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

Background And Aim: 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 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 MgCl. The optimized liquid medium was spray-dried to produce a dehydrated powder for practical application. The encystation efficiency of different 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 MgCl 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 American Type Culture Collection 50739 in glucose-free conditions. Notably, optimal glucose concentrations varied among 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 spp. The medium enhances laboratory research and diagnostic capabilities, paving the way for future advancements in understanding and managing infections.