斑马鱼对垃圾渗滤液胁迫的生物标志物响应

为了评价垃圾渗滤液的潜在生态风险和对水生动物的毒性,以斑马鱼(Danio rerio)为受试动物,探讨垃圾渗滤液对斑马鱼的急性毒性及其在亚致死浓度胁迫下鱼鳃、肝脏中的超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)及Na+,K+-腺苷三磷酸(ATP)酶的变化.结果表明:(1)垃圾渗滤液对斑马鱼的24、48、72、96 h的半致死浓度(LC50)分别为25.58％(体积分数,下同)、24.28％、23.18％和22.51％.(2)斑马鱼肝脏和鱼鳃中的SOD活性、GSH含量和Na+,K+-ATP酶活性的变化可以反映垃圾渗滤液序列间歇式活性污泥法(SBR)出水对斑马鱼的肝脏和鳃都具有损伤作用.(3)斑马鱼鱼体的SOD、GSH和Na+,K+-ATP酶对外界污染物的生物影响是敏感的,可将其作为垃圾渗滤液慢性胁迫的有效生物标志物.     

Toxicity of hexahydro-1,3,5-trinitro-1,3,5-triazine to larval zebrafish (Danio rerio)

Hexahydro-1,3,5-trinitro-1,3,5-triazine, a cyclonitramine commonly known as RDX, is used in the production of military munitions. Contamination of soil, sediment, and ground and surface waters with RDX has been reported in different places around the world. Acute and subacute toxicities of RDX have been relatively well documented in terrestrial vertebrates, but among aquatic vertebrates the information available is limited. The objective of this study was to characterize the acute toxicity of RDX to larval zebrafish. Mortality (LC50) and incidence of vertebral column deformities (EC50) were two of the end points measured in this study. The 96-h LC50 was estimated at 22.98 and 25.64 mgl(-1) in two different tests. The estimated no-observed-effective-concentration (NOEC) values of RDX on lethality were 13.27+/-0.05 and 15.32+/-0.30 mgl(-1); and the lowest-observed-effective-concentration (LOEC) values were 16.52+/-0.05 and 19.09+/-0.23 mgl(-1) in these two tests, respectively. The 96-h EC50 for vertebral deformities on survivors from one of the acute lethality tests was estimated at 20.84 mgl(-1), with NOEC and LOEC of 9.75+/-0.34 and 12.84+/-0.34 mgl(-1), respectively. Behavioral aberrations were also noted in this acute toxicity study, including the occurrence of whirling movement and lethargic behavior. The acute effects of RDX on survival, incidence of deformities, and behavior of larval zebrafish occurred at the high end of the most frequently reported concentrations of RDX in aquatic environments. The chronic effects of RDX in aquatic vertebrates need to be determined for an adequate assessment of the ecological risk of environmental RDX.     

Maternal transfer of brominated flame retardants in zebrafish (Danio rerio)

In many species reproduction and embryonic development have been shown to be sensitive to environmental contaminants. Understanding embryonic exposure to environmental contaminants is thus highly important. In this study concentrations of brominated flame retardants (BFRs) were measured in zebrafish eggs after parental exposure for 42 days via the diet. Zebrafish were exposed to two doses of eleven structurally-diverse BFRs. Eight BFRs were detected in the female zebrafish and maternal transfer to eggs was evident for all eight compounds. The highest concentrations in eggs were observed for hexabromocyclododecane (HBCD) and 2,4,4'-tribromodiphenyl ether (BDE 28), followed by 2,2',3,4,4',5',6-heptabromodiphenyl ether (BDE 183) and tetrabromobisphenol A 2,3-dibromopropyl ether (TBBPA DBPE). Five potential BFR metabolites were tentatively identified in female fish and maternal transfer was observed also for these compounds. The lipid adjusted concentrations in eggs were significantly higher than the concentrations in female fish for several of the BFRs. Further, the results showed a generally higher transfer in the lower exposure level and also indicated a dependency on the physico-chemical properties of the compounds.     

Study on acute toxicity and structure-activity relationship of zebrafish (Danio rerio) exposed to naphthoquinones

In this paper, we evaluated the acute toxicity of five naphthoquinone compounds, expressed as median lethal concentration at 96 h (LC₅₀) using zebrafish as a model. The five compounds were classified as harmful or very toxic. The Gaussian method was used to calculate the quantitative-chemical parameters of each compound. Based on the structure-activity relationship analysis, the interaction between the naphthoquinones and the target is likely mediated by 1-carbonyl and the hydrophobic fraction substituted in α -position of naphthoquinone via hydrogen bonding and hydrophobic interactions. Our results provide a foundation for evaluating the aquatic ecological risk and the mode of action of naphthoquinones in the environment.     

磺胺甲恶唑对斑马鱼胚胎/仔鱼的毒性效应

为研究磺胺甲恶唑对斑马鱼胚胎/仔鱼的早期发育及抗氧化系统的影响,将受精2h的斑马鱼胚胎暴露在不同浓度的磺胺甲恶唑水溶液中,观察斑马鱼胚胎死亡率、孵化率、仔鱼心包囊肿率、心跳数、体长以及仔鱼内抗氧化酶活性、丙二醛(MDA)含量的变化。结果表明,磺胺甲恶唑对斑马鱼胚胎/仔鱼120h的急性毒性半数致死质量浓度(LC50)为1 250.5mg/L;斑马鱼胚胎孵化率随着暴露浓度的增加呈现明显的下降趋势,96h胚胎孵化率抑制的半数有效质量浓度(EC50)为1 299.7mg/L;当暴露质量浓度≥1 500mg/L时,斑马鱼胚胎患心包囊肿率明显升高,呈现剂量—效应关系;经磺胺甲恶唑暴露96h的斑马鱼仔鱼30s心跳数及120h斑马鱼仔鱼体长总体降低;10~(-2)mg/L磺胺甲恶唑染毒96h后,即观察到斑马鱼仔鱼细胞出现明显的氧化损伤,过氧化氢酶的酶活性受到显著抑制,超氧化物歧化酶和谷胱甘肽S-转移酶被显著诱导,MDA含量显著增加。研究表明,磺胺甲恶唑暴露对斑马鱼胚胎细胞产生了明显的损伤,进而可能影响斑马鱼仔鱼的发育。     

1H-NMR-based metabolomic studies of bisphenol A in zebrafish (Danio rerio)

Proton nuclear magnetic resonance (¹H-NMR) spectroscopy was used to study the response of zebrafish (Danio rerio) to increasing concentrations of bisphenol A (4,4′-(propane-2,2-diyl)diphenol, BPA). Orthogonal partial least squares discriminant analysis (OPLS-DA) was applied to detect aberrant metabolomic profiles after 72 h of BPA exposure at all levels tested (0.01, 0.1, and 1.0 mg/L). The OPLS-DA score plots showed that BPA exposure caused significant alterations in the metabolome. The metabolomic changes in response to BPA exposure generally exhibited nonlinear patterns, with the exception of reduced levels of several metabolites, including glutamine, inosine, lactate, and succinate. As the level of BPA exposure increased, individual metabolite patterns indicated that the zebrafish metabolome was subjected to severe oxidative stress. Interestingly, ATP levels increased significantly at all levels of BPA exposure. In the present study, we demonstrated the applicability of ¹H-NMR-based metabolomics to identify the discrete nature of metabolic changes.     

Synergistic effects caused by atrazine and terbuthylazine on chlorpyrifos toxicity to early-life stages of the zebrafish Danio rerio

This study examined the effects of three widely used pesticides that have been previously detected in aquatic systems neighbouring agricultural fields on the early-life stages of the zebrafish Danio rerio. Tests involving single exposures and binary combinations of the s-triazine herbicides (atrazine and terbuthylazine) and the organophosphate insecticide chlorpyrifos were performed. Several endpoints, such as swimming behaviour, morphological abnormalities and mortality, were studied. In addition, the inhibition of acetylcholinesterase (AChE) activity was investigated in order to evaluate the mode of action and toxicity of chlorpyrifos in the presence of these herbicides. Results indicate that both binary mixtures elicited synergistic responses on the swimming behaviour of zebrafish larvae. Moreover, although the herbicides were not effective inhibitors of the AChE on their own, a synergistic inhibition of the enzyme activity was obtained by exposure to mixtures with chlorpyrifos. We observed a correlation between impairment of swimming behaviour of the larvae and inhibition of AChE activity. This study supports previous studies concerning the risk assessment of mixtures since the toxicity may be underestimated when looking only at the single toxicants and not their mixtures.     

Suborganismic and organismic effects of aldicarb and its metabolite aldicarb-sulfoxide to the zebrafish embryo (Danio rerio)

The new European chemical regulation (REACH) requires a short-term fish test for chemicals where the level of production exceeds 10tons per year. For ethical reasons (3R-concept), an alternative to the acute fish test should be introduced to decrease the number of animal testing with fish. The zebrafish embryo (Danio rerio) test became a valuable tool in ecotoxicology and already replaces the acute fish test for the evaluation of wastewater in Germany. Recent efforts are targeted to use this and other fish embryo tests for the effect assessment of chemicals. The toxic effects of the carbamate insecticide aldicarb and its metabolite aldicarb-sulfoxide to zebrafish embryos were analysed using two approaches with different endpoints. Organismic tests were conducted with zebrafish embryos exposed to the pesticides for 48h. In addition, suborganismic effects were examined analysing the enzyme inhibition of cholinesterases and carboxylesterases. On the organismic level, the only sublethal effect seen was the increase of heart rate at low and decrease at higher concentration with the use of aldicarb-sulfoxide but not with aldicarb (concentration range 0.2-300microM). In contrast, analysis of enzyme inhibitions showed high to very high effects caused by the two carbamates. The enzyme inhibition analysis of whole homogenates of exposed embryos may be advantageous for toxicant screening (biomarker of exposure) and might be used to bridge the gap of sensitivity of the (48h old) zebrafish embryos to adult fish when exposed to anti-cholinesterase substances (biomarker of prospective effect).     

Integrated biological responses of zebrafish (Danio rerio) to analyze water quality in regions under anthropogenic influence

This study analyzed water quality in regions around Patos lagoon (Southern Brazil) that are under anthropogenic pressure. Water samples were collected from five different sites, including one used as a source for human consumption (COR) and others known to be influenced by human activities (IP). Danio rerio (Teleostei, Cyprinidae) organisms were exposed for 24 h to these water samples, plus a control group. It was observed that: (1) reactive oxygen species levels were lower in COR and IP than in the control group; (2) glutamate-cysteine ligase (catalytic subunit) expression was higher in COR than in other sites; (3) exposure to all water samples affected long-term memory (LTM) when compared to control group. Thus, some water samples possess the ability to modulate the antioxidant system and to induce a decline in cognitive functions, as measured by LTM. The obtained results indicate that a combination of variables of different organization level (molecular, biochemical and behavioral) can be employed to analyze water quality in impacted regions.     

Environmental impact of sunscreen nanomaterials: ecotoxicity and genotoxicity of altered TiO2 nanocomposites on Vicia faba

Mineral sunscreen nanocomposites, based on a nano-TiO₂ core, coated with aluminium hydroxide and dimethicone films, were submitted to an artificial ageing process. The resulting Altered TiO₂ Nanocomposites (ATN) were then tested in the liquid phase on the plant model Vicia faba, which was exposed 48 h to three nominal concentrations: 5, 25 and 50 mg ATN/L. Plant growth, photosystem II maximum quantum yield, genotoxicity (micronucleus test) and phytochelatins levels showed no change compared to controls. Oxidative stress biomarkers remained unchanged in shoots while in roots, glutathione reductase activity decreased at 50 mg ATN/L and ascorbate peroxidase activity decreased for 5 and 25 mg ATN/L. Nevertheless, despite the weak response of biological endpoints, ICP-MS measurements revealed high Ti and Al concentrations in roots, and X-ray fluorescence micro-spectroscopy revealed titanium internalization in superficial root tissues. Eventual long-term effects on plants may occur.     

Embryo/larval toxicity and transcriptional effects in zebrafish (Danio rerio) exposed to endocrine active riverbed sediments

Environmental Science and Pollution Research - Sediment toxicity plays a fundamental role in the health of inland fish communities; however, the assessment of the hazard potential of contaminated...     

Lethal effects of abamectin on the aquatic organisms Daphnia similis, Chironomus xanthus and Danio rerio

Abamectin is used as an acaricide and insecticide for fruits, vegetables and ornamental plants, as well as a parasiticide for animals. One of the major problems of applying pesticides to crops is the likelihood of contaminating aquatic ecosystems by drift or runoff. Therefore, toxicity tests in the laboratory are important tools to predict the effects of chemical substances in aquatic ecosystems. The aim of this study was to assess the potential hazards of abamectin to the freshwater biota and consequently the possible losses of ecological services in contaminated water bodies. For this purpose, we identified the toxicity of abamectin on daphnids, insects and fish. Abamectin was highly toxic, with an EC₅₀ 48 h for Daphnia similis of 5.1 ng L⁻¹, LC₅₀ 96 h for Chironomus xanthus of 2.67 μg L⁻¹ and LC₅₀ 48 h for Danio rerio of 33 μg L⁻¹.     

Combined toxicity of imidacloprid,acetochlor, and tebuconazole to zebrafish ( Danio rerio ): acute toxicity and hepatotoxicity assessment

Compound pollution refers to two or more kinds of pollutants with different properties, a pollutant from different sources, or the simultaneous existence of two or more different types of pollutants in the same environment. In this study, we aimed to investigate the individual and combined toxicity of the insecticide imidacloprid (IMI), the herbicide acetochlor (ACT), and the fungicide tebuconazole (TBZ) to zebrafish. The acute toxicity test results showed that the 96-h LC50 values of IMI, ACT, and TBZ were 276.84 (259.62–294.35) mg active ingredient (a.i.) L−1, 1.52 (1.34–1.74) mg a.i. L−1, and 8.16 (7.7–8.6) mg a.i. L−1, respectively. The combinations of IMI, ACT, and TBZ with toxicity ratios of 1:2:2, 1:4:4, 2:4:1, and 4:1:4 displayed synergistic toxic effects on zebrafish, while the toxicity ratios of 1:1:1, 1:1:2, 2:1:2, 2:2:1, and 4:2:1 of IMI, ACT, and TBZ, respectively, exhibited antagonistic toxic effects on zebrafish. The following experiments were performed with a toxicity ratio of 1:4:4 (IMI:ACT:TBZ). The activities of four enzyme biomarkers related to oxidative stress in the liver, catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), and malondialdehyde (MDA) content were evaluated in each exposure group on days 7, 14, 21, and 28. Compared with those of the control group, the activities of CAT, SOD, and GST and the MDA content were significantly altered at different time points in the individual and combined exposure groups. Additionally, the activities of CAT, SOD, and GST and the MDA content were significantly altered in the combined group compared with those of the individual group after 14 days or 21 days of exposure. Therefore, it was confirmed that combined toxicity studies are indispensable in risk assessment.     

Effects of penconazole enantiomers exposure on hormonal disruption in zebrafish Danio rerio (Hamilton, 1822)

Environmental Science and Pollution Research - PEN is a widely used triazole fungicide, usually used to control grape white rot. In the process of agricultural use, PEN will be scattered to the...     

Uptake, accumulation and depuration of sodium perchlorate and sodium arsenate in zebrafish (Danio rerio)

In toxicokinetics studies, interactions between chemicals in mixtures has been largely neglected. This study examines a mixture of perchlorate and arsenate because (1) they have the potential to co-occur in contaminated aquatic habitats, and (2) a previous study by the authors found possible toxicological interactive effects. In the present study, zebrafish (Danio rerio) were exposed to two concentrations of sodium perchlorate (10 and 100 mg l(-1)), sodium arsenate (1 and 10 mg l(-1)), and the mixture-sodium perchlorate+sodium arsenate (10+1 mg l(-1) and 100+10 mg l(-1) Na(2)HAsO(4)-high mixture) for 90 d. Their uptake and accumulation by zebrafish was evaluated at 10, 30, 60, and 90 d. In addition, depuration was examined at 1, 3, and 5d after cessation of the exposure. The uptake of either chemical was concentration-dependent, with significantly higher uptake at high concentrations at either exposure interval. In contrast, there was no significant difference in whole body residue between single chemicals and the corresponding mixture except for 100 mg l(-1) sodium arsenate at 90 d. However, there was increasing accumulation over time at the high concentration of either chemical alone and their mixture, and this increasing trend was more pronounced in the single chemical exposures than in the mixture. At the concentrations tested in the current study, both chemicals reduced the uptake but enhanced the depuration of the other chemical from the zebrafish. This study represents the first examination of the interaction of two anions-perchlorate and arsenate with respect to toxicokinetics.     

Cypermethrin has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish (Danio rerio)

Cypermethrin (CYP), a widely used Type II pyrethroid pesticide, is one of the most common contaminants in the freshwater aquatic system. We studied the effects of CYP exposure on the induction of hepatic oxidative stress, DNA damage and the alteration of gene expression related to apoptosis in adult zebrafish. Hepatic mRNA levels for the genes encoding antioxidant proteins, such as Cu/Zn-Sod, Mn-Sod, Cat, and Gpx, were significantly upregulated when zebrafish were exposed to various concentrations of CYP for 4 or 8 days. In addition, the main genes related to fatty acid β-oxidation and the mitochondrial genes related to respiration and ATP synthesis were also significantly upregulated after exposure to high concentrations (1 and 3 μg L⁻¹) of CYP for 4 or 8 days. Moreover, in a comet assay of zebrafish hepatocytes, tail DNA, tail length, tail moment and Olive tail moment increased in a concentration-dependent manner. The significant induction (p < 0.01) of all four parameters observed with CYP concentrations of 0.3 μg L⁻¹ or higher suggests that heavy DNA damage was induced even at low levels. Furthermore, several apoptosis- related genes, such as p53, Apaf1 and Cas3, were significantly upregulated after CYP exposure, and Bcl2/Bax expression ratio decreased, especially in groups treated with 1 and 3 μg L⁻¹ CYP for 8 days. Taken together, our results suggested that CYP has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in zebrafish. This information will be helpful in fully understanding the mechanism of aquatic toxicology induced by CYP in fish.     

The effects of henna (hair dye) on the embryonic development of zebrafish (Danio rerio)

The powder of henna is extensively used as decorative skin paint for nail coloring and as a popular hair dye in Asian countries. Its human health risk is extensive, and it is frequently released as waste into the aquatic environment raising the concerns. Zebrafish (Danio rerio) embryos were employed to study the developmental effects of henna. Normal fertilized zebrafish embryos under standard water were selected for the control and test chambers. Three predetermined sublethal concentrations (100, 200, and 275 μM) of henna in 24-well cell culture plates were tested on 1-h postfertilized embryo (pfe) for 96 h. Observation for rates of survival and mortality was recorded; digital camera was used to image morphological anomalies of embryos with a stereomicroscope; and functional abnormalities at 24, 48, 72, and 96 h were performed. The hatching rates of embryos were reduced significantly when treated with 200 and 275 μM or higher concentrations of henna. Slow blood circulation in the whole body was observed with a median effect on hatching exposed to 200 and 275 μM of henna at 48-h pfe. At 72- and 96-h pfe, blood circulation was ceased in the whole body but still had a heartbeat. At 96-h pfe, pericardial sac edema, yolk sac edema, head deformation, spine crooked malformation, and tail malformation (bent tails or hook-like tails) were observed in the surviving larvae at 100 μM. In summary, exposure to henna at 100, 200, and 275 μM causes some altered morphological and physiological abnormalities including increased mortality, hatching delay, slow blood circulation, pericardial sac edema, yolk sac edema, abnormal body axes, twisted notochord, tail deformation, weak heartbeat, and growth retardation and was also detected in some treated embryos and groups having adverse effects on embryonic development of zebrafish provoking potential human developmental risk studies.     

Comparison of metabolome profiles in zebrafish (Danio rerio) intestine induced by polystyrene microplastics with different sizes

Environmental Science and Pollution Research - Microplastics (MPs) are widespread in aquatic environments. They could induce intestinal toxicity in the fish. However, research on the metabolic...