Parasites and Vectors (2019) 12 (483) - Transfection of Culicoides

Arnab Ghosh, Dane Jasperson, Lee W. Cohnstaedt and Corey L. Brelsfoard (2019)
Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis
Parasites and Vectors 12 (483)
Abstract:
Background
Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression or replacement strategies.
Methods
Culicoides sonorensis cell lines were transfected with a Wolbachia infection using a modified shell vial technique. Infections were confirmed using PCR and cell localization using fluorescent in situ hybridization (FISH). The stability of Wolbachia infections and density was determined by qPCR. qPCR was also used to examine immune genes in the IMD, Toll and JACK/STAT pathways to determine if Wolbachia were associated with an immune response in infected cells.
Results
Here we have transfected two Culicoides sonorensis cell lines (W3 and W8) with a Wolbachia infection (walbB) from donor Aedes albopictus Aa23 cells. PCR and FISH showed the presence of Wolbachia infections in both C. sonorensis cell lines. Infection densities were higher in the W8 cell lines when compared to W3. In stably infected cells, genes in the immune Toll, IMD and JAK/STAT pathways were upregulated, along with Attacin and an Attacin-like anti-microbial peptides.
Conclusions
The successful introduction of Wolbachia infections in C. sonorensis cell lines and the upregulation of immune genes, suggest the utility of using Wolbachia for a population replacement and/or population suppression approach to limit the transmission of C. sonorensis vectored diseases. Results support the further investigation of Wolbachia induced pathogen inhibitory effects in Wolbachia-infected C. sonorensis cell lines and the introduction of Wolbachia into C. sonorensis adults via embryonic microinjection to examine for reproductive phenotypes and host fitness effects of a novel Wolbachia infection.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Full text of article
Database assignments for author(s): Lee W. Cohnstaedt

Research topic(s) for pests/diseases/weeds:
biocontrol - natural enemies
rearing/culturing/mass production
Research topic(s) for beneficials or antagonists:
general biology - morphology - evolution

---

Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Culicoides sonorensis