This agreement details the plan for conducting a study of honeybee pest diversity to support the development of government emergency response plans for non-native honeybee pests

The purpose of this project is to collect and study a range of honeybee pests (focusing primarily on the rapidly spreading brood parasite tropilaelaps mercedesae. And large specialized predatory vespa SPP. Such as the northern giant hornet, vespa mandarinia). This information will allow US to address gaps in our knowledge that prevent US from establishing effective emergency response plans to pest incursion in the US. Such work would also provide information integral to ongoing eradication efforts of current pests and prevention of future incursions. The project detailed herein represents the second year of a multi-year effort. Data collected from this study will support multiple research efforts focusing on NGH and international efforts to better understand tropi mites ultimately contributing to the betterment of honeybee health globally. The specific aims for year 2 of this study are as follows: -obtain and preserve specimens of asian honeybees, their pests, and symbionts from south asia (SA) east asia (EA), and southeast asia (sea) for further study -establish a database of physical and morphometric characters that can be used to distinguish between ostensibly cryptic species in the genera tropilaelaps, varroa, and vespa -improve the quality of previous subpar full-length genomes of t. Mercedesae and vespa mandarinia specifically targeting locations in asia where potential unresolved cryptic species or distinctive haplotypes may be present -continue identifying viruses found in and potentially vectored by tropi mites to all honeybee species confirmed or suspected to serve as hosts (a. Dorsata, a. Mellifera, a. Cerana, and a. Florea) -sequence the genome of the two dwarf honeybee species, apis andreniformis & apis florea -determine the identities of obscure or unidentified hive symbionts in honeybee species which consistently come in close contact with a. Mellifera in asia (i.e. a. Dorsata, a. Florea, a. Cerana, and thus have a heightened chance of making a host shift for year 2 of this project, we will deliver, by september 2023, the following: 1. At least 100 individual specimens of tropi mites from at least 3 different populations in asia. 2. Specimens of at least two asian honeybee species for genetic and viral analysis 3. A database of tropi and asian vespa SPP. Sequence data consisting of standard markers that can be used in identification and population genomics. 4. DNA sequence data with markers that can differentiate between different vespa or tropi subspecies or cryptic species. 5. Full-length genome sequence of two dwarf honeybee species 6. At least 30 specimens of asian vespa species from multiple asian populations beekeepers and farmers will benefit directly from this work as we reduce the capacity of invasive pests to reach the US or establish upon incursion thereby reducing the number of stress factors that our honeybee populations deal with. Honeybees further contribute to global food security by pollinating a range of important crops. As such, a better understanding of the parasites and pests that destabilize their populations will benefit consumers of these crops by ensuring they can be produced and sold at a reasonable price. In this way, beekeepers, farmers, and consumers of produce (and related products) are all stakeholders in this study. A portion of this award will support the salary and health insurance of a post-doctoral researcher at the university of colorado boulder who will focus their time on this work. As such, a sub-award consisting of $57,000 (salary for 1 year) will be allocated to the university of colorado's biofrontier institute to be disbursed to this researcher.