Department of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
Sci Rep. 2020 Jan 24;10(1):1118. doi: 10.1038/s41598-020-57909-8.
The reduction of bumblebee populations has been reported in the last decades, and the microsporidian parasite Nosema bombi is considered as one of the factors contributing to such reduction. Although the decline of bee populations affects both wild plants and human food supply, the effects of Nosema spp. infections are not known because it is difficult to obtain infective spores from wild bees due to their low prevalence. Microscopical observation of fecal samples or midgut homogenates and/or PCR are generally used for N. bombi detection. However, the germination rate of microsporidian spore declines if they are kept at 4 °C for a long time or frozen. It is therefore crucial to minimize the diagnosis and isolation time of infective spores from field-collected samples. Therefore, we performed a loop-mediated isothermal amplification (LAMP) assay for the direct detection of N. bombi in bumblebee midgut homogenates. Using this method, we could detect N. bombi from individuals from which it was visible under the microscope and directly from wild individuals.
在过去的几十年中，报道了熊蜂种群数量的减少，而微孢子虫寄生虫Nosema bombi 被认为是导致这种减少的因素之一。尽管蜜蜂种群的减少会影响野生植物和人类的食物供应，但由于从野生蜜蜂中获得传染性孢子的难度较大（因为其流行率较低），因此尚不清楚 Nosema spp. 感染的影响。通常使用粪便样本或中肠匀浆的显微镜观察和/或 PCR 来检测 N. bombi。然而，如果将微孢子虫孢子在 4°C 下保存很长时间或冷冻，其发芽率会下降。因此，从现场采集的样本中尽可能减少传染性孢子的诊断和分离时间至关重要。因此，我们对熊蜂中肠匀浆中的 N. bombi 进行了环介导等温扩增（LAMP）检测。使用该方法，我们可以从可在显微镜下观察到的个体以及直接从野生个体中检测到 N. bombi。
