Source: PENNSYLVANIA STATE UNIVERSITY submitted to
TRACKING FERAL HONEY BEE HEALTH: USING CITIZEN SCIENCE AND NEXT GENERATION SEQUENCING TO IMPROVE HONEY BEE RESISTANCE TO DISEASE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
ANIMAL HEALTH
Reporting Frequency
Annual
Accession No.
1011873
Grant No.
(N/A)
Project No.
PEN04620
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Dec 13, 2016
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Project Director
Lopez-Uribe, MA, MA.
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802
Performing Department
Entomology
Non Technical Summary
The beekeeping industry is facing serious challenges to maintain the necessary number of colonies to supply the demands for crop pollination. One of the major problems that honey bees have comes from the large cocktail of parasites and pathogens (bacteria, fungi and viruses) that attack this managed pollinator. Currently, most managed honey bee colonies cannot survive the winter without disease treatment, and even with intensive management regimes, beekeepers nationwide are losing on average 40% of their colonies. Some feral (unmanaged) bee populations have been stable through time despite the lack of beekeeper assistance, suggesting that these colonies have natural adaptations to deal with the multiple disease stressors that are currently killing large numbers of managed honey bee colonies. In this project, we propose to study the feral bee population of PA to identify locally-adapted genetic stocks that could be used for selection and breeding programs to generate disease-resistant stocks of bees. First, we will identify the location of feral honey bee colonies in PA though a citizen-science project. Second, we will use next generation sequencing (RNAseq) to characterize the levels of immunity and pathogen loads in feral and managed bees located in the same landscape context. Based on previous evidence, we predict feral colonies will show higher levels of immunocompetence and lower pathogen loads. By identifying the most resilient feral colonies--based on better overall health metrics--, we will provide key information about locally-adapted genetic pools suitable for breeding programs. We will work in partnership with a local queen rearing program to incorporate findings from this project into effective breeding of disease-resistant honey bees.
Animal Health Component
100%
Research Effort Categories
Basic
40%
Applied
60%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31131991130100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3199 - Invertebrates, general/other;

Field Of Science
1130 - Entomology and acarology;
Goals / Objectives
Objective 1: Work with a network of beekeepers in a citizen-science project to identify and track the survival of both feral honey bee colonies and nearby (within ~10 miles) managed colonies across Pennsylvania. Objective 2: Evaluate levels of immunocompetence and pathogen loads in feral and managed honey bee colonies.Objective 3: Work with the PA Queen Improvement Project to incorporate information about locally adapted genetic stocks into breeding programs.
Project Methods
* To map the distribution of feral honey bees in PA, we will establish a citizen science project that will engage beekeepers and the public into this project. Participants will provide information about where unmanaged honey bee coloniesare located. Feral colonies will be monitored throughout the duration of this project. We will also locate managed colonies within 10 miles of the feral bee colonies to compare the health status of bees that are located in the same landscapes.* To evaluatethe health status of feral and managed bees, we will use an RNAseq approach. RNA will be extracted from 30 individuals per colonies, samples will be pooled and sequenced in a MiSeq sequencer. Bioinformatically, we will identify and quantify expression of immune genes and viral pathogens.* To establish the queen rearingprogram, we will work with existing programs led by regional beekeepers. In addition, we will create an online course on queen rearing through the Penn Extension website.

Progress 12/13/16 to 09/30/19

Outputs
Target Audience:We have trained graduate and undergraduate students (3), collaborated with beekeepers across the state in this project (50), and reached out to the public by talking to the media about our project in multiple occasions (5). Since the beginning of this project, we have facilitated over 15 extension talks to a variety of audiences reaching out to over 850 people. At least 3 scientific and extension publications will be publically available in the next 6 months. Changes/Problems:No major problems with the methodology or data analysis were found. The only change to the methodology was the use of qPCR instead of RNAseq to characterize viruses and immune genes. What opportunities for training and professional development has the project provided? Katy Evans has led the field component of this project and has increased her abilities to do extension work with beekeepers. She will be a co-author in all the peer-reviewed and extension publications that come out of this project. Chauncy Hinshaw is an 'Integrated Pollinator Ecology' fellow (Ph.D. student) who joined the project this summer. He has been the lead author on all the research efforts. Ryan Ford is an undergraduate student who has helped with the RNA extractions and qPCR data collection of the project. After the research experience as part of this project, Ryan is currently seeking graduate school opportunities. Romina Russo is a visiting PhD student from Argentina who is working collaboratively with us on a comparative project about feral bees in the US and Argentina. How have the results been disseminated to communities of interest?We have used listservs, facebook, twitter, and articles in beekeeper newsletters to spread the word about the project. We have reached over 850 beekeepers through 15 talks over the course of the 3 years of this project. We will write an article for the Penn State Extension website and the results from this project will be disseminated through a magazine article in American Bee Journal. In addition, we will generate two peer-reviewed publications in the next 6 months. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1: During the duration of the project, we interacted with over 50 citizen scientists who reported the locations of ~60 feral colonies across Pennsylvania. In 2017, we sampled 16 colonies (8 feral and 8 managed) and, in 2018, we sampled a total of 55 colonies (30 feral and 25 managed) twice per year (Spring and Fall). We included colonies from 21 sites for the laboratory analyses with 2 colonies per site (1 managed and 1 feral), sampled twice per year (Spring and Fall) and 3 replicates per colony. Our results indicate that even though feral colonies received no miticide treatments and no supplemental feeding for the winter, ferals successfully overwinter at rates similar than managed colonies in Pennsylvania. Colony overwintering losses were recorded at 44% and 42% for managed and feral colonies (respectively), for the 2017-2018 winter, and 50% and 59% colony loss for managed and feral colonies, for the 2018-2019 winter. Objective 2: Our laboratory analyses if pathogens and immunocompetence included a total of 150 RNA samples (3 replicates from 50 colonies sampled 4 times). From each extraction, we analyzed a total of 10 genes for qPCR analysis (1 endogenous reference, 4 general immunity, 2 antiviral genes, 2 viruses andNosema ceranae). Reactions were run in triplicate, which translates into 216 qPCR reactions per site and a total of 7,560 individual reactions. We found that the expression of the immune genesHymenoptaecinandVagowere significantly correlated with overwintering success, while levels of Deformed wing virus were negatively correlated with survival. These results clearly characterize biomarkers that can be used for honey bee local breeding programs. Objective 3: We have already secured funding from the North American Pollinator Protection Campaign (NAPCC) and a Penn State Extension Multistate & Integrated Program Grant to sequence the genomes of these feral colonies. The next phase of this project is to find the genetic basis of the traits associated with the high overwintering success of feral colonies despite high levels of Deformed Wing Virus. The results from this molecular characterization of the immune genes of relevance will be linked to phenotyping tests that can be used by the PA Queen Improvement program for their breeding lines. In addition, we have a pending application for a USDA CARE grant in collaboration with colleagues from Purdue to test the performance of feral bee genetics in apiaries in Pennsylvania and Indiana.

Publications


    Progress 10/01/17 to 09/30/18

    Outputs
    Target Audience:We have trained graduate and undergraduate students (3), collaborated withbeekeepers across the state in this project (30), and reached out to the public by talking to the media about our project in multiple occasions (5). Since the beginningof this project, we have facilitated 11 extension talks to a variety of audiences. Scientific publications will come out of the lab before the end of the next reporting year. Changes/Problems:We did change our approach from RNAseq to qPCR data collection. No major problems with the methodology or data analysis was found this previous year. What opportunities for training and professional development has the project provided?• Katy Evans has led the field component of this project and has increased her abilities to do extension work with beekeepers. She will be a co-author in all the peer-reviewed and extension publications that come out of this project. • Chauncy Hinshaw is an 'Integrated Pollinator Ecology' fellow (Ph.D. student) who joined the project this summer. He was part of the Fall sampling and he has been working on the RNA extractions of all samples. • Ryan Ford is an undergraduate student who has helped with the RNA extractions and qPCR data collection of the project. How have the results been disseminated to communities of interest?We have used listservs, facebook, twitter, and articles in beekeeper newsletters to spread the word about the project. We have already talked about this project through three talks and we have scheduled other 3 talks about this project in the next 2 weeks. We will also write an article for the PSU Tree Fruit Newsletter this month. What do you plan to do during the next reporting period to accomplish the goals?We will focus on finalizing the data collection for our samples collected in 2018. We plan to continue with the citizen science project to keep a record of the feral colonies and their survival rates. We also will work on a grant proposal to continue working on this project.

    Impacts
    What was accomplished under these goals? We have effectively accomplished objectives 1 and 2 of this project. We interacted with 47 citizen scientists during 2018, and we recorded 50 new feral colonies in Pennsylvania. In 2018, we sampled a total of 55 colonies (30 feral and 25 managed) from 21 sites across the state in the Spring and in the Fall. We are including colonies from 21 sites for the laboratory analyses. This includes 2 colonies per site (1 managed and 1 feral) and 3 replicates per colony. This adds up to atotal of 204 RNA extractions. From each extraction, we have analyzed a total of 12 genes for qPCR analysis (1 endogenous reference, 4 general immunity, 2 antiviral genes, 4 viruses andNosema ceranae). Reactions are run in triplicate, which translates into 216 qPCR reactions per site and a total of 3,671 reactions for the spring sampling. We are currently working on the 204 RNA extractions and 3,671 qPCR reactions of the Fall 2018 samples. So far, our results have elucidated the presence of biomarkers that can be used for future honey bee local breeding programs. In collaboration with colleagues from Purdue, we are currently writing a proposal to continue working on this local honey bee breeding program. You can find updates of the project in this blog:https://lopezuribelab.com/2018/09/11/tracking-health-feral-bees-pa-2018/

    Publications

    • Type: Other Status: Published Year Published: 2018 Citation: Evans KC, L�pez-Uribe MM (2018) Tracking the Health of Feral Bees in Pennsylvania, preliminary results 2017. The Pennsylvania Beekeeper Newsletter. March Issue.


    Progress 12/13/16 to 09/30/17

    Outputs
    Target Audience:The targeted audience of this project is the beekeepers in Pennsylvania. This group of stakeholderscurrently struggles to maintain healthy colonies throughout the year. Historically, annual mortality rates were around 10% but currently, over 40% losses are the norm and beekeepers in different parts of the state even report 90% colony losses. Based on a survey we conducted at the PA State Beekeepers Association meeting in November 2016, we determined that beekeepers are interested in breeding locally adapted, disease resistant bee stocks (which requires identification and assessment of bee stocks and advanced beekeeping techniques such as queen rearing) and identifying new methods for controlling Varroa mites, the most deadly parasite of honey bees (uncontrolled Varroa infestation is the primary factor associated with colony losses in temperate environments). Through theprogram 'Tracking Feral Honey Bee Health: Using Citizen Science and Next Generation Sequencing to Improve Honey Bee Resistance to Disease', we established a citizen science project that we are using to gather information about the location of feral honey bee colonies throughout the state. Citizens report the location of feral colonies through this website:http://lopezuribelab.com/feral-honey-bees-pennsylvania/ We have disseminated information about this project to thousands of beekeepers across the state through our listserv, through talks to local beekeeping clubs, and through the newsletters of the PA State Beekeeper Association. Changes/Problems:We are considering changing the molecular approach from RNAseq to qPCR. Since we are not sure about how large are the differences in immune systems between these feral and managed bees, we will start by screening colonies with targeted qPCR primers for viruses and immune genes. After that preliminary dataset has been collected, we will decide if we can do RNAseq on these samples. What opportunities for training and professional development has the project provided? Katy Evans has led the field component of this project and has increased her abilities to do extension work with beekeepers. She will be a co-author in all the peer-reviewed and extension publications that come out of this project. Nolan Amon, the laboratory technician, has learned how to make libraries for next-generation sequencing, that could be applied to the molecular aspect of the project. Chauncy Hinshaw is an 'Integrated Pollinator Ecology' fellow (Ph.D. student) who joined the project this summer. He was part of the Fall sampling and he has been working on the RNA extractions of all samples. How have the results been disseminated to communities of interest?We have used listservs, facebook, twitter, and articles in beekeeper newsletters to spread the word about the project. We have already talked about this project through three talks and we have scheduled other 3 talks about this project in the next 2 weeks. We will also write an article for the PSU Tree Fruit Newsletter this month. What do you plan to do during the next reporting period to accomplish the goals?This Fall semester, we will focus on the laboratory analyses of the samples that we collected in 2017. Next year, we will have finalized the data collection of the sampling in 2017, and we will have completed the sampling for 2018.

    Impacts
    What was accomplished under these goals? We successfullydeveloped a citizen science project where people can report the locations of feral honey bee colonies in Pennsylvania:http://lopezuribelab.com/feral-honey-bees-pennsylvania/ We have 34 reports of feral colonies. We have engaged in conversations with all the citizens reporting the colonies. We collected 50 foragers from 19 feral and 12 managed colonies (within a 10-mile radius from the feral colony) during Spring and Fall of 2017. All samples were successfully collected in dry ice, brought to the Lopez-Uribe lab, and stored in a -80 freezer. RNA from these samples is being extracted at the moment. Our goal is to complete the extractions by the end of October 2017. We have continued communications with the PA Queen Improvement Program. We attended some of their workshops and have constant communications about the status of our project.

    Publications

    • Type: Other Status: Published Year Published: 2017 Citation: L�pez-Uribe MM. (2017) Tracking the Health of Feral Bees in Pennsylvania. The Pennsylvania Beekeeper Newsletter. February Issue.