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The initial phase of infection with Tyzzer’s organisms in cultured mouse hepatocytes was observed using indirect immunofluorescence (IF) and a plaque assay. The organism, adhered poorly to aldehyde- or acetone-fixed cells, but once adhered to host cells, whether methanol-fixed or unfixed, they were not removed by methanol or acetone. By the IF as well as the plaqlue assay, both of which discriminated intra- and extracellularly located organisms the number of cell-associated organisms increased linearly up to 3 h post-inoculation. The number of intra-cellular organisms increased rapidly in the first 1 h, followed by linear increase at a much lower rate. Similar results were obtained by the plaque assay.
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