Adverse Effects of Glyphosate-Based Herbicide on Hatching Rate, Morphological Alterations, and Acetylcholinesterase (AChE) Expression in Golden Apple Snail Eggs

Abstract

This research investigated the effects of glyphosate herbicide on golden apple snail (Pomacea canaliculata) eggs, focusing on hatching rates, morphological changes, and acetylcholinesterase (AChE) expression to explore its potential as a biomarker of exposure. Additionally, key bioactive components in golden apple snail eggs were characterized. The study demonstrated that glyphosate toxicity increased with both exposure time and concentration. Optical coherence tomography (OCT) analysis revealed adverse morphological effects on eggs and embryos, including changes in shell structure and organ development. SDS-PAGE analysis identified four main protein bands, including Perivitellin 2 in three isoforms (98, 67, and 31 kDa) and Ovorubin (28 kDa). Lipids, phosphorus, and carbohydrates were identified as key components through Sudan Black B, Methyl Green, and Alcian Blue staining. AChE, with a molecular weight of 71 kDa, was further analyzed by Western blot, showing decreased expression with prolonged and higher concentrations of glyphosate exposure. GC-MS analysis identified major bioactive compounds in untreated eggs, including 3-Fluoro-β, 5-dihydroxy-N-methylbenzeneethanamine, 2-Aziridinylethylamine, and dextroamphetamine, which have pharmaceutical properties such as anti-hypertensive, diuretic, anti-diabetic, and anti-inflammatory effects, as well as potential applications in treating attention deficit hyperactivity disorder and narcolepsy. These compounds were present at lower levels in glyphosate-exposed groups, suggesting glyphosate's impact on the eggs' biochemical defense mechanisms. This study highlights the potential effects of glyphosate on golden apple snail eggs, which may have implications for future snail populations and aquatic ecosystems.