A comparative study on chitosan nanoparticle synthesis methodologies for application in aquaculture through toxicity studies

Chitosan nanoparticles (CSNPs) have been recently used for various applications in aquaculture, especially as drug carriers. The aim of this study was to synthesise and investigate a superlative method of CSNP synthesis for application in aquaculture through aquaculture-based toxicology screening methods. Two different methods were analysed: the first a direct ionic gelation method (A) and the other involving a low-molecular-weight chitosan microparticle intermediate method (B). Dynamic light scattering characterisation revealed that the CSNP particle sizes were 192.7 +/- 11.8 and 22.9 nm from methods A and B, respectively. The LC50 values for brine shrimp toxicity were found to be 1.51 and 0.02 ppt in 24 h for methods A and B, respectively. Acute toxicity studies in Litopenaeus vannamei rendered LC50 values of 3235.94 and 2884.03 ppt in 24 h for methods A and B, respectively. Zebrafish toxicity studies revealed mortality rates of 21.67% and 55% at 20 mg/L concentration for methods A and B, respectively, with an increased expression of intracellular reactive oxygen species in method B. From these findings, it can be concluded that a comparatively reduced toxicity of CSNPs derived from ionic gelation method makes it more appropriate for application in aquaculture.