Departments of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California-Davis, Davis, CA, USA.
SpecialtyVETPATH, Seattle, WA, USA.
J Vet Diagn Invest. 2023 Nov;35(6):630-638. doi: 10.1177/10406387231191732. Epub 2023 Aug 16.
The Western honey bee () is economically important as the primary managed pollinator of many agricultural crops and for the production of various hive-related commodities. Honey bees are not classically or thoroughly covered in veterinary pathology training programs. Given their unique anatomic and biological differences from the other species more traditionally evaluated by veterinary pathologists, establishing routine and consistent methods for processing samples for histology ensures accurate diagnostic and research conclusions. We developed and tested several field protocols for the sampling of honey bees. We compared the tissue-quality outcomes for worker bees fixed, collected, and/or softened under the following protocols: 1) routine formalin fixation; 2) softening chitin via exposure to Nair for 2 d or 3) 5 d; 4) shortened times between formalin submersion and trimming of body segments to enhance penetration of formalin into internal tissues; 5) ethanol submersion of specimen prior to formalin fixation; 6) indirect dry ice exposure; and 7) prolonged -80°C storage. Routine formalin fixation, exposure to Nair for 2 d, indirect dry ice exposure, and trimming body segments within 2 h of formalin submersion resulted in the highest quality histologic tissue sections. The poorest quality sections resulted from softening of chitin by exposure to Nair for 5 d, submersion in ethanol for 3 d before formalin fixation, and prolonged storage at -80°C. Our results indicate that routine formalin fixation is adequate, and that immobilizing bees with indirect dry ice exposure aids in sample collection without negatively impacting the quality of histologic sections.
西方蜜蜂 () 在经济上非常重要，因为它是许多农业作物的主要管理传粉媒介，也是各种与蜂箱相关商品的生产来源。兽医病理学培训课程通常不会全面涵盖蜜蜂。鉴于它们与兽医病理学家传统评估的其他物种在解剖学和生物学上存在独特的差异，为确保准确的诊断和研究结论，建立用于组织学处理样本的常规和一致方法至关重要。我们开发并测试了几种用于采集蜜蜂的现场方案。我们比较了以下方案中固定、收集和/或软化的工蜂组织质量结果：1) 常规福尔马林固定；2) 通过暴露于 Nair 来软化几丁质 2 天或 3 天；3) 5 天；4) 缩短福尔马林浸泡和修剪身体部位之间的时间，以增强福尔马林进入内部组织的渗透；5) 在福尔马林固定之前将标本浸泡在乙醇中；6) 间接干冰暴露；和 7) 长时间储存于-80°C。常规福尔马林固定、暴露于 Nair 2 天、间接干冰暴露以及在福尔马林浸泡后 2 小时内修剪身体部位，可获得最高质量的组织学切片。通过暴露于 Nair 5 天、在福尔马林固定前将标本浸泡在乙醇中 3 天以及长时间储存于-80°C，几丁质软化会导致最差的切片质量。我们的结果表明，常规福尔马林固定是足够的，并且使用间接干冰固定蜜蜂有助于收集样本，而不会对组织学切片的质量产生负面影响。
