Isolation of Babesia gibsoni piroplasms from infected erythrocytes of dogs.
To eliminate the influence of anti-dog erythrocyte membrane antibodies in Babesia gibsoni-infected dogs on the immunological investigations of parasites, an attempt was made to purify B. gibsoni piroplasms from infected erythrocytes by N2 cavitation and centrifugation in a Percoll discontinuous density gradient. Although electron microscopic examination revealed that the concentrated piroplasms visible as a band at the interface between 30% and 40% Percoll solutions were contaminated with a small amount of erythrocyte membrane debris, little contamination was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblots. Hence, the isolated parasites may be usable for immunological examinations of parasites.
Adachi
K
K
Department of Veterinary Internal Medicine, Faculty of Agriculture, Miyazaki University, Japan.
Watanabe
T
T
Yamane
S
S
Makimura
S
S
D000818:Animals; D000953:Antigens, Protozoan / Q000097:blood / Q000302:isolation & purification*; D001403:Babesia / Q000302:isolation & purification* / Q000648:ultrastructure; D001404:Babesiosis / Q000469:parasitology*; D015153:Blotting, Western; D002501:Centrifugation, Zonal; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000469:parasitology / Q000648:ultrastructure; D004912:Erythrocytes / Q000469:parasitology*; D008854:Microscopy, Electron; D008970:Molecular Weight; D011205:Povidone; D012822:Silicon Dioxide
D000953:Antigens, Protozoan; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone