Cryptosporidium oocysts

Li, Xunde, Pereira, Maria das Graças Cabral, Larsen, Royce, Xiao, Chengling, Phillips, Ralph, Striby, Karl, McCowan, Brenda & Atwill, Edward R., 2015, Cryptosporidium rubeyi n. sp. (Apicomplexa: Cryptosporidiidae) in multiple Spermophilus ground squirrel species, International Journal for Parasitology: Parasites and Wildlife 4 (3), pp. 343-350 : 344

2.2. Detection of Cryptosporidium oocysts

Detection of Cryptosporidium oocysts in previous studies were conducted by direct immunofluorescent microscopy (IFA) as described previously ( Atwill et al., 2004; Pereira et al., 2010). Similar methods were used for the feces collected in 2011. Briefly, fecal samples were processed within one week after collection. Feces and antibiotic solution were mixed in deionized water with 0.2% Tween 20 to a final volume of 40 ml. The fecal suspension was strained through 4 layers of cotton gauze into a 50 ml centrifuge tube, which was filled with deionized water to a final volume of 50 ml. Tubes were centrifuged at 1500 g for 15 min and supernatant discarded, leaving a 1:1 ratio of pellet to solution volume. This final suspension was homogenized and 10 M l was used for making slides using the Aqua-Glo G/C Direct kit (Waterborne Inc., New Orleans, LA, USA). Slides were examined using a fluorescent microscope (Olympus BX 60) at ×400 magnification.