Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i>
Bluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by <i>Culicoides</i> biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassor...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-02-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/16/2/240 |
| _version_ | 1797296843852349440 |
|---|---|
| author | Molly Carpenter Jennifer Kopanke Justin Lee Case Rodgers Kirsten Reed Tyler J. Sherman Barbara Graham Mark Stenglein Christie Mayo |
| author_facet | Molly Carpenter Jennifer Kopanke Justin Lee Case Rodgers Kirsten Reed Tyler J. Sherman Barbara Graham Mark Stenglein Christie Mayo |
| author_sort | Molly Carpenter |
| collection | DOAJ |
| description | Bluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by <i>Culicoides</i> biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassortment in <i>Culicoides sonorensis</i> midges coinfected with different ratios of BTV-10 and BTV-17. Midges were fed blood containing BTV-10, BTV-17, or a combination of both serotypes at 90:10, 75:25, 50:50, 25:75, or 10:90 ratios. Midges were collected every other day and tested for infection using pan BTV and <i>cox1</i> (housekeeping gene) qRT-PCR. A curve was fit to the ∆Ct values (pan BTV Ct—<i>cox1</i> Ct) for each experimental group. On day 10, the midges were processed for BTV plaque isolation. Genotypes of the plaques were determined by next-generation sequencing. Pairwise comparison of ∆Ct curves demonstrated no differences in viral RNA levels between coinfected treatment groups. Plaque genotyping indicated that most plaques fully aligned with one of the parental strains; however, reassortants were detected, and in the 75:25 pool, most plaques were reassortant. Reassortant prevalence may be maximized upon the occurrence of reassortant genotypes that can outcompete the parental genotypes. BTV reassortment and resulting biological consequences are important elements to understanding orbivirus emergence and evolution. |
| first_indexed | 2024-03-07T22:10:41Z |
| format | Article |
| id | doaj.art-ff7052347fe54cd5a620a5ecd236da3f |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-07T22:10:41Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-ff7052347fe54cd5a620a5ecd236da3f2024-02-23T15:37:35ZengMDPI AGViruses1999-49152024-02-0116224010.3390/v16020240Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i>Molly Carpenter0Jennifer Kopanke1Justin Lee2Case Rodgers3Kirsten Reed4Tyler J. Sherman5Barbara Graham6Mark Stenglein7Christie Mayo8Department of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USADepartment of Comparative Medicine, Oregon Health & Science University, Portland, OR 97239, USADepartment of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USADepartment of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USAWisconsin Veterinary Diagnostic Laboratory, University of Wisconsin-Madison, Madison, WI 53706, USADiagnostic Medicine Center, Colorado State University, Fort Collins, CO 80526, USADepartment of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USADepartment of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USADepartment of Microbiology, Immunology, and Pathology, Colorado State University, 1601 Campus Delivery, Fort Collins, CO 80526, USABluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by <i>Culicoides</i> biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassortment in <i>Culicoides sonorensis</i> midges coinfected with different ratios of BTV-10 and BTV-17. Midges were fed blood containing BTV-10, BTV-17, or a combination of both serotypes at 90:10, 75:25, 50:50, 25:75, or 10:90 ratios. Midges were collected every other day and tested for infection using pan BTV and <i>cox1</i> (housekeeping gene) qRT-PCR. A curve was fit to the ∆Ct values (pan BTV Ct—<i>cox1</i> Ct) for each experimental group. On day 10, the midges were processed for BTV plaque isolation. Genotypes of the plaques were determined by next-generation sequencing. Pairwise comparison of ∆Ct curves demonstrated no differences in viral RNA levels between coinfected treatment groups. Plaque genotyping indicated that most plaques fully aligned with one of the parental strains; however, reassortants were detected, and in the 75:25 pool, most plaques were reassortant. Reassortant prevalence may be maximized upon the occurrence of reassortant genotypes that can outcompete the parental genotypes. BTV reassortment and resulting biological consequences are important elements to understanding orbivirus emergence and evolution.https://www.mdpi.com/1999-4915/16/2/240bluetongue virusco-infection<i>Culicoides</i>next-generation sequencingreassortment |
| spellingShingle | Molly Carpenter Jennifer Kopanke Justin Lee Case Rodgers Kirsten Reed Tyler J. Sherman Barbara Graham Mark Stenglein Christie Mayo Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> Viruses bluetongue virus co-infection <i>Culicoides</i> next-generation sequencing reassortment |
| title | Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> |
| title_full | Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> |
| title_fullStr | Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> |
| title_full_unstemmed | Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> |
| title_short | Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in <i>Culicoides sonorenesis</i> |
| title_sort | assessing reassortment between bluetongue virus serotypes 10 and 17 at different coinfection ratios in i culicoides sonorenesis i |
| topic | bluetongue virus co-infection <i>Culicoides</i> next-generation sequencing reassortment |
| url | https://www.mdpi.com/1999-4915/16/2/240 |
| work_keys_str_mv | AT mollycarpenter assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT jenniferkopanke assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT justinlee assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT caserodgers assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT kirstenreed assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT tylerjsherman assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT barbaragraham assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT markstenglein assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi AT christiemayo assessingreassortmentbetweenbluetonguevirusserotypes10and17atdifferentcoinfectionratiosiniculicoidessonorenesisi |