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Intracellular Pasteurella pseudotuberculosis: Multiplication in Cultured Spleen and Kidney Cells
1Department of Microbiology and Public Health, Michigan State University, East Lansing, Michigan 48823.
Abstract:
Pasteurella pseudotuberculosis multiplied within rabbit cells in primary culture. Spleen cells from rabbits were either dispersed mechanically, exposed to the organism immediately after cell preparation, and grown as a pellet or the cells were dispersed enzymatically and grown as a monolayer for 4 to 6 days before the bacteria were introduced. Intracellular multiplication proceeded at a logarithmic rate for 1 to 2 days, with a generation time of about 70 min in pelleted cells and 4 to 5 hr in monolayered cells. Under the conditions employed, the wild-type virulent strain and an avirulent mutant multiplied at similar rates. Atmosphere, constituents of the medium, and multiplicity of infection influenced intracellular proliferation. The organism also proliferated in kidney cells. Microscopic observation of stained cells indicated limited growth of the pathogen in spleen cells at 37 C. In kidney cells, the pasteurellae localized in compartments; frequently, a single compartment with masses of proliferating organisms almost filled the cytoplasm.
Insights
Pasteurella pseudotuberculosis multiplies within rabbit spleen and kidney cells. Cell preparation methods significantly impact bacterial growth rates, with pelleted cells supporting faster multiplication than monolayers.
Area of Science:
- Microbiology
- Cell Biology
- Infectious Diseases
Background:
- Pasteurella pseudotuberculosis is a bacterial pathogen.
- Understanding its intracellular behavior is crucial for disease pathogenesis.
Purpose of the Study:
- To investigate the intracellular multiplication of Pasteurella pseudotuberculosis in rabbit spleen and kidney cells.
- To determine the effect of cell culture methods on bacterial proliferation.
Main Methods:
- Rabbit spleen cells were cultured as pellets or monolayers.
- Pasteurella pseudotuberculosis was introduced to cultured cells.
- Bacterial growth was monitored over time.
- Microscopic examination of infected cells was performed.
Main Results:
- Pasteurella pseudotuberculosis multiplied intracellularly in both pelleted and monolayered spleen cells.
- Generation time was significantly shorter in pelleted cells (approx. 70 min) compared to monolayers (4-5 hr).
- Both virulent and avirulent strains exhibited similar multiplication rates.
- Bacterial proliferation was also observed in kidney cells, often within cytoplasmic compartments.
Conclusions:
- Cell culture conditions, specifically cell preparation, profoundly influence Pasteurella pseudotuberculosis intracellular multiplication.
- The pathogen can proliferate within rabbit kidney cells, forming large intracellular populations.
