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Published on: September 25, 2014
Propagation of Anaplasma marginale in bovine lymph node cell culture
Abstract:
Anaplasma marginale was propagated in cell cultures derived from bovine lymph node (LN). Treatment of host cells with diethylaminoethyl dextran (DEAE-D) before inoculation and centrifugation of inoculum onto the monolayers resulted in significant numerical increases of A marginale. The direct fluorescent antibody technique (FAT) was used for detection of the organism in culture. The rat was combined with the standard microscopic count procedure to obtain numerical estimates of the organism in cell culture. Infection of LN cells was irregular, with some cells containing many organisms and others containing none. The organisms were dispersed or in inclusions in the cytoplasm of LN cells. Numerical increases of organisms occurred within 6 hours and these were greatest at 12 to 24 hours. After 24 hours, the organisms decreased rapidly, but small numbers of them were observed for at least 7 days. The average generation time in culture was approximately 17.1 hours.
Insights
Researchers successfully cultured Anaplasma marginale in bovine lymph node cells, optimizing growth with diethylaminoethyl dextran and centrifugation. This advancement aids in understanding Anaplasma marginale propagation and developing control strategies.
Area of Science:
- Veterinary Microbiology
- Cell Biology
- Parasitology
Background:
- Anaplasma marginale is a significant cattle pathogen causing anaplasmosis.
- Efficient cell culture methods are crucial for studying Anaplasma marginale.
- Bovine lymph node cells offer a potential host system for Anaplasma marginale propagation.
Purpose of the Study:
- To establish and optimize a cell culture system for Anaplasma marginale.
- To determine the effects of specific treatments on Anaplasma marginale replication in vitro.
- To characterize the growth kinetics and morphology of Anaplasma marginale in cultured cells.
Main Methods:
- Propagation of Anaplasma marginale in bovine lymph node (LN) cell cultures.
- Treatment of host cells with diethylaminoethyl dextran (DEAE-D) and centrifugation for enhanced inoculation.
- Detection and quantification using the direct fluorescent antibody technique (FAT) and microscopic counts.
Main Results:
- Diethylaminoethyl dextran treatment and centrifugation significantly increased Anaplasma marginale numbers.
- Anaplasma marginale exhibited irregular infection patterns within LN cells, appearing dispersed or in cytoplasmic inclusions.
- Optimal organism increase occurred between 12-24 hours post-inoculation, with a detectable presence for up to 7 days.
- The average generation time of Anaplasma marginale in culture was determined to be approximately 17.1 hours.
Conclusions:
- Bovine lymph node cell cultures, enhanced by DEAE-D treatment and centrifugation, provide a viable system for Anaplasma marginale propagation.
- The established culture method allows for the study of Anaplasma marginale growth dynamics and morphology.
- This research contributes to developing better diagnostic and research tools for Anaplasma marginale.
