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An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines
Published on: October 11, 2022
TYPHUS FEVER : V. THE EFFECT OF TEMPERATURE ON THE MULTIPLICATION OF RICKETTSIA PROWAZEKI IN TISSUE CULTURE
1Department of Pathology, Harvard Medical School, Boston.
Abstract:
The temperature at which tissue cultures infected with typhus Rickettsiae are incubated has been shown to exert a marked influence on the intracellular multiplication of Rickettsia prowazeki. At 41 degrees C. the organisms were not found in the cultures histologically on and after the 2nd day in vitro, and the cultures were non-virulent on and after the 3rd day in vitro, in spite of good preservation and growth of the cells. At 37.5 degrees C. organisms were absent from the cultures histologically and the cultures were non-virulent on and after the 11th day in vitro, in spite of good preservation and growth of the cells. At 32 degrees C. good but slow growth of cells occurred and organisms were found in increasing numbers histologically up to about the 21st day in vitro. At this time, 50 to 99 per cent (approximately) of the cytoplasmic volume of the cultures was occupied by Rickettsiae. From the 21st day to the 51st day (the limit to which cultures have been carried successfully) this condition of unrestricted multiplication remained practically unchanged. Inoculation of guinea pigs with single cultures after varying lengths of time in vitro, (up to the 51st day) always resulted in reproduction of typhus in a characteristic manner. At 27 degrees the cell growth was negligible, but many cells remained alive for 10 days or more. Organisms were absent from the cultures histologically and the cultures were non-virulent on and after the 18th day in vitro. The only histological preparations showing unrestricted multiplication of the organisms (infection of the majority of the cells present) were of cultures incubated at 32 degrees C. It is believed that the detrimental effect of the higher temperatures (37.5 degrees and 41 degrees C.) on the multiplication of the organism is exerted indirectly, by stimulation of the defence mechanism of the cells.
Insights
Incubation temperature significantly impacts typhus Rickettsiae (Rickettsia prowazeki) growth in cell cultures. Optimal multiplication occurred at 32°C, while higher temperatures inhibited Rickettsia prowazeki, suggesting a cellular defense mechanism.
Area of Science:
- Microbiology
- Cell Biology
- Infectious Diseases
Background:
- Rickettsia prowazeki is the causative agent of typhus.
- Understanding Rickettsia prowazeki intracellular multiplication is crucial for developing effective treatments.
- The influence of environmental factors on Rickettsia prowazeki growth requires further investigation.
Purpose of the Study:
- To investigate the effect of incubation temperature on Rickettsia prowazeki intracellular multiplication in tissue cultures.
- To determine the optimal temperature for Rickettsia prowazeki growth in vitro.
- To elucidate the relationship between temperature, Rickettsia prowazeki proliferation, and host cell defense mechanisms.
Main Methods:
- Tissue cultures infected with Rickettsia prowazeki were incubated at various temperatures (41°C, 37.5°C, 32°C, and 27°C).
- Histological examination was performed to assess Rickettsia prowazeki presence and multiplication.
- Virulence of infected cultures was evaluated by inoculating guinea pigs.
Main Results:
- At 41°C and 37.5°C, Rickettsia prowazeki multiplication was inhibited, and cultures became non-virulent.
- At 32°C, unrestricted Rickettsia prowazeki multiplication was observed, occupying significant cytoplasmic volume.
- At 27°C, cell growth was negligible, and cultures were non-virulent after 18 days.
Conclusions:
- Incubation temperature critically affects Rickettsia prowazeki intracellular multiplication.
- 32°C supports extensive Rickettsia prowazeki proliferation, while higher temperatures inhibit it.
- Higher temperatures may trigger host cell defense mechanisms, indirectly hindering Rickettsia prowazeki growth.
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