The real-time textile dyes wastewater contains hazardous and recalcitrant chemicals that are difficult to degrade by conventional methods. Such pollutants, when released without proper treatment into the environment, impact water quality and usage. Hence, the textile dye effluent is considered a severe environmental pollutant. It contains mixed contaminants like dyes, sodium bicarbonate, acetic acid. The physico-chemical treatment of these wastewaters produces a large amount of sludge and costly. Acceptance of technology by the industry mandates that it should be efficient, cost-effective and the treated water is safe for reuse. A sequential anaerobic-aerobic plant-microbe system with acclimatized microorganisms and vetiver plants, was evaluated at a pilot-scale on-site. At the end of the sequential process, decolorization and total aromatic amine (TAA) removal were 78.8% and 69.2% respectively. Analysis of the treated water at various stages using Fourier Transform Infrared (FTIR), High Performance Liquid Chromatography (HPLC)) Gas Chromatography-Mass Spectrometry (GC-MS) Liquid Chromatography-Mass Spectrometry (LC-MS) indicated that the dyes were decolourized and the aromatic amine intermediates formed were degraded to give aliphatic compounds. Scanning Electron Microscope (SEM) and Atomic Force Microscopy (AFM) analysis showed interaction of microbe with the roots of vetiver plants. Toxicity analysis with zebrafish indicated the removal of toxins and teratogens.

Due to technical limitations, there have been minimal studies performed on thermal preferences and thermotactic behaviors of aquatic ectotherm species commonly used in ecotoxicity testing. In this work, we demonstrate an innovative, purpose-built and miniaturized electrothermal array for rapid thermal preference behavioral tests. We applied the novel platform to define thermal preferences in multiple invertebrate and vertebrate species. Specifically, Dugesia notogaea (freshwater planarians), Chironomus tepperi (nonbiting midge larvae), Ostracoda (seed shrimp), Artemia franciscana (brine shrimp), Daphnia carinata (water flea), Austrochiltonia subtenuis (freshwater amphipod), Physa acuta (freshwater snail), Potamopyrgus antipodarum (New Zealand mud snail) and larval stage of Danio rerio (zebrafish) were tested. The Australian freshwater water fleas, amphipods, snail Physa acuta as well as zebrafish exhibited the most consistent preference to cool zones and clear avoidance of zones >27 °C out of nine species tested. Our results indicate the larval stage of zebrafish as the most responsive species highly suitable for prospective development of multidimensional behavioral test batteries. We also showcase preliminary data that environmentally relevant concentrations of pharmaceutical pollutants such as non-steroidal anti-inflammatory drug (NSAID) ibuprofen (9800 ng/L) and insecticide imidacloprid (4600 ng/L) but not anti-depressant venlafaxine (2200 ng/L) and (iv) anticonvulsant medications gabapentin (400 ng/L) can perturb thermal preference behavior of larval zebrafish. Collectively our results demonstrate the utility of simple and inexpensive thermoelectric technology in rapid exploration of thermal preference in diverse species of aquatic animals. We postulate that more broadly such technologies can also have added value in ecotoxicity testing of emerging contaminants.

We examined ingestion and retention rates of microplastics (MPs) by two freshwater (Japanese medaka and zebrafish) and two marine fish species (Indian medaka and clown anemonefish) to determine their color preferences and gastrointestinal-tract retention times. In our ingestion experiments, clown anemonefish ingested the most MP particles, followed by zebrafish, and then Japanese and Indian medaka. Next, we investigated color preferences among five MP colors. Red, yellow, and green MP were ingested at higher rates than gray and blue MPs for all tested fish species. To test whether these differences truly reflect a recognition of and preference for certain colors based on color vision, we investigated the preferences of clown anemonefish for MP colors under light and dark conditions. Under dark conditions, ingestion of MP particles was reduced, and color preferences were not observed. Finally, we assessed gastrointestinal-tract retention times for all four fish species. Some individuals retained MP particles in their gastrointestinal tracts for over 24 h after ingestion. Our results show that fish rely on color vision to recognize and express preferences for certain MP colors. In addition, MP excretion times varied widely among individuals. Our results provide new insights into accidental MP ingestion by fishes.

The beta-blocker atenolol (ATE), and the selective serotonin and norepinephrine reuptake inhibitor, venlafaxine (VEN) are frequently detected in municipal wastewater effluents, but little is known about their ecotoxicological effect on aquatic animals. Herein, ATE and VEN were selected to explore their accumulation and global DNA methylation (GDM) in zebrafish tissues after a 30-day exposure. Molecular dynamics (MD) stimulation was used to investigate the toxic mechanism of ATE and VEN exposure. The results demonstrated that ATE and VEN could reduce the condition factor of zebrafish. The bioaccumulation capacity for ATE and VEN was in the order of liver > gut > gill > brain and liver > gut > brain > gill, respectively. After a 30-day recovery, ATE and VEN could still be detected in zebrafish tissues when exposure concentrations were ≥10 μg/L. Moreover, ATE and VEN induced global DNA hypomethylation in different tissues with a dose-dependent manner and their main target tissues were liver and brain. When the exposure concentrations of ATE and VEN were increased to 100 μg/L, the global DNA hypomethylation of liver and brain were reduced to 27% and 18%, respectively. In the same tissue exposed to the same concentration, DNA hypomethylation induced by VEN was more serious than that of ATE. After a 30-day recovery, the global DNA hypomethylations caused by the two drugs were still persistent, and the recovery of VEN was slower than that of ATE. The MD simulation results showed that both ATE and VEN could reduce the catalytic activity of DNA Methyltransferase 1 (DNMT1), while the effect of VEN on the 3D conformational changes of the DNMT1 domain was more significant, resulting in a lower DNA methylation rate. The current study shed new light on the toxic mechanism and potential adverse impacts of ATE and VEN on zebrafish, providing essential information to the further ecotoxicological risk assessment of these drugs in the aquatic environment.

Various microplastics (MPs) are found in the environment and organisms. MP residues in organisms can affect health; however, their impacts on metabolism in aquatic organisms remain unclear. In this study, zebrafish embryos were exposed to polyethylene MPs with sizes ranging from 1 to 4 μm at concentrations of 0, 10, 100, and 1000 μg/L for 7 days. Through qPCR technology, the results indicated that zebrafish exposed to polyethylene MPs exhibited significant change in microbes of the phyla Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobia, etc. Moreover, 16S RNA gene sequencing revealed that there was a significant difference in alpha diversity between the control and 1000 μg/L MP-treated groups. At the genus level, the abundance of Aeromonas, Shewanella, Microbacterium, Nevskia and Methyloversatilis have increased remarkably. Conversely, the abundance of Pseudomonas, Ralstonia and Stenotrophomonas were significant reduction after MPs exposure. In addition, the levels of TG (triglyceride), TCHO (total cholesterol), NEFA (nonesterified fatty acid), TBA (total bile acid), GLU (glucose) and pyruvic acid significantly changed in MP-treated larval zebrafish, indicating that their metabolism was disturbed by MPs. Transcriptional levels of glucose and lipid metabolism-related genes showed a decreasing trend. Furthermore, LC/MS-based nontargeted metabolomics analysis demonstrated that a total of 59 phospholipid-related substances exhibited significant changes in larval fish treated with 1000 μg/L MPs. The mRNA levels of phospholipid metabolism-related genes were also obviously changed. Pearson correlation analysis indicated that the abundance of Aeromonas, Shewanella and Chitinibacter bacteria showed a negative correlation with most phospholipids, while Nevskia, Parvibacter and Lysobacter showed a positive correlation with most phospholipids. Based on these results, it is suggested that 1–4 μm PE-MPs could impact the microbiome and metabolism of larval zebrafish. All of these results indicated that the health risk of MPs cannot be ignored.

In modern agricultural management, the use of pesticides is indispensable. Due to their massive use worldwide, pesticides represent a latent risk to both humans and the environment. In the present study, 1056 frequently used pesticides were screened for oestrogen receptor (ER) agonistic activity by using in silico methods. We found that 72 and 47 pesticides potentially have ER agonistic activity by the machine learning methods random forest (RF) and deep neural network (DNN), respectively. Among endocrine-disrupting chemicals (EDCs), 14 have been reported as EDCs or ER agonists by previous studies. We selected 3 reported and 7 previously unreported pesticides from 76 potential ER agonists to further assess ERα agonistic activity. All 10 selected pesticides exhibited ERα agonistic activity in human cells or zebrafish. In the dual-luciferase reporter gene assays, six pesticides exhibited ERα agonistic activity. Additionally, nine pesticides could induce mRNA expression of the pS2 and NRF1 genes in MCF-7 cells, and seven pesticides could induce mRNA expression of the vtg1 and vtg2 genes in zebrafish. Importantly, the remaining 48 out of 76 potential ER agonists, none of which have previously been reported to have endocrine-disrupting effects or oestrogenic activity, should be of great concern. Our screening results can inform environmental protection goals and play an important role in environmental protection and early warnings to human health.

In toxicology, standard sigmoidal concentration-response curves are used to predict effects concentrations and set chemical regulations. However, current literature also establishes the existence of complex, bimodal concentration-response curves, as is the case for arsenic toxicity. This bimodal response has been observed at the molecular level, but not characterized at the whole organism level. This study investigated the effect of arsenic (sodium arsenite) on post-gastrulated zebrafish embryos and elucidated effects of bimodal concentration-responses on different phenotypic perturbations.Six hour post fertilized (hpf) zebrafish embryos were exposed to arsenic to 96 hpf. Hatching success, mortality, and morphometric endpoints were evaluated both in embryos with chorions and dechorionated embryos. Zebrafish embryos exhibited a bimodal response to arsenic exposure. Concentration-response curves for exposed embryos with intact chorions had an initial peak in mortality (88%) at 1.33 mM arsenic, followed by a decrease in toxicity (~20% mortality) at 1.75 mM, and subsequently peaked to 100% mortality at higher concentrations. To account for the bimodal response, two distinct concentration-response curves were generated with estimated LC10 values (and 95% CI) of 0.462 (0.415, 0.508) mM and 1.69 (1.58, 1.78) mM for the ‘low concentration’ and ‘high concentration’ peaks, respectively. Other phenotypic analyses, including embryo length, yolk and pericardial edema all produced similar concentration-response patterns. Tests with dechorionated embryos also resulted in a bimodal toxicity response but with lower LC10 values of 0.170 (0.120, 0.220) mM and 0.800 (0.60, 0842) mM, respectively. Similarities in bimodal concentration-responses between with-chorion and dechorionated embryos indicate that the observed effect was not caused by the chorion limiting arsenic availability, thus lending support to other studies such as those that hypothesized a conserved bimodal mechanism of arsenic interference with nuclear receptor activation.

Azoxystrobin (AZ) and pyraclostrobin (PY) are strobilurin fungicides that inhibit fungal mitochondrial respiration. In this study, a representative model, zebrafish (Danio rerio), was used as a test species for acute and developmental toxicity. Survival and malformation rates were observed only PY-treated embryos, with an LC₅₀ value of 77.75 ppb accompanied by a dramatic decrease in hatching rate, while AZ did not show great mortality. Morphological changes were observed in PY-treated embryos with the occurrence of pericadial edema at 25 ppb. A delay in growth was observed after treatment with pyraclostrobin at 50 ppb. Use of genetically engineered Tg(cmlc:EGFP) allowed fluorescence observation during heart development. PY interfered with normal heart development via upregulation of the nppa gene responsible for the expression of natriuretic peptides. Heart function was dramatically reduced as indicated by reduced heart rates. Increased expression of the nppa gene was also seen in AZ-treated embryos. The expression level of cyp24a1 was also up-regulated, while ugt1a1 and sult1st6 were down-regulated after treatment of zebrafish embryos with AZ or PY. Overall, strobilurin fungicides might inhibit normal heart formation and function within the range of concentrations tested.

Despite publication of numerous of papers, the effects of fluoxetine on fish behaviour remains mired in controversy and contradiction. One reason for this controversy is that fluoxetine displays distinct and opposing acute and chronic effects. A second reason is that most studies have been limited to two or at the most three concentrations. To address these deficiencies we exposed adult zebrafish, both single females and shoals consisting of one male and two females, to seven fluoxetine concentrations, ranging from 5 ng/L to 5 μg/L and measured their swimming behaviour, and response to a conspecific alarm substance (CAS) at seven, 14 and 28 days. We also measured the light startle response of unexposed F1 larvae at days seven and 28 post-hatch and the response to CAS at day 28. On day 7 fluoxetine decreased swimming speed at concentrations ≥500 ng/L. After addition of CAS fish exposed to 5, 500 and 1000 ng/L decreased swimming, while fish exposed to 10, 500 and 1000 ng/L significantly increased time motionless. On day 14 only fish exposed to 50 ng/L were significantly slower than controls before addition of CAS, but afterwards fish exposed to 5, 50, 1000 and 5000 ng/L showed significant differences from controls. On day 28 fish exposed to 50 and 5000 ng/L had slower average swimming speeds than controls before addition of CAS. After addition all fish except controls and those exposed to 500 ng/L showed decreased average speed. At seven days post-hatch, F1 larvae whose parents were exposed to 100 ng/L showed significantly higher activity than controls and those exposed to 500 ng/L fluoxetine showed lower activity in the light startle response. This study shows that the effects of fluoxetine vary with time and also in a non-monotonic manner. We suggest that the complex nature of the serotonergic system with multilateral effects at the genomic, biochemical and physiological levels interacting with environmental stimuli result in non-linear dose-response behavioural patterns.

It is necessary to understand the interactions between different pesticides in ecotoxicology because pesticides never appear as individual compounds but rather in combinations with other compounds. In this study, we planned to explicate the combined toxic effect of myclobutanil (MYC) and thiamethoxam (THI) on the zebrafish (Danio rerio) by adopting multiple biomarkers. Results unraveled that the 96-h LC₅₀ values of MYC to D. rerio at various life phases ranged from 5.2 to 10.3 mg L⁻¹, which were lower than those of THI ranging from 147 to 246 mg L⁻¹. Combinations of MYC and THI exhibited synergetic toxicity to zebrafish embryos. The activities of antioxidative enzymes (T-SOD, Cu/Zn-SOD and POD) and detoxification enzyme (GST) were obviously varied in most of the MYC, THI and combined exposures compared to the control. The mRNA expressions of eight genes (Cu-sod, cas3, il-8, cxcl, erα, crh, cyp17 and dio1) involved in antioxidation, apoptosis, immunity and endocrine were obviously altered in the combined exposure of MYC and THI compared to their individual exposures. Our findings hinted the threats when YMC and THI co-existed, which would be beneficial for the risk assessments of pesticide mixtures.