A new duplex qPCR assay for the quantification of honey bee ( Apis mellifera ) parasites Nosema ceranae and Nosema apis tested with low dose experimental exposure
A new duplex qPCR assay for the quantification of honey bee ( Apis mellifera ) parasites Nosema ceranae and Nosema apis tested with low dose experimental exposure
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一种新的双工 qPCR 测定法，用于定量蜜蜂 ( Apis mellifera ) 寄生虫 Nosema ceranae 和 Nosema apis，并通过低剂量实验暴露进行测试
DOI：
10.1080/00218839.2022.2083846
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发表时间：
2022
影响因子：
1.9
通讯作者：
Belden, Lisa K.
中科院分区：
文献类型：
--
作者：
Bradford, Emma L.;Gregory, Casey L.;Roman Longoria, Arturo;Jones, Korin Rex;Bueren, Emma K.;Haak, David C.;Fell, Richard;Belden, Lisa K.
在欧洲蜜蜂中引起疾病的蜜蜂小孢子虫和尖小孢子虫。鼻塞感染被认为是养蜂人丧失蜂群的潜在原因。鉴于鼻塞菌感染对菌落死亡率和生产力的重要性，开发新的和改进的检测和量化感染负荷的方法至关重要。我们设计并测试了一种新的双工qPCR方法，用于准确定量这两种n。ceranaeandN。使用TaqMan化学和新的gBlock®方法进行标准。该方法与自然鼻塞感染呈良好的线性关系，与显微孢子数有很强的相关性。该方法灵敏度高，重复性好，可用于蜂房调查和低剂量实验暴露后的血吸虫感染调查。在当地的荨麻疹中，我们发现氮含量相对较低。当然，很少。在弗吉尼亚州布莱克斯堡的三个地点进行的一项调查显示，西弗吉尼亚州的两个蜂场的n。但氮含量一直很低。api。在实验中，来自两个不同蜂巢的笼养蜜蜂被喂食100nosemaspore（或对照溶液）。两天后或五天后收集暴露的蜜蜂，并使用新的检测方法对感染进行量化。由于低剂量，感染水平不是100%，一些暴露的蜜蜂仍然存活。无菌素，而其他人只发生低水平感染。这种低剂量暴露和随后的感染状态（低水平感染或无感染）可以为鼻塞感染的建立提供新的认识。
Nosema ceranaeandNosema apisare microsporidian parasites that cause disease in the European honey bee.Nosemainfection is identified as a potential cause of colony loss by beekeepers. Given the importance ofNosemainfection in colony mortality and productivity, developing new and improved ways of detecting and quantifying infection loads is vital. We designed and tested a new duplex qPCR assay for the accurate quantification of bothN. ceranaeandN. apisutilising TaqMan chemistry and the new gBlock®method for the standards. The assay showed good linearity with naturalNosemainfection, and a strong correlation with microscopic spore counts. This new assay has high sensitivity and repeatability and was used to investigateNosemainfection in hive surveys and following low dose experimental exposure. In local hives, we found relativity low levels ofN. ceranaeand very littleN. apisacross three sites in Blacksburg, VA. A survey of two bee yards in West Virginia showed much higher levels ofN. ceranae,but consistent low levels ofN. apis. For the experiment, caged bees from two different hives were fed 100Nosemaspores (or a control solution). Exposed bees were collected after two or five days, and infection was quantified using the new assay. Given the low dose, infection levels were not 100%, with some exposed bees remainingN. ceranaefree, while others only developed low level infection. This low dose exposure and subsequent infection status (low level infection or infection free) could provide a new understanding ofNosemainfection establishment.