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Goals and Objective: Goal 1
Objective 1.4

Obj 1.4: Develop diagnostic tools for high-throughput detection and monitoring of honey bee diseases

(Evans, Chen, Aronstein)

Rationale and significance
Recent sharp declines in honey bee colonies worldwide are unusual in their severity, geographical distribution, and failure to present recognized symptoms of disease. Domesticated honey bees face numerous pests and pathogens, tempting hypotheses that colony collapses arise from exposure to new or resurgent pathogens.  Indeed early evidence has tied pathogen abundances to an increased risk of colony collapse (Cox-Foster et al., 2007, vanEngelsdorp et al., 2009). Recent studies have also revealed gaps in identification techniques that have led to the under-reporting of viruses (Cox-Foster et al., 2007, Chen and Evans, 2007, Blanchard et al., 2008) and Nosema species (Chen et al., 2007) that are important for bee health.

Due to the shifting distributions of pathogens, and changes over time in the DNA or RNA sequences upon which diagnostics are often based, it is important to refresh the toolkit of genetic diagnostics available for defining disease threats to honey bees. In addition, the ability of pathogens to cause disease and even collapse of honey bee colonies will depend on pathogen levels in individual bees (e.g., Yue et al., 2005) as well as incidence across colony members. It is therefore important to develop an efficient, low-cost method for precisely identifying the pathogens carried by sick and healthy bees, and for quantifying pathogen loads.

We propose an approach based on widely used real-time polymerase chain reaction protocols since 1) this platform provides an economic screening tool that can be used consistently at small or large genetic and diagnostic laboratories, 2) single-target reactions avoid issues related to the swamping of low-abundance targets by those having high abundance, 3) new screens for novel and resurgent pathogens can be added to diagnostic panels and compared with existing data and controls, and 4) extra labor and reagent costs of single-target screens, when compared to multiplex reactions, are small relative to costs of sample collection, processing, and analysis. 

Expected outcomes
Improving the toolkit for disease diagnostics in honey bees will result in

1) a better understanding of field correlations between disease agents and colony decline,

2) a method for assessing novel and established treatments on bee health and pathogen levels, and

3) a streamlined screening system that can provide a warning system for the arrival or threat of pathogens due to bee transport. 

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Summary Statement for Goal 1
This goal constitutes our attempts at understanding the most important morbidity factors at work in North American Apis mellifera. Work in this Goal is characterized by a high degree of interinstitutional linkages within CAP labs, resulting in four topical groups. The Nosema group is comprised of Lee Solter (Univ IL), Tom Webster (KY State Univ), Zach Huang (MI State), Christina Grozinger (Penn State), and Kate Aronstein (ARS Weslaco). The virus group is made up of Jay Evans and Judy Chen (ARS Beltsville) and Lee Solter. There have been cross-group linkages with Greg Hunt (Purdue) who is studying the genetic basis of bee resistance to N. ceranae and Israeli Acute Paralysis Virus (IAPV). A diagnostics group is comprised of Jay Evans, Judy Chen, and Kate Aronstein. The toxicology group is comprised of Marion Ellis (Univ NB), Maryann Frazier, Jim Frazier, and Chris Mullin (Penn State). A sentinel apiary monitoring group, led by Frank Drummond (Univ. of Maine), is comprised of Nancy Ostiguy (Penn State), Marla Spivak (Univ. of Minn.), Kate Aronstein (ARS Weslaco), Sheppard (Univ. of Wash.), Kirk Visscher (Univ. of CA - Riverside); analytic work by Anne Averill (Univ. of Mass.), Nancy Ostiguy (Penn State), and Brian Eitzer (CT Experiment Station) is collecting baseline data on field colonies and factors contributing to bee morbidity. And finally, an IPM adoption group is headed up by Keith Delaplane (Univ GA).

Progress

Methodology, data and analysis of results to date are shared in an annual report to USDA. Papers generated by team members during the time of the CAP are listed and periodically updated below. Beyond the citation of published papers, the consensus of the group is that it would otherwise be unhelpful or possibly misleading to state preliminary results within the context on this web site.

Future steps
Currently, we are pursuing different leads to transfer technology to private industry to develop a new diagnostic tool based on an antigen capture assay that detects Nosema in field honey bee samples. (1) A proposal has been submitted to the USDA Office of Technology Transfer “Agricultural Technology Innovation Partnership” program (ATIP) to enhance the opportunity for private sector partnerships for the development of a Nosema serological test; (2) The proposal has also been presented at the joint ATIP-USDA meeting in fall of 2010. A manuscript describing this research has been published in Journal of Apicultural Research.

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Publications of objective 1.4 principal investigators (Aronstein, Chen and Evans) to date during the CAP

Aronstein K.A, and J. Adamczyk. 2011. Influence of Genomics: The Post Genomic Era in the Honey Bee Research. The Journal of the Texas Beekeepers Association. 11(1): 12-17

Aronstein, K.A., H.E. Cabanillas. (ed. Samataro) Book: ”Honey Bee Colony Health: Challenges and Sustainable solutions” Book chapter 11: Chalkbrood Re-examined . Taylor and Francis, LLC, (accepted, 2011)

Aronstein, K.A. 2009. Detect Nosema Parasite in Time to Save Bee Colonies. Am. Bee J.150 (1): 63-65

Aronstein, K.A., Eduardo Saldivar, T.C. Webster. 2011. Evaluation of Nosema ceranae Spore-specific polyclonal antibodies. Journal of Apicultural Research 50(2): 145-151

Aronstein, K. A., B. Oppert, and M.D. Lorenzen. (ed. Paula Grabowski) 2011. Book: “RNA Processing,” Chapter “RNAi in the Agriculturally Important Arthropods.” ISBN 978-953-307-332-3, InTech.

Boncristiani, H., J.L. Li, J.D. Evans, and Y.P. Chen. 2011. Scientific note on PCR inhibitors in the compound eyes of honey bees, Apis mellifera, Apidologie (in press)

Chen, Y.P. and Z.Y. Huang. 2010. Nosema ceranae, a newly identified pathogen of Apis mellifera in the U.S. and Asia. Apidologie 41: 364-374, DOI: 10.1051/apido/2010021

Chen, Y.P., J.D. Evans, and J.S. Pettis. 2011. The presence of chronic bee paralysis virus
infection in honey bees (Apis mellifera L.) in the USA. J. Apic. Res.,50:85-86

Cornman, R. S., M.C. Schatz, J.S. Johnston, Y.P. Chen, J.S. Pettis, G. Hunt; B. Lanie, C. Elsik, D. Anderson, C.M. Grozinger, and J.D. Evans. 2010. Genomic survey of the ectoparasitic mite Varroa destructor, a major pest of the honey bee Apis mellifera, BMC Genomics 11:602

Dainat, B.D., J.D. Evans, Y.P. Chen, P. Neumann. 2011. Sampling and RNA quality for
diagnosis of honey bee viruses using quantitative PCR, Journal of Virological
Methods, (in press)

Di Prisco, G., F. Pennacchio, C. Emilio, H. Boncristiani, J.D. Evans, Y.P. Chen. 2011. Varroa destructor is an effective vector of Israeli Acute Paralysis Virus in honey bees, Apis mellifera, J. General Virology, 92: 151-155

Eitzer, B., F. Drummond, J.D. Ellis, N. Ostiguy, K. Aronstein, W.S. Sheppard, K. Visscher, D. Cox-Foster, & A. Averill. 2010. Pesticide analysis at the stationary apiaries, American Bee Journal, 150(5):500

Ellis, J., J.D. Evans, J.S. Pettis. 2010. Colony losses, managed colony population decline,
and Colony Collapse Disorder in the United States, J. Apicultural Res, 49(1) 134- 136

Evans, JD. 2006. Beepath: An ordered quantitative-PCR array for exploring honey bee immunity and disease, Journal of Invertebrate Pathology, 93 (2): 135-139

Evans, J.D., M. Spivak. 2010. Socialized Medicine: Individual and communal disease
barriers in honey bees, Journal Invert. Pathol. 103, S62-S72

Genersch, E, J.D. Evans, I. Fries. 2010. Honey bee disease overview, Journal Invert.
Pathol. 103, S2-S4

Holt, H.L., K.A. Aronstein, and C.M. Grozinger.  Genomic analysis of effects of Nosema parasites on honey bee workers (in prep.)

Johnson, R.M., J.D. Evans, G.E. Robinson, M.R. Berenbaum. 2009. Changes in transcript abundance relating to colony collapse disorder in honey bees (Apis mellifera). Proceedings of the National Academy of Sciences of the United States of America 106:14790-14795

Li, J.L., W.J. Peng, J. Wu,  H. Boncristiani, J.P. Strange, and Y.P. Chen. 2011. Cross-species Infection of Deformed Wing Virus Poses a New Threat to Pollinator Conservation.  J. Econ. Entomol.  104(3): 732-739

Paldi, N., E. Glick, M. Oliva, Y. Zilberberg, L. Aubin, J.S. Pettis, Y.P. Chen, J.D. Evans. 2010. Effective gene silencing of a microsporidian parasite associated with honey bee (Apis mellifera) colony declines, Applied Environmental Microbiology, 76:5960-5964

Peng, W.J., J.L. Li, J. Wu, H. Boncristiani, J.P. Strange, and Y.P. Chen. 2011. Host Range Expansion of Honey Bee Black Queen Cell Virus in the Bumble Bee, Bombus huntii. Apidologie (in press).

vanEngelsdorp D, J.D. Evans, C. Saegerman, et al. 2009. Colony collapse disorder: a
descriptive study. PLoS ONE 4 (8)

vanEngelsdorp,  D., N. Speybroeck, J.D. Evans, B.K. Nguyen, C. Mullin, M. Frazier, J. Frazier, D. Cox-Foster, Y.P. Chen, D.R. Tarpy, E. Haubruge, J.S. Pettis, C. Saegerman. 2010.  Identification of risk factors associated with bee Colony Collapse Disorder by classification and regression tree analysis. J. Econ Entomol, 103: 1517-1523

Webster, T and K.A. Aronstein. Nosema ceranae Detection by Microscopy and Antibody
Tests. (ed. Samataro) Honey bee Colony Health: Challenges and Sustainable solutions (ed. Diana Samataro): Book chapter 10: Taylor and Francis, LLC. (accepted, 2011)

Further Background Information
Documentation of CAP progress in general, and of this objective in particular, is available through the following sources:

  1. Bee Health, an eXention initiative for peer-reviewed scientific recommendations
  2. Colony Collapse Disorder Progress Report for 2009
  3. Genetic Toolkits for Bee Health

Updated July 22, 2011.

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