Brief Report: Evaluation of Ziehl-Neelsen Slide Restaining with 0.4% Malachite Green for Identifying Cyclospora cayetanensis Oocysts

Abstract

Cyclospora cayetanensis oocysts are typically identified using Ziehl-Neelsen acid-fast staining. However, slides that have been stored for extended periods often experience color fading, reducing visual clarity and unsuitable for training and educational purposes. This study aimeds to evaluate the effectiveness of re-staining with 0.4% malachite green in enhancing the contrast and visualization of Cyclospora oocysts on faded slides. This study used a quantitative experimental laboratory design. Ten positive slides stored at the Clinical Parasitology Laboratory, Universitas Indonesia, were selected based on the degree of fading and   thickness of staining. Each slide was (destained with methanol, then??) restained with 0.4% malachite green for 30 seconds. After drying, the slides were examined under a light microscope, and oocysts were manually counted.  Restaining restored the background to a bluish-green color, improving visual contrast and facilitating oocyst identification. Restaining Ziehl-Neelsen slides by adding 0.4% malachite green is a simple and effective method for enhancing visualization and accurate counting of C. cayetanensis oocysts.