A novel method for the detection and diagnosis of virus infections in honey bees

Authors: HUANG, SHAOKANG - US Department Of Agriculture (USDA); LI, JIANGHONG - Fuzhou University; ZHANG, YI - US Department Of Agriculture (USDA); LI, ZHIGUO - Fuzhou University; Evans, Jay; ROSE, ROBYN - Animal And Plant Health Inspection Service (APHIS); GILLIGAN, TODD - Animal And Plant Health Inspection Service (APHIS); LEBRUNC, ANNE - Animal And Plant Health Inspection Service (APHIS); HE, NAN - Fuzhou University; ZHENG, TENG - Fuzhou Customs District Pr China; ZHANG, TIYIN - Fuzhou Customs District Pr China; Hamilton, Michele; Chen, Yanping - Judy

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/11/2021
Publication Date: 4/20/2021
Citation: Huang, Shaokang, Li, Jianghong, Zhang, Yi, Li, Zhiguo, Evans, J.D., Rose, Robyn, Gilligan, Todd, Lebrunc, Anne, He, Nan, Zheng, T., Zhang, T., Hamilton, M.C., Chen, Y. 2021. A novel method for the detection and diagnosis of virus infections in honey bees. Journal of Virological Methods. 293:114163. https://doi.org/10.1016/j.jviromet.2021.114163.
DOI: https://doi.org/10.1016/j.jviromet.2021.114163

Interpretive Summary: For infectious diseases caused by a variety of microorganisms, the ability to detect causative agents is the first step on the path to effective disease treatment and prevention. The detection and diagnosis of honeybee diseases often requires the sacrifice of honeybees for genetic material extraction, while also involving the use of volatile and extremely toxic organic solvents. We developed a novel and non-sacrificial method to obtain a small amount of blood from honey bees and then run molecular assay directly. The tiny puncture wound caused by the blood extraction procedure was found to heal automatically, and the lifespan of experimental workers remained unaffected. Using this method, we were able to detect multiple viruses in honey bees. The simplicity and cost-effectiveness of this innovative approach will allow it to be a valuable contribution to bee disease diagnosis and management programs, making it of interest to researchers, graduate students, beekeepers and policymakers involved or invested in this field around the world.

Technical Abstract: Among various abiotic and biotic factors that negatively affect bee populations, viruses have often been linked to colony losses and pose a serious threat to the health and well-being of honey bees. For infectious diseases caused by a variety of microorganisms, the ability to promptly and accurately identify the causative agents is the first step on the path to all effective management of disease infections. Over the years, PCR-based detection methods have provided powerful tools for rapid, specific, and sensitive detection and quantification of difficult-to-grow pathogenic microorganisms such as viruses in honey bees. However, PCR-based methods have required RNA extraction and purification, which can be quite laborious and time consuming and involve the use of organic solvents and chaotropic agents like phenol and chloroform which are volatile and extremely toxic. We developed a novel and non-sacrificial method for detecting viral infections in honey bees. As little as 1µl of hemolymph was collected from adult workers and queens of bee colonies by puncturing the soft inter-tergal integument between the 2nd and 3rd dorsal tergum with a fine glass capillary. The hemolymph was then subjected to RT-PCR analysis directly. The puncture wound caused by the glass capillary was found to heal automatically without trouble and the lifespan of experimental workers remained unaffected. Using this method, we detected multiple viruses including Deformed wing virus (DWV), Black queen cell virus (BQCV), Israeli acute paralysis virus (IAPV) and Sacbrood virus (SBV) in infected bees. Further, expressed transcripts indicating the induction of innate immune response in virus infected bees were also detected in hemolymph samples. The simplicity and cost-effectiveness of this innovative approach will allow it to be a valuable, time-saving, safer, and environmentally friendly contribution to bee disease management programs.