IR-Based Novel Device for Real-Time Online Acquisition of Fish Heart ECG Signals.

We developed an infrared (IR)-based real-time online monitoring device (US Patent No: US 10,571,448 B2) to quantify heart electrocardiogram (ECG) signals to assess the water quality based on physiological changes in fish. The device is compact, allowing us to monitor cardiac function for an extended period (from 7 to 30 days depending on the rechargeable battery capacity) without function injury and disturbance of swimming activity. The electrode samples and the biopotential amplifier and microcontroller process the cardiac-electrical signals. An infrared transceiver transmits denoised electrocardiac signals to complete the signal transmission. The infrared receiver array and biomedical acquisition signal processing system send signals to the computer. The software in the computer processes the data in real time. We quantified ECG indexes (P-wave, Q-wave, R-wave, S-wave, T-wave, PR-interval, QRS-complex, and QT-interval) of carp precisely and incessantly under the different experimental setup (CuSO4 and deltamethrin). The ECG cue responses were chemical-specific based on CuSO4 and deltamethrin exposures. This study provides an additional technology for noninvasive water quality surveillance.

The acetylcholinesterase (AChE) inhibition analysis of medaka (Oryzias latipes) in the exposure of three insecticides.

The continuous effects on Acetylcholinesterase (AChE) activity of medaka (Oryzias latipes) caused by dichlorvos, methomyl and deltamethrin in vivo were investigated, and the trends of AChE activity inhibition due to the influence of these insecticides were discussed. The LC50-24h of dichlorvos, methomyl and deltamethrin on medaka were 2.3 mg/L, 0.2 mg/L, and 2.9×10(-3) mg/L respectively. The result suggested that at the beginning of the exposure, the AChE activity might increase, and the AChE activity in dead individuals was obviously lower than the live individuals. Though the de novo synthesis of AChE in medaka might help the AChE activity recover, the trends during the exposure in different treatments were downward, and it showed both exposure time and concentration dependent. Meanwhile, higher temperature might cause the AChE inhibition earlier due to the higher metabolic rate. Therefore, as a specific biomarker for organophosphate, carbamate pesticides and pyrethroids, the degree of the AChE inhibition with in vivo conditions is a good tool in continuous monitoring of insecticides, which may induce the nerve conduction disorders.

The specification of zebrafish (Danio rerio) heart electrocardiogram index characteristic responses to different types of pollutants.

Water quality was highly affected by common pollutants. Metals, pesticides and small molecules are ubiquitous pollutants. Advancement in engineering technology (computer-based monitoring systems) increased the efficiency of quantifying toxicity of different chemicals in an organism. The cardiovascular system reflects internal and external stress of an organism, and electrocardiogram (ECG) data reliably measure external stress. As ECG data can accurately reflect the physiological conditions of organisms, and zebrafish (Danio rerio) are considered to be good models for cardiovascular research, it is hypothesized that ECG parameters of zebrafish could indicate the toxicity of water-borne chemicals. To achieve this, we treated zebrafish with different concentrations of target chemicals (CuSO4, C10H19O6PS2 and NH4Cl) for 48 h and ECG data were measured. P-wave, R-wave, T-wave, PR-interval, QRS-complex and QT-interval data were the focus of this study. The results of self-organizing maps and Pearson correlation analysis indicate that the QRS-complex can be used as an indicator for CuSO4 stress. The QT-interval could be used to assess the C10H19O6PS2 stress. The QT-interval and P-wave can be used to evaluate the NH4Cl stress. Responses of zebrafish ECG parameters were identical with other vertebrate model, and were specific to toxicant types. It is proved that zebrafish heart ECG index could be used as a potential indicator in early detection of environmental stress.

Synthesis of Nano-Fe@NdFeB/AC magnetic catalytic particle electrodes and application in the degradation of 2,4,6-trichlorophenol by electro-assisted peroxydisulfate process.

The coupling of electrolysis and the peroxydisulfate (PDS) activation was selected in this study to degrade solution-phase 2,4,6-trichlorophenol (TCP). To enhance the PDS activation efficiency and catalytic recycling ratio, a novel magnetic activator, nano iron coated on neodymium iron boron/activated carbon nanocomposite (Nano-Fe@NdFeB/AC), was synthesized and utilized as catalytic particle electrodes. To increase the mass transfer ability, a novel magnetic internal circulation electrolytic reactor (MICE) was established. The results indicated that globular Fe, with sizes ranging from 25 nm to 300 nm, is present on the surface of the catalyst. This catalyst has sufficient magnetism to be separated by the magnetic separation method and its specific saturation magnetization and residual magnetization were 1.48 and 0.26 emu/g, respectively. At the optimal condition of [pH]0 = 9.0, [Na2S2O8]0 = 2.0 mmol/L, [Nano-Fe@NdFeB/AC]0 = 5.0 g/L and I = 50 mA, the TOC percentage of removal could reach 84% after 30 min of reaction. The TCP mineralization follows pseudo-first-order kinetics. The intermediate products of 2,6-dichloro-2,5-cyclohexadiene-1,4-dione, Tetrachloro-hydroquinone, and 2,3,5,6-tetrachloro-p-benzoquinone were found during the reaction. TCP mineralization was confirmed to have a hybrid mechanism involving reductive dechlorination with Fe, •OH addition oxidation and electron capture by SO•4 -. This study provides a new method for the treatment of degradation-resistant pollutants.

Fabrication and characterization of the magnetic separation photocatalyst C-TiO2@Fe3O4/AC with enhanced photocatalytic performance under visible light irradiation.

Congo red (CR) is a kind of refractory contaminant. Conventional biological treatment processes are ineffective for the CR degradation. However, photocatalysis technology could be an alternative for the decomposition of CR because of its high efficiency. In this study, we prepared three kinds of photocatalysts, all of which have advanced visible light excitation characteristics. The magnetic catalyst C-TiO2@Fe3O4/AC was produced by loading C-TiO2 and nano-Fe3O4 onto granular activated carbon (AC). C-TiO2@Fe3O4/AC has a band gap of 2.535 eV, stable magnetic characteristics, and stability toward CR removal. C-TiO2@Fe3O4/AC showed the best performance for CR removal under both simulated sunlight (200-800 nm) and visible light (400-800 nm) irradiation compared to the other catalysts. The CR removal rate reached 92.9% after 30 min of simulated sunlight irradiation, and the reaction rate constant was 0.1776 min-1. Under visible light, the CR removal rate reached 65%. The hydroxyl radical (OH) was detected, and its concentration was determined. Furthermore, the spectral analysis results indicated that the azo bonds and aromatic rings in CR were destroyed. The C-TiO2@Fe3O4/AC has a self-cleaning ability to prevent organic contamination. This study provides a new way of thinking and a simple preparation technique for magnetic visible light catalyst synthesis.

The continuous physiological changes of zebrafish (Danio rerio) based on metabolism under controlled thallium stress.

In this research, the continuous physiological changes of zebrafish (Danio rerio) in 0.1 µg/L thallium (Tl) in 15 days were investigated. The results showed that Tl(I) stress had a significant positive linear correlation with zebrafish ammonia nitrogen excretion (ANE) (p < 0.001), and the mean value of ANE in Tl(I) treatment (435 ±â€¯227 mg/kg/h) was approximately 2 times higher than in the control group (239 ±â€¯168 mg/kg/h), which suggested that ANE was suitable for Tl(I) stress assessment. A substantial difference based on oxygen consumption rate (OCR) between the control group (587 ±â€¯112 mg/kg/h) and Tl(I) treatment (260 ±â€¯88 mg/kg/h) with a high significance p < 0.001 could be observed, and the results indicated that Tl(I) played a negative role in OCR of zebrafish. The characteristics of both ANE and OCR changes under slight Tl(I) stress could be reflected by the ammonia quotient (AQ). It was noteworthy that AQ increased rapidly in first 6 h from 0.66 to 4.50, which was 3 times higher than 1.2, indicating rapid increase in both anaerobic energy utilization and protein metabolism in 0.1 µg/L Tl(I) exposure. It is concluded that the physiological changes of zebrafish based on metabolism can be regarded as a sensitive biological indicator of Tl(I) pollution, which could work as a substitute of potassium that disrupts the normal biological metabolism in the process of transport.

Parental exposure to tris(1,3-dichloro-2-propyl) phosphate results in thyroid endocrine disruption and inhibition of growth in zebrafish offspring.

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is a re-emerging environmental contaminant used as a suitable substitute for brominated flame retardants. The objective of this study was to evaluate the effects of TDCIPP on thyroid disruption and growth inhibition in zebrafish (Danio rerio) offspring after chronic parental exposure, and to examine the possible molecular mechanisms involved. When adult zebrafish (4 months old) were exposed to 5.66, 25.55, or 92.8 µg TDCIPP/L for 90 days, bioconcentration of TDCIPP and its metabolic product [bis(1,3-dichloro-2-propyl) phosphate, BDCIPP] was observed in 7-day postfertilization (dpf) F1 larvae, which suggests the transfer of this compound from adult fish to their offspring. Our results demonstrated that parental exposure to TDCIPP induced thyroid disruption in the offspring, demonstrated by significantly decreased thyroxine (T4) and increased 3,5,3'-triiodothyronine (T3) levels, and disruption of the transcription of several genes and expression of proteins involved in the hypothalamic-pituitary-thyroid (HPT) axis in F1 larvae. Parental exposure to TDCIPP resulted in developmental abnormalities in offspring; the smaller body length that was recorded might be partly the result of the perturbation of the HPT axis. In addition, the results revealed that growth inhibition also resulted from the downregulation of the transcription of genes and expression of proteins involved in the growth hormone/insulin-like growth factor (GH/IGF) axis. Our study provides a new set of evidence showing that parental exposure to TDCIPP can induce thyroid disruption and inhibition of growth in offspring, and that perturbation of the HPT axis and GH/IGF axis contribute to these adverse effects.

The online monitoring and assessment of thallium stress using oxygen consumption rate and carbon dioxide excretion rate of zebrafish (Danio rerio).

An online monitoring and assessment system of metabolism to measure oxygen consumption rate (OC) and carbon dioxide (CO2) excretion rate (CR) of zebrafish (Danio rerio) was used to illustrate changes in stressful states in 15 days' (360 h) 0.1 µg/L Tl exposure. Tl had a significant inhibition on zebrafish OC and CR (p < 0.01). OC was more suitable for Tl stress assessment than CR, considering that the OC response was more stable and discernible from the control comparing with CR. However, CR is a suitable alternative to characterize toxic effects on different metabolic substrates. Both OC and CR were integrated to present the respiratory quotient (RQ) analysis. RQ was efficient in differentiating between CO2 produced by respiration in the control group (RQ less than 0.7) and CO2 used for urination or stored in tissues after Tl exposure (some RQs larger than 1.0). Circadian rhythm was observed in RC and CR in the controls and persisted in 0.1 µg/L Tl treatments. The rhythm was relatively more disordered in CR. OC and CR would be suitable for indicating physiological stress in the online system as sensitive physiological indices.

Behavior responses of zebrafish (Danio rerio) to aquatic environmental stresses in the characteristic of circadian rhythms.

As behavior shows a distinct circadian rhythm, it is hypothesized that circadian rhythms based on zebrafish (Danio rerio) behavior responses could be affected by contaminants in this study, and then the behavior strength of zebrafish exposed to 0.005 mg/L Cadmium chloride (CdCl2), 0.01 mg/L Dibasic Sodium Phosphate (Na2HPO4), 0.002 mg/L deltamethrin, and 0.003 mg/L atrazine for 6 days is used to illustrate the possibility of behavior circadian rhythms as an indicator in the environmental stress assessment. Statistical analysis with p < 0.01 shows that a clear difference between average values of BS during dark period (AVD) and those during light period (AVL) could be observed, and 24 h circadian rhythms do exist in zebrafish behavior responses. Both BS values and circadian rhythms of zebrafish can be affected in the aspect of periodicity with clear time delay, which were 1 h delay in CdCl2, 4 h delay in Na2HPO4, 4 h delay in deltamethrin, and 1 h delay in atrazine. Behavior circadian rhythms were disturbed according to the repetitive cycles after autocorrelation analysis, and the toxic effects of different chemicals could be reflected by the profiles of the Self-Organizing Map (SOM), which indicated the circadian rhythm disorder in different degrees. These results deduced from the statistical analysis, autocorrelation and SOM strongly supported that circadian rhythms based on zebrafish BS could be used as an indicator in the environmental stress assessment.

A novel fluorescent functional monomer as the recognition element in core-shell imprinted sensors responding to concentration of 2,4,6-trichlorophenol.

We have demonstrated a fluorescent functional monomer instead of the traditional functional monomers for molecularly imprinted sensors. The sensors were firstly used to selectively detect 2,4,6-trichlorophenol (2,4,6-TCP) by solid fluorescence detection without a dispersion solution. Moreover, the selectivity and anti-interference ability of the SiO2@dye-FMIPs sensor meet the requirements of a fluorescent sensor. The novel fluorescent monomer introduced into MIP is no longer just a fluorophore without recognizing ability. The fluorescence intensity of SiO2@dye-FMIPs showed a linear response to 2,4,6-TCP concentration in the range of 0-100 nM with a detection limit of 0.0534 nM. We could also demonstrate that such a system can not only get rid of the confines of traditional functional monomers and detection manner, but also improved the applications of MIPs sensors in sensing systems.

Toxic responses of zebrafish (Danio rerio) to thallium and deltamethrin characterized in the electrocardiogram.

The electrocardiogram (ECG) has been widely used to objectively address the physical condition as an index of stress. Though a numerous accounts of investigations on aquatic organisms' ECGs have been made, differentiation of ECG parameters in responding to specific toxic chemicals has not been extensively studied. In this research, it is hypothesized that zebrafish (Danio rerio) ECG parameters would differently respond to different types of pollutants, a heavy metal, thallium (Tl, 0.10 and 13.00 µg/L) and an organic chemical, deltamethrin (DM, 0.52 and 2.00 µg/L). Based on the SOM training and statistical analyses, QRS complex could be specified as an indicator of Tl stress, while QT interval might be used to evaluate DM stress. Pearson correlation analysis indicated that QRS complex and QT interval were significantly associated with Tl stress (r = 0.854, p = 0.0002) and DM stress (r = 0.792, p = 0.001), respectively. QRS complex and QT interval had the highest R2, the minimum of SSE and the lowest AIC value in Tl and DM treatments, respectively. Bases on the current experimental results and previous reports, QRS complex and QT interval could be considered as a specific indicator of Tl and DM disturbances in the environment, respectively.

Zinc oxide nanoparticles induce oxidative DNA damage and ROS-triggered mitochondria-mediated apoptosis in zebrafish embryos.

Zinc oxide nanoparticles (nano-ZnO) are one of the most important nanoparticles in the industry. The objectives of this study were (1) to investigate the effects of nano-ZnO on oxidative damage to DNA and on apoptosis in zebrafish (Danio rerio) embryos, and (2) to identify the underlying molecular mechanism affecting theapoptotic process. In addition to nano-ZnO, we also investigated the toxic effects of the Zn2+ ion. Zebrafish embryos were exposed to 10, 30, 60, 90, or 120mg/L nano-ZnO for 96h postfertilization. Nano-ZnO (at concentrations between 10 and 120mg/L) significantly reduced the rate of embryo hatching. Embryos/larvae exposed to 120mg/L nano-ZnO had significantly higher heart rates. Increased heart rates could be a physiological mechanism compensating for body hypoxia. Embryos/larvae exposed to nano-ZnO exhibited oxidative stress, due to an excessive generation of reactive oxygen species (ROS). Oxidative stress was evidenced by increased levels of superoxide dismutase, by increased lipid peroxidation, and by increased expression of genes related to the antioxidant defense system (sod1, cat, gpx1a, and pparα), which were altered at different degrees. Upon exposure to nano-ZnO, the percentage of apoptotic cells increased in a dose-dependent manner (0.41% to 4.21%). In addition, altered transcriptional regulation of pro-apoptotic genes (bax, puma, and apaf-1) and anti-apoptotic genes (bcl-2) provided further evidence of the activation of apoptosis. In this study, exposure of zebrafish embryos to nano-ZnO triggered an excessive production of ROS, which was followed by several phenomena: the up-regulation of p53, a reduction in the bcl-2/bax ratio,a reduction in the mitochondrial membrane potential (ψm), the release of cytochrome c into the cytosolic fraction, and the activation of caspases 9 and 3. Collectively, our data imply that nano-ZnO induce an excessive production of ROS which then activate the apoptosis pathway mediated by mitochondria and caspases.

Life-cycle exposure to BDE-47 results in thyroid endocrine disruption to adults and offsprings of zebrafish (Danio rerio).

2,2,4',4'-Tetrabromodi-phenyl ether (BDE-47) is predominantly concentrated in humans and wildlife and disturbs thyroid hormone homeostasis. The purpose of this study was to characterize the thyroid endocrine disruption induced by life-cycle exposure to BDE-47 in adults and offspring of zebrafish (Danio rerio). We exposed zebrafish embryos at the blastula stage to different concentrations of BDE-47 (1, 5, and 10µg/L). Exposure duration was 180days until fish reached adulthood. In F0 larvae, exposure decreased survival and increased malformations at 4 dpf. Thyroid hormone concentrations did not differ significantly between the F0 larvae and controls. All exposures significantly up-regulated expression of tshß, pa8, ugt1 and tg and down-regulated ttr. Significant up-regulation of dio2 and crh was observed in the 10µg/L BDE-47 group. There was no significant difference in the growth and somatic index between F0 adults and controls. BDE-47 (10µg/L) significantly decreased whole-body content of thyroxine (T4) but significantly increased triiodothyronine (T3) in both sexes. All exposures up-regulated expression of crh, tshß, pa8, ugt1 and tg and down-regulated ttr. Exposure to 10µg/L BDE-47 significantly up-regulated dio2 and ugt1 in both sexes. BDE-47 exposure (5 and 10µg/L) significantly increased the activity of pethoxy-resorufin-O-deethylase and UDP-glucuronosyl transferase. BDE-47 (10µg/L) significantly increased activity of ethoxy- and methoxy-resorufin-O-deethylase. In F1 offspring without continued BDE-47 (10µg/L) treatment, T4 significantly decreased and T3 increased. T4 was further decreased and T3 was further increased with continued BDE-47 treatment. Continued BDE-47 exposure decreased hatching and increased malformation compared with those without BDE-47 exposure. Expression of crh, tshß, dio2, pa8, ugt1 and tg was significantly up-regulated without BDE-47 exposure and with continued exposure. With continued BDE-47 exposure, dio1 was significantly up-regulated and ttr was significantly down-regulated. All the genes showed clear differences between continued exposure to 10µg/L BDE-47 and without BDE-47 exposure. These results suggest that parental exposure to BDE-47 results in thyroid endocrine disruption in adults and offspring.

[Research advances in eco-toxicological diagnosis of soil pollution].

Soil eco-toxicology provides a theoretical basis for ecological risk assessment of contaminated soils and soil pollution control. Research on eco-toxicological effects and molecular mechanisms of toxic substances in soil environment is the central content of the soil eco-toxicology. Eco-toxicological diagnosis not only gathers all the information of soil pollution, but also provides the overall toxic effects of soil. Therefore, research on the eco-toxicological diagnosis of soil pollution has important theoretical and practical significance. Based on the research of eco-toxicological diagnosis of soil pollution, this paper introduced some common toxicological methods and indicators, with the advantages and disadvantages of various methods discussed. However, conventional biomarkers can only indicate the class of stress, but fail to explain the molecular mechanism of damage or response happened. Biomarkers and molecular diagnostic techniques, which are used to evaluate toxicity of contaminated soil, can explore deeply detoxification mechanisms of organisms under exogenous stress. In this paper, these biomarkers and techniques were introduced systematically, and the future research trends were prospected.