Assessment of toxic effects of imidacloprid on freshwater zooplankton: An experimental test for 27 species.

The neonicotinoid pesticide imidacloprid has been used worldwide since 1992. As one of the most important chemicals used in pest control, there have been concerns that its run-off into rivers and lakes could adversely affect aquatic ecosystems, where zooplankton play a central role in the energy flow from primary to higher trophic levels. However, studies assessing the effects of pesticides at the species level have relied on a Daphnia-centric approach, and no studies have been conducted using species-level assessments on a broad range of zooplankton taxa. In the present study, we therefore investigated the acute toxicity of imidacloprid on 27 freshwater crustacean zooplankton (18 cladocerans, 3 calanoid copepods and 6 cyclopoid copepods). The experiment showed that a majority of calanoid copepods and cladocerans were not affected at all by imidacloprid, with the exception of one species each of Ceriodaphnia and Diaphasoma, while all six cyclopoid copepods showed high mortality rates, even at concentrations of imidacloprid typically found in nature. In addition, we found a remarkable intra-taxonomic variation in susceptibility to this chemical. As many cyclopoid copepods are omnivorous, they act as predators as well as competitors with other zooplankton. Accordingly, their susceptibility to imidacloprid is likely to cause different responses at the community level through changes in predation pressure as well as changes in competitive interactions. The present results demonstrate the need for species-level assessments of various zooplankton taxa to understand the complex responses of aquatic communities to pesticide disturbance.

Enhancing the potential of rapeseed cake as protein-source food by γ-irradiation.

Rapeseed cake serves as a by-product in the oil extraction industry, characterized by its elevated protein content. However, the presence of antinutritional factors limits the utilization of rapeseed cake as a viable protein source. In this study, different doses of γ-irradiation were used to irradiate rapeseed cake and rapeseed protein isolate was extracted through a modified alkaline solution and acid precipitation method from rapeseed cake. The chemical composition and in vivo acute toxicity of rapeseed protein isolate were determined. The protein recovery rate of rapeseed protein isolate was 39.08 ± 3.01% after irradiation, while the content of antinutritional factors was significantly reduced. Moreover, γ-irradiation did not have any experimentally related effects on clinical observations or clinicopathology in mice. Overall, the reduced antinutrients and increased functional properties suggest that the irradiation of rapeseed cake (<9 kGy) could be utilized as a pre-treatment in the development of rapeseed cake-based value-added protein products.

Toxicological effects of acute and repeated doses (180 days) of fruits from Malpighia emarginata (acerola) in rodents.

Malpighia emarginata has a high amount of vitamin C with pharmacological or food preservation potential. However, despite its wide use and application possibilities its toxicity in repeated doses and for a long time (6 months) has not yet been studied. In this context, this study aimed to evaluate the acute toxicity and repeated doses from fruits of this plant. The extract was produced with the pulp (EMe) of the lyophilized fruit and submitted to chromatographic and spectroscopic analysis (HPLC and ESI-IT-MSn). In the acute test, the EMe was administered orally and parenterally to rodents (mice and rats) for 14 days, at a dose of 2000 mg/kg. Subsequently, the repeated dose toxicity test was administered orally for 180 days at doses of 50, 300 or 1000 mg/kg. The HPLC assay revealed a high concentration of vitamin C (16.3%), and spectroscopic analyses pointed to the presence of five other polyphenolic compounds. In the acute test, the plant extract showed no apparent toxicity or lethality in rodents. The LD50 was estimated to be greater than 2000 mg/kg and falls into category 5 (low toxicity). In the repeated dose assay, there was no evidence of toxicity, and no differences were observed in water intake, food, weight development, or behavior of the animals in relation to the vehicle group (water). However, hematological and biochemical evaluations pointed out some nonconformities in the levels of cholesterol, leukocytes, and neutrophils of the male rats, but overall, these results did not reveal significant toxicity. Therefore, the Level of Unobserved Adverse Effects (NOAEL) was 1000 mg/kg. Together, the results suggest that the extract obtained from the fruits of M. emarginata does not present representative toxicity in rodents.

Toxicity of glyphosate and aminomethylphosphonic acid (AMPA) to the early stages of development of Steindachneridion melanodermatum, an endangered endemic species of Southern Brazil.

This study aimed to evaluate glyphosate (GLY) and aminomethylphosphonic acid (AMPA) toxicity at 65, 650, and 6500 µg L-1 to the initial stages of development of Steindachneridion melanodermatum, an endangered endemic species from the Iguaçu River, assessing hatching, survival, total larval length, deformities, oxidative stress biochemical biomarkers, and neurotoxicity. Overall, looking at the sum of responses through the integrated biomarker response, the species was more sensitive to AMPA than GLY, especially at the lower concentration of 65 µg L-1, which induced mortality, deformities, underdevelopment, and oxidative stress. Considering the risk of exposure and the importance of conservation of the highly endemic ichthyofauna of this basin, it is urgent to investigate and regulate both GLY and AMPA levels at the Iguaçu River to protect not only this species, but the entire ecosystem.

Ecotoxicological evaluation of functional carbon nanodots using zebrafish (Danio rerio) model at different developmental stages.

Considering functional carbon nanodots (FCNs) are potential to be applied in many areas, their risk and toxicity to organisms are imperative to be evaluated. Thus, this study conducted acute toxicity test of zebrafish (Danio rerio) at embryonic and adult stage to estimate the toxicity of FCNs. Results show that the toxic effects of FCNs and nitrogen doped FCNs (N-FCNs) at their 10% lethal concentration (LC10) values on zebrafish are expressed in developmental retardation, cardiovascular toxicity, renal damage and hepatotoxicity. There are interactive relationships between these effects, but the main reason should be ascribed to the undesirable oxidative damage induced by high doses of materials, as well as the biodistribution of FCNs and N-FCNs in vivo. Even so, FCNs and N-FCNs can promote the antioxidant activity in zebrafish tissues to cope with the oxidative stress. FCNs and N-FCNs are not easy to cross the physical barriers in zebrafish embryos or larvae, and can be excreted from intestine by adult fish, which proves their biosecurity to zebrafish. In addition, because of the differences in physicochemical properties, especially nano-size and surface chemical property, FCNs show higher biosecurity to zebrafish than N-FCNs. The effects of FCNs and N-FCNs on hatching rates, mortality rates and developmental malformations are dose-dependent and time-dependent. The LC50 values of FCNs and N-FCNs on zebrafish embryo at 96 hpf are 1610 mg/L and 649 mg/L, respectively. According to the Acute Toxicity Rating Scale of the Fish and Wildlife Service, the toxicity grades of FCNs and N-FCNs are both defined as "practically nontoxic", and FCNs are "Relatively Harmless" to embryos because their LC50 values are above 1000 mg/L. Our results prove the biosecurity of FCNs-based materials for future practical application.

Establishment of a human induced pluripotent stem cell derived alveolar organoid for toxicity assessment.

Alveolar epithelial cells (AECs) are vulnerable to injury, which can result in epithelial hyperplasia, apoptosis, and chronic inflammation. In this study, we developed human induced pluripotent stem cell (hiPS) cell-derived AECs (iAECs) and the iAECs based organoids (AOs) for testing AEC toxicity after chemical exposure. HiPS cells were cultured for 14 days with differentiation medium corresponding to each step, and the iAECs-based AOs were maintained for another 14 days. SFTPC and AQP5 were expressed in the AOs, and mRNA levels of SOX9, NKX2.1, GATA6, HOPX, and ID2 were increased. The AOs were exposed for 24 h to nine chemical substances, and IC50 values of the nine chemicals were determined using MTT assay. When the correlations between iAECs 2D culture and AOs 3D culture were calculated using Pearson's correlation coefficient r value, the nine chemicals that caused a significant decrease of cell viability in 3D culture were found to be highly correlated in 2D culture. The cytotoxicity and nitric oxide release in AO cultured with macrophages were then investigated. When AOs with macrophages were exposed to sodium chromate for 24 h, the IC50 value and nitric oxide production were higher than when the AOs were exposed alone. Taken together, the AO-based 3D culture system provides a useful platform for understanding biological characteristics of AECs and modeling chemical exposures.

On the In Vitro and In Vivo Hazard Assessment of a Novel Nanomaterial to Reduce the Use of Zinc Oxide in the Rubber Vulcanization Process.

Zinc oxide (ZnO) is the most efficient curing activator employed in the industrial rubber production. However, ZnO and Zn(II) ions are largely recognized as an environmental hazard being toxic to aquatic organisms, especially considering Zn(II) release during tire lifecycle. In this context, aiming at reducing the amount of microcrystalline ZnO, a novel activator was recently synthetized, constituted by ZnO nanoparticles (NPs) anchored to silica NPs (ZnO-NP@SiO2-NP). The objective of this work is to define the possible hazards deriving from the use of ZnO-NP@SiO2-NP compared to ZnO and SiO2 NPs traditionally used in the tire industry. The safety of the novel activators was assessed by in vitro testing, using human lung epithelial (A549) and immune (THP-1) cells, and by the in vivo model zebrafish (Danio rerio). The novel manufactured nanomaterial was characterized morphologically and structurally, and its effects evaluated in vitro by the measurement of the cell viability and the release of inflammatory mediators, while in vivo by the Fish Embryo Acute Toxicity (FET) test. Resulting data demonstrated that ZnO-NP@SiO2-NP, despite presenting some subtoxic events, exhibits the lack of acute effects both in vitro and in vivo, supporting the safe-by-design development of this novel material for the rubber industry.

Acute toxicological study: EZY-1 with potent therapeutic effects of idiopathic pulmonary fibrosis and its mechanisms.

EZY-1 is an antifibrosis peptide purified from Eucheuma. In this study, we explored the acute toxicology of EZY-1 and the signaling pathways involved in its antifibrotic role. The mouse model of pulmonary fibrosis was induced by bleomycin. Pathological changes in lung tissue could be effectively inhibited by EZY-1. Acute toxicity and cell proliferation tests indicated that EZY-1 had no apparent toxicity to mice and cells. We identified proteins that could bind directly to EZY-1 in vitro on the basis of liquid chromatography-tandem mass spectrometry and bioinformatics analysis. EZY-1 inhibited pulmonary fibrosis via Wnt/ß-catenin, transforming growth factor (TGF)-ß/Smad, phosphoinositide 3-kinase/protein kinase B/ mammalian target of rapamycin, and activator of transcription 3 and Janus kinase 2/signal transducer pathways. A transwell micropore experiment showed that EZY-1 could inhibit cell migration and invasion. Western blotting analysis on transforming growth factor-ß1 (TGF-ß1)-induced A549 pulmonary fibrosis cell model suggested that EZY-1 could downregulate p-Smad3 (Ser423/Ser425), Smad4, ß-catenin, vimentin, and N-cadherin expression. ELISA showed that EZY-1 could inhibit collagen-I secretion. EZY-1 alleviated idiopathic pulmonary fibrosis (IPF) through regulating TGF-ß/Smad pathways, epithelial-mesenchymal transition processes, and collagen secretion, which provides a potential foundation for theoretical development of EZY-1 as a potential drug against IPF. PRACTICAL APPLICATIONS: We isolated a new 16-amino-acid peptide derived from the polypeptide extract of Eucheuma, named EZY-1. In vitro and in vivo assays show peptide EZY-1 is safe. The EZY-1 peptide alleviates IPF at lower doses than pirfenidone. EZY-1 alleviated idiopathic pulmonary fibrosis (IPF) through regulating TGF-ß/Smad pathways, epithelial-mesenchymal transition (EMT) processes, and collagen secretion, which provides a theoretical basis for the development of EZY-1 as a potential drug against IPF.

The onset of active gill respiration in post-embryonic zebrafish (Danio rerio) larvae triggers an increased sensitivity to neurotoxic compounds.

Originally designed as a general alternative to acute fish toxicity testing (AFT), the fish embryo toxicity test (FET) has become subject to concerns with respect to neurotoxic substances. Whereas oxygen uptake in the fish embryo primarily occurs via diffusion across the skin, juvenile and adult fish rely on active ventilation of the gills. As a consequence, substances including, e.g., neurotoxicants which prevent appropriate ventilation of gills ("respiratory failure syndrome") might lead to suffocation in juvenile and adult fish, but not in skin-breathing embryos. To investigate if this respiratory failure syndrome might play a role for the higher sensitivity of juvenile and adult fish to neurotoxicants, a modified acute toxicity test using post-embryonic, early gill-breathing life-stages of zebrafish was developed with chlorpyrifos, permethrin, lindane, aldicarb, ziram and aniline as test substances. Additionally, a comparative study into bioaccumulation of lipophilic substances with logKow > 3.5 and swimbladder deflation as potential side effects of the respiratory failure syndrome was performed with 4 d old skin-breathing and 12 d old gill-breathing zebrafish. With respect to acute toxicity, post-embryonic 12 d larvae proved to be more sensitive than both embryos (FET) and adult zebrafish (AFT) to all test substances except for permethrin. Accumulation of chlorpyrifos, lindane and permethrin was 1.3- to 5-fold higher in 4 d old than in 12 d old zebrafish, suggesting that (intermediate) storage of substances in the yolk might reduce bioavailability and prevent metabolization, which could be a further reason for lower toxicity in 4 d than in 12 d old zebrafish. Whereas ziram and aniline showed no significant effect on the swimbladder, zebrafish exposed to chlorpyrifos, lindane and permethrin showed significantly deflated swimbladders in 12 d old larvae; in the case of aldicarb, there was a significant hyperinflation in 4 d old larvae. Swimbladder deflation in post-embryonic 12 d zebrafish larvae might be hypothesized as a reason for a lack of internal oxygen supplies during the respiratory failure syndrome, whereas in 4 d old embryos cholinergic hyperinflation of the swimbladder dominates over other effects. Regarding acute lethality, the study provides further evidence that the switch from transcutaneous to branchial respiration in post-embryonic zebrafish life-stages might be the reason for the higher sensitivity of juvenile and adult fish to neurotoxic substances.

In vitro and in vivo anti-tumor efficacy of krill oil against bladder cancer: Involvement of tumor-associated angiogenic vasculature.

Krill oil (KO) obtained from Euphausia superba is nutrient-rich and has a positive effect on human health. Here, we explored the efficacy of KO in inhibiting tumor progression and tumor vascularization. KO (100-300 µg/mL) repressed the proliferation of bladder tumor cell lines MBT-2 and T24. Treatment of cells with KO raised cell-cycle arrest at G0/G1-phase via modulation of positive regulators and negative regulators in bladder cancer cells. KO treatment regulated phosphorylation of proteins involved in PI3K/AKT and MAPK signaling pathways, including ERK, JNK, and p38 MAPK. Additionally, KO hampered the invasion and migration of both cell lines via reduction of MMP-9 expression levels by disrupting transcriptional binding of Sp-1, AP-1, and NF-κB motifs. Moreover, in animal studies, KO (150-300 mg/kg) significantly decreased tumor growth in xenograft mice bearing T24 tumor cells. No significant toxic effects were observed in acute toxicity tests, including biological analysis and H&E staining. The reduced level of CD31 expression in KO-treated tumor tissues prompted us to investigate the effect of KO on tumor angiogenesis. KO (5-40 µg/mL) treatment impeded VEGF-induced capillary tube formation and proliferation by inhibiting VEGFR2-modulated eNOS/AKT/ERK1/2 signaling axis in HUVECs. Treatment of HUVECs with KO inhibited VEGF-stimulated migration and invasion by reducing MMP-2 expression level. VEGF-driven sprouting capacity of neo-microvessels was suppressed in the presence of KO (20-40 µg/mL), as determined via an ex vivo aortic ring assay. Our results indicated that KO can regulate both tumor growth and tumor-associated angiogenesis via a novel mechanism. Thus, KO may be a promising antitumor candidate, potentially useful to prevent or treat bladder cancer.

Treatment of electropolishing industrial wastewater and its impact on the immobilisation of Daphnia magna.

The amount of industrial pollution entering the environment and its impact on living organisms is an ongoing concern. At the same time, due to an increasing awareness, new methods of wastewater treatment are being explored that are not only effective but also environmentally acceptable. Meeting environmental standards for permitted concentrations is a necessity, but investigating the effects of wastewater on living organisms is also an important issue. In this paper, the influence of metal ions (Fe(III), Cr(III), Ni(II), Cu(II)) in industrial wastewater from electropolishing of stainless steel on Daphnia magna has been investigated. Daphnids have been exposed to wastewater both before and after treatment (Ca(OH)2 precipitation, sorption with peat). Immobilisation in a 48-h acute toxicity test and EC50 has been determined. In the case of studied industrial wastewater, the organic content (expressed as total organic carbon) of the effluent has a positive impact in terms of the survival of D. magna and increases the range of heavy metal concentrations tolerated by them. The application of a two-stage process with Ca(OH)2 neutralisation followed by sorption with peat allows for the removal of almost 100% of metal ions from the wastewater. The reduction obtained ensured a limited impact on D. magna and a decrease in immobilisation to less than 10%. Proper execution of the wastewater treatment process ensures a reduction of its negative impact on living organisms.

Mechanisms and Safety of Antidepressant-Like Effect of Nutmeg in Mice.

Nutmeg, a dried seed kernel of a tall evergreen Myristicaceae tree, is widely used as a spice and herbal medicine and is known to have antidepressant-like effects. This study evaluates the mechanisms underlying this antidepressant-like effect and safety of nutmeg n-hexane extract (NNE) in mice. Tail suspension and open field tests showed that NNE (10 mg/kg, per OS (p.o.)) significantly decreased the immobility time of mice without effecting their spontaneous locomotor activity. The reduction of immobility time of mice elicited by NNE was significantly inhibited by ketanserin (5-hydroxytryptamine (5-HT)2A/2C receptor antagonist), ondansetron (5-HT3 receptor antagonist), and yohimbine (α2 receptor antagonist). WAY100635 (5-HT1A receptor antagonist) tended to inhibit the effect of NNE but without significance. Testing according to the Organisation for Economic Co-operation and Development Guidelines, no mice died due to administrated NNE (2000 mg/kg, p.o.), and behavioral and weight changes were not seen in the acute toxicity test. In the Ames test, no increase in the number of revertant colonies for each bacterial strain test strains TA98 and TA100 by nutmeg powder was observed either with or without metabolic activity by S9 mix. These results suggest that NNE shows an antidepressant-like effect involving various serotonergic and noradrenergic nervous systems and maybe a highly safe natural preparation.

Severity Classification of Clinical Signs and Defining the Moribund State as an Experimental Endpoint for Acute Toxicity Testing Using Japanese Medaka (Oryzias latipes).

Use of the moribund state as an endpoint for acute toxicity testing in fish is complicated by the lack of consensus about the sublethal signs that define the state or that can predict imminent death. Defining the moribund state in fish requires determination of the association between each observable sublethal sign and transition to imminent death. To establish a criterion for defining the moribund state in Japanese medaka (Oryzias latipes), we determined death/clinical-sign ratios, defined as the proportion of fish with each clinical sign that transitioned to imminent death, for 13 clinical signs observed in individually housed fish exposed to one of two typical toxicants, three pharmaceuticals, two pesticides, and one metal. Also, the time from onset of each clinical sign to transition to imminent death (referred to as the survival time) was determined by continuous monitoring. Three of the observed clinical signs-immobility, lethargy, and immobility at the surface-were found to be indicators of the moribund state, with each of these signs having a death/clinical-sign ratio of 1.0. Evaluation of the survival time after onset of the other 10 signs enabled determination of whether timely euthanasia would be appropriate, thereby providing a means of reducing the suffering of laboratory fish in the period before death. Environ Toxicol Chem 2022;41:1089-1095. © 2022 SETAC.

Species and Life-Stage Sensitivity of Chinese Rare Minnow (Gobiocypris rarus) to Chemical Exposure: A Critical Review.

Chemical production and consumption in Asia are increasing at an unprecedented rate, calling for regulations on chemical management. Under the New Chemical Substance Notification in China, information on ecotoxicological effects of chemicals is mandatory for the simplified registration of chemicals with the requirement that one ecotoxicological test is conducted locally. It is now mandatory to use the native fish species Chinese rare minnow (Gobiocypris rarus). However, its chemical sensitivity compared to that of fathead minnow (Pimephales promelas) or zebrafish (Danio rerio) is still unclear. We performed a holistic literature review on toxicity data with G. rarus from 1997 to 2020. Species sensitivity among G. rarus, P. promelas, and D. rerio and life-stage sensitivity of G. rarus were systematically investigated for various chemicals using both chemical ratio distribution and probabilistic chemical toxicity distribution approaches. Comparatively, the Chinese native fish species G. rarus was more sensitive than D. rerio, particularly to metals. Juvenile and adult G. rarus were more sensitive than its larvae and embryos. The observed lower sensitivity of G. rarus embryo was likely due to the thick embryonic chorion, discrepant methods of collecting embryos, and the paucity of toxicity data, implying the necessity to standardize G. rarus embryo tests and validate the sensitivity with various types of chemicals. This unique review allows us to conclude that G. rarus studies could be used in worldwide registrations and that further investigations are needed to use G. rarus embryos as alternatives to the fish test. Environ Toxicol Chem 2021;40:2680-2692. © 2021 SETAC.

Investigating the Predictive Power of Three Potential Sublethal Endpoints for the Fathead Minnow Fish Embryo Toxicity Test: Snout-Vent Length, Eye Size, and Pericardial Edema.

The fish embryo acute toxicity (FET) test is known to be less sensitive than the fish acute test for some chemicals, including neurotoxicants. Thus, there is an interest in identifying additional endpoints that can improve FET test performance. The goal of this project was to advance alternative toxicity testing methods by determining whether select developmental abnormalities-snout-vent length, eye size, and pericardial area-are linked to adverse alterations in ecologically-relevant behaviors and delayed mortality. Fathead minnow (Pimephales promelas) FET tests were conducted with 3,4-dicholoroaniline, cadmium, and perfluorooctanesulfonic acid (PFOS) and developmental abnormalities were quantified. Surviving eleutheroembryos were reared in clean water to 14 days post fertilization (dpf), during which time behaviors and mortality were evaluated. None of the abnormalities evaluated were predictive of behavioral alterations; however, embryos with ≥14% reductions in length or ≥3.54-fold increases in pericardial area had an 80% chance of mortality by 14 dpf. When these abnormalities were used as markers of mortality, the LC50s for cadmium and PFOS were less than those calculated using only standardized FET test endpoints and similar to those obtained via larval fish tests, indicating that the snout-vent length and pericardial area warrant consideration as standard FET test endpoints.

Integration of environmental and economic performance of Electro-Coagulation-Anodic Oxidation sequential process for the treatment of soluble coffee industrial effluent.

This study examined a sequential Electro-Coagulation-Anodic Oxidation (EC-AO) process, using low-cost electrodes (EC: iron/stainless steel and AO: graphite/stainless steel), as an alternative for the treatment of soluble coffee industrial effluent (a complex mixture of organic macromolecules). Process operational parameters were optimized using experimental design, ANOVA, the response surface methodology, and a constrained nonlinear optimization algorithm. Experimental results allow discussing the role of graphite electrode, the mineralization via hydroxyl radicals and chlorine species (mainly as HClO and ClO- as defined from pH conditions and equilibrium calculations). EC-AO treatment allows achieving ca. 97% decolorization, as well as 72% and 65% of COD and TOC removal, respectively, with total operation costs of 10.81 USD/m3. The Molecular Weight Distribution (MWD) analysis showed that during the EC step, contaminants with molecular weight > 30 kDa were effectively decomposed. In contrast, AO was quite efficient in the mineralization of contaminants with MW in the range of 5 to 10 kDa. A kinetic analysis of EC-AO allowed defining the operation time of ca. 160 min required to meet permissible discharge limits for soluble coffee effluents, established by Colombian legislation; and obtaining COD and TOC degradation rate laws for upcoming studies on process conceptual design. Finally, the EC-AO process yielded an oxidized (AOS = 0.687), biocompatible (BOD5/COD = 0.41) not toxic effluent.

Effects of biochar and thermally treated biochar on Eisenia fetida survival, growth, lysosomal membrane stability and oxidative stress.

Despite its known positive impacts when added to soil, the negative effects of biochar on earthworms are not fully understood. Here, we investigated the toxicity of nine biochars of three feedstock origins, animal (cow dung), plant (corncob) and microorganism (sewage sludge), produced at three pyrolysis temperatures (350 °C, 550 °C and 750 °C) on earthworms. Vermitoxicity was first assessed using acute toxicity test, neutral red retention time (NRRT) assay and oxidative stress response assay. Furthermore, we evaluated whether the thermal treatment of biochars could reduce their vermitoxicity using an acute toxicity assay. We found that, according to LC50 and earthworm weight loss, cow dung biochar was more toxic than corncob or sewage sludge biochar; thus, production feedstock is apparently important to biochar vermitoxicity. Furthermore, NRRTs indicated cow dung biochar disrupted lysosomal membrane stability in earthworm coelomocytes, providing further evidence for the toxicity of this biochar to earthworms. Disturbed antioxidant enzyme activities and elevated malondialdehyde content showed that earthworm suffer oxidative stress, also implying a potential vermitoxicity. However, thermal treatment of cow dung biochar substantially improved its LC50 and decreased earthworm weight loss, implying that the PAHs in this biochar might be damage factors and that heating could reduce the potential toxicity of biochar. Besides, NRRT assay was first used to evaluate the effects of biochar on earthworms and clear dose-effect relationships indicated that NRRT assay might be a useful tool for assessing the potential negative effects of biochar. Overall, given the different effects of various biochars, including toxicity, reported here, our findings will help improve understanding of biochar vermitoxicity mechanisms, serve to improve biochar ecological risk assessments and provide a reference for the proper application of biochar amendments.

Effects of nickel oxide nanoparticles on survival, reproduction, and oxidative stress biomarkers in the marine calanoid copepod Centropages ponticus under short-term exposure.

Excessive use of nickel oxide nanoparticles (NiO NPs) in various industrial and commercial products can lead to various negative effects in human and environmental health due to their possible discharge into the environment. Nerveless, information about their ecotoxicological effects on marine organisms are lacking. Copepods are good ecotoxicological models because of their high sensitivity to environmental stress and their key role in the marine food webs. In this study, 48 h acute tests were conducted on the marine planktonic copepod Centropages ponticus to assess lethal and sublethal toxicities of NiO NPs. The results revealed LC50 (48 h) of 4 mg/L for adult females. Aggregation and settling of NiO NPs were observed at concentrations ≥ 2 mg/L. Exposure to sublethal concentrations (≥ 0.02 mg/L for 48 h) had significant negative effects on reproductive success in C. ponticus. Egg production after 24 h and 48 h decreased by 32% and 46%, respectively at 0.02 mg/L and 70% and 82%, respectively, at 2 mg/L. Hatching success was reduced by 70% and 79% at 2 mg/L for eggs produced after 24 h and 48 h respectively. Antioxidant enzymatic activity increased significantly with NiO NP concentration and time, indicating that NiO NPs can cause oxidative stress in C. ponticus even under short-term exposure, while significant inhibition of acetylcholinesterase activity at 2 mg/L after 48 h suggests neurotoxic effects of NiO NPs.

Toxicity of soil antimony to earthworm Eisenia fetida (Savingy) before and after the aging process.

Information on soil antimony (Sb) toxicity to earthworm Eisenia fetida (Savingy) is limited. This ecotoxicology study was designed to quantify the soil Sb toxicity to earthworm E. fetida before and after aging process, establishing dose-effect relationship between Sb content and mortality. Results of the avoidance test and acute test showed that the values of net avoidance response, escape rate and mortality were generally decreased in aged treatment compared to that in fresh treatment, respectively from 93.33% to 66.67%, 36.67% to 13.33% and 100% to 53.33% (15 d) taking TL800 (treatment level of 800 mg/kg) for example, meanwhile the values of median lethal content (LC50) at 72 h, 7 d and 15 d were respectively increased from 355.27 mg/kg to 2324.55 mg/kg, 322.19 mg/kg and 1743.19 mg/kg and 282.74 mg/kg to 745.94 mg/kg, indicating that aging process could reduce the Sb acute toxicity to earthworm. According to a three-step sequential extraction procedure, the bioavailable Sb ranged from 24.45% to 43.24% and 16.97% to 27.70% in fresh treatment and aged treatment, respectively, and the mortality of earthworm for 24 h decreased with the decrease of the content of mild acid-soluble antimony (which decreased averagely from 23.09% to 14.00%), which was more suited to assess Sb toxicity. This is the first report that confirms the toxicity of soil Sb to earthworm E. fetida as well as the considering of aging process and speciation.

Toxicological assessment and developmental abnormalities induced by butylparaben and ethylparaben exposure in zebrafish early-life stages.

Toxicological effects of butylparaben (BuP) and ethylparaben (EtP) on zebrafish (Danio rerio) early-life stages are not well established. The present study evaluated, using zebrafish embryos and larvae, the toxicity of BuP and EtP through benchmark dose (BMD) approach. BuP was more toxic than EtP to zebrafish larvae. In fact, Lethal Concentration 50 (LC50) values at 96 h post-fertilization (hpf) for BuP and EtP were 2.34 mg/L and 20.86 mg/L, respectively. Indeed, BMD confidence interval (lower bound (BMDL) - upper bound (BMDU) was 0.91-1.92 mg/L for BuP and 10.8-17.4 mg/L for EtP. Zebrafish embryos exposed to 1 mg/L, 2.5 mg/L of BuP and 5 mg/L, 10 mg/L, 20 mg/L, 30 mg/L of EtP showed several developmental abnormalities and teratological effects compared to negative control. Exposed zebrafish developed reduced heartbeat, reduction in blood circulation, blood stasis, pericardial edema, deformed notochord and misshaped yolk sac. Embryos exposed to the highest concentrations of the chemicals (2.5 mg/L of BuP, 10 mg/L, 20 mg/L and 30 mg/L of EtP) showed the developmental abnormalities at 48 hpf while those treated with 1 mg/L of BuP and 10 mg/L of EtP reported behavioral changes at 72 hpf, including trembling of head, pectoral fins and spinal cord. This research identified the lethal and sublethal effects of BuP and EtP in zebrafish early-life stages and could be helpful to elucidate the developmental pathways of toxicity of parabens.