Early Life Stages Of Zebrafish Research Articles

Relationship between miR-203a inhibition and oil-induced toxicity in early life stage zebrafish (Danio rerio)

Dysregulation of microRNA (miRNA, miR) by environmental stressors influences the transcription of mRNA which may impair organism development and/or lead to adverse physiological outcomes. Early studies evaluating the effects of oil on developmental toxicity in early life stages of fish showed that reductions in expression of miR-203a were associated with enhanced expression of downstream mRNAs that predicted altered eye development, cardiovascular disease, and improper fin development. To better understand the effects of miR-203a inhibition as an outcome of oil-induced toxicity in early life stage (ELS) fish, embryonic zebrafish were injected with an miR-203a inhibitor or treated with 3.5 µM phenanthrene (Phe) as a positive control for morphological alterations of cardiovascular and eye development caused by oil. Embryos treated with Phe had diminished levels of miR-203a at 7 and 72 h after injection. Embryos treated with the miR-203a inhibitor and Phe exhibited a reduced heart rate by 48 h post fertilization (hpf), with an increased incidence of developmental deformities (including pericardial edema, altered eye development, and spinal deformities) and reduced caudal fin length by 72 hpf. There were significant reductions in lens and eye diameters in 120 hpf miR-203a-inhibitor and Phe-treated fish, as well as a significantly reduced number of eye saccades, determined by an optokinetic response (OKR) behavioral assay. The expression of vegfa, which is an important activator during neovascularization, was significantly upregulated in embryos receiving miR-203a inhibitor injections by 7 and 72 hpf with increased trends in vegfa expression in 72 hpf larvae treated with Phe. There were decreasing trends in crx, neurod1, and pde6h expression by 72 hpf in miR-203a inhibitor and Phe treatments, which are involved in photoreceptor function in developing eyes and regulated by miR-203a. These results suggest that an inhibition of miR-203a in ELS fish exhibits an oil-induced toxic response that is consistent with Phe treatment and specifically impacts retinal, cardiac, and fin development in ELS fish.

Combined effects of polystyrene microplastics and cadmium on oxidative stress, apoptosis, and GH/IGF axis in zebrafish early life stages

The toxicological interactions of microplastics (MPs) and heavy metals have been paid much attention in aquatic organism. The mechanisms are not fully clear, particularly in fish early life stages. To the end, zebrafish embryos were exposed to 500 μg/L MPs, 5 μg/L cadmium (Cd), and their combination for 30 days. Body weight, adsorption characteristics of Cd onto MPs, Cd accumulation, oxidative stress, apoptosis, and growth hormone/insulin-like growth factor-I (GH/IGF) axis were examined. Exposure to MPs and Cd alone reduced body weight, which was aggravated by co-exposure. An increase in reactive oxygen species (ROS) levels was observed in larvae exposed to Cd or MPs + Cd, suggesting an induction of oxidative stress. Lipid peroxidation levels were not affected by exposure to MPs and Cd alone but dramatically enhanced by co-exposure, which may be explained by the reduction of total antioxidant capacity (TAOC) and activity levels of Mn-superoxide dismutase (Mn-SOD) and catalase (CAT) after co-exposure. Increased apoptotic cells were observed in the vertebral body of larvae exposed to Cd, the esophagus of larvae exposed to MPs, and both organs of larvae exposed to MPs + Cd, which was further confirmed by changes in the activities of Caspase-3, Caspase-8 and Caspase-9. PCR array on the transcription of genes related to growth, oxidative stress and apoptosis was examined, showing that the combined exposure resulted in greater magnitude of changes than MPs and Cd alone. The results indicate that MPs can enhance the negative effects of Cd on growth, oxidative damage and apoptosis in early life stages of zebrafish. However, the adsorption of Cd onto MPs was not observed and the combined exposure did not increase the Cd content in larvae compared to the single Cd exposure, implying that vector role of MPs in Cd uptake is negligible.

Embryotoxicity of Selective Serotonin Reuptake Inhibitors—Comparative Sensitivity of Zebrafish (Danio rerio) and African Clawed Frog (Xenopus laevis) Embryos

Over the past twenty years, the prescription of antidepressant drugs has increased all over the world. After their application, antidepressants, like other pharmaceuticals, are excreted and enter the aquatic environment. They are dispersed among surface waters mainly through waste water sources, typically at very low concentrations— from a tenth up to hundreds of ng/L. Frequently detected antidepressants include fluoxetine and citalopram—both selective serotonin reuptake inhibitors. The aim of our study was to assess the embryotoxicity of fluoxetine hydrochloride and citalopram hydrochloride on the early life stages of zebrafish (Danio rerio) and the African clawed frog (Xenopus laevis). The embryos were exposed to various concentrations of the individual antidepressants and of their mixtures for 96 h. The tested levels included both environmentally relevant and higher concentrations for the evaluation of dose-dependent effects. Our study demonstrated that even environmentally relevant concentrations of these psychiatric drugs influenced zebrafish embryos, which was proven by a significant increase (p < 0.01) in the embryos’ heart rates after fluoxetine hydrochloride exposure and in their hatching rate after exposure to a combination of both antidepressants, and thus revealed a potential risk to aquatic life. Despite these results, we can conclude that the African clawed frog is more sensitive, since exposure to the highest concentrations of fluoxetine hydrochloride (10,000 μg/L) and citalopram hydrochloride (100,000 μg/L) resulted in total mortality of the frog embryos.

Environmentally relevant concentrations of arsenic induces apoptosis in the early life stage of zebrafish

Arsenic (As) in the aquatic environment is a considerable environmental issue, previous studies have reported the toxic effects of low concentrations (≤ 150 μg/L) of As on fish. However, limited information is available regarding the impact of low levels of As on apoptosis. To evaluate this, zebrafish embryos were exposed to different concentrations (0, 25, 50, 75, and 150 μg/L) of As (arsenite [AsIII] and arsenate [AsV]) for 120 h. Our results indicated that low concentrations of AsIII exposure significantly inhibited the survival of zebrafish larvae, and significantly increased the transcription of Caspase-9 and Caspase-3, the ratio of Bax/Bcl-2 transcription, and protein levels of Caspase-3. In contrast, AsV decreased the ratios of Bax/Bcl-2 transcription and protein levels, as well as protein levels of Caspase-3. Our data demonstrated that AsIII and AsV exert different toxic effects, AsIII induced apoptosis via the mitochondrial pathway and the extrinsic pathway, while AsV induced apoptosis only via the mitochondrial pathway.

Neuromotor activity inhibition in zebrafish early-life stages after exposure to environmental relevant concentrations of caffeine

Caffeine (CAF), a neuroactive compound, has been found in surface waters at concentrations ranging from few nanograms up to micrograms and may induce adverse effects in aquatic vertebrates. Thus, the aim of this study was to evaluate the potential of CAF in affecting fish early-life stages in a wide concentration range, including occurring levels in surface waters. Specimens of zebrafish in early-life stages were exposed to CAF for 168 h and survival, developmental alterations, locomotor activity and acetylcholinesterase activity were evaluated. CAF induced mortality in embryos unable to hatch or in larvae after hatching (LC50 − 168 h = 283.2 mg/L). Tail deformities were observed in organisms exposed to concentrations ≥ 40 mg/L, while edemas were found at concentrations of 100 mg/L. CAF also decreased the total swimming time and distance moved of exposed organisms (LOEC = 0.0006 mg/L). Locomotor inhibition may be associated with an acetylcholinesterase inhibition observed at concentration ≥ 0.0088 mg/L. Therefore, the hazard of CAF for fish populations deserves further attention since unexpected effects on neuro-behavioral parameters occurs at concentrations often detected in natural aquatic ecosystems.

Acute exposure to 17-α-ethinylestradiol disrupt the embryonic development and oxidative status of Danio rerio

17-Alpha-ethinylestradiol (EE2) is an estrogen derived from estradiol (E2). This compound and is one of the most widely used drugs both in humans and animals. Numerous studies have reported the ability of EE2 to alter sex determination and delay sexual maturity, but there are toxic effects that need to be explored. In this work, we analyzed the effect of EE2 on embryonic development and oxidative stress biomarkers in Danio rerio. For this effect, zebrafish embryos in the blastula period (2.5 h post fecundation) were exposed to different concentrations of EE2 (36–106 ng L−1) until 96 hpf. Survival, alterations to embryonic development, and teratogenic effects were evaluated using a stereomicroscope. Furthermore, oxidative stress biomarkers: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) activities, lipid peroxidation (LPX), hydroperoxide content (HPX), and protein carbonyl content (POX) were evaluated at 72 and 96 hpf using spectrophotometric methods. LC50 and EC50 of malformations got values of 82 ng L−1 and 57.7 ng L−1, respectively. The main teratogenic effects found were: chorda malformation, body malformation, and developmental delay. These alterations occurred at 86, 96, and 106 ng L−1. Integrated biomarker index showed that the oxidative stress biomarkers that had the most influence on embryos were SOD, CAT, GPX, and LPX. Overall, our results allow us to conclude that low concentrations of EE2 may potentially alter the development and oxidative status in the early life stages of zebrafish. Therefore, this bio-active estrogen can be considered a hazardous substance for fish.

Long-lasting behavioral effects of quinpirole exposure on zebrafish

The human brain matures into a complex structure, and to reach its complete development, connections must occur along exact paths. If at any stage, the processes are altered, interrupted, or inhibited, the consequences can be permanent. Dopaminergic signaling participates in the control of physiological functions and behavioral processes, and alterations in this signaling pathway are related to the pathogenesis of several neurological disorders. For this reason, the use of pharmacological agents able to interact with the dopaminergic signaling may elucidate the biological bases of such disorders. We investigated the long-lasting behavioral effects on adult zebrafish after quinpirole (a dopamine D2/D3 receptor agonist) exposure during early life stages of development (24 h exposure at 5 days post-fertilization, dpf) to better understand the mechanisms underlying neurological disorders related to the dopaminergic system. Quinpirole exposure at the early life stages of zebrafish led to late behavioral alterations. When evaluated at 120 dpf, zebrafish presented increased anxiety-like behaviors. At the open tank test, fish remained longer at the bottom of the tank, indicating anxiety-like behavior. Furthermore, quinpirole-treated fish exhibited increased absolute turn angle, likely an indication of elevated erratic movements and a sign of increased fear or anxiety. Quinpirole-treated fish also showed altered swimming patterns, characterized by stereotypic swimming. During the open tank test, exposed zebrafish swims from corner to corner in a repetitive manner at the bottom of the tank. Moreover, quinpirole exposure led to memory impairment compared to control fish. However, quinpirole administration had no effects on social and aggressive behavior. These findings demonstrate that dopaminergic signaling altered by quinpirole administration in the early life stages of development led to late alterations in behavioral parameters of adult zebrafish.

Tetrabromobisphenol S alters the circadian rhythm network in the early life stages of zebrafish

Tetrabromobisphenol S (TBBPS), an emerging brominated flame retardant (BFR) has been widely detected in the environment, and may potentially pose environmental risks. However, data on the occurrence and toxic effects of TBBPS are limited. Circadian rhythms govern multiple behavioral and physiological processes, and their disruption is closely associated with various pathological conditions. Little is known about the potential for TBBPS to perturb circadian rhythm networks or circadian-driven locomotor behavior. In the present study, behavior assays and gene expression analysis based on circadian rhythm pathways were designed to investigate the potential circadian rhythm impairments and subsequent adverse effects caused in 120 h post-fertilization (hpf) zebrafish larvae by TBBPS. The development of embryos was inhibited by TBBPS exposure even at concentrations below the maximal non-lethal concentration (MNLC, 3.47 mg/L). Our results indicated remarkable alterations in the expression of several key circadian rhythm genes due to TBBPS exposure. Compared to control, the expression of per1a, per1b, per3, cry2, and csnk1da was increased, while the expression of clocka, clockb, cry4, cry1ba, arntl1a, and cank1db was decreased. Significant alterations of the circadian rhythm network could be observed in the zebrafish embryos. TBBPS exposure also significantly affected the behavioral responses of larvae. Our findings suggest the circadian rhythm network could be a potential target of TBBPS. Further study is needed to explore whether the transcriptional alterations in circadian rhythm translate into physiological effects.

Contamination and Toxicity of Surface Waters Along Rural and Urban Regions of the Capibaribe River in Tropical Northeastern Brazil.

The Capibaribe River provides water to a population of 1.7 million people in the Brazilian northeast, while receiving agricultural runoff and industrial and domestic effluents along its 280 km. The present study evaluated the ecotoxicity of surface waters along ten sites in rural and urban areas using zebrafish (Danio rerio) early-life stages and related it to water quality indices and chemical abiotic variables. Lethality rates, delays in embryo-larval development quantified by the general morphology score (GMS), and frequencies of developmental abnormalities were analyzed. A correlation was detected between zebrafish GMS and water quality index (WQI), sensitivity to domestic sewage contamination, and trophic state index, focused on eutrophication. These indices agreed in identifying a spatial pattern of smaller impact in terms of ecotoxicity, domestic sewage contamination, and eutrophication risk at three sites in rural areas (mean GMS 16.9), an intermediate impact at four sites with urban and agricultural influence (mean GMS 16.4), and greatest impacts at three more urbanized sites (mean GMS 14.9). Most frequent developmental abnormalities included noninflation of the swim bladder, delayed hatching, nonprotrusion of the mouth, blood stasis, and nondevelopment of pectoral fins. Toxic NH3 concentrations varied spatially, with higher concentrations in urban sites; and blood stasis correlated positively with NH3 , suggesting a causal relationship. Polycyclic aromatic hydrocarbons were detected in both rural and urbanized sites, contributing to detected toxicity. The present study demonstrates the potential of zebrafish early-life stages as an ecotoxicological model that may contribute to a better understanding of surface water quality and ecotoxicity in tropical river systems. Environ Toxicol Chem 2021;40:3063-3077. © 2021 SETAC.

Ecotoxicological assessment of spent battery extract using zebrafish embryotoxicity test: A multi-biomarker approach

Water environmental pollution caused by spent batteries is a nonignorable environmental issue. In this study, the early life stage of zebrafish was employed to assess the environmental risk of spent batteries after exposure to 0, 1%, 2%, 5% and 10% spent battery extract for 120 h. Our results clearly indicated that spent battery extract can significantly decrease the survival rate, hatching rate and body length and increase heart rate. Moreover, spent battery extract exposure-induced zebrafish larvae generate oxidative stress and inhibit the mRNA transcriptional levels of heat shock protein (HSP70) and metallothionein (MT) genes. These results showed that the spent batteries not only affected the survival and development performance of zebrafish at an early life stage but also caused oxidative stress and interfered with the detoxification of zebrafish. This study provided novel insight into spent battery induced toxicity in the early life stage of fish.

Adverse effects of bisphenol B exposure on the thyroid and nervous system in early life stages of zebrafish

Bisphenol B (BPB), a widely used alternative of bisphenol A (BPA), has been detected in various environmental media and foodstuffs. However, the knowledge of the health risks about BPB is still limited. In this study, the effects of BPB on thyroid hormone homeostasis and neuronal development were evaluated by exposure of embryos 2 h post-fertilization (hpf) to BPB (0, 1, 10, 100 and 1000 μg/L) until 144 hpf. The results showed that 100 and 1000 μg/L BPB exposed larvae exhibited abnormal morphologies in phenotype and brain histological patterns. Significant decline of thyroid hormone thyroxine (T4) content and elevation of 3,5,3′-triiodothyronine (T3) content, along with the up-regulated expression of tg, trhr1, dio1, dio2, thrα, thrβ genes and down-regulated expression of tsh, ttr and trh genes in BPB exposed zebrafish larvae were observed. Moreover, locomotor activity of larvae was decreased, and the transcription of genes (e.g., elavl3, gap43, zn5, α-tubulin, syn2a and mbp) related to neuronal development were inhibited after exposure to BPB. The mechanism of neurotoxicity and thyroid disruption in zebrafish larvae induced by BPB were discussed.

Role of the cardiovascular system in ammonia excretion in early life stages of zebrafish (Danio rerio).

The purpose of this study was to investigate if the cardiovascular system is important for ammonia excretion in the early life stages of zebrafish. Morpholino knockdowns of cardiac troponin T (TNNT2) or vascular endothelial growth factor A (VEGFA) provided morphants with nonfunctional circulation. At the embryonic stage [30-36 h postfertilization (hpf)], ammonia excretion was not constrained by a lack of cardiovascular function. At 2 days postfertilization (dpf) and 4 dpf, morpholino knockdowns of TNNT2 or VEGFA significantly reduced ammonia excretion in all morphants. Expression of rhag, rhbg, and rhcgb showed no significant changes but the mRNA levels of the urea transporter (ut) were upregulated in the 4 dpf morphants. Taken together, rhag, rhbg, rhcgb, and ut gene expression and an unchanged tissue ammonia concentration but an increased tissue urea concentration, suggest that impaired ammonia excretion led to increased urea synthesis. However, in larvae anesthetized with tricaine or clove oil, ammonia excretion was not reduced in the 4 dpf morphants compared with controls. Furthermore, oxygen consumption was reduced in morphants regardless of anesthesia. These results suggest that cardiovascular function is not directly involved in ammonia excretion, but rather reduced activity and external convection may explain reduced ammonia excretion and compensatory urea accumulation in morphants with reduced cardiovascular function.

Accumulation and Distribution of Fluorescent Microplastics in the Early Life Stages of Zebrafish

Accumulation and Distribution of Fluorescent Microplastics in the Early Life Stages of Zebrafish

Accumulation and Distribution of Fluorescent Microplastics in the Early Life Stages of Zebrafish.

As a new type of environmental pollutant, microplastic has been widely found in the aquatic environment and poses a high threat to aquatic organisms. The bioaccumulation of microplastics plays a key role in their toxic effects; however, as a particulate, their bioaccumulations are different from many other pollutants. Described here is a feasible method to visually determine the accumulation and distribution of microplastics in zebrafish embryos or larvae using fluorescent microplastics. Embryos are exposed to different concentrations (0.1, 1, and 10 mg/L) of fluorescent microplastics with a diameter of 500 nm for 120 h. It is shown in the results that microplastics can bioaccumulate in zebrafish embryos/larvae in a concentration-dependent manner. Before hatching, strong fluorescence is found around the embryonic chorion; while in zebrafish larvae, the yolk sac, pericardium, and gastrointestinal tract are the main accumulated sites of microplastics. The results demonstrate the uptake and internalization of microplastics in zebrafish at early life stages, which will provide basis for better understanding the impact of microplastics on aquatic animals.

Invited Perspective: PFAS Bioconcentration and Biotransformation in Early Life Stage Zebrafish and Its Implications for Human Health Protection.

Invited Perspective: PFAS Bioconcentration and Biotransformation in Early Life Stage Zebrafish and Its Implications for Human Health Protection.

A Roadmap to the Structure-Related Metabolism Pathways of Per- and Polyfluoroalkyl Substances in the Early Life Stages of Zebrafish (Danio rerio).

Background:Thousands of per- and polyfluoroalkyl substances (PFAS) with diverse structures have been detected in the ambient environment. Apart from a few well-studied PFAS, the structure-related toxicokinetics of a broader set of PFAS remain unclear.Objectives:To understand the toxicokinetics of PFAS, we attempted to characterize the metabolism pathways of 74 structurally diverse PFAS samples from the U.S. Environmental Protection Agency’s PFAS screening library.Methods:Using the early life stages of zebrafish (Danio rerio) as a model, we determined the bioconcentration factors and phenotypic toxicities of 74 PFAS. Then, we applied high-resolution mass spectrometry–based nontargeted analysis to identify metabolites of PFAS in zebrafish larvae after 5 d of exposure by incorporating retention time and mass spectra. In vitro enzymatic activity experiments with human recombinant liver carboxylesterase (hCES1) were employed to validate the structure-related hydrolysis of 11 selected PFAS.Results:Our findings identified five structural categories of PFAS prone to metabolism. The metabolism pathways of PFAS were highly related to their structures as exemplified by fluorotelomer alcohols that the predominance of or taurine conjugation pathways were primarily determined by the number of hydrocarbons. Hydrolysis was identified as a major metabolism pathway for diverse PFAS, and perfluoroalkyl carboxamides showed the highest in vivo hydrolysis rates, followed by carboxyesters and sulfonamides. The hydrolysis of PFAS was verified with recombinant hCES1, with strong substrate preferences toward perfluoroalkyl carboxamides.Conclusions:We suggest that the roadmap of the structure-related metabolism pathways of PFAS established in this study would provide a starting point to inform the potential health risks of other PFAS. https://doi.org/10.1289/EHP7169

Developmental toxicity of Zishen Guchong Pill on the early life stages of Zebrafish

Background: Zishen Guchong Pill (ZGP), a traditional oriental herbal medicine, has been applied broadly in the treatment of threatened or recurrent miscarriages in clinics, but little is known about its developmental toxicity.Aim of the study: This study aimed to investigate the potential toxicity of ZGP and its mechanisms.methods: We investigated the developmental toxicity of ZGP using a zebrafish model. Zebrafish embryos at 4 hours post-fertilization (hpf) were exposed to 25, 50, 100, and 200 µg/mL ZGP until 120 hpf. The zebrafish morphology, organ development, reactive oxygen species (ROS) content, oxidative stress-related enzyme activity, and mRNA levels of oxidative stress and apoptosis-related genes were measured.Results: Our results demonstrated that ZGP had no toxicity at 50 µg/mL ZGP but induced phenotypic defects and decreased the hatching rate and body length at more than 50 µg/mL ZGP (high dose). High doses of ZGP caused severe heart failure, including a significant increase in the sinus venosus (SV) and bulbus arteriosus (BA) distance and pericardial area, and reduced heart rate and numbers of red blood cells. In addition, high doses of ZGP caused liver atrophy and decreased the length of dopamine ganglia and neurovascular. High doses of ZGP elicited the generation of ROS and elevated malondialdehyde levels, but reduced the activity of superoxide dismutase (SOD) and catalase (CAT). Real-time PCR data showed the downregulation of oxidative stress-related genes (sod and cat) and the activation of Nrf2 (nrf2, keap1, ho-1, nqo1, gclc, and gclm) and P53 (p53, bcl-2, and bax) signal pathway genes after high ZGP exposure.Conclusions: Taken together, our results indicate that ZGP has no developmental toxicity under 50 µg/mL ZGP but induced developmental toxicity in zebrafish embryos at more than 50 µg/mL ZGP, and oxidative stress contributed to the toxic response.•ZGP is safe and reliable in clinical use.•ZGP (high does) caused an increase in ROS, which induced oxidative stress, leading to apoptosis and developmental toxicity.•The Nrf2 signaling pathway is impaired during the early stages of ZGP-induced developmental toxicity.

Suspended solids-associated toxicity of hydraulic fracturing flowback and produced water on early life stages of zebrafish (Danio rerio)

Hydraulic fracturing flowback and produced water (HF-FPW), which contains polyaromatic hydrocarbons (PAHs) and numerous other potential contaminants, is a complex wastewater produced during the recovery of tight hydrocarbon resources. Previous studies on HF-FPW have demonstrated various toxicological responses of aquatic organisms as consequences of combined exposure to high salinity, dissolved organic compounds and particle/suspended solids-bound pollutants. Noteworthy is the lack of studies illustrating the potentially toxic effects of the FPW suspended solids (FPW-SS). In this study, we investigated the acute and sublethal toxicity of suspended solids filtered from six authentic FPW sample collected from two fracturing wells, using a sediment contact assay based on early-life stages of zebrafish (Danio rerio). PAHs profiles and acute toxicity tests provided initial information on the toxic potency of the six samples. Upon exposure to sediment mixture at two selected doses (1.6 and 3.1 mg/mL), results showed adverse effects in larval zebrafish, as revealed by increased Ethoxyresorufin-O-deethylase (EROD) activity. Transcriptional alterations were also observed in xenobiotic biotransformation (ahr, pxr, cyp1a, cyp1b1, cyp1c1, cyp1c2, cyp3a65, udpgt1a1, udpgt5g1), antioxidant response (sod1, sod2, gpx1a, gpx1b) and hormone receptor signaling (esr1, esr2a, cyp19a1a, vtg1) genes. The results demonstrated that even separated from the complex aqueous FPW mixture, FPW-SS can induce toxicological responses in aquatic organisms' early life stages. Since FPW-SS could sediment to the bottom of natural wetland acting as a continuous source of contaminants, the current findings imply the likelihood of long-term environmental risks of polluted sediments on aquatic ecosystems due to FPW spills.

Immunotoxicity of F[sbnd]53B, an alternative to PFOS, on zebrafish (Danio rerio) at different early life stages

As an alternative to perfluorooctane sulfonate (PFOS), 6:2 chlorinated polyfluorinated ether sulfonate (F53B) has emerged in the Chinese market in recent years and has been frequently detected in the aquatic environment, but its ecological risk assessment is limited. In this study, zebrafish embryos and larvae were separately exposed to F53B, and their 96-h LC50 values were estimated to be 15.1 mg/L and 2.4 mg/L, respectively, suggesting that embryos were more resistant to F53B than larvae. The bioconcentration factor in larvae was basically higher than that of embryos, and the body growth of larvae was significantly affected by F53B rather than embryos, indicating that F53B may cause more severe toxicity to larvae. In addition to the excessive production of ROS and NO, the expression of many immune-related genes was increased in both embryos and larvae, but the number of dysregulated genes in larvae was more than that in embryos. Finally, the results of Point of Departure (PoD) indicated that the immunotoxicity of F53B was more sensitive to larvae than embryos at the molecular level. Our findings revealed the ecological risk of F53B by exploring the adverse effects of immunoregulation at different early life stages of zebrafish and indicated that the zebrafish larvae were more sensitive than embryos.

Toxicological effects of nanoplastics and phenanthrene to zebrafish (Danio rerio)

Despite the known hazards to wildlife, plastic waste to waterways and oceans continues to increase at alarming rates every year. Nanoplastics are a breakdown product of plastic debris in the environment (0.1–100 nm). Their small size renders them bioavailable, and they are likely to accumulate/adsorb persistent organic pollutants, which could increase their toxicity. In this study, we evaluated the effects of nanoplastics and phenanthrene on the early life stages of zebrafish. Zebrafish embryos were exposed to fluorescent nanoparticles and observed with fluorescence microscopy. Polystyrene nanoplastics (80 nm) attached to the embryonic chorionic membrane and were ingested when zebrafish reached 144 h after fertilization, the point at which they first open their mouth, as observed by the fluorescence in their intestines. Exposure alone or in combination with phenanthrene resulted in zebrafish pericardial edema, spinal curvature, increased embryonic heart rate, shortened body length, and increased mortality. Co-exposure to 5 mg/L nanoplastics and 0.1 mg/L phenanthrene caused more negative effects on length, teratogenicity and mortality of zebrafish embryos than the corresponding concentrations of single exposure. This study provides clear evidence that nanoplastics can be ingested by zebrafish larvae, thus future work to assess the toxicological effects is important to marine wildlife management.