Related Experiment Videos
Isolation of spotted fever group rickettsias from triturated ticks using a modification of the centrifugation-shell
Abstract:
A modification of the centrifugation-shell vial technique was used to isolate spotted fever group (SFG) rickettsias from triturated ticks which had previously been stored at -80 degrees C for up to 7 months. SFG rickettsias were successfully isolated in Vero cells from all 7 haemolymph positive ticks (Amblyomma hebraeum) used in the experiment. Attempts were also made to use these tick triturates to establish SFG rickettsial infections in tissue culture. Vero cells were used for 5 of the tick triturates and human embryonic lung cells (HEL) for the remaining 2. Infections were successfully established in 4 of the 5 Vero cell cultures, but in none of the HEL fibroblast cultures.
Insights
This study successfully isolated spotted fever group (SFG) rickettsias from frozen ticks using a modified centrifugation-shell vial technique. Vero cells proved effective for isolating SFG rickettsias and establishing infections from tick samples.
Area of Science:
- Veterinary Microbiology
- Tick-borne Diseases
- Cell Culture Techniques
Background:
- Spotted fever group (SFG) rickettsias are significant tick-borne pathogens.
- Effective isolation methods are crucial for studying SFG rickettsias and their transmission.
- Tick storage conditions can impact pathogen viability and isolation success.
Purpose of the Study:
- To evaluate a modified centrifugation-shell vial technique for isolating SFG rickettsias from frozen ticks.
- To assess the efficacy of Vero cells and human embryonic lung (HEL) cells for SFG rickettsia isolation and infection establishment.
Main Methods:
- A modified centrifugation-shell vial technique was employed to process tick triturates.
- Ticks (Amblyomma hebraeum) were stored at -80°C for up to 7 months prior to processing.
- Isolation attempts were performed in Vero cells and human embryonic lung (HEL) fibroblast cells.
Main Results:
- SFG rickettsias were successfully isolated from all 7 haemolymph-positive ticks using the modified technique.
- Infections were established in 4 out of 5 Vero cell cultures inoculated with tick triturates.
- No infections were established in the 2 HEL fibroblast cultures inoculated with tick triturates.
Conclusions:
- The modified centrifugation-shell vial technique is effective for isolating SFG rickettsias from ticks stored at -80°C.
- Vero cells are a suitable host cell line for both isolation and establishing SFG rickettsial infections from tick samples.
- HEL fibroblast cells were less effective for establishing SFG rickettsial infections in this study.