Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830)

Bell‐Sakyi, Lesley; Jaafar, Fauziah Mohd; Monsion, Baptiste; Luu, Lisa; Denison, Eric; Carpenter, Simon; Attoui, Houssam; Mertens, Peter P. C.

doi:10.3390/microorganisms8060825

Cited by 8 publications

(9 citation statements)

References 63 publications

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“…for cultures inoculated with flea organs, tick cells were resuspended by pipetting and cytocentrifuge smears were prepared from small aliquots (~50 µL) of cell suspension, stained with Giemsa and examined at ×500–1000 magnification. Wolbachia -infected BME/CTVM23 cells were prepared for transmission electron microscopy as described previously [ 61 ].…”

Section: Methodsmentioning

confidence: 99%

Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines

et al. 2020

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Wolbachia are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although Wolbachia DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not Wolbachia can infect and grow in tick cells, cell lines from three tick species, Ixodes scapularis, Ixodes ricinus and Rhipicephalus microplus, were inoculated with Wolbachia strains wStri and wAlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an I. scapularis cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of Wolbachia genes. The wStri strain infected Ixodes spp. cells and was maintained through 29 passages. The wAlbB strain successfully infected Ixodes spp. and R. microplus cells and was maintained through 2–5 passages. A novel strain of Wolbachia belonging to the supergroup F, designated wCfeF, was isolated in I. scapularis cells from a pool of Ctenocephalides sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a Wolbachia strain from a flea and the first isolation of any Wolbachia strain outside the “pandemic” A and B supergroups. The study demonstrates that tick cells can host multiple Wolbachia strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of Wolbachia.

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Section: Methodsmentioning

confidence: 99%

Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines

et al. 2020

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“…Many different cell lines derived from C. sonorensis exist, and their susceptibility to a number of BTV and epizootic hemorrhagic disease virus (EHDV) serotypes are known [ 22 ]. Additionally, cell lines derived from C. nubeculosus have recently been established [ 82 ]. As an initial step to investigate virus inhibitory effects induced by Wolbachia infections or transgene-based genetic modifications, the rate of viral proliferation in modified cells can be compared to that reported in previous studies.…”

Section: Research Gaps Concerning the Use Of Novel Control Approachesmentioning

confidence: 99%

Next-generation tools to control biting midge populations and reduce pathogen transmission

et al. 2021

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Biting midges of the genus Culicoides transmit disease-causing agents resulting in a significant economic impact on livestock industries in many parts of the world. Localized control efforts, such as removal of larval habitat or pesticide application, can be logistically difficult, expensive and ineffective if not instituted and maintained properly. With these limitations, a population-level approach to the management of Culicoides midges should be investigated as a means to replace or supplement existing control strategies. Next-generation control methods such as Wolbachia- and genetic-based population suppression and replacement are being investigated in several vector species. Here we assess the feasibility and applicability of these approaches for use against biting midges. We also discuss the technical and logistical hurdles needing to be addressed for each method to be successful, as well as emphasize the importance of addressing community engagement and involving stakeholders in the investigation and development of these approaches.

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“…After several subcultures, cells were passaged into sealed, 25-cm 2 culture flasks. Cells were cryopreserved in medium with 20% FBS and 10% DMSO and resuscitated as described previously for Culicoides nubeculosus cell lines [30]. Primary C. pipiens and P. papatasi cultures and young cell lines were periodically examined by preparation of Giemsa-stained cytocentrifuge smears as described previously [30].…”

Section: Pipiens and P Papatasi Cell Line Generationmentioning

confidence: 99%

“…Cells were cryopreserved in medium with 20% FBS and 10% DMSO and resuscitated as described previously for Culicoides nubeculosus cell lines [30]. Primary C. pipiens and P. papatasi cultures and young cell lines were periodically examined by preparation of Giemsa-stained cytocentrifuge smears as described previously [30]. Absence of contaminating Mycoplasma in all young cell lines was confirmed using two commercial tests, the Mycoalert Mycoplasma Detection Kit (Lonza, Fisher Scientific, Loughborough, UK) and the PCR Mycoplasma Test Kit (Promocell, VWR, Lutterworth, UK), following the manufacturers' instructions.…”

Section: Pipiens and P Papatasi Cell Line Generationmentioning

confidence: 99%

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Isolation in Natural Host Cell Lines of Wolbachia Strains wPip from the Mosquito Culex pipiens and wPap from the Sand Fly Phlebotomus papatasi

Bell‐Sakyi

Beliavskaia

Hartley

et al. 2021

Insects

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Endosymbiotic intracellular bacteria of the genus Wolbachia are harboured by many species of invertebrates. They display a wide range of developmental, metabolic and nutritional interactions with their hosts and may impact the transmission of arboviruses and protozoan parasites. Wolbachia have occasionally been isolated during insect cell line generation. Here, we report the isolation of two strains of Wolbachia, wPip and wPap, during cell line generation from their respective hosts, the mosquito Culex pipiens and the sand fly Phlebotomus papatasi. wPip was pathogenic for both new C. pipiens cell lines, CPE/LULS50 and CLP/LULS56, requiring tetracycline treatment to rescue the lines. In contrast, wPap was tolerated by the P. papatasi cell line PPL/LULS49, although tetracycline treatment was applied to generate a Wolbachia-free subline. Both Wolbachia strains were infective for a panel of heterologous insect and tick cell lines, including two novel lines generated from the sand fly Lutzomyia longipalpis, LLE/LULS45 and LLL/LULS52. In all cases, wPip was more pathogenic for the host cells than wPap. These newly isolated Wolbachia strains, and the novel mosquito and sand fly cell lines reported here, will add to the resources available for research on host–endosymbiont relationships, as well as on C. pipiens, P. papatasi, L. longipalpis and the pathogens that they transmit.

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Continuous Cell Lines from the European Biting Midge Culicoides nubeculosus (Meigen, 1830)

Cited by 8 publications

References 63 publications

Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines

Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines

Next-generation tools to control biting midge populations and reduce pathogen transmission

Isolation in Natural Host Cell Lines of Wolbachia Strains wPip from the Mosquito Culex pipiens and wPap from the Sand Fly Phlebotomus papatasi

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