Development of Carvacrol-Loaded Albumin Nanoparticles: Evaluation of Antimicrobial Activity Against Aquaculture Pathogens and Controlled Release Propertiese Pathogens
Abstract
The increasing prevalence of bacterial infections in aquaculture highlights the growing need for
environmentally friendly and effective antimicrobial strategies. This study aimed to develop and evaluate
Carvacrol-loaded human serum albumin nanoparticles (CRV-Al@NPs) as a controlled-release system with
antibacterial activity against aquaculture pathogens. CRV-Al@NPs were successfully prepared using a
modified desolvation method, achieving high encapsulation efficiency (81.7±2.1%) and loading capacity
(56.6±2.3%). The nanoparticles exhibited uniform spherical morphology, narrow size distribution (172 nm,
PDI 0.17), and moderate electrostatic stability (zeta potential –27 mV). In vitro release studies demonstrated
pH-dependent and controlled release of Carvacrol, with slower release under alkaline conditions, confirming
that the albumin matrix effectively modulates compound diffusion. Stability tests conducted over 48 h at pH
6.5–8.5 showed that the nanoparticles maintained their size, polydispersity, and colloidal stability.
Antibacterial activity was evaluated against Aeromonas salmonicida and Listonella anguillarum, yielding
inhibition zones of 17.3 mm and 15.4 mm, and MIC values of 37.5 μg/mL and 18.75 μg/mL, respectively.
Blank nanoparticles and solvent controls showed no activity, indicating that the antibacterial effect is
attributed to Carvacrol. These findings demonstrate that Carvacrol retains its antibacterial efficacy after
encapsulation and that CRV-Al@NPs can provide controlled and prolonged activity against the target
pathogens. Overall, this study presents a novel nanoparticle system capable of pH-responsive controlled
release, and the combination of natural antimicrobial compounds with nanoparticle-based delivery offers a
promising and environmentally sustainable approach for disease management in aquaculture.
Keywords
Carvacrol , albumin nanoparticles , aquaculture pathogens , controlled release , pH-responsive
References
- Abdelkarim, E.A., Elsamahy, T., El Bayomi, R.M., Hussein, M.A., Darwish, I.A., El-Tahlawy, A.S., Alahmad, W., Darling, R.J., Hafez, A.E.S.E., Sobhi, M., Abdelfattah, A., & Sobhy, M. (2025). Nanoparticle-driven aquaculture: transforming disease management and boosting sustainable fish farming practices. Aquaculture International, 33, 288. https://doi.org/10.1007/s10499-025-01952-7
- Alizadeh, F., & Khodavandi, A. (2026). Evaluation of synergistic effects of carvacrol-loaded bovine serum albumin nanoparticles and visible light on mixed biofilms of Candida tropicalis and Pseudomonas aeruginosa. Journal of Photochemistry and Photobiology B: Biology, 276, 113385. https://doi.org/10.1016/j.jphotobiol.2026.113385
- Bibi, A., Din, F., Anwar, Y., Alkenani, N.A., Zari, A.T., Mukhtiar, M., Abu Zeid, I.M., Althubaiti, E.H., Nazish, H., Zeb, A., Ullah, I., Khan, G.M., & Choi, H.G. (2022). Cilostazol-loaded solid lipid nanoparticles: bioavailability and safety evaluation in an animal model. Journal of Drug Delivery Science and Technology. 74, 103581. https://doi.org/10.1016/j.jddst.2022.103581
- Bondad-Reantaso, M.G., MacKinnon, B., Karunasagar, I., Fridman, S., Alday-Sanz, V., Brun, E., Le Groumellec, M., Li, A., Surachetpong, W., Karunasagar, I., Hao, B., Dall'Occo, A., Urbani, R., & Caputo, A. (2023). Review of alternatives to antibiotic use in aquaculture. Reviews in Aquaculture, 15(4), 1421-1451. https://doi.org/10.1111/raq.12786
- Cacciatore, F.A., Maders, C., Alexandre, B., Barreto Pinilla, C.M., Brandelli, A., & da Silva Malheiros, P. (2022). Carvacrol encapsulation into nanoparticles produced from chia and flaxseed mucilage: Characterization, stability and antimicrobial activity against Salmonella and Listeria monocytogenes. Food microbiology, 108, 104116. https://doi.org/10.1016/j.fm.2022.104116
- CLSI (Clinical and Laboratory Standards Institute). (2006). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard. 7th ed. Document M7-A7.Wayne, PA: Clinical and Laboratory.
- CLSI (Clinical and Laboratory Standards Institute). (2016). Performance standards for antimicrobial standards institute susceptibility testing, 26 th ed. CLSI supplement M100S. Wayne, PA: Clinical and Laboratory.
- Erdal Altıntaş, Ö. (2025). Fabrication of levan/gellan gum nanocomposite hydrogels containing dexamethasone-loaded albumin nanoparticles for controlled release application. Journal of Drug Delivery Science and Technology. 113, 107350. https://doi.org/10.1016/j.jddst.2025.107350
- Hao, X., Yan, W., Sun, Z., Yang, J., Bai, Y., Qian, H., Chowwanonthapunya, T., & Zhang, D. (2022). pH-responsive allicin-based coatings with antibacterial and antifouling effects in marine environments. Frontiers in Materials, 9, 852731. https://doi.org/10.3389/fmats.2022.852731
- Jaiswal, J., Srivastav, A.K., Rajput, P.K., Yadav, U.C.S., & Kumar, U. (2023). Integrating synthesis, physicochemical characterization, and in silico studies of cordycepin-loaded bovine serum albumin nanoparticles. Journal of Agricultural and Food Chemistry, 71(32), 12225- 12236. https://doi.org/10.1021/acs.jafc.3c03608
