Zebrafish Embryo Toxicity Test Research Articles

Green Engineered CuO/SnO2/HNT Composites: Illuminating the Photocatalytic Path for Organic Pollutant Remediation and Zebrafish Embryo Toxicity Evaluation.

Photocatalysis stands out as a promising technique for treating organic contaminants, yet the quest for visible light active composite materials, crafted through cost-effective, eco-friendly, and uncomplicated processes, poses formidable challenges. Here, we introduce a successful endeavor, the synthesis of a CuO/SnO2 (CS) composite via a microwave method, employing Carissa edulis fruit extract as a reducing as well as capping agent. Various loadings of CS-HNT composites were prepared by ultrasonically incorporating CS onto nanotubular halloysite (HNT) clay. Employing a suite of types of characterization, including XRD, XPS, FT-IR, FE-SEM, HR-TEM, ζ potential, UV-vis-DRS, TGA, BET, and EIS, we meticulously explored the morphology, structure, stability, surface area, electrochemical, and optical properties of the developed CS-HNT composites. HR-TEM observations unveiled the formation of a heterojunction between cubic CuO and spherical SnO2 on the HNT clay surface. Optical and EIS analysis highlighted that the 20CS-HNT composite displayed significant absorption in the visible region, efficient electron-hole pair separation, and enhanced interfacial charge transport relative to other loadings. Photocatalytic evaluations and optimization studies revealed that the 20CS-HNT photocatalyst achieved notable removal efficiencies, eliminating 85% of Congo red (CR) and 80% of tetracycline (TC) within 90 min and 74% of ATZ in 3 h under visible light conditions. Scavenging investigations, the fluorescence probe method, and a NBT transformation study underscored the pivotal roles of hydroxyl and superoxide radicals in pollutant removal. The reusability trials highlighted the exceptional stability and recyclability of the photocatalyst, even after five cycles. In addition, zebrafish embryo toxicity tests revealed improved survival and hatching rates in photocatalyst-treated samples compared to those of controls. Moderate toxicity was observed in treated TC, while the treated CR sample showed non-lethal toxicity. In essence, this study unveils a straightforward and efficacious approach for developing photocatalysts for large-scale wastewater treatment. Furthermore, it proposes the adoption of safe clay-based bimetal oxide photocatalysts in diverse environmental applications.

Lab scale evaluation of biomineralization-assisted decolourization of reactive blue 4 and toxicity analysis

The process of microbial mineral precipitation has proven to be highly effective in addressing a wide range of pollutants in different environments. The objective of this study is to utilize the mineralization abilities of microbial strains found in the Noyyal River basin, which is home to a significant textile center in India, in order to both remove the color from reactive blue 4 and promote biomineralization. Promising results were obtained from the microbial isolates Streptomyces sp., Rhodococcus ruber, and Bacillus sp., and their consortium which demonstrated the ability to induce biomineralization and decolorize reactive blue 4. Characterization of the obtained biominerals through X-ray diffraction and FT-IR analysis showed the presence of well-crystallized polymorphs of calcium carbonate, along with other organic compounds. SEM-EDX analysis of biominerals formed during biomineralization-assisted decolourization of RB4 revealed distinct morphologies and elemental composition of the biominerals produced by microbial interactions with dye molecules. Further, to assess the safety and efficacy of these biominerals, zebrafish embryo toxicity tests were conducted by comparing biominerals produced during dye decolourization with the pure reactive blue 4 dye. Zebrafish embryos treated with biominerals exhibited normal developmental patterns, whereas those exposed to reactive blue 4 displayed mortality and abnormal hatching rates. The heart rates of the Zebrafish embryos were normal for controls and biomineral groups, but lower for RB4-exposed embryos. In conclusion, this study underscores the potential of biomineralization in remediating dye-contaminated wastewater, offering circular solutions for waste reduction and resource utilization.

Sensitivity Variations in Developmental Toxicity of Imidacloprid to Zebrafish Embryos at Different Neurodevelopmental Stages.

Neonicotinoids are ubiquitous in global surface waters and pose a significant risk to aquatic organisms. However, information is lacking on the variations in sensitivity of organisms at different developmental stages to the neurotoxic neonicotinoids. We established a spectrum of toxicity to zebrafish embryos at four neurodevelopmental stages (1, 3, 6, and 8 h post fertilization [hpf]) and dechorionated embryos at 6 hpf based on external and internal exposure to imidacloprid as a representative neonicotinoid. Embryos at the gastrula stage (6 and 8 hpf) were more sensitive to imidacloprid than embryos at earlier developmental stages. Dechorionated embryos were more sensitive to imidacloprid than embryos with a chorion, suggesting that the chorion offers protection against pollutants. Nine sublethal effects were induced by imidacloprid exposure, among which uninflated swim bladder (USB) was the most sensitive. Water depth and air availability in the exposure chambers were critical factors influencing the occurrence of USB in zebrafish larvae. Internal residues of metabolites accounted for <10% of imidacloprid, indicating that imidacloprid was metabolized in a limited fashion in the embryos. In addition, acute toxicity of the main metabolite 5-hydroxy-imidacloprid was significantly lower than that of imidacloprid, indicating that the observed toxicity in embryos exposed to imidacloprid was mainly induced by the parent compound. Our research offers a fresh perspective on choosing the initial exposure time in zebrafish embryo toxicity tests, particularly for neurotoxicants. Environ Toxicol Chem 2024;43:2398-2408. © 2024 SETAC.

Comparative Evaluation of Cytotoxicity of Zinc Oxide Nanoparticles Mixed With Herbal Extract as an Intracanal Medicament: A Zebrafish Model Study.

Objective In this study, zebrafish embryos are used to study the cytotoxic effects of a novel intracanal medication (ICM) based on zinc oxide nanoparticles (ZnO NPs) loaded with polyherbal extracts (Azadirachta indica and Solanum xanthocarpum). Material and methods In the present study, a green and sustainable method was employed for the synthesis of ZnO NPs mixed with bark and seed extracts of Azadirachta indica and Solanum xanthocarpum to be used as a polyherbal ICM. Formulation of ZnO NPs was confirmed with color change in mixture produced upon dissolving zinc acetate dihydrate in distilled water followed by slow addition of sodium hydroxide solution and herbal extracts. The effects of these green synthesized ZnO NPs were evaluated through a zebrafish embryo toxicity test. Embryos were exposed to different concentrations (25, 50, and 100 µg/mL) of synthesized experimental doses of ZnO NP and compared with the control embryos. Toxicological endpoints, such as the zebrafish embryo's survival rate, hatching rate, and heart rate, were noted and described. Results A concentration-dependent increase in mortality rate and hatching delay followed by declined heart rate was observed in green synthesized ZnO NP-treated embryos. The maximum toxicity was observed with an increase in the concentration of 100 µg/mL of the experimental dose, and at a low concentration of 25 µg/mL, it does not effectively show any developmental alteration in zebrafish embryos. Conclusion A novel polyherbal ICM loaded with ZnO NPs exhibited a dose-dependent effect on the heart rate, hatching, and mortality rate of the embryos. At optimal concentrations, the medication demonstrated minimal developmental malformations and cytotoxic effects, indicating its safety for use. However, increasing concentrations of the medication resulted in severe developmental malformations.

Assessing the Efficiency of Microalgae in the Removal of Salicylic Acid from Contaminated Water: Insights from Zebrafish Embryo Toxicity Tests

Microalgae-based water treatments for the removal of different types of pollution have been gaining considerable attention during the last few decades. However, information about microalgae potential in the removal of pharmaceuticals and personal care products (PPCPs) and the ecotoxicological outcomes is still limited. Therefore, in this work, we aimed at investigating salicylic acid removal from water by three different microalgae strains, namely Chlorella sorokiniana, Chlorella vulgaris and Scenedesmus obliquus. For such a purpose, photobioreactors were operated under batch and semi-continuous mode. Apart from determining the reduction in the concentration of salicylic acid attained by each strain, we used zebrafish embryo bioassays to assess the efficiency of microalgae to reduce its toxicity effects. S. obliquus was the strain that achieved the most significant decrease in the concentration and toxic effects of salicylic acid. Indeed, S. obliquus was able to rescue mortality and reduce abnormalities at practically 100%. The efficiency of C. sorokiniana and, especially, that of C. vulgaris were not so remarkable, indicating that the removal of SA and its toxic effects from water by microalgae is markedly strain dependent. The obtained results proved the importance of considering toxic effects for a more comprehensive evaluation of microalgae efficiency in the removal of PPCPs in view of an adequate selection for water treatment.

Toxicology of milrinone by zebrafish embryotoxicity test.

Milrinone, a phosphodiesterase III inhibitor with contractile and vasodilatory effects, is widely used in acute decompensated heart failure and medically refractory end-stage heart failure (HF). The adverse reactions of milrinone have been extensively explored clinically, but its possible toxicities and underlying molecular mechanisms in embryo development need further understanding as its clinical applications increase. Herein, we assessed the milrinone toxicity using the zebrafish embryotoxicity test (ZET), with a view of providing evidence and guidancefor gravidas medicine. We carried out ZET by exposing embryos to a milrinone culture with a series concentration gradients since 1.5 hours post fertilization (hpf) and observed and assessed mortality and hatching rates of drug-treated zebrafishes at 24, 48, 72, and 96 hpf. No significant lethal effect was found in milrinone-treated zebrafish, but hatching rate of eggs at 48 hpf was up-regulated with the increase of milrinone concentration. The impact of milrinone on embryogenesis was assessed through body length, eye area, yolk sac area, swim bladder inflation area, pericardial area and venous congestion area at 96hpf. 150 μg/mL or higher milrinone treatment showed significant effects in the indicators. Organ disorders including enlarged pericardium, liver atrophy and decreased blood vessels were observed in dysplasia individuals versus controls. TUNEL assay suggested the ability of milrinone to induce apoptosis in malformation embryos. Quantitative real-time PCR showed aberrant expressions of transcription factors associated with heartdevelopment and genes related to liver development and apoptosis regulation. Therefore, ZET is feasible for the milrinone toxicity test, and high-dose milrinone causes harm to the embryonic development of zebrafish, especially in embryonic carcinogenesis, vasculogenesis, and hepatogenesis.

A Review on Unlocking Toxicity: How Zebrafish embryos Help Ensure Safe Chemicals

Aquatic ecotoxicity testing has benefited greatly from zebrafish embryo toxicity testing (ZFET) because of its high throughput, low cost, sensitivity to a variety of toxicants, and ethical issues. This study highlights the benefits of ZFET over conventional fish toxicity testing, such as its capacity to detect teratogenic defects and possible long-term impacts. The limitations of ZFET are also addressed, notably its emphasis on acute dosages and difficulties with interspecies extrapolation. In addition to the generation of in vitro alternatives utilizing zebrafish cell lines, recent developments in ZFET technology are also highlighted, including the use of transgenic zebrafish lines and high-throughput screening techniques. The paper's conclusion includes a discussion of potential prospects for ZFET research. These include improving interspecies extrapolation methods, optimizing chronic toxicity assessment procedures, and integrating ZFET with other ecotoxicity testing strategies. ZFET is a great instrument for environmental protection and a crucial component in avoiding chemical contamination of aquatic ecosystems, so long as it is acknowledged for what it is and is open to future improvement

Indigo dyes: Toxicity, teratogenicity, and genotoxicity studies in zebrafish embryos

Wastewater released by textile dyeing industries is a major source of pollution. Untreated wastewater released from indigo dyeing operations affects aquatic ecosystems and threatens their biodiversity. We have assessed the toxicity of natural and synthetic indigo dye in zebrafish embryos, using the endpoints of teratogenicity, genotoxicity, and histopathology. The zebrafish embryo toxicity test (ZFET) was conducted, exposing embryos to ten concentrations of natural and synthetic indigo dyes; the 96-hour LC50 values were approximately 350 and 300 mg/L, respectively. Both dyes were teratogenic, causing egg coagulation, tail detachment, yolk sac edema, pericardial edema, and tail bend, with no significant difference in effects between the natural and synthetic dyes. Both dyes were genotoxic (using comet assay for DNA damage). Real-time RT-PCR studies showed upregulation of the DNA-repair genes FEN1 and ERCC1. Severe histological changes were seen in zebrafish larvae following exposure to the dyes. Our results show that indigo dyes may be teratogenic and genotoxic to aquatic organisms, underscoring the need for development of sustainable practices and policies for mitigating the environmental impacts of textile dyeing.

Integrated mRNA- and miRNA-sequencing analyses unveil the underlying mechanism of tobacco pollutant-induced developmental toxicity in zebrafish embryos

Tobacco pollutants are prevalent in the environment, leading to inadvertent exposure of pregnant females. Studies of these pollutants' toxic effects on embryonic development have not fully elucidated the potential underlying mechanisms. Therefore, in this study, we aimed to investigate the developmental toxicity induced by cigarette smoke extract (CSE) at concentrations of 0.25, 1, and 2.5% using a zebrafish embryo toxicity test and integrated transcriptomic analysis of microRNA (miRNA) and messenger RNA (mRNA). The findings revealed that CSE caused developmental toxicity, including increased mortality and decreased incubation rate, in a dose-dependent manner. Moreover, CSE induced malformations and apoptosis, specifically in the head and heart of zebrafish larvae. We used mRNA and miRNA sequencing analyses to compare changes in the expression of genes and miRNAs in zebrafish larvae. The bioinformatics analysis indicates that the mechanism underlying CSE-induced developmental toxicity was associated with compromised genetic material damage repair, deregulated apoptosis, and disturbed lipid metabolism. The enrichment analysis and RT-qPCR show that the ctsba gene plays a crucial function in embryo developmental apoptosis, and the fads2 gene mainly regulates lipid metabolic toxicity. The results of this study improve the understanding of CSE-induced developmental toxicity in zebrafish embryos and contribute insights into the formulation of novel preventive strategies against tobacco pollutants during early embryonic development.

Synergistic induction of apoptosis in lung cancer cells through co-delivery of PLGA phytol/α-bisabolol nanoparticles.

This study explored the potential of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles to enhance the effectiveness of anticancer treatments through combination therapy with phytol and α-bisabolol. The encapsulation efficiency of the nanoparticles was investigated, highlighting the role of ionic interactions between the drugs and the polymer. Characterization of PLGA-Phy+Bis nanoparticles was carried out using DLS with zeta potential and HR-TEM for size determination. Spectrophotometric measurements evaluated the encapsulation efficiency, loading efficiency, and in vitro drug release. FTIR analysis assessed the chemical interactions between PLGA and the drug actives, ensuring nanoparticle stability. GC-MS was employed to analyze the chemical composition of drug-loaded PLGA nanocarriers. Cytotoxicity was evaluated via the MTT assay, while Annexin V-FITC/PI staining and western blot analysis confirmed apoptotic cell death. Additionally, toxicity tests were performed on L-132 cells and in vivo zebrafish embryos. The study demonstrates high encapsulation efficiency of PLGA-Phy+Bis nanoparticles, which exhibit monodispersity and sizes of 189.3±5nm (DLS) and 268±54 nm (HR-TEM). Spectrophotometric analysis confirmed efficient drug encapsulation and release control. FTIR analysis revealed nanoparticle structural stability without chemical interactions. MTT assay results demonstrated the promising anticancer potential of all thethree nanoparticle types (PLGA-Phy, PLGA-Bis, and PLGA-Phy+Bis) against lung cancer cells. Apoptosis was confirmed through Annexin V-FITC/PI staining and western blot analysis, which also revealed changes in Bax and Bcl-2 protein expression. Furthermore, the nanoparticles exhibited non-toxicity in L-132 cells and zebrafish embryo toxicity tests. PLGA-Phy+Bis nanoparticles exhibited efficient encapsulation, controlled release, and low toxicity. Apoptosis induction in A549 cells and non-toxicity in healthy cells highlight their clinical potential.

Unsymmetrical dimethylhydrazine (UDMH) exposure to zebrafish embryos causes lethal and severe teratogenic effects

The assessment of lethal and teratogenic effects of unsymmetrical dimethylhydrazine (UDMH) using zebrafish embryo toxicity test for 72 h was performed. Following UDMH concentrations were studied for embryotoxic effects: 0.64M, 0.13M, 0.06M, 0.04M, 5x10-3M and 1,7x10-3M after 3 h embryo exposure to UDMH at 7-10 hpf. It was shown that high UDMH concentrations (0,64М, 0,13М, and 0,06М) possess significant embryotoxic action to D.rerio embryos followed by concentration-dependent embryo lysis. 0.04M UDMH and vapors caused severe teratogenic effects and growth retardation prior to hatching resulted in embryo mortality to 48 hpf and 72 hpf, respectively. Zebrafish embryos exposure to low UDMH concentrations of 5x10-3M and 1,7x10-3M caused no significant lethal, but pronounced teratogenic effect (in latter case accumulated from 24 to 72 hpf), with observed malformations of spinal cord and tail tip, yolk sac and pericardial edemas, hypopigmentation. Thus short term 3 h. exposure of D.rerio embryos to UDMH at gastrulation stage induces alterations that further revealed themselves throughout embryonic development causing dose-dependent lethal and teratogenic effects.

Using Electroporation to Improve and Accelerate Zebrafish Embryo Toxicity Testing.

Zebrafish have emerged as a useful model for biomedical research and have been used in environmental toxicology studies. However, the presence of the chorion during the embryo stage limits cellular exposure to toxic elements and creates the possibility of a false-negative or reduced sensitivity in fish embryo toxicity testing (FET). This paper presents the use of electroporation as a technique to improve the delivery of toxic elements inside the chorion, increasing the exposure level of the toxins at an early embryo stage (<3 h post-fertilization). A custom-made electroporation device with the required electrical circuitry has been developed to position embryos between electrodes that provide electrical pulses to expedite the entry of molecules inside the chorion. The optimized parameters facilitate material entering into the chorion without affecting the survival rate of the embryos. The effectiveness of the electroporation system is demonstrated using Trypan blue dye and gold nanoparticles (AuNPs, 20-40 nm). Our results demonstrate the feasibility of controlling the concentration of dye and nanoparticles delivered inside the chorion by optimizing the electrical parameters, including pulse width, pulse number, and amplitude. Next, we tested silver nanoparticles (AgNPs, 10 nm), a commonly used toxin that can lower mortality, affect heart rate, and cause phenotypic defects. We found that electroporation of AgNPs reduces the exposure time required for toxicity testing from 4 days to hours. Electroporation for FET can provide rapid entry of potential toxins into zebrafish embryos, reducing the time required for toxicity testing and drug delivery experiments.

Evaluating the in vitro developmental toxicity potency of a series of petroleum substance extracts using new approach methodologies (NAMs)

The present study evaluates the in vitro developmental toxicity and the possible underlying mode of action of DMSO extracts of a series of highly complex petroleum substances in the mouse embryonic stem cell test (mEST), the zebrafish embryotoxicity test (ZET) and the aryl hydrocarbon receptor reporter gene assay (AhR CALUX assay). Results show that two out of sixteen samples tested, both being poorly refined products that may contain a substantial amount of 3- to 7-ring polycyclic aromatic compounds (PACs), induced sustained AhR activation in the AhR CALUX assay, and concentration-dependent developmental toxicity in both mEST and ZET. The other samples tested, representing highly refined petroleum substances and petroleum-derived waxes (containing typically a very low amount or no PACs at all), were negative in all assays applied, pointing to their inability to induce developmental toxicity in vitro. The refining processes applied during the production of highly refined petroleum products, such as solvent extraction and hydrotreatment which focus on the removal of undesired constituents, including 3- to 7-ring PACs, abolish the in vitro developmental toxicity. In conclusion, the obtained results support the hypothesis that 3- to 7-ring PACs are the primary inducers of the developmental toxicity induced by some (i.e., poorly refined) petroleum substances and that the observed effect is partially AhR-mediated.

The Biocompatibility of a Ginger-Containing Herbal Toothpaste on Developing Zebrafish Embryos

The biocompatibility of toothpaste in an oral cavity should be approved by clinical trials. Nowadays, herbal toothpaste is increasing in popularity due to its natural ingredients. Being genetically similar to humans, zebrafish are used in potential toxicity testing. The zebrafish embryotoxicity test is a fast and straightforward method to study chemical toxicity during embryogenesis. Objective: This study aimed to evaluate if there was any biocompatibility of the toothpaste on zebrafish embryos. Methods: Adult AB strain zebrafish were used according to Institutional Animal Care and Use Committee protocols. Normally dividing, spherical embryos were exposed to herbal toothpaste with ginger (Gumgumix, Turkey) (50 mg/L) and conventional toothpaste (Signal, Expert Protection, Bulgaria) (50 mg/L) in well plates containing 20 embryos, having four replicates. Developmental effects, mortality, and hatching rates were evaluated for 72h. Results: Zebrafish embryos exposed to conventional toothpaste had a higher mortality rate than those exposed to herbal toothpaste; they hatched later and delayed in development. There was no difference between herbal toothpaste and the control group regarding mortality and hatching rates (p &gt; 0.005). Conclusion: The herbal toothpaste showed higher biocompatibility on zebrafish embryos compared to the conventional toothpaste under the condition of this study.

Advancing the use of new approach methodologies for assessing teratogenicity: Building a tiered approach

Many New Approach Methodologies (NAMs) have been developed for the safety assessment of new ingredients. Research into reproductive toxicity and teratogenicity is a particularly high priority, especially given their mechanistic complexity. Forty-six non-teratogenic and 39 teratogenic chemicals were screened for teratogenic potential using the in silico DART model from the OECD QSAR Toolbox; the devTox quickPredict™ (devTox assay) test and the Zebrafish Embryotoxicity Test (ZET). The sensitivity and specificity were 94.7% and 84.1%, respectively, for the DART tree (83 chemicals), 86.1% and 35.6% for the devTox (81 chemicals) and 77.8% and 76.7% for the ZET (57 chemicals). Fifty-three chemicals were tested in all three assays and when results were combined and based on a “2 out of 3 rule”, the sensitivity and specificity were 96.0% and 71.4%, respectively. The specificity of the devTox assay for a sub-set of 43 chemicals was increased from 26.1% to 82.6% by incorporating human plasma concentrations into the assay interpretation. When all 85 chemicals were assessed in a decision tree approach, there was an excellent predictivity and assay robustness of 90%. In conclusion, all three models exhibited a good sensitivity and specificity, especially when outcomes from all three were combined or used in “2 out of 3” or a tiered decision tree approach. The latter is an interesting predictive approach for evaluating the teratogenic potential of new chemicals. Future investigations will extend the number of chemicals tested, as well as explore ways to refine the results and obtain a robust Integrated Testing Strategy to evaluate teratogenic potential.

In Silico and In Vitro Methods in the Characterization of Beta-Carotene as Pharmaceutical Material via Acetylcholine Esterase Inhibitory Actions.

Molecular docking is widely used in the assessment of the therapeutic potential of pharmaceutical agents. The binding properties of beta-carotene (BC) to acetylcholine esterase (AChE) proteins were characterized using the molecular docking method. The mechanism of AChE inhibition was assessed by an experimental in vitro kinetic study. In addition, the role of BC action was tested by the zebrafish embryo toxicity test (ZFET). The results of the docking ability of BC to AChE showed significant ligand binding mode. The kinetic parameter, i.e., the low AICc value shown as the compound was the competitive type of inhibition of AChE. Further, BC also showed mild toxicity at a higher dose (2200 mg/L) in ZFET assessment with changes in biomarkers. The LC50 value of BC is 1811.94 mg/L. Acetylcholine esterase (AChE) plays a pivotal role in the hydrolysis of acetylcholine, which leads to the development of cognitive dysfunction. BC possesses the regulation of acetylcholine esterase (AChE) and acid phosphatase (AP) activity to prevent neurovascular dysfunction. Therefore, the characterization of BC could be used as a pharmaceutical agent for the treatment of cholinergic neurotoxicity-associated neurovascular disorders such as developmental toxicity, vascular dementia, and Alzheimer's disease due to its AChE and AP inhibitory actions.

A metabolomics approach to reveal the mechanism of developmental toxicity in zebrafish embryos exposed to 6-propyl-2-thiouracil

A crucial component of a substance registration and regulation is the evaluation of human prenatal developmental toxicity. Current toxicological tests are based on mammalian models, but these are costly, time consuming and may pose ethical concerns. The zebrafish embryo has evolved as a promising alternative model to study developmental toxicity. However, the implementation of the zebrafish embryotoxicity test is challenged by lacking information on the relevance of observed morphological alterations in fish for human developmental toxicity. Elucidating the mechanism of toxicity could help to overcome this limitation. Through LC-MS/MS and GC-MS metabolomics, we investigated whether changes to the endogenous metabolites can indicate pathways associated with developmental toxicity. To this aim, zebrafish embryos were exposed to different concentrations of 6-propyl-2-thiouracil (PTU), a compound known to induce developmental toxicity. The reproducibility and the concentration-dependence of the metabolome response and its association with morphological alterations were studied. Major morphological findings were reduced eye size, and other craniofacial anomalies; major metabolic changes included increased tyrosine, pipecolic acid and lysophosphatidylcholine levels, decreased methionine levels, and disturbance of the ‘Phenylalanine, tyrosine and tryptophan biosynthesis’ pathway. This pathway, and the changes in tyrosine and pipecolic acid levels could be linked to the mode of action of PTU, i.e., inhibition of thyroid peroxidase (TPO). The other findings suggested neurodevelopmental impairments. This proof-of-concept study demonstrated that metabolite changes in zebrafish embryos are robust and provide mechanistic information associated with the mode of action of PTU.

Green synthesis of photocatalytic TiO2/Ag nanoparticles for an efficient water remediation

This work reports a green method to produce efficient photocatalytic nanopowders. Titanium dioxide nanoparticles were easily synthetized by an extract of Thymus vulgaris, and then decorated with commercial silver nanoparticles. A detailed morphological, structural, and chemical characterization was performed by scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) adsorption–desorption of N2, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), Raman spectroscopy, transmission electron microscopy (TEM), and UV–Vis spectroscopy. The synthetized TiO2 nanoparticles showed a mean size of 10–15 nm, and had the anatase crystallographic phase. The photocatalytic efficiency of the TiO2 nanoparticles was demonstrated by the photo-degradation under UV-light irradiation of different pollutants in aqueous solution (methylene blue, diclofenac, and sodium dodecyl sulfate). TiO2 was coupled to Ag nanoparticles with different percentage in weight (from 0.25 to 3%), to investigate an eventual improvement of its photocatalytic efficiency. The best photocatalytic composite among the tested compositions was the one with the lowest Ag percentage (0.25%). Zebrafish embryo toxicity tests ruled out the toxicity of the synthetized nanoparticles. The present results offer a new, green, and easy method to prepare TiO2/Ag nanoparticles with high photocatalytic efficiency. The proposed materials are therefore promising for applications in photocatalysis, especially for wastewater remediation and reuse.

Intergrading Water Quality Parameters, Benthic Fauna and Acute Toxicity Test for Risk Assessment on an Urban-Rural River

Climate change, river pollution and loss of biodiversity are increasing and becoming global environmental concerns. The Yellow River is China’s mother river, providing water for about 114 million residents in towns and cities along its route. Yet in 2012, the Yellow River received 4.474 × 109 tons of sewage containing a large number of exogenous pollutants, posing a huge ecological and public health threat. Water quality safety is not only a matter of ecosystem health but also of human survival and social development. Therefore, the effects of pollutants on water quality safety should be carefully studied, which is important to ensure the sustainability of the Yellow River and the surrounding cities and towns. In this study, water and sediment samples from the Jishan River of the Juancheng, a typical city in the lower reaches of the Yellow River, were collected and evaluated by integrating the traditional physicochemical water indicators, benthic Index of Biological Integrity (B-IBI) and zebrafish embryotoxicity test. The results showed that water dissolved oxygen, pH, total nitrogen and total phosphorus were strongly correlated with zebrafish embryonic teratogenicity, lethality, abundance index and Shannon Winner index. A total of 21 benthic species were collected, including mollusks, arthropods and annelids, with the upper reaches having the largest biomass and B-IBI values indicating the urban reaches have better biological integrity than rural reaches. The teratogenic rate of zebrafish embryos in raw water was greatest in rural rivers and was significantly different from the negative control (p &lt; 0.05). When exposed to 100 mg/mL sediment samples, embryo hatching rates were inversely correlated with teratogenic rates, with lethal rates reaching over 96% in all rural reaches. The results showed that the water quality safety at the rural farms in the Yellow River transfer-type towns is poor, and they especially recommended that the river near the farms in rural areas should be monitored with emphasis.

The role of receptor-mediated activities of 4- and 5-ring unsubstituted and methylated polycyclic aromatic hydrocarbons (PAHs) in developmental toxicity.

The present study evaluated the aryl hydrocarbon receptor (AhR), estrogen receptor-α (ER-α), and retinoic acid receptor (RAR) mediated activities of nine 4- and 5-ring unsubstituted and monomethylated polycyclic aromatic hydrocarbons (PAHs) using a series of Chemical-Activated LUciferase gene eXpression (CALUX) assays. The potential role of these aforementioned receptors in relation to the developmental toxicity of these PAHs was further assessed in the zebrafish embryotoxicity test (ZET). The results show that all nine tested PAHs were AhR agonists, benz[a]anthracene (BaA) and 8-methyl-benz[a]anthracene (8-MeBaA) were ER-α agonists, and none of the tested PAHs induced ER-α antagonistic or RAR (ant)agonistic activities. In the AhR CALUX assay, all the methylated PAHs showed higher potency (lower EC50) in activating the AhR than their respective unsubstituted PAHs, implying that the addition of a methyl substituent on the aromatic ring of PAHs could enhance their AhR-mediated activities. Co-exposure of zebrafish embryos with each individual PAH and an AhR antagonist (CH223191) counteracted the observed developmental retardations and embryo lethality to a certain extent, except for 8-methyl-benzo[a]pyrene (8-MeBaP). Co-exposure of zebrafish embryos with either of the two estrogenic PAHs (i.e., BaA and 8-MeBaA) and an ER-α antagonist (fulvestrant) neutralized embryo lethality induced by 50 μM BaA and the developmental retardations induced by 15 μM 8-MeBaA. Altogether, our findings suggest that the observed developmental retardations in zebrafish embryos by the PAH tested may partially be AhR- and/or ER-α-mediated, whereas the RAR seems not to be relevant for the PAH-induced developmental toxicity in the ZET.