How Much Is Enough? an Analysis of the 5' Nontranslated Region of the Deformed Wing Virus in Honeybees

Honeybees are an essential component of today¿s agricultural system because of their role as pollinators. However, viruses, including a member of the Picornavirales order known commonly as Deformed Wing Virus (DWV), are compromising the health of honeybee colonies. Many picornaviruses, such as poliovirus, have been studied in depth because of their relation to human disease, but also because of their use of an Internal Ribosome Entry Site (IRES) to initiate translation. The primary goal of this thesis was to determine if the 5¿ Non-Translated Region (NTR) of Deformed Wing Virus (DWV) functions as an IRES. A secondary goal was to determine if there are specific parts of that 5¿ NTR that are important to IRES function. Six plasmids were constructed by inserting three different sections of the 5¿ NTR of DWV, in both sense and antisense directions, between two reporter genes. These plasmids, along with several control plasmids, were transfected into Sf9 cells, and post-transfection luciferase assays were conducted. Results were inconclusive. This could have been due to an inability of the plasmids to be expressed in Sf9 cells, an error in the construction of the plasmids, or a mechanical error in the assay procedure. At this time it appears most likely that the 5¿ NTR of DWV may be cell-type or species specific, and the next step would be to transfect the plasmids into a recently developed cultured honeybee cell line.

“What Happens When Pollinators Get Sick? Behavioral Impacts of Honeybee Viruses”

Like all organisms on the planet, honeybees (Apis mellifera) are susceptible to infection with a wide variety of viruses. These viruses may produce infections with no visible symptoms or may have devastating consequences on both the individual bee and the entire hive. Deformed Wing Virus, a member of the Iflavirus group of viruses, has an RNA genome and has had a particularly important impact on bee health. It can be spread between bees in a several ways – bees can infect each other during feeding or grooming activities, drones can pass the virus to the queen during mating and queens can lay infected eggs. The primary and most devastating way that these viruses are transmitted within and between hives involves a parasitic mite, an animal known ominously as Varroa destructor. The talk will discuss the effect that viruses have on the health and behavior of honeybees and will outline the collaborative research activities of Drs. Evans and Pizzorno over the last 7 years.

Localization of Deformed Wing Virus (DWV) in the Brains of Apis mellifera (European Honey Bees)

The purpose of the current research project is to design a successful in-situ hybridization to identify regions within the brains of honeybees where DWV replicates. The localization of the virus in the brains of the bees can draw a connection between CCDand DWV.In conclusion, these results demonstrate that in bees infected with DWV the virus replicates actively in very important regions of the brain, including neuropils that are responsible for vision and olfaction. This means that the virus could adversely affect the vision and olfaction of the honeybees making it difficult for bees to behave normally.