Fecal, spleen and the infected cell culture samples were processed for EM observation as previously described[7 (link)]. Briefly, fecal and spleen samples were processed in 1:10 (w/v) with 10 mM, pH7.2 PBS, and homogenized before centrifuging at 8000 × g for 20 min at 4°C. The infected cells were frozen and thawed for 3 times before centrifugation as described above. The supernatants were then incubated with 1% phosphotungstic acid, and the viruses were examined using electron microscope (JEM-2200FS/CR )(JEOL, Tokyo, Japan).
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