Culicoides 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 Culicoides nubeculosus, 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 cytochrome oxidase 1 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 2n = 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 Culicoides-borne pathogens.

中文翻译：

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来自欧洲咬蠓蠓的连续细胞系 (Meigen, 1830)。

库蠓蠓（双翅目：蠓科）的兽医或医学重要性传输虫媒病毒，包括蓝舌病毒（BTV）和施马伦贝格病毒，以及造成严重刺激牲畜和人类。节肢动物细胞系是分离和传播媒介传播病原体以及研究宿主-媒介-病原体相互作用的重要实验室研究工具。在这里，我们报告了从Culicoides nubeculosus胚胎中建立两个连续细胞系，CNE/LULS44 和 CNE/LULS47，这是一种分布在整个西古北地区的蠓。通过聚合酶链反应 (PCR) 扩增和细胞色素氧化酶 1片段的测序确认培养细胞的物种来源基因，并且细菌 16S rRNA PCR 证实没有细菌污染。两条线都已成功冷冻保存和复苏。在这两个细胞系中检查的大多数细胞具有 2 n = 6的预期二倍体染色体数。CNE/LULS44 细胞的透射电子显微镜显示在不同种群的细胞内存在大线粒体，而病毒样颗粒阵列则没有看到了。CNE/LULS44 细胞支持 BTV 血清型 1 菌株的复制，但不支持未知昆虫传播的血清型 26 菌株的复制。这些新的细胞系将扩大研究范围的库蠓-borne病原体。