Uninflated Swim Bladder Research Articles

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.

Dldhcri3 zebrafish exhibit altered mitochondrial ultrastructure, morphology, and dysfunction partially rescued by probucol or thiamine.

Dihydrolipoamide dehydrogenase (DLD) deficiency is a recessive mitochondrial disease caused by variants in DLD, the E3 subunit of mitochondrial α-keto (or 2-oxo) acid dehydrogenase complexes. DLD disease symptoms are multisystemic, variably manifesting as Leigh syndrome, neurodevelopmental disability, seizures, cardiomyopathy, liver disease, fatigue, and lactic acidemia. While most DLD disease symptoms are attributed to dysfunction of the pyruvate dehydrogenase complex, the effects of other α-keto acid dehydrogenase deficiencies remain unclear. Current therapies for DLD deficiency are ineffective, with no vertebrate animal model available for preclinical study. We created a viable Danio rerio (zebrafish) KO model of DLD deficiency, dldhcri3. Detailed phenotypic characterization revealed shortened larval survival, uninflated swim bladder, hepatomegaly and fatty liver, and reduced swim activity. These animals displayed increased pyruvate and lactate levels, with severe disruption of branched-chain amino acid catabolism manifest as increased valine, leucine, isoleucine, α-ketoisovalerate, and α-ketoglutarate levels. Evaluation of mitochondrial ultrastructure revealed gross enlargement, severe cristae disruption, and reduction in matrix electron density in liver, intestines, and muscle. Therapeutic modeling of candidate therapies demonstrated that probucol or thiamine improved larval swim activity. Overall, this vertebrate model demonstrated characteristic phenotypic and metabolic alterations of DLD disease, offering a robust platform to screen and characterize candidate therapies.

Embryotoxicity Induced by Triclopyr in Zebrafish (Danio rerio) Early Life Stage.

Triclopyr, an auxin-like herbicide that is widely employed for managing weeds in food crops and pastures, has been identified in various environmental settings, particularly aquatic ecosystems. Limited understanding of the environmental fate of this herbicide, its potential repercussions for both the environment and human health, and its insufficient monitoring in diverse environmental compartments has caused it to be recognized as an emerging contaminant of concern. In this study, we have investigated how triclopyr affects zebrafish, considering a new alternative methodology. We focused on the endpoints of developmental toxicity, neurotoxicity, and behavior of zebrafish embryos and larvae. We determined that triclopyr has a 96 h median lethal concentration of 87.46 mg/L (341.01 µM). When we exposed zebrafish embryos to sublethal triclopyr concentrations (0.5, 1, 5, 10, and 50 μM) for up to 144 h, we found that 50 µM triclopyr delayed zebrafish egg hatchability. Yolk sac malabsorption was significant at 0.5, 1, 5, and 10 µM triclopyr. In zebrafish larvae, uninflated swim bladder was significant only at 50 µM triclopyr. Furthermore, zebrafish larvae had altered swimming activity after exposure to 10 µM triclopyr for 144 h. In summary, these comprehensive results indicate that even low triclopyr concentrations can elicit adverse effects during early zebrafish development.

Time-course transcriptome analysis discloses PFDMO2OA (C8 HFPO-TA)-induced developmental malformations and cardiovascular toxicities in zebrafish

PFDMO2OA (C8 HFPO-TA), a novel substitute for perfluorooctanoic acid (PFOA), has been frequently detected in surface waters. However, information on its toxicity remains scarce. In the present study, zebrafish embryos were exposed to varying concentrations of PFDMO2OA, ranging from 80 to 800 mg/L, until 120 h post-fertilization (hpf) to explore its potential developmental toxicities. The LC50 value for mortality was 505.9 mg/L, comparable to that of PFOA (over 500 mg/L), suggesting a lack of safety of PFDMO2OA compared to PFOA. At 120 hpf, PFDMO2OA exposure led to various malformations in embryos, including uninflated swim bladder, yolk sac oedema, spinal deformation, and pigmentation changes, with pericardial oedema being prominent. Analysis using O-dianisidine stain indicated a decline in erythrocytes over time. Transcriptome analysis further revealed the cardiovascular toxicity caused by PFDMO2OA at the molecular level. Time-course differential analysis pointed to the apoptosis dependent on disrupted mitochondrial function as a significant contributor to erythrocyte disappearance, as confirmed by the TUNEL stain. Therefore, the present findings suggest that PFDMO2OA induces developmental malformations and cardiovascular toxicities in zebrafish embryos, demonstrating a toxic potency comparable to that of PFOA. The results further highlight the significance of evaluating the health risks associated with PFDMO2OA.

Single and mixture toxicity of cadmium and copper to swim bladder in early life stages of Japanese medaka (Oryzias latipes).

Toxicity observed in aquatic ecosystems often cannot be explained by the action of a single pollutant. Likewise, evaluation standards formulated by a single effect cannot truly reflect the environmental quality requirements. The study of mixtures is needed to provide environmental relevance and knowledge of combined toxicity. In this study, the embryos of Japanese medaka (Oryzias latipes) were treated with individual and binary mixture of copper (Cu) and cadmium (Cd) until 12days post-fertilization (dpf). Hatching, mortality, development, histology and gene expression were assessed. Our results showed that the highest concentration mixture of Cd (10mg/L) and Cu (1mg/L) affected survival, hatching time and hatching success. Occurrence of uninflated swim bladder was the highest (value) with exposure to 10mg/L Cd. Swim bladder was commonly over-inflated in a mixture (0.1mg/L Cd + 1.0mg/L Cu) exposure. Individuals exposed to the mixture (0.1 Cd + 1.0 Cu mg/L) showed up to a 7.69% increase in swim bladder area compared to the control group. The mixtures containing 0.1 or 10mg/L Cd, each with 1.0mg/L Cu resulted in significantly increased of Pbx1b expression, higher than any Cd or Cu alone (p < 0.01). In the co-exposure group (0.1/10 Cd + 1.0 Cu mg/L), Pbx1b expression was found at 12dpf but not 7dpf in controls. Higher concentrations of Cd may progressively reduce Pbx1b expression, potentially explaining why 75% of individuals in the 10mg/L Cd group failed to inflate their swim bladders. Additionally, the swim bladder proved to be a valuable bio-indicator for biological evaluation.

Tralomethrin causes cardiovascular toxicity in zebrafish (Danio rerio) embryos.

Tralomethrin, a synthetic pyrethroid insecticide used to control a wide range of pests in agriculture and public health, is highly toxic to aquatic organisms. However, data regarding the toxicity and underlying mechanisms of tralomethrin in aquatic organisms are limited. Thus, this study aimed to investigate the toxicity of tralomethrin in zebrafish. Zebrafish embryos were exposed to tralomethrin at different concentrations (16.63, 33.25, and 49.88 μg/L). Results showed that tralomethrin exposure caused cardiovascular dysplasia and dysfunction, including developmental abnormalities (pericardial edema, delayed yolk absorption, and uninflated swim bladder), elevated heart rate, and erythrogenesis disorders. Moreover, the expression patterns of crucial genes responsible for cardiovascular development (alas2, gata1a, hbbe2, nkx2.5, myl7, and myh6) also exhibited dysregulation in response to tralomethrin exposure. Oxidative stress occurred in embryos after exposure to tralomethrin. Collectively, our data suggest that exposure to tralomethrin induces cardiovascular and developmental toxicity in zebrafish. These findings are instrumental for evaluations of the environmental risk of tralomethrin in aquatic ecosystems in the future.

Comparison of the effects of empagliflozin and sotagliflozin on a zebrafish model of diabetic heart failure with reduced ejection fraction

The sodium-glucose cotransporter 2 (SGLT2) inhibitor empagliflozin (EMPA) and dual SGLT1/2 inhibitor sotagliflozin (SOTA) are emerging as heart failure (HF) medications in addition to having glucose-lowering effects in diabetes mellitus (DM). However, the precise mechanism underlying this cardioprotective effect has not yet been elucidated. Here, we evaluated the effects of EMPA and SOTA in a zebrafish model of DM combined with HF with reduced ejection fraction (DM-HFrEF). To compare the effects of the two drugs, survival, locomotion, and myocardial contractile function were evaluated. The structural binding and modulating effects of the two medications on sodium-hydrogen exchanger 1 (NHE1) were evaluated in silico and in vitro. DM-HFrEF zebrafish showed impaired cardiac contractility and decreased locomotion and survival, all of which were improved by 0.2–5 μM EMPA or SOTA treatment. However, the 25 μM SOTA treatment group had worse survival rates and less locomotion preservation than the EMPA treatment group at the same concentration, and pericardial edema and an uninflated swim bladder were observed. SOTA, EMPA and cariporide (CARI) showed similar structural binding affinities to NHE1 in a molecular docking analysis and drug response affinity target stability assay. In addition, EMPA, SOTA, and CARI effectively reduced intracellular Na+ and Ca2+ changes through the inhibition of NHE1 activity. These findings suggest that both EMPA and SOTA exert cardioprotective effects in the DM-HFrEF zebrafish model by inhibiting NHE1 activity. In addition, despite the similar cardioprotective effects of the two drugs, SOTA may be less effective than EMPA at high concentrations.

Deep Learning-Enabled Morphometric Analysis for Toxicity Screening Using Zebrafish Larvae.

Toxicology studies heavily rely on morphometric analysis to detect abnormalities and diagnose disease processes. The emergence of ever-increasing varieties of environmental pollutants makes it difficult to perform timely assessments, especially using in vivo models. Herein, we propose a deep learning-based morphometric analysis (DLMA) to quantitatively identify eight abnormal phenotypes (head hemorrhage, jaw malformation, uninflated swim bladder, pericardial edema, yolk edema, bent spine, dead, unhatched) and eight vital organ features (eye, head, jaw, heart, yolk, swim bladder, body length, and curvature) of zebrafish larvae. A data set composed of 2532 bright-field micrographs of zebrafish larvae at 120 h post fertilization was generated from toxicity screening of three categories of chemicals, i.e., endocrine disruptors (perfluorooctanesulfonate and bisphenol A), heavy metals (CdCl2 and PbI2), and emerging organic pollutants (acetaminophen, 2,7-dibromocarbazole, 3-monobromocarbazo, 3,6-dibromocarbazole, and 1,3,6,8-tetrabromocarbazo). Two typical deep learning models, one-stage and two-stage models (TensorMask, Mask R-CNN), were trained to implement phenotypic feature classification and segmentation. The accuracy was statistically validated with a mean average precision >0.93 in unlabeled data sets and a mean accuracy >0.86 in previously published data sets. Such a method effectively enables subjective morphometric analysis of zebrafish larvae to achieve efficient hazard identification of both chemicals and environmental pollutants.

Toxic effects of isofenphos-methyl on zebrafish embryonic development

Isofenphos-methyl (IFP) is widely used as an organophosphorus for controlling underground insects and nematodes. However, excessive use of IFP may pose potential risks to the environment and humans, but little information is available on its sublethal toxicity to aquatic organisms. To address this knowledge gap, the current study exposed zebrafish embryos to 2, 4, and 8 mg/L IFP within 6–96 h past fertilization (hpf) and measured mortality, hatching, developmental abnormalities, oxidative stress, gene expressions, and locomotor activity. The results showed that IFP exposure reduced the rates of heart and survival rate, hatchability, and body length of embryos and induced uninflated swim bladder and developmental malformations. Reduction in locomotive behavior and inhibition of AChE activity indicated that IFP exposure may induce behavioral defects and neurotoxicity in zebrafish larvae. IFP exposure also led to pericardial edema, longer venous sinus-arterial bulb (SV-BA) distance, and apoptosis of the heart cells. Moreover, IFP exposure increased the accumulation of reactive oxygen species (ROS) and the content of malonaldehyde (MDA), also elevated the levels of antioxidant enzymes of superoxide dismutase (SOD) and catalase (CAT), but decreased glutathione (GSH) levels in zebrafish embryos. The relative expressions of heart development-related genes (nkx2.5, nppa, gata4, and tbx2b), apoptosis-related genes (bcl2, p53, bax, and puma), and swim bladder development-related genes (foxA3, anxa5b, mnx1, and has2) were significantly altered by IFP exposure. Collectively, our results indicated that IFP induced developmental toxicity and neurotoxicity to zebrafish embryos and the mechanisms may be relevant to the activation of oxidative stress and reduction of acetylcholinesterase (AChE) content.

Cabotegravir Exposure of Zebrafish (Danio rerio) Embryos Impacts on Neurodevelopment and Behavior.

As most new medications, Cabotegravir (CAB) was recently approved as an antiretroviral treatment of HIV infection without in-depth safety information on in utero exposure. Although no developmental toxicity in rats and rabbits was reported, recent studies demonstrated that CAB decreases pluripotency of human embryonic stem cells. CAB exposure effects during development were assessed in zebrafish embryos by the Fish Embryo Toxicity test after exposure at subtherapeutic concentrations up to 25× the human Cmax. Larvae behavior was assessed by the light-dark locomotion test. The expression of factors involved in neurogenesis was evaluated by whole-mount in situ hybridization. CAB did not cause gross morphological defects at low doses, although pericardial edema, uninflated swim bladder, decreased heartbeats, growth delay, and decreased hatching rate were observed at the highest concentrations. Decreased locomotion was observed even at the subtherapeutic dose, suggesting alterations of nervous system integrity. This hypothesis was supported by the observation of decreased expression of crucial factors involved in early neuronal differentiation in diencephalic and telencephalic dopaminergic areas, midbrain/hindbrain boundary, and craniofacial ganglia. These findings support CAB effects on neurogenesis in zebrafish embryos and suggest long-term follow-up of exposed infants to provide data on drug safety during pregnancy.

Developmental Phenotypic and Transcriptomic Effects of Exposure to Nanomolar Levels of 4-Nonylphenol, Triclosan, and Triclocarban in Zebrafish (Danio rerio).

Triclosan, triclocarban and 4-nonylphenol are all chemicals of emerging concern found in a wide variety of consumer products that have exhibited a wide range of endocrine-disrupting effects and are present in increasing amounts in groundwater worldwide. Results of the present study indicate that exposure to these chemicals at critical developmental periods, whether long-term or short-term in duration, leads to significant mortality, morphologic, behavioral and transcriptomic effects in zebrafish (Danio rerio). These effects range from total mortality with either long- or short-term exposure at 100 and 1000 nM of triclosan, to abnormalities in uninflated swim bladder seen with long-term exposure to triclocarban and short-term exposure to 4-nonylphenol, and cardiac edema seen with short-term 4-nonylphenol exposure. Additionally, a significant number of genes involved in neurological and cardiovascular development were differentially expressed after the exposures, as well as lipid metabolism genes and metabolic pathways after exposure to each chemical. Such changes in behavior, gene expression, and pathway abnormalities caused by these three known endocrine disruptors have the potential to impact not only the local ecosystem, but human health as well.

Failure to gulp surface air induces swim bladder adenomas in Japanese medaka (Oryzias latipes).

In order to elucidate the effects of swim bladder inflation failure on swim bladder carcinogenesis, we investigated the sequential histopathological changes of swim bladders at 13, 24, 35, and 53 days post-hatch (dph) in medakas with an uninflated swim bladder, which was experimentally induced by denying access to the air–water interface between 0 and 6 dph. The reactive oxygen species (ROS) levels were measured at 24 dph. An uninflated swim bladder was induced in 47.3% of the fish denied access to the air–water interface (the denied group). The total incidence of swim bladder adenoma was 54.1% in the denied group; however, these tumors were observed in all fish with an uninflated swim bladder. In fact, these tumors were observed from 13 dph and onwards. The TBARS levels of the juveniles showed a 2.6-fold increase in fish with an uninflated swim bladder in the denied group compared to that in the control group. It is speculated that swim bladder inflation failure has some effects on the gas gland to produce ROS, leading to DNA damage in the gas glandular epithelium, which develops into swim bladder adenomas. Consequently, it is concluded that denying access to the air-water interface between 0 and 6 dph in medaka is an easy method of inducing swim bladder tumors in a short-term period, and is a useful method for producing tumor-bearing fish.

Toxicity of Weathered Sediment-Bound Dilbit to Early Life Stages of Zebrafish (Danio rerio).

Due to high viscosity, bitumen extracted from the Alberta oil sands is diluted with natural gas condensates to form diluted bitumen (dilbit) to facilitate transport through pipelines. Dilbit that is spilled into or near a waterbody is subject to environmental weathering processes such as evaporation and interaction with sediments. This is the first study that assessed the toxicity of weathered sediment-bound dilbit (WSD) to fish early life stages. Exposure of zebrafish (Danio rerio) embryos to water-soluble fractions (WSFs) or water-accommodated fractions (WAFs) of WSD from 30 min to 120 h postfertilization resulted in pericardial edema, yolk sac edema, and incidences of uninflated swim bladder. The presence of oil-mineral aggregates (OMAs) in the WAFs greatly increased toxicity, despite all fractions having similar concentrations of dissolved polycyclic aromatic hydrocarbons (PAHs). There were greater cyp1a mRNA abundances in larvae exposed to WAFs, suggesting that there were differences in bioavailability of PAHs between fractions. However, there was little evidence that embryotoxicity was caused by oxidative stress. Results suggest that evaporation and sediment interaction do not completely attenuate toxicity of dilbit to zebrafish early life stages, and OMAs in exposures exacerbate toxicity.

Lethal and sublethal toxicity of perfluorooctanoic acid (PFOA) in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow)

Perfluoroalkyl substances (PFAS), including perfluorooctanoic acid (PFOA), are industrial chemicals that are of concern due to their environmental presence, persistence, bioaccumulative potential, toxicity, and capacity for long-range transport. Despite a large body of research on environmental exposure, insufficient chronic aquatic toxicity data exist to develop water quality targets for clean-up of federal contaminated sites in Canada. Thus, our objective was to assess the aqueous toxicity of PFOA in chronic tests with Hyalella azteca (amphipod) and early-life stage tests with Pimephales promelas (fathead minnow). Toxicity data were analyzed based on measured PFOA concentrations. Amphipod exposures were 42 d (0.84–97 mg/L) and examined survival, growth, and reproduction. Fathead minnow exposures were 21 d (0.010–76 mg/L), which encompassed hatching (5 d) and larval stages until 16 d post-hatch; endpoints included hatching success, deformities at hatch, and larval survival and growth. Amphipod survival was significantly reduced at 97 mg/L (42-d LC50 = 51 mg/L), but growth and reproduction were more sensitive endpoints (42-d EC50 for both endpoints = 2.3 mg/L). Fathead minnows were less sensitive than amphipods, exhibiting no significant effects in all endpoints with the exception of uninflated swim bladder, which was significantly higher at 76 mg/L (15%) than controls (0%). Maximum concentrations of PFOA are generally in the ng/L range in global surface waters, but can reach the μg/L range in close proximity to major source inputs; therefore, environmental concentrations are well below those that caused toxicity in the current study. Our data will provide valuable information with which to assess the risk of PFOA at contaminated sites, and to set a target for site remediation.

Bisphenol-A induced antioxidants imbalance and cytokines alteration leading to immune suppression during larval development of Labeo rohita.

Recently, the oxidative stress and immunotoxicity biomarkers have been extensively used in embryotoxicity using fish embryos as promising models especially after exposure to chemical-like environmental estrogens. Bisphenol-A (BPA) is an estrogenic endocrine disruptor and is ubiquitous in the aquatic environment. Larvae of Labeo rohita were exposed to low concentrations of BPA (10, 100, 1000μg/l) for 21days. Innate immune system, antioxidants parameters, and developmental alterations were used as biomarkers. Exposure to BPA caused developmental abnormalities including un-inflated swim bladder, delayed yolk sac absorption, spinal curvature, and edema of pericardium. Lipid peroxidation increased and activity of catalase (p < 0.05), superoxide dismutase (p < 0.05), and glutathione peroxidase (p < 0.01) decreased after exposure to BPA. Level of reduced glutathione also decreased (p < 0.05) in BPA-exposed group. Lower expression of tumor necrosis factor-α (p < 0.05) and interferon-γ (p < 0.001) was observed in BPA-exposed groups while expression of interleukin-10 increased (p < 0.05) in larvae exposed to 10μg/l BPA. Moreover, exposure of BPA caused a concentration-dependent increase in expression of heat shock protein 70 (p < 0.05). The present study showed that the exposure to BPA in early life stages of Labeo rohita caused oxidative stress and suppress NF-κB signaling pathway leading to immunosuppression. The results presented here demonstrate the cross talk between heat shock protein 70 and cytokines expression.

Lethal and Sub-lethal Effects of Nitrofurantoin on Zebrafish Early-Life Stages

Antibiotics are among the most extensively used pharmaceuticals worldwide. They are natural or synthetic drugs with the capacity to kill or inhibit the growth of microorganisms. Several antibiotics have been detected in aquatic environments, but little is known about their effects on non-target organisms, especially fish. The aim of this study was to evaluate the effects of the antibiotic nitrofurantoin (NTF) using zebrafish embryos as model organisms. To assess mortality and development effects, the embryos were exposed to 0, 4, 9, 44, 100, 223 and 500 mg/L of NTF. A sub-lethal range of concentrations (0, 0.001, 0.02, 0.32, 5.62 and 100 mg/L) was used for biomarker analyses, namely cholinesterase, lactate dehydrogenase, glutathione S-transferase and catalase. The results indicated low toxicity of NTF to zebrafish, with a 168 h-LC50 value of 129.2 mg/L. The main effect on development was the loss of equilibrium related to the uninflated swim bladder (168 h-EC50 = 96.72 mg/L). Biomarker activity was induced in concentrations as low as 0.02 mg/L (cholinesterase, lactate dehydrogenase, glutathione S-transferase). Exposure to NTF induced no significant effects on zebrafish larvae behaviour. In summary, short-term exposure of zebrafish embryos to NTF induced developmental alterations only at high concentrations. However, biochemical changes occurred at lower levels of exposure, suggesting long-term effects on fish populations.

Adverse effect of cylindrospermopsin on embryonic development in zebrafish (Danio rerio).

Eutrophication of freshwater bodies increases the occurrence of toxic cyanobacterial blooms. The cyanobacterial toxin cylindrospermopsin (CYN) is receiving great interest due to its increasing presence in waterbodies. However, the toxic effects of CYN on zebrafish development are poorly understood, especially the toxicological mechanism, which is still unclear. In this study, we examined the adverse effects of CYN on embryonic development in zebrafish. CYN (2-2000 nM) exposure decreased embryos survival rate, hatching rate, body length and eye size in a concentration-dependent manner and caused abnormalities in embryo morphology, including pericardial edema, spinal curvature, tail deformity, uninflated swim bladder, cardiac and vascular defects. CYN at concentrations of 20 nM or higher significantly increased ROS level and promoted cell apoptosis in zebrafish embryos. To preliminarily elucidate the potential mechanism of zebrafish developmental toxicity caused by CYN, we examined the expression of oxidative stress- and apoptotic-related genes. CYN could promote the expression of oxidative stress-related genes (SOD1, CAT and GPx1) and induce changes in transcriptional levels of apoptotic-related genes (p53, Bax and Bcl-2). Taken together, CYN induced adverse effects on zebrafish embryos development, which may associate with oxidative stress and apoptosis. These outcomes will advance our understanding of CYN toxicity, environmental problems and health hazards caused by climate changes and eutrophication.

Osteological development and deformities in hatchery-reared longtooth grouper (Epinephelus bruneus ): Vertebral column, dorsal-fin supports and caudal-fin skeleton

Skeletal deformities are one of the serious problems in hatchery-reared longtooth grouper (Epinephelus bruneus). In this study, seed production of longtooth grouper was carried out in four large hatchery-grade tanks. Osteological development of the vertebral column, caudal-fin skeleton and dorsal-fin supports in larvae and juveniles was described. The vertebral ontogeny started at 4.8 mm total length (TL) with the formation of neural spines 2 and 3, and the vertebral centra began forming at 5.3 mm TL. By 16 mm TL, all vertebral bony elements were formed except for the dorsal ribs. The onsets of the ontogeny of the caudal-fin skeleton and dorsal-fin supports occurred at approximately 5.1 mm and 3.9 mm TL, respectively, and the completion of these bony elements occurred at approximately 20 mm TL. The incidence of deformity was examined; some type of deformity was observed in 57%–68% of the specimens. These deformities mainly occurred as three types: lordosis, around vertebrae 1–5; saddleback-like syndrome, around vertebrae 7–11; and vertebrae fusion, around vertebrae 22–24. The incidence of prehaemal lordosis was significantly higher among individuals with an uninflated swim bladder than among those with an inflated swim bladder.

Developmental toxicity of synthetic phenolic antioxidants to the early life stage of zebrafish

Synthetic phenolic antioxidants (SPAs) have gained high concerns due to their extensive usages and unintended environmental release via various routes. Their contamination in water system could pose potential threat to aquatic organisms, therefore, the studies on the aquatic toxicology of this kind of chemicals are of high importance. In this research, the developmental toxicities of four commonly used SPAs, including butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tert-butyl hydroquinone (TBHQ), and 2,2′-methylenebis (6-tert-butyl-4-methylphenol) (AO2246) were investigated using the zebrafish embryo toxicity test (ZFET). The results showed that these four SPAs exerted different acute toxicities to zebrafish, and the toxic order, based on their 96 h LC50 values, was AO2246 > TBHQ > BHA > BHT, and decreased hatching rates were induced for the embryos in BHA, TBHQ and AO2246 exposure groups. Non-lethal exposures of BHA (≤20 μM), TBHQ (≤20 μM), BHT (≤200 μM) and AO2246 (≤2 μM) decreased the heart rates and body lengths of zebrafish in exposure concentration-dependent manners. Diverse morphological deformities, including uninflated swim bladder, pericardial edema, spinal curvature, severe yolk deformation, or abnormal pigmentation, were induced in zebrafish larvae upon SPA treatments. The transcriptional levels of the related genes, examined by quantitative PCR, indicated that the interferences of SPAs with hypothalamic-pituitary-thyroid axis (HPT axis), GH/PRL synthesis and Hedgehog (hh) pathway contributed to their developmental toxicities in zebrafish. The up-regulation of pluripotency biomarker, Oct4, caused the developmental retardation during the early stages of zebrafish embryos in BHA and TBHQ exposure groups. The results obtained herein provided important information on the developmental toxicity of SPAs, which could be very helpful in guiding the risk assessment on their aquatic toxicology.

Toxicological Assessment of a Lignin Core Nanoparticle Doped with Silver as an Alternative to Conventional Silver Core Nanoparticles.

Elevated levels of silver in the environment are anticipated with an increase in silver nanoparticle (AgNP) production and use in consumer products. To potentially reduce the burden of silver ion release from conventional solid core AgNPs, a lignin-core particle doped with silver ions and surface-stabilized with a polycationic electrolyte layer was engineered. Our objective was to determine whether any of the formulation components elicit toxicological responses using embryonic zebrafish. Ionic silver and free surface stabilizer were the most toxic constituents, although when associated separately or together with the lignin core particles, the toxicity of the formulations decreased significantly. The overall toxicity of lignin formulations containing silver was similar to other studies on a silver mass basis, and led to a significantly higher prevalence of uninflated swim bladder and yolk sac edema. Comparative analysis of dialyzed samples which had leached their loosely bound Ag+, showed a significant increase in mortality immediately after dialysis, in addition to eliciting significant increases in types of sublethal responses relative to the freshly prepared non-dialyzed samples. ICP-OES/MS analysis indicated that silver ion release from the particle into solution was continuous, and the rate of release differed when the surface stabilizer was not present. Overall, our study indicates that the lignin core is an effective alternative to conventional solid core AgNPs for potentially reducing the burden of silver released into the environment from a variety of consumer products.