Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis
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 suppr...
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| Format: | Article |
| Language: | English |
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BMC
2019-10-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-019-3716-0 |
| _version_ | 1818348635165818880 |
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| author | Arnab Ghosh Dane Jasperson Lee W. Cohnstaedt Corey L. Brelsfoard |
| author_facet | Arnab Ghosh Dane Jasperson Lee W. Cohnstaedt Corey L. Brelsfoard |
| author_sort | Arnab Ghosh |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T17:53:11Z |
| format | Article |
| id | doaj.art-56f94a282e41428ebcf5d4cf42bd7cb9 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-13T17:53:11Z |
| publishDate | 2019-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-56f94a282e41428ebcf5d4cf42bd7cb92022-12-21T23:36:27ZengBMCParasites & Vectors1756-33052019-10-0112111010.1186/s13071-019-3716-0Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientisArnab Ghosh0Dane Jasperson1Lee W. Cohnstaedt2Corey L. Brelsfoard3Department of Biological Sciences, Texas Tech UniversityUSDA-ARS Arthropod Borne Animal Disease Research UnitUSDA-ARS Arthropod Borne Animal Disease Research UnitDepartment of Biological Sciences, Texas Tech UniversityAbstract 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.http://link.springer.com/article/10.1186/s13071-019-3716-0Culicoides sonorensisWolbachia pipientisPopulation replacementPopulation suppressionBiting midge |
| spellingShingle | Arnab Ghosh Dane Jasperson Lee W. Cohnstaedt Corey L. Brelsfoard Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis Parasites & Vectors Culicoides sonorensis Wolbachia pipientis Population replacement Population suppression Biting midge |
| title | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
| title_full | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
| title_fullStr | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
| title_full_unstemmed | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
| title_short | Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis |
| title_sort | transfection of culicoides sonorensis biting midge cell lines with wolbachia pipientis |
| topic | Culicoides sonorensis Wolbachia pipientis Population replacement Population suppression Biting midge |
| url | http://link.springer.com/article/10.1186/s13071-019-3716-0 |
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