Progress 10/01/08 to 09/30/12
Outputs
OUTPUTS: Poster: Int. Symbiosis Society 2009. The Presence of Actinobacteria within Honey Bee (Apis mellifera) Hives and their Floral Food Sources: Implications for a New Symbiosis. PARTICIPANTS: K. Grubbs, PhD student; C. Currie PI <p> </p> Poster: Perlman Symposium on Antibiotic Discovery and Development 2010. An Actinobacterium from Apis mellifera Hives Produces an Antibiotic that Specifically Inhibits American Foulbrood PARTICIPANTS: K. Grubbs, PhD student; C. Currie PI <p> </p> Poster: Arthropod Genomics Symposium 2010: The genome sequence of the leaf-cutter ant Atta cephalotes. PARTICIPANTS: K. Grubbs, PhD student; C. Currie PI <p> </p> Oral presentation: Arthropod Genomics Symposium 2011. Transcriptional response to manipulation of fungal symbiosis in the leaf-cutter ant Atta cephalotes. PARTICIPANTS: K. Grubbs, PhD student; C. Currie PI <p> </p> Oral presentation: Entomological Society of America 2012. Component wise variance in community profiles of Apis mellifera hives. PARTICIPANTS: Kirk Grubbs, PhD student; C. Currie PI <p> </p> A couple hundred strains of antibiotic-producing Actinobacteria have been isolated from both honey bee hives and some flowers that are pollinated by honey bees. Further exploration of one of these isolates yielded the novel compound Apinimycin. PARTICIPANTS: Kirk Grubbs, PhD student; Cameron Currie PI <p> </p> Kirk Grubbs, who was supported by this Hatch, helped analyze the draft genome for the leaf-cutter ant Atta cephalotes. This provided the training necessary for Kirk to work on the honeybee microbiome. PARTICIPANTS: K. Grubbs <p> </p> We generated a membrane lipid profiling of the microbial community from honeybee colonies. PARTICIPANTS: K. Grubbs <p> </p> We conducted an experiment to look at the effect chemical pesticides have on the microbiota of honeybees. Six colonies were selected, 3 receiving Chlorothalonil (a fungicide used in WI), and 16S rRNA pyrotag sequencing of the microbiota was conducted. This work provides insights into the general microbiota of honeybee colonies, as well as the impact of a fungicide on this community. PARTICIPANTS: K. Grubbs <p> </p> Three students have received training. PhD candidate Kirk Grubbs has received training in microbiology, entomology, and molecular ecology. Undergraduate Robert Dugenske was trained in basic microbiology and phylogenetic methods (2009-2012). James Estevez received training in microbiology and entomology working as an undergraduate assistant (2010). <p> </p> Summary: One graduate and two undergraduate students received training in microbiology and symbiosis. A novel antibiotic was discovered from an Actinobacterium isolated from a honeybee colony. This antibiotic is highly active against the most significant bacterial disease of honeybees, American Foulbrood (AFB). Thus, it has the potential to be a used to help treat AFB by apiculturists. PARTICIPANTS: PhD candidate Kirk Grubbs has received training in microbiology, entomology, and molecular ecology. He conducted research, mentored undergrad students, conducted fieldwork, and performed pyrotag sequencing. Undergraduate Robert Dugenske and James Estevez were trained in basic microbiology and phylogenetic methods. They made microbiological media, helped maintain honeybee colonies, helped conduct DNA isolation and Sangar sequencing of isolates PI Cameron Currie oversaw progress of Kirk Grubbs' graduate training and mentoring. TARGET AUDIENCES: Genomics community, Insect genomics community, Apiculturists, Symbiosis researchers, Entomological Society of America, Int. Symb. Society. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We conducted the first lipid based (fatty acid methyl ester and phospholipid-derived fatty acid) microbial community characterization of honeybees and their hive components (i.e., bees, comb, propolis, honey, and stored pollen). Given that the different components within hives can be physically separated and are nutritionally variable, in this work we explored the hypothesis that unique microbial communities occur within the different microenvironments of honey bee colonies. Overall microbial community richness was found to vary from lowest to highest in honey, comb, pupae, pollen, adults and propolis, respectively. Finally, microbial community lipid profiles were more similar when compared by component than by hive, location or sampling year. Specifically, we found that individual hive components typically exhibited several dominant lipids and that these dominant lipids differ between components. As part of this project, we conducted extensive isolation for the presence of Actinobacteria inside the hives of honeybee colonies. We isolated X colony forming units of Actinobacteria from honey bees and their hive components. Specifically, the majority of these strains belong in the genus Streptomyces, most commonly from pollen and adult bees. Together these findings suggest that Actinobacteria are maintained within the hive and that at least some of the strains are acquired during pollen foraging. One specific strain was found to produce a novel secondary metabolite, Apinimycin, which has antibiotic activity specific to the common hive pathogen, Paenibacillus larvae (American Foulbrood). To characterize the microbial community (microbiome) of honeybees we have conducted 16S rRNA pyrosequencing. Further, we explored the impact of a common crop pesticide (Chlorothalonil) on the normal microbial community associated with hive. Furthermore, this study will provide further detail on the microbial profiles of individual hive components. In this work, we have gained valuable insights into the microbiology of honeybees. Some of the main findings we have generated are as follows. First, we have shown that the community of bacteria is most diverse in adult bees, and least diverse in pupae. Second, we have shown that the community of microbes undergoes significant shifts over time. We have found that the community is dominated by Proteobacteria and Firmicutes, as well as Bacteriodetes and Actinobacteria. We have also found a shift in the microbial community associated with hives based on our Chlorothalonil treatment. <p> </p> Summary: Through pyrosequencing and lipid-based microbial community profiling we have characterized the microbial community associated with honeybees and their hive components. This includes showing that distinct hive components (e.g., brood, pollen, workers, etc.) appear to have specialized and distinct communities of microbes. Finally, we have discovered one novel antibiotic from a strain of Actinobacteria associated with hives and shown that this compound has high activity against the major bacterial pathogen of honeybees.
Publications
- Kirk J. Grubbs, Peter H.W. Biedermann, Garret Suen, Sandra M. Adams, Joseph A. Moeller, Jonathan L. Klassen, Lynne A. Goodwin, Tanja Woyke, A. Christine Munk, David Bruce, Chris Detter, Roxanne Tapia, Cliff S. Han and Cameron R. Currie. The Complete Genome Sequence of Streptomyces cf. griseus (XylebKG-1), an Ambrosia Beetle-Associated Actinomycete. J. Bact. 2011; Published ahead of print 1 April 2011, doi: 10.1128/ JB.00330-11
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: We have isolated bacteria from colonies of honeybees, specifically actinobacteria in the genus Streptomyces. We have also conducted membrane lipid profiling of the microbial community from honeybee colonies. Finally, Kirk Grubbs who is supported by this Hatch helped with analyses of the draft genome for Atta cephalotes. PARTICIPANTS: Kirk Grubbs TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We have found the presence of antibiotic producing bacteria inside the hives of honeybee colonies. From one of these colonies, we have discovered a novel small molecule (antibiotic), which has high activity against a bacteria pathogen of honeybees. Further, our lipid analysis suggestion that different hive components have distinct microbial communities associated with them. We have also generated a draft genome for Atta cephalotes. Analysis of this genome suggests genetic modifications that reflects its obligate dependence on the fungal cultivar for nutrients.
Publications
- Suen, G., Teiling, C., Li, L., Holt, C., Abouheif, E., Bornberg-Bauer, E., Bouffard, P., Caldera, E.J., Cash, E., Cavanaugh, A., Denas, O., Elhaik, E., Fave, M-J., Gadau, J., Gibson, J.D., Graur, D., Grubbs, K.J., Hagen, D.E., Helmkampf, M., Hu, H., Johnson, B.R., Kim, J., Marsh, S.E., Moeller, J.A., Munoz-Torres, M.C., Murphy, M.C., Naughton, M.C., Nigam, S., Overson, R., Rajakumar, R., Reese, J.T., Scott, J.J., Smith, C.R., Tao, S., Tsutsui, N.D., Viljakainen, L., Wissler, L., Yandell, M.D., Zimmer, F., Harkins, T.T., Taylor, J., Slater, S.C., Clifton, S.W., Warren, W.C., Elsik, C.G., Smith, C.D., Weinstock, G.M., Gerardo, N.M. & Currie, C.R. 2010. (in press) The Genome Sequence of the Leaf-cutter Ant Atta cephalotes Reveals Insights into its Obligate Symbiotic Lifestyle. Public Library of Science Genetics.
- Smith, C.R., Smith, C.D., Robertson, H.M., Helmkampf, M., Zimin, A., Yandell, M., Holt, C., Hu, H., Abouheif, E., Benton, R., Cash, E., Croset, V., Currie, C.R., Elhaik, E., Elsik, C.G., Fave, M.J., Ferandes, V., Gibson, J.D., Graur, D., Gronenber, W., Grubbs, K.J., Hagen, D., Vinniegra, A.S.I., Johnson, B.R., Johnson, R., Khila, A., Kim, J.W., Mathis, K.A., Munoz-Torres, M.C., Murphy, M.C., Mustard, J.A., Nakamura, R., Neihuis, O., Nigham, S., Overson, R., Placek, J., Rajakumar, R., Resse, J.T., Suen, G., Shu, T., Torres, C.W., Tsutsui, N.D., Viljakainen, L., Wolschin, F. and J. Gadau. 2010. A Draft Genome of the Red Harvester Ant, Pogonomyrmex barbatus: a Model for Reproductive Division of Labor and Social Complexity. Proceedings of the National Academy of Sciences USA. (in press).
- Smith, C.D., Abouheif, E., Benton, R., Croset, V., Currie, C.R., Elhaik, E., Elsik, C.G., Fave, M.J, Fernandes, V., Gadau, J., Gibson, J.D., Graur, D., Hagen, D.E., Helmkampf, M., Holt, C., Hu, H., Johnson, B.R., Johnson, R.M., Abderrahman, K., Kim, J.K., Mathis, K.A., Munoz-Torres, M.C., Murphy, M.C., Nakamura, R., Nigam, S., Overson, R., Placek, J., Rajakumar, R., Reese, J.T., Robertson, H.M., Smith, C.R., Suen, G., Tao, S., Torres, C.W., van Wilgenburg, E., Viljakainen, L., Vinniegra, A.S. I., Walden, K.K.O., Yandell, M.D., Zimin, A. and N.D. Tsutsui. 2010. The Genome of the Globally Widespread and Invasive Argentine Ant (Linepithema humile). Proceedings of the National Academy of Sciences USA. (in press).
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