Live and dead qPCR detection demonstrates that feeding of <i>Nosema ceranae</i> results in infection in the honey bee but not the bumble bee

As the honey bee and bumble bee may suffer from the same or related microbial pathogens, cross contamination from commercially reared <i>Bombus</i> spp. to honey bees and wild bumble bees and vice versa is a major concern. Honey bee-collected pollen to feed commercially reared <i>Bombus</i> spp. is a potential risk. <i>Nosema</i> spp. is a fungal pathogen in bees. In this study, we developed new quantitative detection tools based on the detection of RNA using a TaqMan-based RT-qPCR for <i>Nosema ceranae</i> and <i>Nosema apis</i>, with extraction controls based on the actin gene of honey bees and bumble bees, respectively. These tools were subsequently applied to study the epidemiology of <i>N. ceranae</i>, a main disease in honey bees. We screened gamma radiation and cold treatment sterilisation for their efficacy to kill <i>N. ceranae</i> spores fed in sugar water and in pollen to honey bees and bumble bees, respectively. <i>N. ceranae</i> infection in adult bumble bees was checked. Spores passing the inter-alimentary track were found but no infection was observed. <i>N. ceranae</i> spores were fed to honey bees. Their presence and multiplication were demonstrated, showing the spores were both viable and infectious. Our results indicate that <i>N. ceranae</i> found in honey bees cannot infect commercially reared bumble bees (<i>Bombus terrestris</i>) and, that gamma radiation effectively kills <i>N. ceranae</i>. The highly specific and sensitive molecular assays developed, were exploited to detect <i>N. ceranae</i> in pollen and faeces, which would allow more comprehensive epidemiological studies on this important pathogen.鉴于蜜蜂与熊蜂可能感染相同或相关的微生物病原，商业化饲养的熊蜂属（<i>Bombus</i> spp.）种群与蜜蜂、野生熊蜂之间的双向交叉污染已成为学界高度关注的问题。用于饲喂商业化饲养熊蜂的蜜蜂采集花粉，存在潜在的病原传播风险。微孢子虫属（<i>Nosema</i>）是一类侵染蜜蜂的真菌性病原体。本研究针对雅氏微孢子虫（<i>Nosema ceranae</i>）与蜜蜂微孢子虫（<i>Nosema apis</i>），开发了基于TaqMan探针的实时荧光定量逆转录聚合酶链反应（TaqMan-based RT-qPCR）的RNA检测新型定量检测技术，内参提取对照分别采用蜜蜂与熊蜂的肌动蛋白基因。随后，利用该技术开展了雅氏微孢子虫的流行病学研究——该病原是引发蜜蜂主要病害的关键致病菌。我们分别针对饲喂蜜蜂的糖水与饲喂熊蜂的花粉中的雅氏微孢子虫孢子，评估了γ射线辐照与低温处理两种灭菌手段的杀孢子效能。我们检测了成年熊蜂的雅氏微孢子虫感染情况，结果虽检出可通过消化道的孢子，但未观察到感染现象。我们将雅氏微孢子虫孢子饲喂给蜜蜂，随后证实了孢子的存在与增殖情况，证明该孢子兼具活性与侵染性。本研究结果表明：蜜蜂体内检出的雅氏微孢子虫无法侵染商业化饲养的地熊蜂（<i>Bombus terrestris</i>），且γ射线辐照可有效杀灭雅氏微孢子虫孢子。本研究开发的高特异性、高灵敏度分子检测技术，可用于检测花粉与粪便中的雅氏微孢子虫，为该重要病原的更全面流行病学研究提供了技术支撑。

搜集汇总

背景与挑战

背景概述

该数据集聚焦于通过开发新型RT-qPCR检测工具，研究真菌病原体Nosema ceranae在蜜蜂和熊蜂中的感染差异。研究发现，N. ceranae能通过喂养感染蜜蜂并繁殖，但不能感染商业饲养的熊蜂（Bombus terrestris），同时验证了γ辐射灭菌的有效性。这些工具可用于检测花粉和粪便中的病原体，为蜜蜂健康管理和流行病学调查提供技术支持。