Toxicity assessment of carvacrol and its acetylated derivative in early staged zebrafish (Danio rerio): Safer alternatives to fipronil-based pesticides?

Fipronil is a broad-spectrum pesticide presenting high acute toxicity to non-target organisms, particularly to aquatic species. Natural compounds stand out as promising alternatives to the use of synthetic pesticides such as fipronil. Thus, our study aimed to compare the toxicity of carvacrol (natural), acetylcarvacrol (semisynthetic), and fipronil (synthetic) to early staged zebrafish. We conducted a series of toxicity assays at concentrations ranging from 0.01 µM to 25 µM for fipronil and 0.01 µM to 200 µM for carvacrol and acetylcarvacrol, depending on the assay, after 7-days post-fertilization (dpf). The potency (EC50) of fipronil was ∼1 µM for both deformities and mortality at 7 dpf, whereas EC50 was >50 µM for carvacrol and >70 µM for acetylcarvacrol. Fipronil at 0.1 and 1 µM caused a decrease in body length and swim bladder area of larvae at 7dpf, but no difference was observed for either carvacrol or acetylcarvacrol. Based upon the visual motor response test, fipronil induced hypoactivity in larval zebrafish at 1 µM and acetylcarvacrol induced hyperactivity at 0.1 µM. Anxiolytic-type behaviors were not affected by any of these chemicals. All chemicals increased the production of reactive oxygen species at 7 dpf, but not at 2 dpf. Genes related to swim bladder inflation, oxidative stress, lipid metabolism, and mitochondrial activity were measured; only fipronil induced upregulation of atp5f1c. There were no changes were observed in oxygen consumption rates of fish and apoptosis. Taken together, our data suggest that carvacrol and its derivative may be safer replacements for fipronil due to their lower acute toxicity.

Molecular and behavioral toxicity assessment of tiafenacil, a novel PPO-inhibiting herbicide, in zebrafish embryos/larvae.

Tiafenacil is a newly registered herbicide and a protoporphyrinogen IX oxidase inhibitor. However, sub-lethal effects of PPO-inhibitors in aquatic species are unknown. Embryos or larvae were exposed to 0.1 µg/L up to 10 mg/L tiafenacil for 7-days post-fertilization. Decreased survival (> 50%) and deformities were noted at concentrations > 1 mg/L. Potency (EC50) of tiafenacil for 5- and 7-day larvae were 818.1 µg/L and 821.7 µg/L, respectively. Pericardial and yolk sac edema were the most frequent deformities observed. Heartbeat frequency at 3 dpf was decreased in zebrafish exposed to > 10 µg/L tiafenacil, coinciding with increased reactive oxygen species. Oxygen consumption rates were not affected by tiafenacil, nor did we detect differences in indicators of apoptosis. The abundance of eighteen transcripts related to oxidative stress and mitochondrial complexes I through V were unchanged. Larval activity was decreased with exposure to 1000 µg/L tiafenacil. These data contribute to risk assessment for a new class of herbicide.

Developmental and mitochondrial toxicity assessment of perfluoroheptanoic acid (PFHpA) in zebrafish (Danio rerio).

The potential toxicity of several perfluoroalkyl and polyfluoroalkyl substances (PFASs) to aquatic species are not well understood. Here, we assessed the sub-lethal toxicity potential of perfluoroheptanoic acid (PFHpA) to developing zebrafish. PFHpA was not acutely toxic to fish up to 50 µM and there was > 96% survival in all treatments. Exposure to 200 µM PFHpA decreased ATP-linked respiration of embryos. There was no evidence for ROS induction in 7-day-old larvae fish exposed to 0.1 µM or 1 µM PFHpA. Twenty-four transcripts related to mitochondrial complexes I through V were measured and atp06, cox4i1, and cyc1 levels were decreased in larval zebrafish in a concentration-dependent manner by PFHpA exposure. Locomotor activity was reduced in fish exposed to 0.1 µM PFHpA based on a visual motor response test. Anxiolytic-type behaviors were not affected by PFHpA. This study contributes to environmental risk assessments for perfluorinated chemicals.