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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
Spotted Fever Group Rickettsia Infection and Transmission Dynamics in Amblyomma maculatum
Chanakan Suwanbongkot1, Ingeborg M Langohr1, Emma K Harris1
1Vector-Borne Disease Laboratories, Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, USA.
Abstract:
Tick vectors are capable of transmitting several rickettsial species to vertebrate hosts, resulting in various levels of disease. Studies have demonstrated the transmissibility of both rickettsial pathogens and novel Rickettsia species or strains with unknown pathogenicity to vertebrate hosts during tick blood meal acquisition; however, the quantitative nature of transmission remains unknown. We tested the hypothesis that if infection severity is a function of the rickettsial load delivered during tick transmission, then a more virulent spotted fever group (SFG) Rickettsia species is transmitted at higher levels during tick feeding. Using Amblyomma maculatum cohorts infected with Rickettsia parkeri or "Candidatus Rickettsia andeanae," a quantitative PCR (qPCR) assay was employed to quantify rickettsiae in tick salivary glands and saliva, as well as in the vertebrate hosts at the tick attachment site over the duration of tick feeding. Significantly greater numbers of R. parkeri than of "Ca Rickettsia andeanae" rickettsiae were present in tick saliva and salivary glands and in the vertebrate hosts at the feeding site during tick feeding. Microscopy demonstrated the presence of both rickettsial species in tick salivary glands, and immunohistochemical analysis of the attachment site identified localized R. parkeri, but not "Ca Rickettsia andeanae," in the vertebrate host. Lesions were also distinct and more severe in vertebrate hosts exposed to R. parkeri than in those exposed to "Ca Rickettsia andeanae." The specific factors that contribute to the generation of a sustained rickettsial infection and subsequent disease have yet to be elucidated, but the results of this study suggest that the rickettsial load in ticks and during transmission may be an important element.
Insights
This study investigated Rickettsia transmission by ticks. Higher Rickettsia parkeri loads in tick saliva correlated with more severe disease in hosts, suggesting rickettsial load is key to spotted fever group infections.
Area of Science:
- Veterinary Entomology
- Infectious Diseases
- Microbiology
Background:
- Ticks transmit various Rickettsia species, causing diseases with differing severity.
- The quantitative aspect of Rickettsia transmission during tick feeding is not well understood.
- Spotted fever group (SFG) Rickettsia species vary in pathogenicity.
Purpose of the Study:
- To test if infection severity correlates with the rickettsial load transmitted by ticks.
- To quantify Rickettsia transmission levels from infected ticks to vertebrate hosts.
- To compare the transmission and pathogenicity of Rickettsia parkeri and "Candidatus Rickettsia andeanae".
Main Methods:
- Used Amblyomma maculatum ticks infected with Rickettsia parkeri or "Candidatus Rickettsia andeanae".
- Employed quantitative PCR (qPCR) to measure rickettsial loads in tick salivary glands, saliva, and host tissues.
- Utilized microscopy and immunohistochemistry to identify rickettsial presence in hosts.
Main Results:
- Significantly higher loads of Rickettsia parkeri were quantified in tick saliva, salivary glands, and host attachment sites compared to "Candidatus Rickettsia andeanae".
- Microscopy confirmed Rickettsia parkeri, but not "Candidatus Rickettsia andeanae", localized in host tissues.
- Vertebrate hosts exposed to Rickettsia parkeri developed more severe lesions.
Conclusions:
- Rickettsial load during tick transmission appears to be a critical factor in disease severity.
- Rickettsia parkeri is transmitted at higher levels and causes more severe pathology than "Candidatus Rickettsia andeanae".
- Further research is needed to elucidate factors driving sustained Rickettsia infections and disease.
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