Rapid imaging, detection, and quantification of Nosema ceranae spores in honey bees using mobile phone-based fluorescence microscopy

Abstract

Recent declines in honey bee colonies in the United States have put increased strain on agricultural pollination. Nosema ceranae and Nosema apis, are microsporidian parasites that are highly pathogenic to honey bees and have been implicated as a factor in honey bee losses. While traditional methods for quantifying Nosema infection have high sensitivity and specificity, there is no field-portable device for field measurements by beekeepers. Here we present a field-portable and cost-effective smartphone-based platform for detection and quantification of chitin-positive Nosema spores in honey bees. The handheld platform, weighing only 374 g, consists of a smartphone-based fluorescence microscope, a custom-developed smartphone application, and an easy to perform sample preparation protocol. We tested the performance of the platform using samples at different parasite concentrations and compared the method with manual microscopic counts and qPCR quantification. We demonstrated that this device provides results that are comparable with other methods, having a limit of detection of 0.5 × 10⁶ spores per bee. Thus, the assay can easily identify infected colonies and provide accurate quantification of infection levels requiring treatment of infection, suggesting that this method is potentially adaptable for diagnosis of Nosema infection in the field by beekeepers. Coupled with treatment recommendations, this protocol and smartphone-based optical platform could improve the diagnosis and treatment of nosemosis in bees and provide a powerful proof-of-principle for the use of such mobile diagnostics as useful analytical tools for beekeepers in resource-limited settings.