Rising Tide Seminar

Enhancing Genetic Diversity in the U.S. Honey Bee Gene Pool Susan Cobey Endemic Range

Apis mellifera Diversity carnica macedonica

Honey bees are (1 (2 (2 6 1 4 Endemic to Europe, da d da ys ay ys Africa & Central Asia ) s) )

28 recognized subspecies

The first honey bees imported to the US were the “dark” bee of Northern Europe, Apis mellifera mellifera

Colonies were established in Virginia by 1622.

Dispersed and established widely in the countryside ahead of human colonial expansion. Historical Importations of Italian bees

1859 – First importation of A. m. ligustica. Sourced from Johann Dzierzon’s apiary in Germany.

1860 –A. m. ligustica imported directly from Italy to the U.S. Samual Parsons with beekeeping assistance from L. L. Langstroth.

A subset of these subspecies were brought to North America

And were established to form the basis for current honey bee populations in the United States

1922 Honey Bee Act

Restricted further importations in attempt to keep out tracheal mites California Queen & Package Bee Industry

Supplier of Queens & Bees to Re-Stock the Industry Nationwide

Garvey Also Responsible for …

Selection & Maintenance of Genetic Material Increasing Challenges… Loss of Diverse Forage Intensive Agricultural Practices Exposure to Chemical Residues Pests, Parasites & Pathogens Reduced Genetic Diversity

Garvey Increasing Challenges… Malnutrition- Lack of Adequate and Diverse Forage

Monoculture Crops Habitat Degradation Climate Change Sub-lethal Effects Of Chemical Exposure Queen Effects: Mating, Signaling Egg production/laying Pre-mature supersedure Brood / Nutritional Effects: Inhibits bee bread microorganisms Delayed development Growth abnormalities Reduced longevity Increased morality Neurological Effects: Impaired visual sensory Reduced memory & learning performance Communication difficulties

Increasing Challenges…

Multiple Resident Pathogens Co-Exist In Colonies

More Pathogenic In Association with “Stressors” Varroa, Nosema, Malnutrition, Chemical Residues Varying Tolerance / Susceptibility Among Colonies Changing Combinations of Pathogens & Conditions Crisis Management Production Demands, “Quick Fix” Treatments

Changes In Commercial U.S. Honey Bee Stocks High Colony Losses Mid 1980’s - Tracheal Mites 1990’s & Continuing – Varroa Mites 2000’s - CCD, Nosema ceranae, Viruses

WSU Survey Of Commercial Queen Producers Schiff & Sheppard , 1995 & 1996. Delaney, Meixner, Schiff & Sheppard, 2009 25% Loss of Alleles in 10 Yrs, 1993-4 to 2004-5 Gain of 10% - Russian, Australian, Africanized

USDA- AIA Survey Winter Loss of Managed Colonies 21.7% Winter Loss 2011-2012 30.0% Winter Loss 2010-2011 33.8% Winter Loss 2009-2010 Garvey 28.6 % Winter Loss 2008-2009 35.8 % Winter Loss 2007-2008 31.8 % Winter Loss 2006-2007 26 Old World Honey Bee Subspecies Recognized A small subset introduced to establish Domestic US populations Honey Bee Act of 1922, Banned Importation

Garvey Garvey Genetic Bottlenecks of US Honey Bee Populations

Founding Population Feral Population Consolidation by Commercial A small subset sampling Eliminated by Queen Production of several subspecies Parasitic mites 400-500 Mothers produce introduced before the 1.5 million queens annually. 1922 Honey Bee Act A Diverse Workforce Increases Colony Fitness

Duong & Schneider, 2008 Mattila & Seeley, 2007 Jones, et.al., 2004 Page et al, 1995 Oldroyd, et.al., 1992,

A Diverse Workforce Reduces Prevalence, Severity of Disease & Pests

Tarpy & Seeley, 2007, Seeley & Tarpy, 2007 Tarpy, 2003, Fuchs & Schade. 1994

Insemination Quality Affects Queen Performance

MDI vs SDI Queen More Attractive Larger Retinue Response

Mating Quality Changes: Queen Physiology, Pheromone Profiles, Social Interactions

Richard, Tarpy, Grozinger, 2007 Mating Behavior Encourages Intra-Colony Genetic Diversity

Greater Microbial Communities Reduce Pathogen Loads Bacteria transform collected pollen into Bee Bread Garvey High intra-colony genetic diversity : Greater microbial diversity, Reduced pathogen loads. Increased abundance helpful bacteria

Mattila et al 2012

Garvey Genetic Diversity & Behavioral Mechanisms Compensate for Genome Deficiencies

Few Genes for Immunity Increased Risk to Pests and Pathogens Few Genes for Detoxification Vulnerable to Pesticides

Mechanisms of Resistance Ability to Detect and Remove Disease & Pests Propolis -Collection of plant resins with antimicrobial activity UCD & WSU Collaborative Project Genetic Enhancement of Commercial Stocks Importation of Germplasm

Transport of Honey Bee Semen Insemination to Domestic Queens

Goals for Germplasm Importation

•Enhance the genetic diversity of US honey bee populations • Provide new “raw material” for selection and breeding efforts •Reintroduce a previously available strain - the Caucasian honey bee APHIS Importation Permit- Honey Bee Semen

USDA-APHIS Quarantine Established WSU Ecological Preserve Smoot Hill Prairie

3 Year Permit 2008-2010 2011-2013 A.m.ligustica From Italy A.m.carnica From Germany and Slovenia A.m.caucasica From Rep. of Georgia

Instrumental Insemination

USDA-APHIS Quarantine Apiary WSU Ecological Preserve

Virus screening of semen & daughter workers by Beltsville USDA lab

Released on a case by case basis by USDA- APHIS

Propagation & Distribution to Cooperating Queen Producers Importations from Italy Bari- 2008, 2009 Bologna -2010 Various Locations - 2012

New World Carniolan 1981-Present Page-Laidlaw Closed Population Breeding Program Selection for General Performance

Productivity Brood Viability

Over-Wintering Temperament Selection for Specific Traits of Resistance

Hygienic Behavior Assay Select Multiple Traits

4. Following Spring Final Selection: Spring Buildup Production 1. Spring Varroa Growth Queen Rearing Brood Diseases &Insemination 2. Fall Pre-selection: Brood Viability Temperament Pollen/Honey Stores Disease/Parasites Hygienic Behavior 3. Over-wintering

Traditional Beekeeping in Slovenia Apis mellifera carnica Apis mellifera carnica Semen Collection

Re-Establishing Apis mellifera caucasica from the Republic Of Georgia

Propolis - Resin Collection

Social Immunity, Behavioral defense Reduces immune response Self medication

Simone-Finstrom & Spivak, 2012 Evans & Spivak, 2010 The Road to Mestia The Traffic Roadside Beekeeping Drone Collection Resistance Mechanisms

Variable Among Colonies Indicates Potential Highly Complex Partially Understood Constantly Changing Dependent Upon Nutritional Factors Management Practices Variation in Virulence of Varroa Auto Alo Variation in Susceptibility Combination with Associated Pathogens Drawings By O. Environmental Conditions Boecking

Cost vs Benefit Selection for Resistance vs Productivity

Resistant Strains Less Productive Reduced Brood Production High Swarming Tendency Un-Selected Increased Costs Increased Labor Chemical Residues

Mechanisms of Resistance Involve Behavioral, Physiological & Beekeeper Management

Olfactory Sensitivity - Detection & Removal Hygienic & Grooming Behavior Development Time Post-capping Stage Duration & Breaks In the Brood Cycle Re-Queening Dividing Colonies Inhibition of Varroa Reproduction Cuticular Hydrocarbons

Garvey Partnering With California Queen Producers Collaborative Efforts Between Industry and Research

Partnering with the Ca. Tech Transfer Team Tech Transfer Specialized Classes To Promote Stock Improvement

Instrumental Insemination

Queen Rearing Cryopreservation of Imported Semen

Brandon Hopkins, WSU Cryopreservation Techniques

Multiple Generations of Queens Produced From Cyropreserved Semen in One Season

Hopkins et al, 2012 Instrumental Insemination

• Semen Thawed • Straws fit directly into the insemination system • Each queen inseminated with 5 μL semen Benefits of Germplasm Repository • Simplify Stock Maintenance • Conservation of “Top Tier” Stocks and Threatened Subspecies & Ecotypes • Selection Across Time • Rapid Stock Improvements

Long Term Goal .. National Honey Bee Germplasm Repository

Conservation of Honey Bee Stocks

Honey bee spermatozoa Acknowledgements California Almond Board

California State Beekeepers Assoc.

National Honey Board

American Beekeeping Federation Foundation for the Preservation of Honey Bees

California Bee Breeders Association Garvey Cooperating Researchers Acknowledgements Steve Sheppard, WSU Brandon Hopkins, WSU Cooperating Queen Producers Judy Chen, USDA Beltsville, Md. Valerie Severson Michelle Flenniken, U of MT Pat Heitkem Marla Spivak, U of MN Jackie Burris-Park The CA. Tech Transfer Team Rick Schubert Glenda & Shannon Wooten Pat & Bonnie Stayer Frank & Sheri Pendel Tom & Suki Glenn Koehnen & Sons, Inc. Leonard Pankrast Buzz Landon

Special Thanks to

Elizabeth Frost Brian Fishback Tylan Selby

Visiting Scholars Dr. Roger Beleen, Universidade Federal de Alagoas, Brazil Dr. Jakub Gabka, Warsaw University , Poland Dr. John Pollard Dr. Claire Plante, GeneSafe Technologies, Canada

Kathy Garvey, Photography

Questions ?