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.

Dietary supplementation of curcumin-loaded chitosan nanoparticles stimulates immune response in the white leg shrimp Litopenaeus vannamei challenged with Vibrio harveyi.

Pathogen infections in shrimps trigger the release of reactive oxygen species (ROS) as a part of immune response. The excessive accumulation of ROS causes the production of oxidative stress, which leads to oxidative damage of the biomolecules in the host cells. The inclusion of dietary antioxidants is known to mitigate oxidative stress and stimulate immunity. Curcumin, a potential antioxidant was encapsulated in chitosan nanoparticles to surge its bioavailability and was administered orally to Vibrio harveyi challenged and non-challenged Litopenaeus vannamei. The non-challenged shrimps fed with curcumin-loaded chitosan nanoparticles (Cur-CSNPs) showed a significant increase (p ≤ 0.05) in the specific growth rate, daily growth coefficient and survival rate. A significant increase (p ≤ 0.05) in the phenoloxidase activity, total hemocyte count and superoxide dismutase activity was observed in both the challenged and non-challenged shrimps fed with Cur-CSNPs. Additionally, a significant increase (p ≤ 0.05) in the relative mRNA expression of lysozyme, cMnSOD and lectin was observed in the Cur-CSNPs fed shrimps. The findings of this research suggest that Cur-CSNPs reinforce the immune system of L. vannamei against V. harveyi infection. Moreover, the non-challenged shrimps showed improvement in the growth parameters in addition to immunostimulation. Thereby a routine inclusion of dietary Cur-CSNPs could mitigate the oxidative damage caused by the incidence of environmental or pathogen-mediated oxidative stress.

Curcumin loaded chitosan nanoparticles fortify shrimp feed pellets with enhanced antioxidant activity.

Chitosan nanoparticles (CSNPs) have been recently explored as a potential drug carrier to enhance the bioavailability and aqueous solubility of drugs. Curcumin, an antioxidant with a remarkable antiradical scavenging activity was encapsulated in CSNPs to revamp its bioavailability. While changes in the optimal farming condition can induce oxidative stress in the animals, curcumin loaded chitosan nanoparticles (Cur-CSNPs) were amalgamated into shrimp feed pellets to ameliorate its antioxidant content in an attempt to bolster the organisms against oxidative stress. Cur-CSNPs were synthesized in two different concentrations of curcumin as Cur-CSNPs A and B. Characterization of the synthesized Cur-CSNPs revealed asymmetrical nanoparticles with semispherical geometry and a zeta potential ˃50 mV. HPLC studies substantiated encapsulation efficiencies of 77.53% and 80.35% for Cur-CSNPs A and B respectively. DPPH, ABTS and FRAP assays manifested a significant enhancement in the antioxidant property of the Cur-CSNPs fortified feed pellets. This is the first study to investigate and demonstrate the ability of Cur-CSNPs to enhance the antioxidant property of aquaculture feed pellets. These findings substantiate that Cur-CSNPs fortified feed may be applied to reinforce aquaculture animals against oxidative stress.