Purification of Merozoites from In Vitro Cultured Babesia caballi-Infected Equine Erythrocytes

Abstract

A method for purifying merozoites of Babesia caballi was developed. Infected erythrocytes prepared from in vitro culture was enriched to at least 60% by Percoll-gradient centrifugation. Enriched infected erythrocytes were lysed with 0.83% ammonium chloride and then the lysate was subjected to centrifugation in a Percoll density gradient. Free merozoites were recovered from a band formed at the interface of 1.018 and 1.053 g/ml Percoll solutions. These merozoites were revealed to be morphologically intact by the electron microscopic observation. The merozoites were not contaminated with erythrocyte membrane protein as detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.