Effect of Acute Toxicity of 2,4-D Dimethylammonium on Rice Field Crab (Esanthelphusa dugasti): Toxicity Testing, Hemocyte Abnormality, Neurotoxicity, and Alteration of Tissue
DOI:
https://doi.org/10.48048/tis.2026.12658Keywords:
Acetylcholinesterase, Aquatic toxicology, Hemocyte morphology, Optical coherence tomography, Rice field crab, 2,4-D dimethylammoniumAbstract
This study investigated the acute and sub-lethal toxicity of 2,4-D dimethylammonium on the rice field crab (Esanthelphusa dugasti), an ecologically important crustacean in Southeast Asian rice-field ecosystems. Crabs were exposed to 0 - 3 mg/L of 2,4-D dimethylammonium for up to 96 h, and toxicity was assessed using mortality, hematological parameters, cholinesterase activity, protein expression, histopathology, immunohistochemistry, and Optical Coherence Tomography (OCT). Mortality increased in a concentration- and time-dependent manner, with the median lethal concentration (LC50) values decreasing from 4.67 mg/L at 24 h to 2.98 mg/L at 96 h. Significant reductions in total hemocyte counts and marked hemocyte deformities were observed at multiple exposure levels (p < 0.05). Neurotoxicity was evidenced by progressive inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with AChE activity declining from 149.45 ± 0.95 nmol/min/mg protein in controls to 119.66 ± 2.02 nmol/min/mg protein at 3 mg/L after 96 h, accompanied by reduced AChE expression. Histopathological analysis revealed concentration-dependent damage in gill and hepatopancreas tissues, including vacuolation, lumen distension, lamellar collapse, and hemocyte infiltration (p < 0.0001). Notably, OCT enabled non-destructive, depth-resolved visualization of internal tissue disruption, corroborating histological findings and providing three-dimensional insight into organ-level pathology. Unlike previous studies on 2,4-D dimethylammonium toxicity in crabs that relied on conventional endpoints, this study demonstrates the novel application of OCT as a complementary imaging tool linking molecular neurotoxicity with structural tissue damage. These findings highlight the ecological risk of 2,4-D dimethylammonium to rice-field crabs and support the use of OCT in advanced aquatic ecotoxicological assessment.
HIGHLIGHTS
- 2,4-D dimethylammonium causes dose- and time-dependent mortality in the rice field crab (Esanthelphusa dugasti).
- AChE and BChE activities significantly decrease with increasing herbicide concentration and exposure duration.
- Hemocyte counts and morphology are altered, indicating the immunotoxic effects of. 2,4-D dimethylammonium exposure.
- Histological and immunohistochemical (IHC) analyses reveal progressive damage in the gill and hepatopancreas tissues.
- Non-destructive OCT imaging effectively detects organ-level damage, which is consistent with the biochemical and histological findings.
GRAPHICAL ABSTRACT
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