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https://hdl.handle.net/2142/68231
Description
Title
In Vitro Cultivation of Anaplasma Marginale
Author(s)
Kessler, Raul Henrique
Issue Date
1980
Department of Study
Veterinary Medical Science
Discipline
Veterinary Medical Science
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Veterinary Science
Language
eng
Abstract
Virulent Anaplasma marginale propagated in culture of bovine erythrocytes showed an increasing rate of replication in a sequence of three experiments. The changes in percentage of parasitized erythrocytes (PPE) were monitored by Giemsa staining and the organism was identified by the direct fluorescent antibody (FA) test and transmission electron microscopy. The PPE increased more than 3 times in the first experiment during a period of 14 days. Regression analysis of the data showed that the trend of growth was significantly (P = 0.05) higher after the third passage of the organism in culture. Serologic and ultrastructural studies demonstrated that the in vitro propagated organism was similar to A. marginale isolated from naturally infected bovine blood. Organisms recovered from 13- and 33-day cultures of the first and second experiments were infective for susceptible cattle.
Ovine erythrocytes infected with the attenuated A. marginale were co-cultured with either normal ovine erythrocytes or normal bovine erythrocytes for 42 days. In both systems, the organism showed an initial period of rapid growth followed by a gradual decrease in PPE and accompanied by cyclic peaks. The presence of bovine erythrocytes in infected ovine erythrocytes resulted in a higher degree of parasitization. Transmission of the organism from infected ovine erythrocytes to normal bovine erythrocytes was demonstrated by use of a two-step FA method which allowed for differentiation between cells and identification of the organism. It is concluded that under in vitro conditions A. marginale is directly transmitted from erythrocyte to erythrocyte without any intermediate cell type.
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