Bifenthrin, a third-generation synthetic pyrethroid, is widely used as an agricultural insecticide. However, it can flow into surface and groundwater, leading to adverse consequences such as immunotoxicity, hepatotoxicity, hormone dysregulation, or neurotoxicity. Nevertheless, the entire range of its neurotoxic consequences, particularly in aquatic organisms, remains unclear. In this study, we conducted an extensive examination of how exposure to bifenthrin affects the behavior and nervous system function of aquatic vertebrates, using a zebrafish model and multiple-layered assays. We exposed wild-type and transgenic lines [tg(elavl3:eGFP) and tg(mbp:mGFP)] to bifenthrin from <3 h post-fertilization (hpf) to 120 hpf. Our findings indicate that bifenthrin exposure concentrations of 103.9 and 362.1 μg/L significantly affects the tail-coiling response at 24 hpf and the touch-evoked responses at 72 hpf. Moreover, it has a significant effect on various aspects of behavior such as body contact, distance between subjects, distance moved, and turn angle. We attribute these effects to changes in acetylcholinesterase and dopamine levels, which decrease in a concentration-dependent manner. Furthermore, neuroimaging revealed neurogenesis defects, e.g., shortened brain and axon widths, and demyelination of oligodendrocytes and Schwann cells. Additionally, the transcription of genes related to neurodevelopment (e.g., gap43, manf, gfap, nestin, sox2) were significantly upregulated and neurotransmitters (e.g., nlgn1, drd1, slc6a4a, ache) was significantly downregulated. In summary, our data shows that bifenthrin exposure has detrimental effects on neurodevelopmental and neurotransmission systems in the zebrafish embryo/larvae model.

Acrylamide has neurotoxic and/or cardiotoxic effects on humans however available information regarding the neuro- and cardiotoxicity currently is very limited for freshwater organism models. Using three distinct techniques, thus, we investigated the neuro- and cardiotoxic effects of acrylamide in the freshwater invertebrate model, Daphnia magna. We exposed D. magna to acrylamide at concentrations of 0.3, 2.7, and 11.1 mg/L for 48 h alongside a control group. We then conducted physiological (thoracic limb activity and heart rate) and behavioral tests (including distance moved, velocity, turn angle, moving duration, the distance between subjects, and body contact frequency), as well as gene transcription analyses (related to cardiomyopathy, the serotonergic synapse, neuroactive ligand-receptor interactions, the GABAergic synapse, and acetylcholine receptors). After acrylamide exposure, the thoracic limb activity and heart rates of D. magna showed time- and dose dependent inhibition. From low to high exposure concentrations, both heart rates and thoracic limb activity were decreased. Additionally, the distance between subjects and body contact frequencies was significantly reduced. At the gene transcription level, acrylamide significantly altered the transcription of five genes related to cardiomyopathy and eight genes related to the serotonergic synapse, neuroactive ligand-receptor interactions, and the GABAergic synapse. The signs of hindered neural and cardiac functions were shown in D. magna. This suggests that acrylamide exposure leads to cardiotoxicity and neurobehavior defects in D. magna. Because cardiotoxicity and neurobehavioral changes may cause an ecological imbalance via predation of D. magna, acrylamide may also be considered a threat to freshwater ecosystem.

The zebrafish, a diurnal vertebrate, is commonly used in circadian rhythm studies due to its genetic and neurological similarities to humans. Circadian rhythms, which regulate sleep, neurotransmitter, behavior, and physiological responses to environmental changes, can be disrupted by various environmental factors. Phthalic acid esters (PAEs) are pervasive endocrine disruptors that individuals are frequently exposed to in daily life. However, the impact of PAEs on circadian rhythms during early development remains poorly understood. This study aimed to investigate the effects of exposure to diethyl phthalate (DEP) and butyl benzyl phthalate (BBzP) on the behavior and circadian rhythms of developing zebrafish larvae using a series of layered assays. Zebrafish larvae were exposed to the two PAEs from less than 2 hour post-fertilization (hpf) until 96 hpf. The results demonstrated a concentration-dependent reduction in tail coiling (TC), touch-evoked response (TER), and locomotor activity, alongside an increase in sleep time and alterations in sleep bouts and sleep latency during both 24-hour and Light1/Dark/Light2 (7/10/7-hour) periods. Additionally, exposure to BBzP led to increased acetylcholinesterase (AChE) and dopamine (DA) levels, and a decrease in 5-hydroxytryptamine (5-HT) levels. Gene expression analysis revealed that DEP and BBzP exposure increased the expression of circadian rhythm and light-response-related genes. In conclusion, exposure to these PAEs disrupts the circadian rhythm of zebrafish larvae, providing novel insights into the developmental impact of these common environmental contaminants. Further research is needed to understand the broader implications of these findings for human health and environmental safety.

Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder characterized by impaired social interaction, communication deficits, and repetitive behaviors. The rising prevalence of ASD necessitates intensified research. 2-Ethylhexanol, is a synthetically produced branched-chain alcohol used in plasticizer synthesis. However, its role in ASD-like symptoms and potential neurotoxic effects remains largely unexplored. This study employed a multimodal neurotoxicity testing approach to evaluate the adverse effects of 2-ethylhexanol on zebrafish neurobehavior and neurodevelopment. Wild-type and transgenic zebrafish lines (tg(elavl3: eGFP) and tg(mbp:mGFP)) were exposed to 2-ethylhexanol for 120 hours post-fertilization (hpf). Significant disruptions were observed in early motor activities, such as tail coiling and touch-evoked responses, which aligned with later locomotor impairments, including reduced distance traveled and increased turn angle. These behavioral changes were accompanied by decreased levels of acetylcholinesterase (AChE) and dopamine (DA). Deficits in social behavior (e.g., reduced body contact) were identified, potentially linked to altered transcription of autism-associated genes (adsl, eif4a1, mbd5, vps13b, and tsc1b). Abnormalities in neurogenesis, including reduced brain and spinal cord size, and demyelination of oligodendrocytes and Schwann cells, were evident. Additionally, transcriptional changes related to neurodevelopment (gap43, manf, sox2) and neurotransmitter signaling (drd1, mao, htr1bd) were observed. Our findings provide compelling evidence that 2-ethylhexanol exposure leads to neurodevelopmental impairments and behavioral alterations reminiscent of ASD. This research highlights the importance of further investigations to assess the potential risks of 2-ethylhexanol exposure and develop prevention and mitigation strategies.

Safe endocrine-disrupting alternatives for bisphenol A (BPA) are needed because its adverse health effects have become a public concern. Some bisphenol analogues (bisphenol F and S) have been applied, but their endocrine-disrupting potential is either not negligible or weaker than that of BPA. However, the endocrine-disrupting potential of bisphenol AP (BPAP), another BPA alternative, has not yet been fully assessed. Hence, we evaluated the thyroid hormone (TH)-disrupting potency of BPAP because THs are essential endocrine hormones. Zebrafish embryos were exposed to BPAP (0, 18.2, 43.4, or 105.9 μg/L) for 120 h, and TH levels, the transcription of 16 TH-related genes, the transcriptome, development, and behavior were evaluated. In our study, a decrease in T4 level was observed only at the maximum nonlethal concentration, but significant changes in the T3 and TSHβ levels were not detected. BPAP did not cause significant changes in transcription and gene ontology enrichment related to the TH system. Developmental and behavioral changes were not observed. Despite T4 level reduction, other markers were not significantly affected by BPAP. These might indicate that BPAP has weak or negligible potency regarding TH disruption as a BPA alternative. This study might provide novel information on the TH-disrupting potential of BPAP.

Polyhexamethylene guanidine-phosphate (PHMG-P), a guanidine-based cationic antimicrobial polymer, is an effective antimicrobial biocide, potent even at low concentrations. Due to its resilient bactericidal properties, it has been used extensively in consumer products. It was safely used until its use in humidifiers led to a catastrophic event in South Korea. Epidemiological studies have linked the use of PHMG-P as a humidifier disinfectant to pulmonary fibrosis. However, little is known about its harmful impacts other than pulmonary fibrosis. Thus, we applied a zebrafish embryo/larvae model to evaluate developmental and cardiotoxic effects and transcriptome changes using RNA-sequencing. Zebrafish embryos were exposed to 0.1, 0.2, 0.3, 0.4, 0.5, 1, and 2 mg/L of PHMG-P from 3 h to 96 h post fertilization. 2 mg/L of PHMG-P resulted in total mortality and an LC₅₀ value at 96 h was determined at 1.18 mg/L. Significant developmental changes were not observed but the heart rate of zebrafish larvae was significantly altered. In transcriptome analysis, immune and inflammatory responses were significantly affected similarly to those in epidemiological studies. Our qPCR analysis (Itgb1b, TNC, Arg1, Arg2, IL-1β, Serpine-1, and Ptgs2b) also confirmed this following a 96 h exposure to 0.4 mg/L of PHMG-P. Based on our results, PHMG-P might induce lethal and cardiotoxic effects in zebrafish, and crucial transcriptome changes were linked to immune and inflammatory response.