Vet World   Vol.18   January-2025  Article - 13 

Research Article

Veterinary World, 18(1): 110-121

https://doi.org/10.14202/vetworld.2025.110-121

Development of a novel encystment medium: Enhancing diagnostic potential of Acanthamoeba spp.

Julalak Chuprom1, Suthinee Sangkanu2, Watcharapong Mitsuwan3, Rachasak Boonhok4, Alok K. Paul5, Sonia M. Rodrigues Oliveira6, Maria L. Pereira7, Tajudeen O. Jimoh8, Mohammed Rahmatullah9, Polrat Wilairatana10, Christophe Wiart11, Ajoy K. Verma12, and Veeranoot Nissapatorn2
1. Department of General Education, School of Languages and General Education, Walailak University, 222 Thaiburi, Thasala, Nakhon Si Thammarat, Thailand, 80160.
2. School of Allied Health Sciences, Southeast Asia Water Team (SEA Water Team) and World Union for Herbal Drug Discovery (WUHeDD), Walailak University, Nakhon Si Thammarat, Thailand.
3. Akkhraratchumari Veterinary College, Walailak University, Nakhon Si Thammarat, Thailand.
4. Department of Medical Technology, School of Allied Health Sciences, Research Excellence Center for Innovation and Health Products, Walailak University, Nakhon Si Thammarat, Thailand.
5. School of Pharmacy and Pharmacology, University of Tasmania, Hobart, TAS, Australia.
6. Faculty of Dental Medicine, the Catholic University of Portugal.
7. Department of Medical Sciences, CICECO-Aveiro Institute of Materials, University of Aveiro, Aveiro, Portugal.
8. Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, USA.
9. Department of Biotechnology and Genetic Engineering, University of Development Alternative, Lalmatia, Dhaka, Bangladesh.
10. Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
11. The Institute for Tropical Biology and Conservation, Kota Kinabalu, Sabah, Malaysia.
12. Department of Microbiology, National Institute of Tuberculosis and Respiratory Diseases, New Delhi, India. 
13. Futuristic Science Research Center, School of Science, Walailak University, Nakhon Si Thammarat, Thailand.

Background and Aim: Acanthamoeba spp. are pathogenic microorganisms linked to severe infections in humans and animals, requiring a deeper understanding of their encystation process for effective diagnostics and research. This study focused on developing a novel encystment medium to induce synchronized encystation of Acanthamoeba spp. efficiently and rapidly.

Materials and Methods: The study employed response surface methodology with a central composite design to optimize the encystment medium formulation. The key components included Tris-HCl, NaCl, glucose, and MgCl2. The optimized liquid medium was spray-dried to produce a dehydrated powder for practical application. The encystation efficiency of different Acanthamoeba strains was assessed using hemocytometry and fluorescence microscopy.

Results: The optimized medium, comprising 3.152 g/L Tris-HCl, 5.55 g/L NaCl, 8% (w/v) glucose, and 5.0 mM MgCl2 at pH 9.0, demonstrated exceptional encystation efficiency with rates ranging from 99% to 100%. A spray-dried powdered version of this medium was equally effective, achieving a 98.77% encystation rate for A. castellanii American Type Culture Collection 50739 in glucose-free conditions. Notably, optimal glucose concentrations varied among Acanthamoeba strains, with certain strains reaching maximum encystation at 6–8% glucose.

Conclusion: This study successfully developed an innovative encystment medium that promotes rapid and efficient cyst production in Acanthamoeba spp. The medium enhances laboratory research and diagnostic capabilities, paving the way for future advancements in understanding and managing Acanthamoeba infections.

Keywords: Acanthamoeba, diagnostic tools, encystation, medium optimization, response surface methodology.

How to cite this article: Chuprom J, Sangkanu S, Mitsuwan W, Boonhok R, Paul AK, Oliveira SMR, Pereira ML, Jimoh TO, Rahmatullah M, Wilairatana P, Wiart C, Verma AK, and Nissapatorn V (2025) Development of a novel encystment medium: Enhancing diagnostic potential of Acanthamoeba spp., Veterinary World, 18(1): 110-121.

Received: 2024-07-23    Accepted: 2024-12-12    Published online: 2025-01-22

Corresponding author: Veeranoot Nissapatorn    E-mail: nissapat@gmail.com

DOI: 10.14202/vetworld.2025.110-121

Copyright: Chuprom, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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