High-yield preparation of purified Anaplasma marginale from infected bovine red blood cells.
Purification of Anaplasma marginale from infected bovine RBC was achieved through enzyme treatment and density-gradient centrifugation. A relative yield of 41.6% was obtained by dividing the number of organisms in the final purified preparation by the number of A marginale-infected RBC. Purified parasites were verified as A marginale by light microscopy, electron microscopy, and immunologic tests. The purified parasites reacted positively with calf and rabbit anti-A marginale sera in interfacial and slide agglutination tests. Anti-bovine RBC serum did not agglutinate purified A marginale, indicating absence of any contaminating RBC stroma. Anaplasma marginale was antigenic, but did not cause infection when the preparation was inoculated into a susceptible calf. The density of A marginale was determined to be 1.19 g/ml and cell diameters ranged from 0.25 to 0.63 micron. This method provided procedures for obtaining A marginale free of bovine RBC antigens for accurate biochemical assays and vaccine production.
D000372:Agglutination Tests / Q000662:veterinary; D000709:Anaplasma / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure; D000712:Anaplasmosis / Q000382:microbiology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D001428:Bacterial Vaccines; D001431:Bacteriological Techniques / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D002499:Centrifugation, Density Gradient; D004912:Erythrocytes / Q000382:microbiology; D008297:Male
D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines