Screening of selected compounds against Enterocytozoon hepatopenaei (EHP) in shrimps: an in silico and in vitro approach

Abstract

Shrimp aquaculture plays an essential role in meeting the growing global demand for seafood, supporting nutritional security and economic livelihoods. However, its sustainability is increasingly jeopardized by Enterocytozoon hepatopenaei (EHP), a microsporidian parasite responsible for Hepatopancreatic microsporidiosis, which leads to growth retardation in farmed shrimp. Existing disease management strategies remain largely ineffective, lacking targeted therapeutics and relying heavily on labour-intensive practices. This study aimed to identify a feed-based intervention through a combined in silico and in vitro evaluation of three compounds, sulphadimethoxine, benzophenone and sorbitol, targeting the β-tubulin protein in shrimp. Molecular dynamics simulations over 200 ns confirmed the structural stability of the sulphadimethoxine β-tubulin complex, with root mean square deviation stabilizing at approximately 50 ns and maintaining a value of ~ 0.21 nm. The root mean square fluctuation analysis revealed minimal fluctuations (~ 0.2–.3 nm) at critical residues, indicating strong local stability. Hydrogen bond analysis showed sulphadimethoxine formed up to 4 bonds during the simulation, suggesting moderate yet sustained interaction, in contrast to benzophenone (0–2 bonds) and sorbitol (0–3 bonds). The in vitro study revealed that sulphadimethoxine supplementation at 3–5% concentrations significantly enhanced shrimp growth from 18.4 ± 0.9 g to 26.1 ± 0.1 g in weight and from 12.6 ± 0.6 to 16.0 ± 0.5 cm in length over 45 days. The shrimps treated with the sulphadimethoxine reported the EHP negative. These findings highlight sulphadimethoxine as a promising, feed-based candidate for EHP control and growth enhancement in shrimp aquaculture. Integrating computational modelling with experimental validation presents a novel and sustainable alternative to traditional pond management, offering scalable benefits to shrimp producers worldwide.