Visualization of hydatid elements: Comparison of several techniques

Abstract

Some techniques available at our laboratory were tested for their ability to aid in the morphological diagnosis of hydatid elements (Echinococcus granulosus ['Taenia echinococcus']) isolated from cysts in humans and sheep. Unstained, methanol-fixed hooklets were fluorescent, most starkly so under violet light (excitation filter wavelength, 405 nm; long- pass filter wavelength, 495 nm). Auramine-rhodamine and Gram procedures failed to stain hooklets. Ziehl-Neelsen stain yielded indifferent results when organisms were viewed under transmitted light but resulted in a surprisingly intense red fluorescence when organisms were viewed under green light (excitation, 546 nm; long pass, 590 nm). Wheatley trichrome stain gave better and more uniform results than fuchsin. Ryan trichrome blue stain was the best under transmitted light; hooklets stained uniformly and intensely and were easily distinguishable from the background. Very satisfactory results were also obtained with a much simpler procedure (modified Baxby technique: no fixation, steaming hot 1% safranin for 2 min, and malachite green for 30 s). Therefore, Ryan and modified Baxby stains are recommended for the examination of E. granulosus under transmitted light. For fluorescence microscopy, Ziehl-Neelsen stain under green excitation light, or violet light with no staining, is also very useful. Epifluorescence microscopy is especially convenient for examining samples concentrated by filtration, as it renders the filter pores inconspicuous.