Yolk Sac Absorption Research Articles

Risk assessment of developmental and neurotoxicity by the flavoring agent perillaldehyde: NAC (N-acetylcysteine) mitigation of oxidative stress-mediated inhibition of the Nrf2 pathway.

Perillaldehyde (PAE), a prevalent flavoring agent, has raised safety concerns due to conflicting evidence regarding its toxicity. This study provides a comprehensive assessment of the developmental and neurotoxic effects of PAE in zebrafish, elucidating the underlying mechanisms of its toxicity. Results showed that PAE affected the viability and hatching rate of zebrafish at 96 h postfertilization with the 50 % lethal concentration (LC50) of 7.975 mg/L. Furthermore, exposed‌ to a non-lethal concentration of 4 mg/L PAE induced a spectrum of morphological abnormalities, such as pericardial edema, delayed yolk sac absorption, reduced body length, and microphthalmia. Behavioral observations revealed that PAE reduced motor ability, and was accompanied by an increase in spontaneous turning angle and angular velocity. Using the TG(elav13:EGFP) transgenic model, we observed the number of newborn neurons was reduced, indicating that PAE induced obvious neurotoxic effects. Additionally, this concentration facilitated the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), concomitantly decreasing the activity of antioxidant enzymes. QRT-PCR analysis revealed that PAE down-regulated Nestin and Neurogenin1 gene expression, up-regulated Glipr1a and Nox1 gene expression, and inhibited the Nrf2/HO-1 pathway. Notably, co-administration of N-acetylcysteine (NAC), an inhibitor of oxidative stress, mitigated oxidative stress levels and partially ameliorated the neurotoxicity. These findings suggest that oxidative stress is the primary mediator of PAE-induced neurotoxicity. This study provides crucial insights for the safe application of PAE.

Gene expression in the hypothalamic-pituitary-thyroid axis in Seriola rivoliana early larvae development at different temperatures

We analyzed the expression of genes involved in the hypothalamic-pituitary-thyroid axis (HPT-axis) in the longfin yellowtail Seriola rivoliana early larva, including temperature effects (22, 26 and 28 °C) and days of development (day one, day two, and day six after hatching). We aimed to determine if egg and larval incubation at different temperatures could disrupt this critical endocrine axis, which, in an aquaculture context, it could provoke mortality during early metamorphosis. There was a significant interaction between temperature and developmental timing on the relative expression of thyrotropin releasing hormone (trh). Larvae at 22 °C was the longest and increased more trh expression than larvae at higher temperatures. Interestingly, thyrotropin stimulating hormone (tsh) was highly expressed after hatching. Subsequently, it was downregulated at any temperature at least until day four, suggesting a temporal inhibition of the HPT axis. Therefore, we suggest that tsh-binding (tshr) to follicles should have occurred from hatching, creating a further “cascade effect” of upregulation of larval thyroglobulin (tg) from day two in a temperature-dependent manner. Consequently, new thyroid hormones should have been produced after yolk sac absorption. The above may indicate a narrow window of larval survival, where the larval transition from endogenous to exogenous feeding would depend on the correct timing to synthesize tg. Temperature significantly affected the expressions of deiodinase 1 (dio1-downregulated) and deiodinase 2 (dio2-upregulated) after hatching. The expressions of thyroid receptors alpha (trα) and beta (trβ) remained constant after hatching without significant effects of temperature and days of development. Then, the differential expression on day six showed that all HPT-axis transcripts increased their expressions as larvae developed, which suggested a functional HPT. Finally, there was no evidence that any temperature would disrupt the endocrine’s larval axis, which indicated that the longfin yellowtail has a wide temperature adaption. Nevertheless, based on tg upregulation, we suggest that larvae should be maintained around 25–26 °C after hatching for a better chance of survival and development.

Ontogeny and Growth of Early Life Stages of Barred Spiny Eel Macrognathus pancalus (Hamilton, 1822)

Background: The objective of the study was to examine the growth and development of M. pancalus larvae in captive conditions, including pigmentation patterns, yolk and oil globule utilization, growth rates and metamorphosis, as well as to identify suitable live food for larviculture. Methods: The larvae, produced through induced breeding, were kept unfed until the yolk-sac absorption and then fed with freshwater rotifer, mixed zooplankton and Tubifex worms at different stages of development. The larvae were reared in indoor glass tanks and outdoor reinforced plastic tanks to document their morphological and chronological development. Result: The findings showed that the larvae exhibited typical characteristics of eel. Notably, dentition was observed before yolk resorption, indicating the highly predatory nature of the larvae. The bands of melanophores, which were prominent at the beginning, disappeared between 5-7 days post-hatch. Additionally, the study identified distinct features of spiny eels, such as a trunk-like rostral projection at 14 days post-hatch and erectile dorsal spines at 21 days post-hatch. The growth pattern, morphological changes and adaptation to benthic burrowing habits indicated that the larvae metamorphosed into the fry stage between 28-45 days post-hatch. These results provide valuable insights into the ontogenetic development and rearing of M. pancalus and can be used to support future aquaculture and conservation programs.

Development of the cranial lateral line system of Brook Trout, Salvelinus fontinalis (Teleostei: Salmonidae): Evolutionary and ecological implications.

The mechanosensory lateral line (LL) system of salmonid fishes has been the focus of comparative morphological studies and behavioral and physiologicalanalyses of flow sensing capabilities, but its morphology and development have not been studied in detail in any one species. Here, we describe the post-embryonic development of the cranial LL system in Brook Trout, Salvelinus fontinalis, using vital fluorescent staining (4-Di-2-ASP), scanning electron microscopy, µCT, and clearing and staining to visualize neuromasts and the process of cranial LL canal morphogenesis. We examined the relationship between the timing of LL development, the prolonged life history of salmonids, and potential ecological implications. The LL system is composed of seven canals containing canal neuromasts (CNs) and four lines of superficial neuromasts (SNs) on the skin. CNs and SNs increase in number and size during the alevin (larval) stage. CN number stabilizes as canal morphogenesis commences, but SN number increases well into the parr (juvenile) stage. CNs become larger and more elongated than SNs, but the relative area occupied by sensory hair cells decreases during ontogeny in both types of neuromasts. Neuromast-centered canal morphogenesis starts in alevins (yolk sac larvae), as they swim up into the water column from their gravel nests (~4 months post-fertilization), after which yolk sac absorption is completed and exogenous feeding begins. Canal morphogenesis proceeds asynchronously within and among canal series and is not complete until ~8 months post-fertilization (the parr stage). Three characters in the LL system and associated dermal bones were used to identify their homologs in other actinopterygians and to consider the evolution of LL canal reduction, thus demonstrating the value of salmonidsfor the study of LL evolution. The prolonged life history of Brook Trout and the onset of canal morphogenesis at swim-up are predicted to have implications for neuromast function at these critical behavioral and ecological transitions.

Prediction of developmental toxic effects of fine particulate matter (PM2.5) water-soluble components via machine learning through observation of PM2.5 from diverse urban areas

The global health implications of fine particulate matter (PM2.5) underscore the imperative need for research into its toxicity and chemical composition. In this study, zebrafish embryos exposed to the water-soluble components of PM2.5 from two cities (Harbin and Hangzhou) with differences in air quality, underwent microscopic examination to identify primary target organs. The Harbin PM2.5 induced dose-dependent organ malformation in zebrafish, indicating a higher level of toxicity than that of the Hangzhou sample. Harbin PM2.5 led to severe deformities such as pericardial edema and a high mortality rate, while the Hangzhou sample exhibited hepatotoxicity, causing delayed yolk sac absorption. The experimental determination of PM2.5 constituents was followed by the application of four algorithms for predictive toxicological assessment. The random forest algorithm correctly predicted each of the effect classes and showed the best performance, suggesting that zebrafish malformation rates were strongly correlated with water-soluble components of PM2.5. Feature selection identified the water-soluble ions F− and Cl− and metallic elements Al, K, Mn, and Be as potential key components affecting zebrafish development. This study provides new insights into the developmental toxicity of PM2.5 and offers a new approach for predicting and exploring the health effects of PM2.5.

Reduction of methyltestosterone concentration in feed during masculinization of Nile tilapia (Oreochromis niloticus) in biofloc system

Most larviculture of Nile tilapia is carried out in ponds with a large presence of live food, with temperature variations throughout the day and year, and feed leftoovers due to dissipation in the water. As a consequence, the masculinization process needs to be done with a high concentration of 17α-methyltestosterone (MT) in the diet. Production in closed systems, such as biofloc technology (BFT), allows controlling the water temperature, uses a smaller volume of water and generates less effluent, being an alternative system for the masculinization process. This study evaluated the masculinization rates of Nile tilapia in the BFT, with a controlled temperature (∼28 °C), using concentrations of MT in the feed lower than those recommended for masculinization in ponds and BFT (60 mg ∙ Kg−1 of feed). Tilapia larvae (post yolk sac absorption) were raised in BFT (2 larvae ∙ L−1 or 100 larvae ∙ tank−1) and submitted to different concentrations of MT in the diet (0, 30, 40, 50 and 60 mg ∙ Kg−1 of feed), which was offered five times a day for a period of 28 days. The experimental design was entirely randomized. After the hormonal treatment, the water in the tanks was completely replaced with new water, free of hormonal residues, where the fry remained until they reached a viable size for sexing. It was also tested to maintain the original water used during the hormonal treatment period, during the post-experimental period, where 30 mg ∙ Kg−1 of feed was used. The water quality variables (temperature, pH, dissolved oxygen, settleable solids, total suspended solids, alkalinity, nitrogen compounds, salinity, phosphate and organic carbon) and growth performance variables (final body weight and length, specific growth rate, condition factor and survival) did not differ between MT concentration in the diet (p > .05). The treatment without hormone (control) presented 73.6% of males and the treatments that received hormone showed masculinization rates >99%, significantly higher than control (p < .05), but did not differ from each other (p > .05). The percentage of male individuals in the control treatment was high and this may have occurred because of the temperature of 28 °C throughout the experiment. However, this percentage of males is not considered high enough for production, which requires higher rates close to 100% as possible. The MT was not detected in the water 12 h after the last feeding with feed containing hormone. Thus, it is feasible to use a lower concentration (30 mg of MT ∙ Kg−1 of feed) in BFT. Our study has a direct impact on the global tilapia production scenario, offering a new environmentally sustainable masculinization protocol, using less water and hormones in the process and avoiding the release of hormonal residues.

Synergistic developmental effects of zebrafish exposed to combined perfluorooctanoic acid and atrazine

Perfluorooctanoic acid (PFOA) and atrazine are two endocrine disruptors that are widely found in waters. Negative effects of PFOA and atrazine have been studied individually, but few data have focused on their combined effects. Here, zebrafish embryos were used as model to investigate the combined toxicity of PFOA and atrazine. The acute toxicity of atrazine (11.9 mg/L) to zebrafish embryos was much higher than that of perfluorooctanoic acid (224.6 mg/L) as shown by the 120h-LC50 value. Developmental effects, including delayed yolk sac absorption, spinal curvature, and liver abnormalities, were observed in both one- and two-component exposures. Notably, the rate of embryonic malformations in the co-exposure group was more than twice as high as that of single component exposure in the concentration range of 1/8-1/2 EC50, which indicated a synergistic effect of the binary mixture. The synergistic effect of PFOA-atrazine was further validated by combinatorial index (CI) modeling. In addition, changes of amino acid metabolites, reactive oxygen species and superoxide dismutase indicated that oxidative stress might be the main pathway for enhanced toxicity under co-exposure condition. Overall, co-exposure of PFOA and atrazine resulted in stronger developmental effects and more complicated amino acid metabolic response toward zebrafish, compared with single component exposure.

Dimethyl fumarate induces cardiac developmental toxicity in zebrafish via down-regulation of oxidative stress

Dimethyl fumarate (DMF) is an immunosuppressant commonly used to treat multiple sclerosis and other autoimmune diseases. Despite known side effects such as lymphopenia, the effect of DMF on cardiac development remains unclear. To assess this, we used zebrafish to evaluate the cardiac developmental toxicity of DMF. Our study showed that DMF reduced the survival rate of zebrafish embryos, with those exposed to 1, 1.3, and 1.6 mg/L exhibiting heart rate reduction, shortened body length, delayed yolk sac absorption, pericardial edema, increased distance from sinus venous to bulbus arteriosus, and separation of cardiomyocytes and endocardial cells at 72 hpf. Heart development-related genes showed disorder, apoptosis-related genes were up-regulated, and the oxidative stress response was down-regulated. Treatment with cysteamine ameliorated the heart development defects. Our study demonstrates that DMF induces cardiac developmental toxicity in zebrafish, possibly by down-regulating oxidative stress responses. This study provides a certain research basis for further study of DMF-induced cardiac developmental toxicity, and provides some experimental evidence for future clinical application and study of DMF.

Comparison of cardiotoxicity induced by alectinib, apatinib, lenvatinib and anlotinib in zebrafish embryos

Four tyrosine kinase inhibitors, alectinib, apatinib, lenvatinib and anlotinib, have been shown to be effective in the treatment of clinical tumors, but their cardiac risks have also raised concerns. In this study, zebrafish embryos at 6 h post fertilization (hpf) were exposed to the four drugs at concentrations of 0.05–0.2 mg/L until 72 hpf, and then the development of these embryos was quantified, including heart rate, body length, yolk sac area, pericardial area, distance between venous sinus and balloon arteriosus (SV-BA), separation of cardiac myocytes and endocardium, gene expression, vascular development and oxidative stress. At the same exposure concentrations, alectinib and apatinib had little effect on the cardiac development of zebrafish embryos, while lenvatinib and anlotinib could induce significant cardiotoxicity and developmental toxicity, including shortened of body length, delayed absorption of yolk sac, pericardial edema, prolonged SV-BA distance, separation of cardiomyocytes and endocardial cells, and downregulation of key genes for heart development. Heart rate decreased in all four drug treatment groups. In terms of vascular development, alectinib and apatinib did not inhibit the growth of embryonic intersegmental vessels (ISVs) and retinal vessels, while lenvatinib and anlotinib caused serious vascular toxicity, and the inhibition of anlotinib in vascular development was more obvious. Besides, the level of reactive oxygen species (ROS) in the lenvatinib and anlotinib treatment groups was significantly increased. Our results provide reference for comparing the cardiotoxicity of the four drugs.

Hepatic effects of acetochlor chiral isomers in zebrafish and L02 cells

The pesticide acetochlor (ACT) is a chiral isomer commonly detected in the global environment, yet its specific impacts on liver function remain poorly understood. We utilized zebrafish and L02 cells as research models to comprehensively investigate how ACT and its chiral isomers affect the liver. Our investigations unveiled that the R, Rac, and S isomers of ACT disrupt hepatic lipid transport, catabolism, and synthesis, leading to delayed yolk sac absorption and the accumulation of lipids in zebrafish embryos. These isomers induce oxidative stress in the liver of zebrafish embryos, reducing antioxidant levels and enzyme activity. The accumulated lipids in the liver render it susceptible to oxidative stress, further exacerbating hepatocyte damage. Hepatocyte damage manifests as extensive vacuolization of liver cells and alterations in liver morphology, which are induced by R, Rac, and S. Furthermore, we elucidated the molecular mechanisms underpinning the disturbance of hepatic lipid metabolism by R, Rac, and S in L02 cells. These compounds stimulate lipid synthesis through the upregulation of the AMPK/SREBP-1c/FAS pathway while inhibiting lipolysis via downregulation of the PPAR-α/CPT-1a pathway. Remarkably, our results highlight that S exhibits significantly higher hepatotoxicity in comparison to R. This study provides valuable insights into the hepatic effects of ACT chiral isomers.

Growth performance, reproductive status, and chromosomal instability in triploid Nile tilapias.

Reproductive control is one of the biggest challenges in tilapia production and triploidy was developed as an alternative to sterilization. In general, polyploids present chromosomal instability but for triploid Nile tilapia it has yet to be reported. This study evaluated the chromosomal instability from juveniles to adulthood, growth performance and gonadal status of tilapia hatched from eggs submitted or not to heat shock for triploid induction. Nile tilapia oocytes were fertilized (1,476 oocytes), half of the eggs were subjected to a four-minute shock in 41 °C water four minutes after fertilization and the other half were not (Control group). The eggs were incubated (at 27°C) and 160 larvae from the treated group hatched and survived after yolk sac absorption. The determination of ploidy was performed by flow cytometry at 85th (juveniles) and 301st (adults) days of age post yolk sac absorption. At the time of the first cytometry analysis there were 73 surviving juveniles from the treated group, and only 14 were confirmed triploid. However, at the analysis of adult ploidy, one out of 8 surviving adult tilapias from the 14 confirmed triploid juveniles remained triploid. Gonadal histology showed that the non-remaining triploids continued to produce gametes. The growth performance of triploid tilapia was initially superior to that of diploid tilapia during the juvenile phase, but similar in adults. Once the chromosome sets are lost and the tilapias become diploid again, at least in tissues with a high proliferation rate, such as the hematopoietic tissue that was analyzed (and possibly in gonads), all possible advantages of triploids are probably lost. Thus, our results suggest that, due to genomic instabilities, the triploid generation of tilapia has low efficiency.

Effects of cold stress on physiologic metabolism in the initial phase and performance of broiler rearing

This study aimed to investigate the effects of 8 h of cold stress (18 °C) every day in broiler chicks during the first 7 days of rearing on crop filling analysis, yolk sac consumption, digestive and immune organs weights, and physiological metabolism at seven days and performance between 1 and 35 days. Cobb500 male broiler chickens (n = 274) were randomly assigned to two treatments. The treatments consisted of varying environmental temperatures during the first week post-housing. Chicks were reared at a thermoneutral temperature (32 °C) or under cold stress (18 °C) for 8 h/day during the first week, and both groups were subsequently reared at a thermoneutral temperature for 8–35 days. The thermoneutral group reached 90% full crop after 48 h of housing (P < 0.05), while the cold-stressed group had more empty crops at 2 h and 48 h after housing (P < 0.05). The chick cloacal temperature was not affected by the treatments (P > 0.05). Additionally, the treatment did not affect serum amylase and corticosterone levels, feed intake, body weight gain, or feed conversion ratio (P > 0.05, while the cold-stressed group had elevated heterophil/lymphocyte count at day 7 (P < 0.05). The thermoneutral group showed higher viability (%) at 7 and 35 days and a higher production factor at 35 days (P < 0.05). Broiler chickens under cyclic cold stress experienced decreased yolk sac absorption during the first week and increased feed intake and feed conversion ratio after 35 days of rearing. Viability was also lower in the cold-stressed group. An appropriate strategy to minimize these adverse effects is to rear the chicks in a thermoneutral environment during the first week.

Apoptotic mechanism of development inhibition in zebrafish induced by esketamine

Esketamine, a widely used intravenous general anesthetic, is also employed for obstetric and pediatric anesthesia, and depression treatment. However, concerns regarding esketamine abuse have emerged. Moreover, the potential in vivo toxicity of esketamine on growth and development remains unclear. To address these concerns, we investigated the effects of esketamine exposure on developmental parameters, cell apoptosis, and gene expression in zebrafish. Esketamine exposure concentration-dependently decreased the heart rate and body length of zebrafish embryos/larvae while increasing the hatching rate and spontaneous movement frequency. Developmental retardation of zebrafish larvae, including shallow pigmentation, small eyes, and delayed yolk sac absorption, was also observed following esketamine treatment. Esketamine exposure altered the expression of apoptosis-related genes in zebrafish heads, primarily downregulating bax, caspase9, caspase3, caspase6, and caspase7. Intriguingly, BTSA1, a Bax agonist, reversed the anti-apoptotic and decelerated body growth effects of esketamine in zebrafish. Collectively, our findings suggest that esketamine may hinder embryonic development by inhibiting embryonic apoptosis via the Bax/Caspase9/Caspase3 pathway. To the best of our knowledge, this is the first study to report the lethal toxicity of esketamine in zebrafish. We have elucidated the developmental toxic effects of esketamine on zebrafish larvae and its potential apoptotic mechanisms. Further studies are warranted to evaluate the safety of esketamine in animals and humans.

A Comprehensive Biochemical Characterization of Hybrid Grouper Larvae (Epinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂) during Yolk-Sac Larval Development.

To investigate the shifts in the biochemical composition of hybrid grouper during the early larval stages, we collected samples at various developmental milestones, spanning from newly hatched larvae (stage I) to 4 days after hatching (stage V). Our findings revealed several notable trends: (1) The total length of hybrid grouper larvae exhibited a significant increase as the yolk-sac absorption progressed from stage I to V. Concurrently, there was a marked decrease in yolk volume and oil volume during the transition from stage I to III, followed by a gradual decline from stage III to V. (2) Dry weight and total lipid content displayed a rapid reduction throughout the larval development period, while the total protein content exhibited a declining trend. (3) The concentrations of triacylglycerols and wax esters/steryl esters decreased considerably, particularly at stage V. However, no differences were observed among the contents of ketones, hydrocarbons, and sterols. (4) As yolk-sac larvae developed from stage I to V, a significant reduction was observed in the levels of essential amino acids (EAAs), such as leucine, valine, isoleucine, phenylalanine, glycine, alanine, serine, proline, and tyrosine. This trend was also observed for non-EAAs and total amino acids, with fluctuations in the content of other amino acids. (5) There was a significant decrease in the levels of specific fatty acids, including C16:0, saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), C18:0, 18:1n-9, and C20:4n-6. In contrast, the contents of C22:6n-3, polyunsaturated fatty acids (PUFAs), n-3 PUFA, n-6 PUFA, and the combination of docosahexaenoic acid (DHA) + eicosapentaenoic acid (EPA), as well as the DHA/EPA ratio, remained stable from stage I to III but declined thereafter. (6) During the early developmental stages, the utilization sequence of fatty acids followed a pattern of prioritizing SFAs, followed by MUFAs, n-6 PUFA, and n-3 PUFA. These findings provide further insights into the nutritional priorities of hybrid grouper larvae during their early development, with a particular emphasis on lipids and fatty acids as vital energy sources. Additionally, our results highlight variations in the efficiency of utilization among different types of fatty acids, while protein utilization remained relatively stable, characterized by the selective consumption of amino acid content.

Benzophenone induces cardiac developmental toxicity in zebrafish embryos by upregulating Wnt signaling

Benzophenone (BP) is found in many popular consumer products, such as cosmetics. BP potential toxicity to humans and aquatic organisms has emerged as an increased concern. In current study, we utilized a zebrafish model to assess BP-induced developmental cardiotoxicity. Following BP exposure, zebrafish embryos exhibited developmental toxicity, including increased mortality, reduced hatchability, delayed yolk sac absorption, and shortened body length. Besides, BP exposure induced cardiac defects in zebrafish embryos, comprising pericardial edema, reduced myocardial contractility and rhythm disturbances, and altered expression levels of cardiac developmental marker genes. Mechanistically, BP exposure disturbed the redox state and increased the level of apoptosis in zebrafish cardiomyocytes. Transcriptional expression levels of Wnt signaling genes, involving lef1, axin2, and β-catenin, were upregulated after BP treatment. Inhibition of Wnt signaling with IWR-1 could rescue the BP-induced cardiotoxicity in zebrafish. In summary, BP exposure causes cardiotoxicity via upregulation of the Wnt signaling pathway in zebrafish embryos.

The larval attachment organ of the bowfin Amia ocellicauda Richardson, 1836 (Amiiformes: Amiidae) and its phylogenetic significance.

Larval attachment organs (LAOs) are unicellular or multicellular organs that enable the larvae of many actinopterygian fishes to adhere to a substrate before yolk-sac absorption and the free-swimming stage. Bowfins (Amiiformes) exhibit a sizable LAO on the snout, which was first described in the late 19th and early 20th centuries. In this study, we document the LAO of Amia ocellicauda (Richardson, 1836) using a combination of scanning electron microscopy (SEM) and light microscopy, and histochemistry. We examined material representing three stages with SEM ranging in size from 5.8 to 11.2 mm in notochord length and one stage histochemically. We compare the LAO of A. ocellicauda to that of the lepisosteid Atractosteus tropicus Gill, 1863 and show that although the LAOs of A. ocellicauda and A. tropicus are both super-organs, the two differ in the ultrastructure of the entire organ. A. ocellicauda possesses two distinct lobes, with the organs arranged on the periphery with none in the middle, whereas A. tropicus also possesses two lobes, but with the organs scattered evenly across the super-organ. The individual organs of A. ocellicauda possess adhesive cells set deep to support cells with the adhesive substance released through a pore, whereas A. tropicus possesses both support cells and adhesive cells sitting at a similar level, with the adhesive substance released directly onto the surface of the organ. We additionally provide a table summarizing vertebrate genera in which attachment organs have been documented and discuss the implications of our study for hypotheses of the homology of attachment organs in the Holostei.

Immunotoxicity and transcriptome analysis of zebrafish embryos exposure to Nitazoxanide

Nitazoxanide (NTZ) is a broad-spectrum immunomodulatory drug, and little information is about the immunotoxicity of aquatic organisms induced by NTZ. In the present study, reduced body length and decreased yolk sac absorption in the NTZ-treated group were observed. Meanwhile, the number of innate immune cells and adaptive immune cells was substantially reduced upon NTZ exposure, and the migration and retention of macrophages and neutrophils in the injured area were inhibited. Following NTZ stimulation, oxidative stress levels in the zebrafish increased obviously. Mechanistically, RNA-seq, a high-throughput method, was performed to analyze the global expression of differentially expressed genes (DEGs) in zebrafish embryos treated with NTZ. 531 DEGs were identified by comparative transcriptome analysis, including 121 up-regulated and 420 down-regulated genes in zebrafish embryos after NTZ exposure. The transcriptome sequences were further subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) and analysis, showing phototransduction and metabolic pathway, respectively, and were most enriched. In addition, some immune-related genes were inhibited after NTZ exposure. RNA-seq results confirmed by qRT-PCR were used to verify the expression of the 6 selected genes. The other immune-related genes such as two pro-inflammatory cytokines (IL-1β, tnfα) and two chemokines (CXCL8b.3, CXCL-c1c) were further confirmed and were differentially regulated after NTZ exposure. In summary, NTZ exposure could lead to immunotoxicity and increased ROS in zebrafish embryos, this study provides valuable information for future elucidating the molecular mechanism of exogenous stimuli-induced immunotoxicity in aquatic ecosystems.

Combined Toxicities of Di-Butyl Phthalate and Polyethylene Terephthalate to Zebrafish Embryos.

The increasing concern for the ecological risks of microplastics (MPs) as carriers of hydrophobic organic contaminants is evident. Di-butyl phthalate (DBP) is extensively utilized as an additive in plastic products, and both DBP and MPs are widespread in the environment. However, the combined toxicity of these substances remains uncertain. In this study, zebrafish embryos were employed to assess the toxic effects of polyethylene terephthalate (PET, MPs) and DBP, with a focus on the DBP toxicities influenced by PET. The embryonic chorion was partially covered by PET particles, and PET led to a delayed hatching of zebrafish embryos without inducing death or teratogenesis. On the other hand, exposure to DBP considerably inhibited the hatching of embryos, leading to severe lethal and teratogenic effects. The most common phenotypes induced by DBP exposure were delayed yolk sac absorption and pericardial edema. The mortality increased in co-treatment with 100 particles/mL PET and 2 mg/L DBP at 24 hpf and 48 hpf. The malformation phenotype, bent notochord, and delayed yolk sac absorption became more severe in 1 mg/L DBP exposition with the co-exposure of 100 particles/mL PET at 72 hpf. PET might act as a carrier that enhances the bioavailability of ambient DBP.

Determination of Hatching Performance, Yolk-Sac Absorption, and Larval Growth Rates in Abant Trout (Salmo trutta abanticus), Brook Trout (Salvelinus fontinalis), and their Hybrids

In this study, hatching performance, yolk-sac absorption, and growth rates of Abant trout (Salmo trutta abanticus), brook trout (Salvelinus fontinalis), and their hybrids were investigated. In this contex, relationships between duration (days) and total larvae length, total larvae wet weight, dry larvae body weight and yolk-sac weights, and larval growth were determined. At the end of the study, eye pigmentation, hatching, and larval survival rates were defined as 99.50%, 97.68%, 98.97% for Abant trout, 87.35%, 68.97%, 71.54% for brook trout, 96.12%, 64.18%, 56.82% for S.t.a. ♀ x S.f. ♂ hybrid and for S.f. ♀ x S.t.a. ♂ hybrid 14.98%, 0.45%, respectively. The results indicated there were significant linear relationships between degree-day and length, total wet weight, dry body weight, and dry yolk-sac weight in Abant trout, brook trout, and their hybrids. This relationship has been demonstrated as decreasing in the yolk-sac and as increasing in others. As a result of crossing, it was observed that the hatching efficiency of hybrid (S.t.a.♀ x S.f.♂) was similar to brook trout, and growth performance was similar to Abant trout. The results will help further studies and will contribute to the management practices of the stocks of Abant trout, brook trout, and their hybrids.

Lethal and Sublethal Toxicity of Un-Ionized Ammonia to Early-Life Stages of Danio rerio.

Un-ionized ammonia (NH3) is a prevalent contaminant found in aquatic ecosystems, frequently associated with complex mixtures of other toxic contaminants. Early embryo-larval stages of zebrafish Danio rerio became an important model for water quality monitoring, and it is necessary to characterize its susceptibility to NH3 exposure. Fertilized eggs were exposed to NH3 concentrations ranging from 0.02 to 5.23 mg NH3 L-1 until 168 h postfertilization (hpf). The lethal concentration to 50% of exposed zebrafish during 96 h was 2.07 mg NH3 L-1, 25% above the median value reported values for early developmental stages of fishes. Sublethal toxicity endpoints indicated the lowest observed effect concentrations for slow blood circulation at 24 hpf, decreased heart ventricular contractions at 72 hpf, incomplete yolk sac absorption at 96 hpf, failure in swim bladder inflation at 96 hpf, developmental retardation at 96 hpf, decreased total length, decreased swimming speed, altered trajectories, and acetylcholinesterase inhibition at 168 hpf of 0.85, 0.06, 0.15, 0.06, 0.15, 0.61, 1.35, 0.35, and 0.85 mg NH3 L-1, respectively. Environmentally relevant NH3 concentrations can affect zebrafish's early development and larval viability, and our results help discriminate NH3 contribution to the toxicity of complex environmental mixtures when zebrafish is used in water quality monitoring.