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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Main Authors: Lesley Bell-Sakyi, Fauziah Mohd Jaafar, Baptiste Monsion, Lisa Luu, Eric Denison, Simon Carpenter, Houssam Attoui, Peter P. C. Mertens
Format: Article
Language:English
Published: MDPI AG 2020-05-01
Series:Microorganisms
Subjects:
Online Access:https://www.mdpi.com/2076-2607/8/6/825
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spelling 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
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