Source: MONTANA STATE UNIVERSITY submitted to
NOSEMA CERANAE INFECTIONS AND HONEY BEE HEALTH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0213543
Grant No.
(N/A)
Project No.
MONB00020
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2007
Project End Date
Sep 30, 2009
Grant Year
(N/A)
Project Director
Cramer, R. A.
Recipient Organization
MONTANA STATE UNIVERSITY
(N/A)
BOZEMAN,MT 59717
Performing Department
Immunology & Infectious Diseases
Non Technical Summary
Honey bees are dying across the United State at unprecedented rates, a phenomenon termed Colony Collapse Disorder (CCD). Currently, no causal agent has been identified. However, recent data has identified a potentially new pathogen in honey bee colonies across the USA. This pathogen, Nosema ceranae, is a fungal-like organism that can cause mortality in honey bee colonies. In this proposal, we will examine the incidence of Nosema ceranae infections in honey bee colonies in Montana and possibly neighboring states by utilizing a molecular biology based technique, PCR, that allows quick and accurate diagnosis of infection. Our second objective is to understand how Nosema infections affect honey bee health by examining the immune response to Nosema infections in honey bees. One possible hypothesis is that Nosema infections severely compromise the honey bee's immune system, making them more susceptible to other pathogens and environmental stresses.
Animal Health Component
20%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2113010104040%
2113010109040%
2113010110220%
Goals / Objectives
Nosema ceranae infections are an emerging agricultural problem in honey bee health. While it is not definitive, a potential correlation between the appearance of N. ceranae infections and colony collapse disorder (CCD) has been hypothesized. In this proposal, we will explore two objectives. First, we will develop a polymerase chain reaction (PCR) based assay for the detection of Nosema infections in honey bees. This will allow us to assess the incidence of Nosema infections in honey bees in Montana and establish a possible correlation with CCD. Second, we will attempt to asses the honey bee immune response to Nosema infections utilizing DNA microarray technology. These studies will provide data on a possible correlation between Nosema infections and CCD.
Project Methods
For objective 1, honey bee samples from around the state of Montana will be provided by Dr. Jerry Bromenshenk at the University of Montana Missoula. DNA samples will be extracted from groups of honey bees from specific bee keeping operations and from hives within the individual operations. PCR primers based on the 16S rRNA gene that are capable of differentiating between N. ceranae and N. apis will be utilized to test for the presence of Nosema infections. Overall colony health data will be provided by Dr. Bromenshenk, and possible correlations with Nosema presence or absence and the health of the colony will be assessed. In objective 2, we will utilize the publicly available honey bee genome DNA microarrays from the University of Illinois. Honey bees will be infected with Nosema spores using published procedures. At defined time points total RNA will be extracted and processed with standard amino-allyl Cy-dye labeling techniques. Data analysis will be conducted with the TIGR microarray software suite and differentially expressed genes between uninfected and Nosema infected honey bees identified with Significance Analysis of Microarray (SAM) algorithm.

Progress 10/01/07 to 09/30/09

Outputs
OUTPUTS: Activities: The major completed activity of this project was the establishment of honeybee research infrastructure at Montana State University in Bozeman, Montana. In collaboration with Dr. Jerry Bromenshenk at the University of Montana-Missoula, and Dr. Kevin Wanner at MSU-Bozeman, we have established a small apiary near the MSU campus that will now facilitate honeybee research at MSU-Bozeman. Establishing these hives provided training opportunities for graduate students and laboratory technicians to become familiar with honeybee management practices. A graduate student and laboratory technician were hosted by Dr. Bromenshenk and his research group up in Missoula to learn how to establish and maintain these colonies. Dr. Bromenshenk and his research assistant spent a day training members of our laboratory in how to conduct cage-trials of honeybees at MSU. Thus, the project has contained important teaching, mentoring and collaborative activities important for establishing honey bee research at MSU-Bozeman. These interactions have facilitated the research progress involved with this project. Importantly, this project has allowed us to make contact with beekeepers across the state of Montana. Some are now actively helping in our honeybee research activities by providing samples and access to their operations. Honeybee colonies from across the Pacific Northwest and upper midwest regions have now been screened for Nosema infections completing objective 1 of this project. Events: Dr. Cramer attended and gave an oral presentation at the California State Beekeepers Association Conference in November 2008. Dr. Cramer participated in a workshop in Bozeman Montana at the Bioneers Conference in October entitled "Be Aware, Bee Happy, Be involved: What Bees are dealing with, and how it affects Montana" Services: Dr. Cramer has consulted with numerous beekeepers across the country and in Montana about potential alternative treatments for Nosema ceranae. Dr. Cramer's laboratory technician attended the 2009 Montana State Beekeepers convention and gave a presentation on our research progress. A graduate student and a laboratory technician were involved in teaching and mentoring activities at the University of Montana with Dr. Bromenshenk during the summer of 2009. Dr. Bromenshenk and colleagues also spent a day at MSU during the summer of 2009 helping setup cage trials. Products: We have provided diagnostic services to beekeepers across the country and in Montana for detection of bee pathogens by PCR and light microscopy. Dissemination: Invited lectures at the above conferences and publication of article on the topic of sporicidal compounds in the trade magazine Bee Culture. PARTICIPANTS: University of Montana-Missoula and Bee Alert Technologies - Dr. Jerry Bromenshenk and Scott Debnam. Montana State University - Veterinary Molecular Biology - Julie Elser, laboratory technician, Peggy Lehmann, technician, Joanna Gress, Ph.D. student, and Dr. Kevin Wanner Department of Plant Sciences MSU-Bozeman. TARGET AUDIENCES: The target audience for our project is beekeepers. PARTICIPANTS: Individuals: Dr. Robert Cramer Jr., Montana State University - Bozeman, MT (PI). Julie Elser, Laboratory Technician Cramer Laboratory MSU-Bozeman. Peggy Lehmann, Laboratory Technician Cramer Laboratory, MSU-Bozeman. Partner Organizations: University of Montana - Missoula, BeeAlert Technologies, Missoula, MT, Department of Plant Sciences Montana State University, Bozeman, MT. California State BeeKeepers Association (grant support in 2008). Collaborators: Dr. Jerry Bromenshenk, University of Montana, Missoula; Dr. Kevin Wanner, Department of PLant Sciences MSU-Bozeman, Joanna Gress, Ph.D. Student Plant Sciences MSU-Bozeman, Randy Oliver, Beekeeper California, Scott Debnam BeeAlert Technologies, Missoula, MT, Dave Baumbauer, Plant Sciences, MSU Bozeman. Training and Professional Development: Beekeeping practice training for graduate students and laboratory technicians, training in honeybee pathogen diagnostics, basic molecular biology training. TARGET AUDIENCES: Target Audience: Beekeepers Efforts: Oral Presentations at conference attended by researchers and beekeepers as outlined in Outcome portion of this report. PROJECT MODIFICATIONS: The major change with this project was that Objective 2 was not started due to timing and funding constraints. Thus, the major effort was focused on completing Objective 1 which was successfully completed. In addition, while not written into the original proposal as a specific objective, the overall objective of this proposal was to establish honeybee research infrastructure at MSU-Bozeman and this was successfully accomplished as evidenced by the established collaborations with other honeybee researchers, Montana and other Pacific Northwest beekeepers, and the development of a small apiary at MSU-Bozeman. These results will enable the completion of objective 2 and additional honeybee research at MSU-Bozeman in future years.

Impacts
Change in knowledge: The financial resources made available to us in this animal health project have resulted in greater awareness of the occurrence of Nosema ceranae infections in honeybees in the pacific northwest and have led to promising leads on potential compounds that may be used to disinfect honeybee combs to prevent the occurrence and spread of Nosema ceranae infections. Identification of other pathogens in combination with Nosema ceranae, such as viruses, have also given us a new understanding of the pathogen challenges faced by honeybees in Montana and the Pacific Northwest. Change in Action: Based on our research and the research of others, beekeepers are now more aware of the need to monitor their colonies for Nosema levels and levels of other important bee pathogens. Ideally, this knowledge will lead to a change in beekeeping practices to minimize the impact of these pathogens on overall honeybee health. Change in Conditions: Funds from this project enabled the development of a small apiary at Montana State University - Bozeman that will not greatly facilitate honeybee research on this campus. These funds have also enabled links with beekeepers across the state of Montana and with honeybee researchers at the University of Montana in Missoula.

Publications

  • No publications reported this period


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: Activities: In collaboration with the University of Montana-Missoula, honeybee colonies across the Pacific Northwest and upper midwest regions were screened for Nosema infections. In addition, numerous compounds were tested for their ability to kill Nosema ceranae spores in the laboratory Events: I attended the California State Beekeepers Association Conference in November 2008 to report our research findings. I participated in a workshop in Bozeman Montana at the Bioneers Conference in October entitled "Be Aware, Bee Happy, Be involved: What Bees are dealing with, and how it affects Montana" Services: I have consulted with numerous beekeepers across the country about potential alternative treatments for Nosema ceranae Dissemination: Invited lectures at the above conference and publication of article on the topic of sporicidal compounds in the trade magazine Bee Culture PARTICIPANTS: University of Montana-Missoula and Bee Alert Technologies - Dr. Jerry Bromenshenk and Scott Debnam. Montana State University - Veterinary Molecular Biology - Julie Elser, laboratory technician TARGET AUDIENCES: The target audience for our project is beekeepers. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Change in knowledge. The financial resources made available to us in this animal health project have resulted in greater awareness of the occurrence of Nosema ceranae in the pacific northwest and have led to promising leads on potential compounds that may be used to disinfect honeybee combs to prevent the occurrence and spread of Nosema ceranae. Importantly, we applied state of the art flow cytometry technique, not previously employed to examine Nosema spores, to determine the effect of various compounds on spore viability. These results should lead to a publication in 2009.

Publications

  • Cramer, R.A. 2008. Heating treatment kills Nosema spores. From The Speedy Bee, Vol. 37, No. 4
  • Cramer, R.A. 2009. Stopping Nosema. Bee Culture Magazine, January 2009