Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830)
<i>Culicoides</i> biting midges (Diptera: Ceratopogonidae) transmit arboviruses of veterinary or medical importance, including bluetongue virus (BTV) and Schmallenberg virus, as well as causing severe irritation to livestock and humans. Arthropod cell lines are essential laboratory resea...
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doaj-5f9800fec715450484a8855efbacedef2020-11-25T03:34:08ZengMDPI AGMicroorganisms2076-26072020-05-01882582510.3390/microorganisms8060825Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830)Lesley Bell-Sakyi0Fauziah Mohd Jaafar1Baptiste Monsion2Lisa Luu3Eric Denison4Simon Carpenter5Houssam Attoui6Peter P. C. Mertens7Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKUMR1161 Virologie, INRAE, Ecole Nationale Vétérinaire d’Alfort, ANSES, Université Paris-Est, 94700 Maisons-Alfort, FranceUMR1161 Virologie, INRAE, Ecole Nationale Vétérinaire d’Alfort, ANSES, Université Paris-Est, 94700 Maisons-Alfort, FranceInstitute of Infection, Veterinary and Ecological Sciences, University of Liverpool, 146 Brownlow Hill, Liverpool L3 5RF, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UKUMR1161 Virologie, INRAE, Ecole Nationale Vétérinaire d’Alfort, ANSES, Université Paris-Est, 94700 Maisons-Alfort, FranceSchool of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, LE12 5RD, UK<i>Culicoides</i> biting midges (Diptera: Ceratopogonidae) transmit arboviruses of veterinary or medical importance, including bluetongue virus (BTV) and Schmallenberg virus, as well as causing severe irritation to livestock and humans. Arthropod cell lines are essential laboratory research tools for the isolation and propagation of vector-borne pathogens and the investigation of host-vector-pathogen interactions. Here we report the establishment of two continuous cell lines, CNE/LULS44 and CNE/LULS47, from embryos of <i>Culicoides nubeculosus</i>, a midge distributed throughout the Western Palearctic region. Species origin of the cultured cells was confirmed by polymerase chain reaction (PCR) amplification and sequencing of a fragment of the <i>cytochrome oxidase</i> <i>1</i> gene, and the absence of bacterial contamination was confirmed by bacterial 16S rRNA PCR. Both lines have been successfully cryopreserved and resuscitated. The majority of cells examined in both lines had the expected diploid chromosome number of 2<i>n</i> = 6. Transmission electron microscopy of CNE/LULS44 cells revealed the presence of large mitochondria within cells of a diverse population, while arrays of virus-like particles were not seen. CNE/LULS44 cells supported replication of a strain of BTV serotype 1, but not of a strain of serotype 26 which is not known to be insect-transmitted. These new cell lines will expand the scope of research on <i>Culicoides</i>-borne pathogens.https://www.mdpi.com/2076-2607/8/6/825CulicoidesCeratopogonidaeMonoculicoidescell linevectormidge |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lesley Bell-Sakyi Fauziah Mohd Jaafar Baptiste Monsion Lisa Luu Eric Denison Simon Carpenter Houssam Attoui Peter P. C. Mertens |
spellingShingle |
Lesley Bell-Sakyi Fauziah Mohd Jaafar Baptiste Monsion Lisa Luu Eric Denison Simon Carpenter Houssam Attoui Peter P. C. Mertens Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) Microorganisms Culicoides Ceratopogonidae Monoculicoides cell line vector midge |
author_facet |
Lesley Bell-Sakyi Fauziah Mohd Jaafar Baptiste Monsion Lisa Luu Eric Denison Simon Carpenter Houssam Attoui Peter P. C. Mertens |
author_sort |
Lesley Bell-Sakyi |
title |
Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) |
title_short |
Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) |
title_full |
Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) |
title_fullStr |
Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) |
title_full_unstemmed |
Continuous Cell Lines from the European Biting Midge <i>Culicoides nubeculosus </i>(Meigen, 1830) |
title_sort |
continuous cell lines from the european biting midge <i>culicoides nubeculosus </i>(meigen, 1830) |
publisher |
MDPI AG |
series |
Microorganisms |
issn |
2076-2607 |
publishDate |
2020-05-01 |
description |
<i>Culicoides</i> biting midges (Diptera: Ceratopogonidae) transmit arboviruses of veterinary or medical importance, including bluetongue virus (BTV) and Schmallenberg virus, as well as causing severe irritation to livestock and humans. Arthropod cell lines are essential laboratory research tools for the isolation and propagation of vector-borne pathogens and the investigation of host-vector-pathogen interactions. Here we report the establishment of two continuous cell lines, CNE/LULS44 and CNE/LULS47, from embryos of <i>Culicoides nubeculosus</i>, a midge distributed throughout the Western Palearctic region. Species origin of the cultured cells was confirmed by polymerase chain reaction (PCR) amplification and sequencing of a fragment of the <i>cytochrome oxidase</i> <i>1</i> gene, and the absence of bacterial contamination was confirmed by bacterial 16S rRNA PCR. Both lines have been successfully cryopreserved and resuscitated. The majority of cells examined in both lines had the expected diploid chromosome number of 2<i>n</i> = 6. Transmission electron microscopy of CNE/LULS44 cells revealed the presence of large mitochondria within cells of a diverse population, while arrays of virus-like particles were not seen. CNE/LULS44 cells supported replication of a strain of BTV serotype 1, but not of a strain of serotype 26 which is not known to be insect-transmitted. These new cell lines will expand the scope of research on <i>Culicoides</i>-borne pathogens. |
topic |
Culicoides Ceratopogonidae Monoculicoides cell line vector midge |
url |
https://www.mdpi.com/2076-2607/8/6/825 |
work_keys_str_mv |
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