Assessing the toxicity posed by mixtures of unknown chemicals to aquatic organisms is challenging. In this study, water samples from six cross-sections along the Xitiaoxi River Basin (XRB) were monthly or bimonthly collected in 2014. The year-period physiochemical parameters as well as one-month-water sample based acute biotoxicity tests showed that the river water quality of the year was generally in a good status. High performance liquid chromatography (HPLC) screening based on one-month-water samples suggested that the organic pollutants might be non-to-moderately-polar chemicals in very low concentrations. One-month-water sample based RNA-seq was performed to measure the mRNA differential expression profile of zebrafish larvae to furtherly explore the potential bioeffect and the spatial water quality change of the river. Result indicated that the number of deferentially expressed genes (DEGs) tended to increase along the downstream direction of the river. Gene ontology (GO) enrichment analysis implied that the key pollutants might mainly be the function disruptors of biological processes. Principle components analysis (PCA) combining with transcripts and one-month-water sample based physiochemical parameters indicated that the pollution might be similar at TP, DP and CTB sites while pollution homology existed on some extent between YBQ and JW sites. Although the water quality of the river had a complex time-space alternation during the year, and the one-month-data based RNA-seq could not reflex the whole year-water quality of a watershed, the gene expression profile via RNA-seq provided an alternative way for assessing integrated biotoxicity of surface water, and it was relatively fit for early-warning of water quality of a watershed with unobservable acute toxicity. However, the identification of detail toxicants and the links between DEGs and pollution level as well as physiological-biochemical toxicity needed further investigation.

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.

Cyclophosphamide (CP) and Ifosfamide (IF) are two nitrogen mustard drugs widely prescribed in cancer therapy. They are continuously released via excreta into hospital and urban wastewaters reaching wastewater treatment plants. Although CP and IF, their metabolites and transformation products (TPs) residues have been found in the aquatic environment from few ng L⁻¹ to tens of μg L⁻¹, their environmental toxic effects are still not well known. The present study aimed to investigate the acute and chronic ecotoxicity of CP and IF and their commercially available human metabolites/TPs, i.e. carboxy-CP, Keto-CP and N-dechloroethyl-CP on different organisms of the aquatic trophic chain. The experiments were performed using the green alga Pseudokirchneriella subcapitata, the rotifer Brachionus calyciflorus and the crustaceans Thamnocephalus platyurus and Ceriodaphnia dubia. Moreover, to assess the treatment conditions in regards to parent compound removal and formation of new TPs, CP and IF were UV- irradiated for 6 h, 12 h, 24 h, 36 h and 48 h, followed by toxicity evaluation of treated samples by algae, rotifers and crustaceans. Between the parent compounds, IF resulted as more toxic drug under tested conditions, exerting both acute and chronic effects especially on C. dubia (LC50:196.4 mg L⁻¹, EC50:15.84 mg L⁻¹). Among the tested metabolites/TPs, only carboxy-CP inhibited the reproduction in the rotifer. However, LOEC and NOEC values were calculated for CP and IF for all organisms. In addition, despite a low degradation of CP (28%) and IF (36%) after 48 h UV-irradiation, statistically significant effect differences (p < 0.05) from not-irradiated and irradiated samples were observed in both acute and chronic assays, starting from 6 h UV-irradiation. Our results suggest that the toxic effects found in the aquatic organisms may be attributable to interactions between the parent compounds and their metabolites/TPs.

Microplastics (<5 mm, MP) are ubiquitously distributed in the environment, causing increasing concern regarding their potential toxicity to organisms. To date, most research has focussed on the impacts of MPs on marine and estuarine organisms, with fewer studies focussing on the effects of microplastics on freshwater ecosystems, especially under different environmental conditions. In the present study, the sensitivity of two temperate Cladoceran species, Daphnia magna and Daphnia pulex, and a smaller tropical species Ceriodaphnia dubia, to primary microplastics (PMP) and secondary (weathered) microplastics (SMP) was assessed. A prolonged acute toxicity assay (up to 72 or 96 h) was performed at 18°, 22°, and 26 °C, to determine the influence of temperature as an additional stressor and survival data were analysed using toxicokinetic-toxicodynamic (TK-TD) model. Acute sensitivity of D. magna and D. pulex to both PMP and SMP increased sharply with temperature, whereas that of C. dubia remained relatively stable across temperatures. C. dubia was the most sensitive species at 18 °C, followed by D. pulex and D. magna, which were of comparable sensitivity. However, this ranking was reversed at 26 °C as could be seen from the No Effect Concentration (NEC) estimates of the TK-TD model. In addition, SMP and PMP had a similar effect on D. magna and D. pulex, but PMP was more toxic to C. dubia. Effects on survival were strongly time-dependent and became substantially more severe after the standard 48 h test period. Our results indicate that sensitivity to microplastics may differ between species for different types of microplastics, and could be drastically influenced by temperature albeit at high exposure concentrations.

At present, the majority of investigations concerning nanotoxicology in the nematode C. elegans address short-term effects. While this approach allows for the identification of uptake pathways, exposition and acute toxicity, nanoparticle-organism interactions that manifest later in the adult life of C. elegans are missed. Here we show that a microhabitat composed of liquid S-medium and live bacteria in microtiter wells prolongs C. elegans longevity and is optimally suited to monitor chronic eNP-effects over the entire life span (about 34 days) of the nematode. Silver (Ag) nanoparticles reduced C. elegans life span in concentrations ≥10 μg/mL, whereas nano ZnO and CeO₂ (1–160 μg/mL) had no effect on longevity. Monitoring of locomotion behaviors throughout the entire life span of C. elegans showed that Ag NPs accelerate the age-associated decline of swimming and increase of uncoordinated movements at concentrations of ≥10 μg/mL, whereas neuromuscular defects did not occur in response to ZnO and CeO₂ NPs. By means of a fluorescing reporter worm expressing tryptophan hydroxylase-1::DsRed Ag NP-induced behavioral defects were correlated to axonal protein aggregation and neurodegeneration in single serotonergic HSN as well as sensory ADF neurons. Notably, serotonergic ADF neurons represented a sensitive target for Ag NPs in comparison to GABAergic neurons that showed no signs of degeneration under the same conditions. We conclude that due to its analogy to the jellylike boom culture of C. elegans on microbe-rich rotting plant material liquid S-medium culture in spatially confined microtiter wells represents a relevant as well as practical tool for comparative identification of age-resolved nanoparticle effects and vulnerabilities in a significant target organism. Consistent with this, specifically middle-aged nematodes showed premature neuromuscular defects after Ag NP-exposure.

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.

A pesticide monitoring study including 80 and 60 active ingredients (in surface waters and sediments, respectively) was carried out in a river basin in Costa Rica during 2007–2012. A special emphasis was given on the exceptional ecological conditions of the tropical agro-ecosystem and the pesticide application strategies in order to establish a reliable monitoring network. A total of 135 water samples and 129 sediment samples were collected and analyzed. Long-term aquatic ecotoxicological risk assessment based on risk quotient in three trophic levels was conducted. Short-term risk assessment was used to calculate the toxic unit and prioritization of sampling sites was conducted by the sum of toxic units in both aquatic and sediment compartments. Dimethoate (61.2 μg/L), propanil (30.6 μg/L), diuron (22.8 μg/L) and terbutryn (4.8 μg/L) were detected at the highest concentrations in water samples. Carbendazim and endosulfan were the most frequently detected pesticides in water and sediment samples, respectively. Triazophos (491 μg/kg), cypermethrin (71.5 μg/kg), permethrin (47.8 μg/kg), terbutryn (38.7 μg/kg), chlorpyrifos (18.2 μg/kg) and diuron (11.75 μg/kg) were detected at the highest concentrations in sediment samples. The pesticides carbendazim, diuron, endosulfan, epoxyconazole, propanil, triazophos and terbutryn showed non-acceptable risk even when a conservative scenario was considered. Sum TUsite higher than 1 was found for one and two sampling sites in water and sediment compartments, respectively, suggesting high acute toxicity for the ecosystem.Exceptional ecological conditions of the tropical agro-ecosystem affect the fate of pesticides in water and sediment environment differently than the temperate one.

Dinotefuran, as the latest generation of neonicotinoid insecticide, has broad application prospects around the world. However, dinotefuran is easily metabolized and the two main metabolites are 1-methyl-3-(tetrahydro-3-furylmethyl) urea (UF) and 1-methyl-3-(tetrahydro-3-furylmethyl) guanidium dihydrogen (DN). In the present study, the risks of UF and DN in soil on typical non-target species earthworm were investigated. In the same time, the degradation in soil and accumulation in earthworms of UF and DN were monitored. The present results showed that the toxicities of UF and DN were similar in their effect on earthworms and they were supertoxic pollutants to earthworms. The degradation trend in soil and accumulation trend in earthworms of UF and DN were consistent throughout the whole exposure period. At 1.0 mg/kg and 2.0 mg/kg, UF and DN could induce the excess production of ROS, resulting in oxidative stress effects in earthworm cells. The excess ROS induce changes in antioxidant enzyme activities, damage in biomacromolecules, and abnormal expression of function genes. The present results showed that UF and DN may have high risks for earthworms.

Fluralaner is a novel isoxazoline insecticide which shows high insecticidal activity against parasitic, sanitary and agricultural pests, but there is little information about the effect of fluralaner on non-target organisms. This study reports the acute toxicity, bioconcentration, elimination and antioxidant response of fluralaner in zebrafish. All LC50 values of fluralaner to zebrafish were higher than 10 mg L⁻¹ at 24, 48, 72 and 96 h. To study the bioconcentration and elimination, the zebrafish were exposed to sub-lethal concentrations of fluralaner (2.00 and 0.20 mg L⁻¹) for 15 d and then held 6 d in clean water. The results showed medium BCF of fluralaner with values of 12.06 (48 h) and 21.34 (144 h) after exposure to 2.00 and 0.20 mg L⁻¹ fluralaner, respectively. In the elimination process, a concentration of only 0.113 mg kg⁻¹ was found in zebrafish on the 6th day after removal to clean water. After exposure in 2.00 mg L⁻¹ fluralaner, the enzyme activities of SOD, CAT, and GST, GSH-PX, CarE and content of MDA were measured. Only CAT and CarE activities were significantly regulated and the others stayed at a stable level compared to the control group. Meanwhile, transcriptional expression of CYP1C2, CYP1D1, CYP11A were significantly down-regulated at 12 h exposed to 2.00 mg L⁻¹ of fluralaner. Except CYP1D1, others CYPs were up-regulated at different time during exposure periods.Fluralaner and its formulated product (BRAVECTO®) are of low toxicity to zebrafish and are rapidly concentrated in zebrafish and eliminated after exposure in clean water. Antioxidant defense and metabolic systems were involved in the fluralaner-induced toxicity. Among them, the activities of CAT and CarE, and most mRNA expression level of CYPs showed fast response to the sub-lethal concentration of fluralaner, which could be used as a biomarker relevant to the toxicity.

Stability of silver sulfide nanoparticle (Ag₂S-NP) in the environment has recently drawn considerable attention since it is associated with environmental risk. Although the overestimated stability of Ag₂S-NP in aqueous solution has already been recognized, studies on transformation of Ag₂S-NP in environmental water are still very scarce. Here we reported that Ag₂S-NP could undergo dissolution by manganese(IV) oxide (MnO₂), an important naturally occurring oxidant in the environment, even in environmental water, although the dissolved silver would probably be adsorbed onto the particles (>0.45 μm) in environmental water, mitigating the measurable levels of dissolved silver. The extent and rate of Ag₂S-NP dissolution rose with the increasing concentration of MnO₂. In addition, environmental factors including natural organic matter, inorganic salts and organic acids could accelerate the Ag₂S-NP dissolution by MnO₂, wherein an increase in dissolution extent was also observed. We further documented that Ag₂S-NP dissolution by MnO₂ was highly dependent on O₂ and it was an oxidative dissolution, with the production of SO₄²⁻. Finally, dissolution of Ag₂S-NP by MnO₂ affected zebra fish (Danio rerio) embryo viability, showing significant reduction in embryo survival and hatching rates, compared to embryos exposed to Ag₂S-NP, MnO₂ or dissolved manganese alone. These findings would further shed light on the stability of Ag₂S-NP in the natural environment - essential for comprehensive nano risk assessment.