crucial physiological process in arthropods development, growth, and adult reproduction, where the chitinolytic enzyme chitobiase (CB) and the apoptosis process (caspase-3 activity) play crucial roles. Both molecular endpoints have been observed to be affected by different toxics that may be present in aquatic environments. However, the role of ultraviolet radiation (UVR) in the molting process remains with poor evidence and the possible effect of the previous exposure on F1 generation is unknown. Here, we conducted laboratory experiments with chronic UVR exposure to test the effect on the molting process of Daphnia commutata. Our results showed a clear negative effect of the UVR that affected the molting process with a reduction in individual growth. This trend was also observed in CB and caspase-3 activities. Our results also suggest that the UV dose received by the mother and eggs has an additive effect with the dose received by the offspring. These results imply that the cumulative impact of small UVR doses (2 h per day, daily dose: 2520 J m⁻² of 340 nm) on mothers and eggs (which cannot be discriminated in our experiments) can have an additive or synergistic effect along with the generations through a potential increase in lethal effect. Finally, the observed desynchronization in the molting process by UVR will affect the fitness of individuals and population dynamics.

The aim of this study was to examine the effects of arbuscular mycorrhizal (AM) fungi, biochar (BC) addition and phosphorus (P) fertilizer applications on the mycorrhizal response, biomass and elemental uptake of Trifolium repens in cadmium (Cd)-polluted soils. The results showed that mycorrhizal colonization were significantly decreased by 100 mg P kg⁻¹ fertilizer input. Moreover, AM fungi, BC addition and P fertilizer significantly increased shoot biomass accumulation at all treatments. In the absence of BC, the nitrogen (N), potassium (K), calcium (Ca) and magnesium (Mg) contents in the shoots were not affected by AM fungi after P fertilizer application, but the P content in the shoots significantly increased in response to AM fungi. In the absence of BC, both AM fungi and P fertilizer significantly reduced the Cd concentrations in the plant tissues as well as the soil diethylenetriaminepentaacetic acid (DTPA)-Cd concentration. These results indicated that the translocation factors (TFs) were influenced only by BC addition and that the roots could accumulate greater amounts of Cd than the shoots. On the basis of the hygienic standard for feed in China, the shoot Cd concentration in white clover was below the maximum permitted Cd concentration (1 μg g⁻¹) across all treatments. Therefore, it is suggested that no negative mycorrhizal-white clover symbiotic relationships were observed and T. repens could be a suitable forage species for planting in soils with low concentrations of Cd contamination when BC and P fertilizer are applied.

The combined toxicity of mixed chemicals is usually evaluated according to several specific endpoints, and other potentially toxic effects are disregarded. In this study, we provided a metabolomics strategy to achieve a comprehensive understanding of toxicological interactions between mixed chemicals on metabolism. The metabolic changes were quantified by a pseudotargeted analysis, and the types of combined effects were quantitatively discriminated according to the calculation of metabolic effect level index (MELI). The metabolomics strategy was used to assess the combined effects of polycyclic aromatic hydrocarbons (PAHs) and short-chain chlorinated paraffins (SCCPs) on the metabolism of human hepatoma HepG2 cells. Our data suggested that exposure to a combination of PAHs and SCCPs at human internal exposure levels could result in an additive effect on the overall metabolism, whereas diverse joint effects were observed on various metabolic pathways. The combined exposure could induce a synergistic up-regulation of phospholipid metabolism, an additive up-regulation of fatty acid metabolism, an additive down-regulation of tricarboxylic acid cycle and glycolysis, and an antagonistic effect on purine metabolism. SCCPs in the mixture acted as the primary driver for the acceleration of phospholipid and fatty acid metabolism. Lipid metabolism disorder caused by exposure to a combination of PAHs and SCCPs should be an important concern for human health.

A combination of multiple stressors may be linked to global amphibian declines. Of these, pesticides and UVB radiation co-exposures were examined on the African clawed frog (Xenopus laevis) to provide information that may be useful for amphibian conservation. The independent action model and inferential statistics were used to examine interactions between pesticides (malathion, endosulfan, α-cypermethrin, or chlorothalonil) and environmentally relevant UVB exposures. UVB radiation alone caused 35–68% mortality and nearly 100% of malformations. Pesticides and UVB had additive effects on larval mortality; however, several non-additive effects (antagonistic and synergistic interactions) were observed for total body length. Insecticides mainly affected axial development, whereas UVB radiation caused high incidence of edema, gut malformations, and abnormal tail tips. These results suggest that sublethal developmental endpoints were more sensitive for detecting joint effects. This work has implications for amphibian risk assessments for ecosystems where pesticides and high UVB radiation may co-occur.

The negative health effects of simulated acid mists and fluoranthene on juvenile Japanese red pine were investigated, and the methods of protection from these pollutants were examined. The needle gas exchange, chlorophyll fluorescence, chemical contents and visual damage to needles caused by acid mist applied alone or its conjunction with fluoranthene were investigated over 60 d and 20 d, respectively. Acid mist at pH 2 and 3 caused physiological and visual damage, which was enhanced by the addition of fluoranthene to the mist. However, fluoranthene and acid mist at pH 4 and 5 showed only minor effects. These findings indicate that acid mist may be more harmful to pine trees if it occurs in conjunction with polycyclic aromatic hydrocarbons. Moreover, suppression of the singular and additive effects of these compounds was achieved using mannitol, which may be widely applicable to suppression of reactive oxygen species-mediated plant damage.