Single-cell analysis reveals the effects of glutaraldehyde and formaldehyde on individual Nosema bombycis spores
Single-cell analysis reveals the effects of glutaraldehyde and formaldehyde on individual Nosema bombycis spores
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单细胞分析揭示戊二醛和甲醛对单个家蚕微孢子虫孢子的影响
DOI：
10.1039/c8an02425a
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发表时间：
2019-05-07
期刊：
影响因子：
4.2
通讯作者：
Wang, Guiwen
中科院分区：
文献类型：
--
作者：
Miao, Zhenbin;Zhang, Pengfei;Wang, Guiwen
家蚕微孢子虫（Nb）是引起家蚕微粒子病的病原体。醛类是蚕业中常用的有效消毒剂。然而，它们对Nb孢子作用的精确机制仍不清楚。在此，我们利用激光镊子拉曼光谱研究戊二醛和甲醛对单个Nb孢子的影响，并利用相差显微镜成像监测单个处理孢子的萌发动态，以更深入地了解醛类的作用机制，为建立有效的蚕业疾病控制策略提供理论参考。在处理过程中，拉曼峰的位置保持不变。随着处理时间的增加，拉曼强度增强，孢子的萌发率显著降低。单个孢子开始萌发的时间T - lag和完全萌发的时间T - germ随着处理强度的增加而增加。萌发时间（T - germ）与未处理孢子相比无显著差异。显示出异质性，这与Nb孢子对醛类的抗性有关。结果表明，戊二醛和甲醛不会破坏孢子壁和质膜，不会导致细胞内成分泄漏，可能也不会损坏挤出装置。醛类对Nb孢子的作用主要在孢子外壳上。它们可能阻断刺激孢子萌发的外部因素。基于新型光学技术的单细胞分析实时揭示了化学杀菌剂对微孢子虫孢子的作用，并解释了细胞抗逆性的异质性。这些新技术的应用为传统消毒剂提供了新的见解。
Nosema bombycis (Nb) is the pathogen that causes pebrine in silkworms. Aldehydes are effective disinfectants commonly used in sericulture. However, the precise mechanism of their action on Nb spores remains unclear. Here, we used laser tweezers Raman spectroscopy to investigate the effects of glutaraldehyde and formaldehyde on individual Nb spores, as well as phase contrast microscopy imaging to monitor the germination dynamics of individual treated spores, to acquire a deeper understanding of the mechanism of action of aldehydes and to provide a theoretical reference for establishing an effective strategy for disease control in sericulture. The positions of the Raman peaks remained constant during treatment. The Raman intensity was enhanced and the germination rate of the spores significantly decreased with treatment time. T-lag, the time when individual spores begin to germinate, and T-germ, the time for complete germination, increased with enhanced treatment. The germination time (T-germ) showed no significant difference from that for untreated spores. Heterogeneity was shown, which is relevant to the resistance of Nb spores to aldehydes. The results indicate that glutaraldehyde and formaldehyde do not destroy the spore wall and plasma membrane, do not cause the leakage of intracellular components, and might not damage the extrusion apparatus. The effects of aldehydes on Nb spores are mainly on the spore coat. They may block the external factors that stimulate spore germination. Single-cell analysis based on novel optical techniques reveals the action of chemical sporicides on microsporidia spores in real time and explains the heterogeneity of cell stress resistance. These applications of new techniques offer new insight into traditional disinfectants.