2013 Annual Report Project Narratives

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Lintilhac Foundation
Reproductive Differentiation in Plants: A Structural and Mathematical Analysis .

Principal Investigator: J. Barrington

Accomplishments & Outputs:
This basic research is an extension of previous work on the factors governing the transition from vegetative growth to reproductive growth in plants. The purpose of the study is to isolate the factors specifically responsible for the differentiation of the first reproductive cells in plants. This is a problem unique to plants where there is no reserved-germ-line as there is in animals. We will develop new methods for the study of plant cells grown in culture under conditions where we can manipulate the physical environment of growing cells in order to alter their developmental fate. Previous work in this lab has focused on the biophysics of cell enlargement and its role in the control of morphogenesis. Our most recent contributions include a new paradigm for plant cell wall stress relaxation, which has fostered a new way of modeling all plant growth.

Publications:
Lintilhac, P.M. (2013). The problem of morphogenesis: Unscripted biophysical control systems in plants. Protoplasma, DOI 10.1007/s00709-013-0522-y.

Lintilhac, P.M. (2012). Plant Physics by Karl J. Niklas and Hanns-Christof Spatz. A review in: Am. J. Phys. 80:842

1

Hatch
Enhancing farmers capacity to produce high quality organic bread wheat in Vermont.

Principal Investigator: H. Darby

Accomplishments & Outputs:
The demand for local organic food continues to rise throughout Vermont and the Northeast. Vermont millers and bakers cannot find enough locally grown organic wheat to meet their increasing demand, and that which is available often does not meet the higher quality standards for bread production. Our overall project goal was to enhance the capacity of farmers to produce high quality organic wheat through the following objectives: Objective 1: Develop viable organic bread wheat production strategies for Vermont farmers by addressing key production constraints. Sub-objective 1a: Determine the impact of spring wheat cultivar selection and planting date on weed biomass, Fusarium Head Blight (FHB) incidence and grain yield and quality. Sub-objective 1b: Determine the impact of winter wheat cultivar selection and harvest timing on grain yields and end-use quality. Through this project we determined that mid to late April planting dates produced significantly higher yields compared to mid to end of May planting dates. We also identified that varieties with taller stature competed better with weeds. Weed biomass was significantly higher as planting dates became later. Therefore taller varieties should be planted especially if planting dates must be delayed past the optimum time. We found that harvest dates also influenced yields. Highest yields were obtained at wheat physiological maturity (30% moisture). However, a commercial combine is unable to harvest wheat at this high moisture. Interestingly as wheat harvest was delayed the yield and test weight declined. Therefore it was determined that best yields could be obtained when wheat was harvested as soon as it reaches acceptable moisture for a combine. Vomitoxin (DON) levels appeared to be more influenced by climatic conditions during the growing season rather than specific planting or harvest dates. However, more research needs to be done to confirm our results. Research results were summarized and posted online via the UVM Extension Crops and Soils website (www.uvm.edu/extension/cropsoil) and the Northern Grain Growers Association website (www.northerngraingrowers.org). In addition hard copies of these reports were printed and disseminated at our on-farm field days, farm shows, and conferences. A field day held in Alburgh, VT highlighted the research trials and disseminated results to over 250 stakeholders. Research results were also presented to an audience of 125 at the Northern Grain Growers Annual Conference.

Publications:
Darby, H. and Cummings, E. 2012. Time to Plant Spring Grains! Agriview Volume 76, Number 6. Available online: http://www.vermontagriculture.com/Agriview/2012/4-13-2012Agriview.pdf

Darby, H., Cummings, E., Harwood, H., Madden, R., and Monahan, S. 2013. 2012 Organic Spring Wheat Planting Date Trial Report. Available online: www.uvm.edu/extension/cropsoil and www.northerngraingrowers.org

Darby, H. and Cummings, E. 2013. Time to Start Thinking Spring Grains! Northern Grain Growers Association Newsletter, Winter 2013 Edition.

1

Hatch
Best Agronomic Practices for Oilseed Production in Vermont .

Principal Investigator: H. Darby

Accomplishments & Outputs:
This project sought to provide technical assistance for oilseed crops in the region, helping established growers and facilitating adoption of a new crop for prospective growers. Goals included development of best management practices for growing oilseed crops in our region. As a result of this project, growers can now access regionally-specific agronomic research on varietal selection, planting dates, seeding rates, weed control, and fertility management of oilseeds. In 2012, research results were consolidated and reported to the public via UVM Extension Northwest Crops and Soils website and printed reports. Five sunflower planting dates were assessed in 2012 (18-May, 25-May, 1-Jun, 8-Jun, and 15-Jun). Bird damage was significantly lower in later planted sunflowers. There were no significant differences in seed-boring insect damage by planting date. The highest seed and oil yields were obtained when sunflowers in planted in mid-June. Four winter canola planting dates between the end of August and the third week of September were evaluated. There was significantly better winter survival and greater seed and oil yields in early-planted winter canola planted earliest. All 2012-2013 winter canola planting date trials failed due to winter desiccation. However, canola planted 29-Aug was more successful at overwintering than canola planted earlier or later. Five sunflower seeding rates were examined in 2012. There was no significant difference in white mold (Sclerotinia sclerotium) incidence or bird damage by seeding rate. Plants at higher seeding rates were significantly taller, with narrower heads. Seed yields were highest in sunflowers planted at low seeding rates (20,000 seeds per acre). The manual Oilseed Production in the Northeast: A Guide for Growers of Sunflower and Canola was published in March, 2013. Results from the previous three years of research were compiled and used to provide agronomic guidelines to growers on crop production, pest management, and best harvesting and processing strategies. The manual was distributed at the annual Oilseed Producers Meeting in March, as well as being posted online. Project reports and summaries were distributed as hard copies at outreach events in 2013. An hour-long oilseeds workshop at the annual NOFA-VT Conference on 16-Feb attracted about two dozen attendees (mostly potential growers). On 19-Mar, the annual Oilseed Producers Meeting brought 26 growers together to discuss animal nutrition, pest management, small-scale equipment, culinary oil production, and agronomic management of oilseed crops. A field day at Borderview Research Farm on 1-Aug hosted 173 people (including 75 farmers). At this event, tours of the research plots included an emphasis on sunflower planting dates, bird management, interseeding and reduced tillage, and ongoing insect monitoring projects. Future research will focus on evaluating winter canola and practices that contribute to successful overwintering. With an increasing interest in local food production, next steps in research will assess post harvest practices that lead to high quality culinary oil.

Publications:
Darby, H. P. Halteman, H. Harwood. March 2013. Oilseed production in the northeast: A guide for growers of sunflower and canola. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/OilseedManualFINAL.pdf (verified 29 October 2013).

Darby, H., H. Harwood, E. Cummings, R. Madden, S. Monahan. February 2013. 2012 Winter canola planting date trial. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/2012WinterCanolaPD-final.pdf (verified 29 October 2013).

Darby, H., H. Harwood, E. Cummings, R. Madden, S. Monahan. March 2013. 2012 Sunflower planting date trial. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/2012-Sunflower-Planting-Date.pdf (verified 29 October 2013).

Darby, H., H. Harwood, E. Cummings, R. Madden, S. Monahan. March 2013. 2012 Sunflower population and nitrogen rate trial. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/2012-Sunflower-POPxNR.pdf (verified 29 October 2013).

Darby, H., H. Harwood, E. Cummings, S. Monahan. July 2013. 2012-2013 Winter canola survival. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/2012-2013-Winter-Canola-Survival.pdf (verified 29 October 2013).

Harwood, H., H. Darby. February 2013. 2012 National sunflower survey. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/2012NationalSunflowerSurvey_summaryfinal.pdf (verified 29 October 2013).

Harwood, H., H. Darby. January 2013. Impacts of population and nitrogen application rate on stand characteristics and yields of sunflower. Poster presented at the annual research symposium at the National Sunflower Association, Fargo, ND.

Harwood, H., S. Lewins, H. Darby. October 2013. Sunflower insects: Scouting and identification. Univ. of Vermont Ext. report. Burlington, VT. Available at: http://www.uvm.edu/extension/cropsoil/wp-content/uploads/SunflowerInsects.pdf (verified 29 October 2013).

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Hatch
Assessing the Potential for Carbon Neutral Food Systems for Chittenden County, Vermont .

Principal Investigator: J. Farley

Accomplishments & Outputs:
Society must increase agriculture production to feed a growing population while reducing agriculture's global environmental impact or risk potentially catastrophic consequences. The goal of this project is to model a food system compatible with an 80% reduction in net greenhouse gas emissions, with a case study of Burlington VT. The grant funded a masters thesis (Farr, 2013) that estimated social and ecological boundaries for the food system, and assessed policies for remaining within them. The study found that with current agricultural technologies it would not be possible to feed 7 billion people without exceeding planetary boundaries for nitrogen, greenhouse gasses, and biodiversity loss, particularly in an economic system that allocates food based on preferences weighted by purchasing power rather than needs. A closely related journal article is currently under revision for Ecological Economics, and research results contributed to 3 conference presentations and 5 plenary or keynote addresses. This coming year we will develop a Bayesian neural network model of carbon footprints and food production from different agroecosystems to determine the minimum level of greenhouse gas emissions compatible with meeting the nutritional requirements for Burlington, Vermont.

Publications:
De Groot, D., J. Aronson, J. Blignaut, T. Elmquist, J. Farley (2013) Evidence from the Field Shows that Investing in Ecosystem Restoration Pays. Conservation Biology. DOI number (10.111/cobi.12158)

Muradian, R., M. Arsel, L. Pellegrini, F. Adaman, B. Aguilar, B. Agarwal, E. Corbera, D. Ezzine-de Blas, J. Farley, G. Froger, E. Garcia-Frapolli, E. Gomez, J. Gowdy, N. Kosoy, J. F. Le Coq, P. Leroy, P. May, P. Meral, P. Mibielli and R. Norgaard (2013). Markets for biodiversity and ecosystem services and the fatal attraction of win-win solutions. Conservation Letters 6(4): 274-279.

Alvez, J., A. Schmitt F., J. Farley, G. Alarcon, A. Fantini (2012) The Potential for Agroecosystems to Restore Ecological Corridors and Sustain Farmer Livelihoods: The Evidence From Brazil. Ecological Restoration. 30(4) 288-290.

Costanza, R., G. Alperovitz, H. Daly, J. Farley, C. Franco, T. Jackson, I. Kubiszewski, J. Schor, P. Victor and R. Colman (2013) Vivement 2050 ! Programme Pour Une Economie Soutenable Et Desirable (English title: Building a Sustainable and Desirable Economy-in-Society-in-Nature). Translated by Valerie Denot. Institut Veblen pour les reformes economiques: Paris

Farley, J. and S. Perkins (2013) Economics of Information in a Green Economy. In R. Robertson (ed.) Building a Green Economy. Michigan State University Press, Lansing pp.

Farley, J. (2013) The Economics of Sustainability in U. Diwekar and H. Cabezas (ed.) Sustainability: Multi-Disciplinary Perspectives. Bentham Open E-books.

Costanza, R., G. Alperovitz, H. Daly, J. Farley, C. Franco, T. Jackson, I. Kubiszewski, J. Schor and P. Victor (2013). Building a Sustainable and Desirable Economy-in-Society-in-Nature. State of the World. W. W. staff. Washington, DC, Island Press pp. 126-142

Schmitt Filho, A., J. Farley, G. Alarcon, J. Alvez and P. Rebollar (2013). Integrating Agroecology and PES in Santa Catarina's Atlantic Forest. R. Muradian and L. Rival (eds.) Governing the provision of environmental services. Springer Verlag pp. 333-356

Richnau, G., P. Angelstam, S. Valasiuk, L. Zahvoyska, R. Axelsson, M. Elbakidze, J. Farley, I. Jonsson, I. Soloviy. (2013), Multifaceted Value Profiles of Forest Owner Categories in South Sweden: The River Helge a Catchment as a Case Study. AMBIO: A Journal of the Human Environment 42(2): 188-200.

Farley, J. (2013) Economia para um Planeta Pos-carbono. Revista Nucleo de Estudos de Economia Catarinense. 1 (2) pp. 52-68

Farley, J. and T. Farley (2013) The Economics of Health Care. Solutions. 4 (3) 48

1

Hatch
Climate Change Best Management Practices on Vermont Farms .

Principal Investigator: S. Hurley

Accomplishments & Outputs:
This project seeks to better understand land management by Vermont farmers in the context of climate change adaptation and mitigation strategies and implementations, i.e. climate change best management practices, or CCBMPs. Our transdisciplinary approach integrates methodologies including agroecological analysis, greenhouse gas sampling, and alternative futures scenario visualization. This year we have interfaced with a related project funded by UVM's Food Systems Research Spire to create a final list of CCBMPs (n=20), many of which are explored in the context of this research project. We have made connections with several farms at which we will sample greenhouse gases, starting in Fall 2013, throughout the seasons for the next two years. We have also photo-documented nearly a dozen different farms who employ (or may in the future choose to employ) the identified CCBMPs, created five sets of photosimulations illustrating existing conditions and proposed conditions with CCBMPs in place, and we have developed a visual library for creating additional photosimulations in the near future. We have participated in two field days with farmers and planned outreach workshops for the coming year,

Publications:
none

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Department of Agriculture USDA
Applied Master's Degree in Food Systems .

Principal Investigator: J. Kolodinsky

Accomplishments & Outputs:
The development of a transdisciplinary graduate degree in Food Systems was possible through support of a USDA Higher Education Challenge grant. Food Systems Program Bylaws govern the Food Systems program and state that in the program, students learn about food systems through a transdisciplinary curriculum. The program bridges social science, humanities, and natural science approaches to understanding complex and interdependent food systems of varying scope and scale. The University of Vermont welcomed its first cohort of students in the fall of 2012. The FS MS includes two possible tracks: 1) the professional track, a one year program, during which the students work with a Faculty Advisor to design a final project based on applied skills and desired interests or 2) the research track, a two year program, during which the students work with a Faculty Advisor to complete a year-long research-based project, while continuing to take courses during the second year. The FS MS Program solicited applications for its first cohort in the spring of 2012. About ten applications were received and five students started the program in the fall of 2012. The five entering students had undergraduate degrees in anthropology (2), English literature, psychology and, biology. Students also varied in terms of professional experience: food systems non-profit, journalism, food retail, health care and science teacher. Applications for the second cohort were solicited in the spring of 2013, 30 applications were received, 12 students were accepted, 7 students are starting in the fall of 2013 and 1 deferred. The seven entering students received undergraduate degrees in nutrition and food sciences, anthropology/environmental thought and practice, cognitive science, environmental soil and water science, critical/womens studies, mathematics, sociology and anthropology. Their professional experience include: natural resources management (3), food justice, farm workers (4), food retail, restaurant, food and nutrition education and writer. The first five students were all in the research track. After the completion of the first year of the program in Spring 2013,the Center for Rural Studies conducted a program evaluation. The evaluation of the FS MS program was conducted using a process and outcome based approach. The approach incorporated the collection of primary and secondary data, including: 1) interview of Food Systems Faculty members, 2) focus group of Food Systems graduate students, 3) information from student applications to the program, 4) faculty department and areas of interest and 5) application and acceptance rate to the program. The program received positive evaluations from students and faculty. The interest in both tracks of the Food Systems program is still high; there have been over 50 inquiries and 12 campus visits by prospective students for enrollment in fall of 2014.

Publications:
none

1

Hatch
Feeding stearidonic acid-rich oils to enhance animal health and nutritional attributes of bovine milk .

Principal Investigator: J. Kraft

Accomplishments & Outputs:
Because stearidonic acid, an omega-3 fatty acid, has been recently recognized as a lipid of high biological value that may impart health benefits, it represents a promising feed ingredient for dairy cows to improve the healthfulness of bovine milk fat (value-added milk). The purpose of this project is to establish a sustainable feeding strategy (using rumen-protected stearidonic acid-containing oils) to enhance nutritional attributes of bovine milk fat through raising milk and tissue levels of stearidonic acid and its biological active metabolites. Two experiments were conducted to test and evaluate a rumen-protected seed oil derived from a sustainable terrestrial plant (echium). Echium oil was protected via microencapsulation; and in the first experiment, we determined the resistance to ruminal degradation. In the second experiment, we assessed the extent of intestinal digestion of the rumen-protected oil. The evaluation of an additional stearidonic acid-rich plant oil is currently under way. Results from this project will be disseminated further along in the research.

Publications:
Experiments in progress; no publication to report yet.

1

Hatch
Enhancing the nutritional attributes of bovine milk to achieve a more desirable product tailored to provide health-promoting benefits .

Principal Investigator: J. Kraft

Accomplishments & Outputs:
The project is designed to develop a feeding strategy to enhance the nutritional quality of bovine milk for human consumption. The activities for the past reporting period included establishing the transfer efficiency of omega-3 fatty acids from a rumen-protected plant-oil, fed at 1.5 percent and 3 percent of dry matter, respectively, into milk and determining the change in fatty acid composition and content. We demonstrated that the treatment had no effect on the dry matter intake, milk yield, milk components (fat, protein, and lactose), or feed efficiency of the cow. Omega-3 fatty acid concentrations increased in the milk of both treatment groups. The omega-6-to-omega-3 ratio decreased in milk of supplemented cows relative to the control group. However, the transfer efficiency of the supplemented omega-3 fatty acids was only 4 percent, much lower than predicted. Hence, the next step will be the examination of the postruminal release of the omega-3 fatty acids from the rumen-protected product. Laboratory analyses are currently in progress to evaluate differences in fatty acid distributions among serum lipid classes. The results will provide insight in partitioning of the omega-3 fatty acid into the plasma lipid classes and thus, their metabolism. Results have been disseminated through presentations.

Publications:
Experiments in progress; no publication to report yet.

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University of Delaware
Farm Transfer and Business Succession Education for New England Farmers .

Principal Investigator: R. Parsons

Accomplishments & Outputs:
This project which started in July 2014 will provide educational programs on the areas of farm transfer and transition including estate planning, retirement, wills, estate taxes, transferring assets, health care issues, and business organization. We are planning to conduct Farm Transition and Transfer Education programs during winter 2014 in Connecticut, Maine, New Hampshire, and Vermont. The programs will be directed to farm owners, farm operators, farm families, and potential new farmers. The goal will be to empower farm families planning the future of their farmland and farm businesses to enhance the preservation of Vermont's Working Landscape.

Publications:
none

1

National Institute of Food and Agriculture/Department of Agriculture
Costs and Challenges Associated with Developing and Implementing a Community-Wide Biosecurity Plan .

Principal Investigator: J. Smith

Accomplishments & Outputs:
Livestock farmers and surrounding communities need cost-effective and reasonable approaches to prevent disease and minimize the indirect costs of disease control in the event of a highly contagious animal disease emergency such as foot-and-mouth disease. Engaging with farmers and communities about a hypothetical disaster is difficult when they are facing more imminent threats (e.g., Tropical Storm Irene and other weather challenges and higher grain and fuel costs). Open-ended interviews have been successful in identifying perceptions and possible solutions to monitoring access of farms. Initial results of cost benefit analysis indicate that losses would put a number of farms out of business, resulting in substantial losses to the local economy. In this context, we are creating a website that will serve as a resource for local farms and governments in developing plans to respond to a highly contagious disease emergency while reducing economic costs and losses.

Publications:
none

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Auburn University
Development of Biosecurity Materials for eXtension Website .

Principal Investigator: J. Smith

Accomplishments & Outputs:
A sub-team of the Extension Disaster Education Network eXtension Community of Practice was formed to create web-content on the topic of biosecurity. The team defined the term biosecurity as referring to "strategies and management practices that mitigate biological risk". With the assistance of summer interns at UVM, over 70 articles on eXtension related to animal diseases or biosecurity were created, revised, or updated. Overall the organization of content was improved. The summer interns also created and recorded public service announcements on staying healthy at the fair. The Community of Practice is committed to ongoing maintenance of links and revision of content as needed.

Publications:
none

1a

Hatch
A Statewide Assessment of the Health Needs of Hispanic Farmworkers in Vermont .

Principal Investigator: D. Baker

Accomplishments & Outputs:
The project has four objectives: 1) Identify the health status and needs of Vermonts dairy farmworkers, with an emphasis on Hispanic migrant dairy workers. 2) Understand the challenges Vermonts health care providers face as they seek to meet the needs of Hispanic dairy workers. 3) Facilitate improved coordination, strategic planning and policy development between Vermont farmers, health care and other service providers who work with Hispanic migrant dairy workers. 4) Track changes in public opinion toward Hispanic migrant workers through annual statewide surveying. In 2013 the P.I. and project partners met with staff from Sen. Leahys office to provide data on current migrant labor issues facing Vermont dairy farmers. Dr. Baker co-presented with Erin Shea, director of UVM Extensions Vermont Migrant Program at the 2013 Northeast Organic Farming Associations (NOFA) annual conference on current agricultural labor visa issues, trends and policy. The P.I. also surveyed residents statewide through the Vermonter Poll on their perceptions of migrant dairy labor, contributing to a longitudinal study of public opinion. The research was disseminated to a broad regional audience in an article published by the Federal Reserve Bank of Boston. New research was conducted on to the practice of self-care, self-prescription and traditional health practices by Hispanic migrant farmworkers in Vermont. This is a qualitative study that included interviews with 34 participants on 32 dairy farms located in 10 Vermont counties. Twenty nine participants were male and five were female. Preliminary results from the projects research on self-care by migrant workers found that workers had been on their current farms ranged from over 7 years to two months. An estimated 50 percent of participants have requested medications and vitamins from their country of origin. Our statewide survey of Vermont residents found support for migrant dairy labor remains strong and awareness is high. We found in 2013 that 65 percent of respondents (n=622) said that they had personally spoken with a farmer or family member, similar to what we found in 2012. When asked in 2013 how they felt about a bias free policing policy, 84.5 percent (n=576) were in favor, up from 72 percent who said they favored the adoption of such a policy in Vermont in 2012. Support for development of a 3-year guest worker program also rose in 2013 to 89.7 percent (n=609), an increase from 83 percent (n=699) in 2012.

Publications:
Baker, D. (2013) Latino Dairy Workers in Vermont. Communities & Banking 24: 2, 5-7. Federal Reserve Bank of Boston.

1a

Hatch
Adjusting to Vermont:Stress Among Latino Farmworkers & State Policy Development .

Principal Investigator: D. Baker

Accomplishments & Outputs:
Vermont is a new Latino destination where many of these Spanish-speaking immigrants have work on dairy farms. The health status of these workers has been a concern and recent research in Vermont found that barriers to health care of Hispanic dairy farmworkers include fear of immigration law enforcement, language issues and lack of transportation. Relatively few workers reported health problems, of those who did a surprisingly large percentage reported experiencing depression or anxiety. This finding highlighted how little is known about the mental health impacts of stress and isolation on these farmworkers. This project seeks to conduct a rapid assessment of depression and anxiety among Latino dairy farm workers, building on the research teams experience working with this population. The results will be shared with health care providers in Vermont through an annual statewide meeting and work with agencies providing services to these workers. The development of Vermont state policies to address the mental and physical health issues faced by Latino dairy workers will also be investigated. A 1-year no-cost extension was received for this project.

Publications:
none

1a

Hatch
Food Safety from Milking to Marketing for Vermont Artisanal Cheese Makers .

Principal Investigator: . Barlow

Accomplishments & Outputs:
The objective of this project is to support Vermont's growing artisanal cheese production regional food chain with food safety microbiology, epidemiology and consumer research, and extension activities directly linked to human health concerns. The proposed project addresses food safety research and extension gaps for Vermont's artisanal cheese makers. First, food safety pathogen epidemiology and detection research is addressed across the entire on-farm cheese production chain. Second, research on a novel pathogen detection technology that may be scalable to all size dairy operations, including small-scale on-farm artisanal cheese makers is proposed. The integration of these detection technologies with an improved understanding of pathogen epidemiology will fill gaps in our current abilities to mitigate food safety risk on dairy farms producing cheese or other raw milk products. Third, proposed research on consumer beliefs, attitudes, and practices brings the project full circle from milk production to marketing and back to production again by providing artisan cheese makers with knowledge on consumer issues. Finally, extension efforts which will help disseminate research results and knowledge to key stakeholders are integrated into this proposal at multiple levels, and form the foundation for establishing future research and collaborations on artisanal cheese production and food safety.

Publications:
none

1a

Agricultural Research Service/Department of Agriculture
Holstein Cattle Major Histocompatibilty (MHC) Gene Diversity and Tracking the Immune Response to an Adenoviral Vectored Foot and Mouth Disease Vaccine Using MHC Tetramer Technology .

Principal Investigator: J. Barlow

Accomplishments & Outputs:
The purpose of this project is to determine the allele diversity and frequency of the major histocompatibility complex (MHC) class I genes in pure-bred herd of Holstein dairy cattle. These genes are critical in the T cell mediated immune response to intracellular pathogens including viruses. We have adapted molecular methods to determine the class I allele types of 136 Holstein cattle from a UVM Dairy Center of Excellence herd. Phylogenetic analysis of published MHC class I alleles has been completed and demonstrates clustering of alleles within putative gene clusters. Allele typing has identified new allele sequences and allele combinations which have not been reported previously. We conclude from these results that MHC genetic structure may be more complex than previously presumed. In the next year we will develop high-throughput next-generation sequence-based MHC allele typing for Holstein cattle. We will compare results of this high-throughput typing method to the established sequence specific typing and sub-cloning of expressed genes. We will use these data to develop assays to evaluate how MHC allele-type influences the immune response to Foot and Mouth Disease Virus vaccination. We have developed 6 recombinant bovine MHC class I and in the next year will develop 2 class II molecules. For class I molecules we have determined epitope binding affinity and stability in an in vitro system, and these results have been used to inform the predictive algorithm NetMHCpan. These data will be used to predict putative antigenic peptides from the Foot and Mouth Disease Virus (FMDV) capsid proteome. These synthetic MHC molecules will be used as immunologic tools to evaluate T cell immune responses of Holstein cattle following vaccination or natural infection with FMDV. Results of this work will be disseminated to the research community by presentations at international meetings and through submission of manuscripts for publication in peer-reviewed scientific journals.

Publications:
none

1a

National Science Foundation
BREAD: A Modern Approach for the Development of Cattle Vaccines for Critical Bovine Diseases Impacting Smallholder Farmers in Sub-Saharan Africa .

Principal Investigator: J. Barlow

Accomplishments & Outputs:
An international team of scientists from the United States, Kenya, and Denmark have established a collaborative research program to examine the use of new technologies in immunology with the goal of accelerating vaccine development for two critical cattle diseases that impact smaller holder farmers in sub-Saharan Africa. The objectives of the project are to establish and advance gene typing methods for the major histocompatibility (MHC), also known as bovine leukocyte antigens (BoLA), class I and II genes of cattle. We have successfully identified methods and initiated typing of Holstein cattle in a University of Vermont Dairy Center of Excellence herd. In collaboration with Washington State University, we have evaluated the haplotypes of approximately 80 animals for the MHC Class II DRB3 alleles by the polymerase chain reaction restriction fragment length polymorphism method. For typing MHC Class I genes we have adapted PCR based methods of Ellis et al. and developed extended methods for allele specific PCR using a bio-informatics approach to assay design. Collaborators have completed synthesis of a panel of synthetic recombinant bovine MHC molecules which will be used as immunologic tools to evaluate T cell immune responses of Holstein cattle following vaccination or natural infection. Experiments have been completed describing the binding affinity of these molecules. These results are expanding our understanding of BoLA gene expression and diversity in Holstein cattle. Results of this work will be disseminated to the research community by presentations at international meetings and through submission of manuscripts for publication in peer-reviewed scientific journals.In the next years we plan to demonstrate the use of a panel of bovine class II MHC-tetramer molecules in studies of Holstein cattle immune responses.

Publications:
none

1a

Agricultural Research Service/Department of Agriculture
Duration of Immunity to Vaccination of Cattle with Human Ad5FMDV Vacine .

Principal Investigator: J. Barlow

Accomplishments & Outputs:
The goals of this research are to evaluate the duration of immunity and protection from viral challenge following vaccination of Holstein cattle with the newly approved Foot and Mouth Disease Virus vaccine. We have established a group of Holstein steers to initiate trials in the first year of the project. In the next year we will vaccinate a total of 88 animals and evaluate their immune response at 3, 6, 9 and 12 months following vaccination. Results of this work will be disseminated to the research community by presentations at international meetings and through submission of manuscripts for publication in peer-reviewed scientific journals.

Publications:
none

1a

Hatch/Multistate
Mastitis Resistance to Enhance Dairy Food Safety .

Principal Investigator: J. Barlow

Accomplishments & Outputs:
The multi-state project NE-1048 objectives include: Objective 1) Characterization of host mechanisms associated with mastitis susceptibility and resistance; Objective 2) Characterization and manipulation of virulence factors of mastitis pathogens for enhancing host defenses; Objective 3) Assessment and application of new technologies that advance mastitis control, milk quality and dairy food safety. The Barlow lab will address objective 2 and the sub-objectives i) characterization of pathogen virulence factors, ii) antimicrobial resistance, and iii) molecular epidemiology and diagnostic tools. We will examine potential associations between antimicrobial use practices, carriage of antimicrobial resistance genes and response to antimicrobial therapy among different strain types of Staphylococcus aureus and Streptococcus uberis. We will characterize and evaluate the impact of putative virulence factors in vitro and the examine association of these factors with DNA-based strain types for S. aureus. We will evaluate the use of molecular epidemiologic tools to describe transmission dynamics of contagious mastitis pathogens and antimicrobial resistance. Our work will focus on the epidemiology of S. aureus and Coxiella burnetii in emerging sectors of the dairy industry in Vermont including niche dairy farms (e.g. on-farm artisanal cheese producers and organic dairy farms) and dairy farms adopting automated milking technologies. We have completed a pilot study of contagious mastitis pathogen transmission in a herd milking cows in robotic systems. We have also initiated a 12 month study of milk quality on 30 organic dairy farms and have established a library of S. aureus isolates from these farms. Our goal is to combine molecular and mathematical epidemiological approaches to unravel the complex problems of chronic intramammary infections and antimicrobial resistance emergence and transmission in cattle populations. In the next year we will conduct multilocus sequence typing and in vitro studies of virulence for S. aureus isolates from our library. We will also evaluate antimicrobial treatment practices and conduct in vivo studies of mastitis treatments. Outputs will include peer reviewed publications, presentations at national and local meetings, and contributions to bacterial isolate collections with epidemiological data.

Publications:
Barlow, J.W., Zadoks, R.N., and Schukken, Y.H. 2013 Effect of Lactation Therapy on Staphylococcus aureus transmission dynamics in two commercial dairy herds. BMC Veterinary Research 9: 28-40.

1a

Hatch/Multistate
Northeast Pasture Consortium .

Principal Investigator: J. Gorres

Accomplishments & Outputs:
Rotational grazing is regarded as an environmentally friendly and sustainable way to feed livestock for the production of dairy products. Water quality issues such as nutrient runoff and leaching is lower than in silage corn because pasture has continuous and complete soil cover. Water quality benefits may be jeopardized by the high herd densities in rotationally grazed paddocks. Compaction in particular may reduce pasture quality potentially causing vegetation loss. It is thus important that compaction caused by livestock hoof fall is mitigated. The primary objective of this study is to investigate alternative ways to loosen pasture soil compaction. Methods presently include subsurface plowing along hydrologically active lines and tillage radishes Tillage radishes are a.k.a. daikon and have deep roots and broad crowns. The taproots can grow 1.5 feet deep with diameters in excess of 3 inches. Both methods create macropores which loosen the soil improving soil drainage. We demonstrated that keyline plowing decreases soil moisture of wet fields by approximately 30%. This is a significant finding because cows can return to the pasture sooner if saturation is avoided. What we were not able to confirm was that organic matter increases with this management practice. In both the radish and the keyline pasture treatments the number of earthworms was several fold greater than in untreated pastures. This is a significant finding because earthworms are an indicator of soil health, and more importantly, of more organic resources in the soil. While we have no evidence that above ground productivity is increased, soil productivity seems to support more earthworms. There are additional objectives relevant to other agricultural settings were developed from this work. Findings from these studies will feedback into our pasture method. In flood-prone fields used for vegetable production, flooding can diminish soil fertility by either chemical reactions or through compacting soils. The objective is to investigate how small elevation changes (a proxy for length of inundation) affect soil fertility and how losses in fertility can be remedied by cover crops with deep tap roots. Even small differences in elevation impact soil fertility. Soil between the growing radish roots becomes compacted, but after winter kill of the radish but we expect that the decomposing roots leave large pores that will improve drainage. We expect that these findings will inform pasture management on flood-prone pastures. Even when feeding cows on pastures, herds can produce significant amounts of manure. Little is known about compost quality for pasture management. An experiment that evaluates blends of high quality vermicompost and windrow compost that includes food scraps showed that blends develop greater electrical conductivity, i.e. fertility, than the two pure blend components. This is a significant finding because blends cost less than the pure vermicompost and because it promises improved food scrap compost quality. Results from this study may inform management strategies for organic waste processing required by Act 148.

Publications:
Melnichuk, R and J. Gorres. Greenhouse Gas Emissions from Forest Soil Emissions. SSSA Annual Meetings, Cincinnati, Oh, October 21-24, 2012

J. Gorres, R. Melnichuk and K. Belliturk. Development of an Amynthas Agrestis (Goto and Hatai) Populationin a Sugar Maple (Acer Saccharum Marsh) Stand in the Champlain Valley, Vermont. SSSA Annual Meetings, Cincinnati, Oh, October 21-24, 2012.

J. Gorres. Earthworms as Agents of Change in Our Forests. Chester, VT, November 13, 2012.

Gorres, J. and R. Gilker. Earthworms Count: Using Keyline Plows and Tillage Radishes to AddressPasture Compaction. 18th Annual VT Grazing & Livestock Conference. Lake Morley Resort, VT., January 17- 18, 2013.

Hilshey, B., S. Bosworth, and J. Gorres. Raw Waste Milk as a Pasture Amendment. Poster Presentation, Northeast Pasture Consortium, Manchester, February 7 - 8, 2013.

Gilker, R., J. Gorres. Possible Tools for Pasture Soil Quality: Keyline Plowing and Forage Radishes. Poster Presentation, Northeast Pasture Consortium, Manchester, February 7 - 8, 2013.

Gorres, J. Vermicomposting. Presentation at the Vermont Organics Recycling Summit - Managing Organics as a Resource: Vermont Takes Up the Challenge Randolph Center, Vermont, March 28, 2013.

Gorres, J. Earthworms: Gardners' Little Helpers, Foresters' Stealthy Foes. Marlboro College Technical Center, Brattleboro, VT April 24th, 2013.

Gorres, J. Earthworm Invasions in Vermont's Forest. NorthWoods Stewardship Center, East Charleston, Vt, June 15, 2013.

Gorres, J. ACT 148 AND THE EARTHWORM AMYNTHAS AGRESTIS, AN UNDERESTIMATED FOREST PEST. Fact sheet for Vermont Composters. September, 2013.

1a

National Institute of Food and Agriculture/Department of Agriculture
Effects of Variation in Pathogen Detection and Signaling Pathways on Resistance to Bovine Mastitis .

Principal Investigator: D. Kerr

Accomplishments & Outputs:
Experiments are being conducted to evaluate the hypothesis that genetic or epigenetic differences in genes associated with pathogen detection and response contribute to differences between cows in their innate resistance to mastitis. We are using dermal fibroblasts as a model cell type to evaluate between-animal differences in the innate response to an E. coli cell wall component (LPS) and to a Gram-positive cell wall component (PamsCSK4). We have also established a relationship between our in vitro results and in vivo results for experiments in both calves and lactating cows. Fibroblast cultures have been established from 15 calves sampled at approximately 5 and 11, and 16 months of age. Responses of the fibroblasts to LPS reveal substantial between-animal variation in IL-8 production. This molecule is key to recruiting immune cells to the site of infection and is being used as a marker of the strength of the innate immune response. In addition, there is a marked increase in response of cells collected at 15 versus 5 months of age. The age related difference in response is not genetic, as the cells were established from the same animals. Rather, we have preliminary evidence that epigenetic mechanisms are responsible. Epigenetic mechanisms modify the ability of genes to be expressed. We are now using cutting edge Next-generation sequencing techniques to locate age-related differences in the epigenetic mechanism known as DNA methylation. We have similarly established and challenged fibroblast cultures from 50 lactating cows. The in vitro responses were used to rank the cows from high to low responders. Five high- and five low-responding cows were subsequently challenged with an intramammary infusion of a mastitis-causing strain of S. aureus. All cows developed clinical mastitis. All five high responder became chronically infected, while two of the low responders cleared the infection at least temporarily. Thus, high responding animals may be at greater risk for developing chronic mastitis. In this experiment we also observed higher concentrations of BSA and IL-8 in milk of the high responder cows in concordance with their fibroblast-based ranking. Experiments are currently in progress to evaluate genetic and epigenetic differences between the high- and low-responding fibroblast cultures. These differences may serve as markers of the response potential that could be used to genetically select superior animals for the dairy herd.

Publications:
Kandasamy, S., and D.E. Kerr. 2012. Genomic analysis of between-cow variation in dermal fibroblast response to lipopolysaccharide. Journal of Dairy Science. 95: 3852-3864.

1a

Animal Health
Effects of genetic variation in host and pathogen on severity of bovine mastitis .

Principal Investigator: D. Kerr

Accomplishments & Outputs:
We have identified an association between strain type of Staphylococcus aureus isolates and transmission rates within dairy herds. Multilocus sequence typing (MLST) was used to identify strain types causing mastitis on 3 dairy farms. Bovine host-adapted strain types (ST151) had higher rates of transmission compared to human host-adapted strains (ST8). Human host adapted strains (ST8) were more likely to carry antimicrobial resistance gene (blaZ) and demonstrate a beta-lactam resistant phenotype, yet bovine host-adapted strains were more-likely to cause chronic infections which were not responsive to antimicrobial treatment. In the coming year we will complete whole genome sequencing of selected strains which differ in phenotypic behavior and have been associated with bovine and human hosts. We have also investigated the in vitro responsiveness of milk-derived mammary epithelial cells in culture to model the ability of cows to respond to S. aureus mastitis. Mammary cells were isolated and successfully cultured from milk of cows. The initial isolation procedure is quite labor intensive, and only five of eight cell lines could be recovered from cryopreservation. These cells produce substantial quantities of a neutrophil attracting chemokine (IL-8) in response to the Gram-negative E. coli endotoxin (LPS), but do not respond to live or heat-killed S. aureus (Gram-positive) of bovine or human adapted strains. They do respond to a synthetic component of the Gram-positive cell wall, PAM2CSK4, but not to the inflammatory cytokine, interleukin-1B (IL-1B). This lack of response to IL-1B is quite interesting in that mammary cells are known to produce this factor in response to infection and it is a potent stimulator of IL-8 production by fibroblasts isolated from the same animals. This suggests that a co-culture system of fibroblasts and mammary epithelial cells would be a more informative model to studying genetic variation in the innate response to mastitis. However, the milk-derived cells are not reliably obtained from study animals. An alternative source is to obtain the cells from mammary tissue collected at slaughter.

Publications:
Barlow, J.W., Zadoks, R.N., and Schukken, Y.H. 2013 Effect of Lactation Therapy on Staphylococcus aureus transmission dynamics in two commercial dairy herds. BMC Veterinary Research 9: 28-40.

1a

Hatch/Multistate
Mastitis Resistance to Enhance Dairy Food Safety .

Principal Investigator: D. Kerr

Accomplishments & Outputs:
Genetic causes for differences between animals in their response to mastitis causing pathogens are being evaluated with a fibroblast cell model system. In this system, fibroblast cells from different cows are obtained from skin biopsies, cultured in the laboratory, and then stimulated with pathogens, or pathogen toxins. The cell's production of inflammatory cytokines is then measured in the culture media. We have made progress in identifying sets of cows whose fibroblasts differ dramatically in their response to stimulation with E. coli lipopolysaccharide (LPS). These fibroblasts are from differing breeds of cattle; one being a dairy breed (Holstein), and one being a beef breed (Angus). This novel finding has great potential to identify genetic polymorphisms in cattle genes that affect the innate immune response that is the key defense against mastitis. We have also compared fibroblasts from Holstein animals demonstrating mild versus severe responses to naturally occurring E. coli mastitis. Preliminary analysis of these samples has not identified substantial differences between groups. However, the Angus-Holstein difference is one that we feel has great potential to identify genetic and/or epigenetic effects on the bovine immune system. We have now submitted samples for next generation sequencing technologies (RNA-seq and Methyl DNA-seq) to examine breed differences in the entire set of expressed genes (the transcriptome) or the global regions of methylated DNA (the methylome), respectively. The RNA-seq data may lead to identification of polymorphisms in the bovine DNA code, while the Mehtyl DNA-seq will allow examination of epigenetic causes for differences in gene expression. DNA methylation is one type of epigenetic effect that appears to cause differences in gene expression and appears to be caused by environmental effects affecting the developing calf either in utero or in early life. This breed difference work has come through an on-going collaboration with Dr. B. Crooker at the University of Minnesota. Bioinformatics support is being conducted in collaboration with Dr. Stephanie McKay from our department, while the next-generation sequencing is being conducted at the UVM core facility.

Publications:
none

1a

Walker (George) Milk Research Fund
Vermont Cattle Health Improvement Program .

Principal Investigator: J. Smith

Accomplishments & Outputs:
The Vermont Cattle Health Improvement Program (Vt CHIP) oversees the state's voluntary Johne's disease control program. Johne's disease is a chronic, primarily intestinal infection of ruminants, and has delayed onset of subclinical and clinical signs, which result in reduced productivity and longevity. With funding from the Walker Foundation, the state Johne's committee determined how the program would continue after the expiration of current funds. Previous federal and state funding are no longer available. Online certification and re-certification modules were made available to veterinarians working with herds in the program. A series of four workshops for dairy producers, students and agricultural service providers was conducted in November 2012 with guest speaker Dr. Franklyn Garry. Updated materials were posted to UVM Extension's Johne's disease page. The program will continue as a herd health program administered by private veterinarians.

Publications:
Smith, J. M. and R. H. Whitlock. 2012. CaseReport: Mycobacterium avium subspecies paratuberculosis infection in three generations of beef cattle. Bovine Practitioner 46:32-35.

1a

Vermont Agency of Agriculture, Food, and Markets
Characterization of Methane-Producing Microorganisms in Dairy Cow Waste Anaerobic Digesters .

Principal Investigator: A. Wright

Accomplishments & Outputs:
Anaerobic digestion on farm involves the decomposition of manure into effluent and methane. The production of methane is often the rate limiting step of anaerobic digestion of organic wastes due to the fast growth of bacteria, which produce all the substrates for the slow-growing methanogens. However, high concentrations of volatile fatty acids, salts, ammonia, and heavy metals can also be inhibitory for methanogens and are the most common reasons for digesters to fail, or not reach their full potential. Similarly, within the anaerobic digesters sulfate reducing bacteria compete with methanogens for hydrogen and, in the process, produce hydrogen sulfide, which is a major problem in these reactors. Reducing the numbers and/or activity of the sulfate reducing bacteria may increase methane production and reduce hydrogen sulfide and subsequent corrosion within these digesters. Specific Aims are to: (a) examine the molecular diversity of bacteria and methane-producing archaea (i.e. methanogens) within each of three digesters (Green Mountain Dairy, Blue Spruce Farms, Chaput Family Farms) to assure a good representation of the microbiome; (b) exan1ine the molecular diversity of the sulphate reducing bacteria within each digester to identify the bacteria responsible for hydrogen-sulfide corrosion; (c) determine the density of the methanogenic archaea, protozoa and bacteria in a variety of locations and depths within each digester using real time PCR; (d) correlate the gas production in vitro to the abundance and/or activity of the methanogens and other key microbes found; and (e) identify probable synergistic relationships within the anaerobic digesters and suggest a possible strategy to increase methane production.

Publications:
St-Pierre, B. & A-D.G. Wright. 2013. Comparative metagenomic analysis of bacterial populations in three full-scale mesophilic anaerobic manure digesters. Appl. Microbiol. Biotechnol. In Press

St-Pierre, B. & A-D.G. Wright. 2013. Metagenomic analysis of methanogen populations in three full-scale mesophilic anaerobic manure digesters operated on dairy farms in Vermont, USA. Bioresource Technology. 138: 277-284.

1a

Central Vermont Public Service Corporation
Study of the Mechanisms of Methane Production in Farm Based Anaerobic Digesters by Analyzing Microbial Community Gene Expression .

Principal Investigator: A. Wright

Accomplishments & Outputs:
The purpose of our project is to study the mechanisms of methane production in farm based anaerobic digesters by analyzing microbial community gene expression. We propose to identify the genes that are expressed or used by methanogens and their competitors from the sampled digesters. Methane producing organisms have approximately 2,000 genes, and each gene is responsible for a specific function that allows the microorganism to either grow, maintain itself, produce specific chemicals, protect itself against unfavorable conditions or change its environment. A microorganism only expresses or uses a gene if it needs its function, otherwise the gene is not expressed. Therefore, identifying the genes that are expressed in a cell can reveal a great deal of information about it: 1. if it is under favorable conditions or under stress, 2. what chemicals it is using or producing, 3. how it is responding to its current conditions, and 4. how it is changing its environment With high-throughput sequencing and computing technology, it is possible to analyze not just the genes that are expressed in a specific type of cell, but also in an entire community of microorganisms, such as those found in a manure digester. Therefore, combining the identification of the microbes that are in a digester with the identification of the genes that they use will tell us, which microbes are desirable in a digester, and which should be eliminated, and what conditions are favorable or unfavorable for each type of microbe. This information should provide us with specific short term and long term strategies to improve digester efficiency. Pyrosequencing of the samples has resulted in 200 million sequences which are currently being analyzed.

Publications:
Data are currently being processed and analyzed.

1a

Central Vermont Public Service Corporation
Bench Top Reactors to be used in the Study of Methaneogens .

Principal Investigator: A. Wright

Accomplishments & Outputs:
The dominant methanogens in manure digesters are not well documented, nor are the interactions between the protozoa (major hydrogen suppliers), bacteria (hydrogen suppliers and hydrogen utilizers - i.e. methanogen competitors) and the methanogens (methane producers). The overarching goal of the current project is to better understand these synergistic relationships, and to use the knowledge gained of the methanogen population dynamics and diversity to enhance, to enrich for, and/or to seed anaerobic digesters with more potent methane producing strains of methanogens to increase gas production. Specific Aims are to: (a) examine the molecular diversity of bacteria and methane-producing archaea (i.e. methanogens) within each of three digesters (Green Mountain Dairy, Blue Spruce Farms, Chaput Family Farms) to assure a good representation of the microbiome; (b) exan1ine the molecular diversity of the sulphate reducing bacteria within each digester to identify the bacteria responsible for hydrogen-sulfide corrosion; (c) determine the density of the methanogenic archaea, protozoa and bacteria in a variety of locations and depths within each digester using real time PCR; (d) correlate the gas production in vitro to the abundance and/or activity of the methanogens and other key microbes found; and (e) identify probable synergistic relationships within the anaerobic digesters and suggest a possible strategy to increase methane production. We purchased six state-of-the-art bench 5-Litre top bench-top anaerobic digesters to carry out some of this important work. Six digesters will allow us to use actual effluent from each of our three partner farms, with backup (replication). They also provide ns with the versatility to design control and treatment groups where we can manipulate different variables (e.g. temperature, feed stuff, and microbial manipulations) to see their effect on methane production, and on the composition and density of the microbial population.

Publications:
Research is in progress

1a

Hatch
Examination of the methanogen microbiome in five breeds of lactating dairy cows .

Principal Investigator: A. Wright

Accomplishments & Outputs:
Holsteins and Jerseys are the most commonly used breeds in the US Dairy Industry. They differ in performance for various traits, such as body size, milk production and composition. With increased concerns about the impact of agriculture on climate change, potential differences for other traits such as methane emission also need to be further characterized from Holsteins and Jerseys, as well as other popular dairy breeds. Since methane is produced in the rumen by methanogens, preliminary data from our laboratory suggest that differences in the methanogen population exist between Jerseys and Holsteins. Here, we propose a more in depth study to confirm these findings and to include milking Short-horns, Linebacks, and Brown Swiss. Pyrosequences of rumen methanogens from each breed will be generated and analyzed to identify rumen methanogens and to determine if their population structures vary significantly. Real-time PCR will also be performed to estimate the mean methanogen density for each breed. If significant differences are found, then head boxes may be used to measure methane production. These data will make a significant contribution to our understanding of the effect of breed differences on the rumen microbiota, which may lead to new strategies for the US Dairy Industry. The objectives are to: (1)identify the methane producing methanogenic archaea (methanogens) in Holsteins, Jerseys, Holstein-Jersey crosses, Milking short-horns, and Brown Swiss using pyrosequencing; and to (2) identify and assess the diversity and structure of the methanogen population in in Holsteins, Jerseys, Holstein-Jersey crosses, Milking short-horns, and Brown Swiss lactating dairy cows.

Publications:
Research is in progress

1a

Hatch
Dairy Pharming: Production of Pharmaceutically Active Human Proteins in Bovine Milk .

Principal Investigator: F. Zhao

Accomplishments & Outputs:
The overall goal of this project is to develop innovative ways to generate transgenic cows which produce human insulin in milk at a minimum concentration of 1 mg/ml at a much reduced cost and with an improved success rate. The major accomplishment during the reporting period is to test our transgene construct by generation of transgenic mice which produce human insulin in milk.Specifically, 1) Transgenic mice expressing human proinsulin in milk were generated by inserting a human proinsulin cDNA into a pBC1 vector controlled by the mammary gland-specific goat beta-casein promoter. Three females (#5, #12, and #15) were identified by regular PCR and quantitative PCR. 2) Milk samples from 3 female founder mice, #5, #12, and #15 were boiled in SDS protein sample buffer and separated by SDS-PAGE. The milk proteins were visualized by staining the gel with coomassie blue, and the bands with size around 10 kDa were analyzed by LC-MS/MS). Obtained tandem mass spectra were searched against a concatenated forward and reverse mouse and human NCI protein database, respectively. The results indicated that human proinsulin protein is identified only when mass spectra were searched against human protein database.3) To test human proinsulin expression in transgenic milk, milk samples were analyzed by western blotting. Milk from wild-type (WT) mice was used as a negative control, and a commercial recombinant human proinsulin protein was used as a positive control. All transgenic milk samples showed a strong signal at 10 kDa, indicating high-level expression of human proinsulin in milk; no band was observed in the negative control. 4) Milk samples from 3 transgenic lines were collected at early, middle and late lactation for ELISA analysis. The concentration of human proinsulin in transgenic milk was relatively low at early lactation but increased dramatically at middle lactation followed by an appreciable decrease at late lactation. The highest expression level of human proinsulin was up to 10.2 µg/µl. 5) mRNA expression level of human proinsulin in the mammary gland tissue at different stages were monitored by qRT-PCR. The results indicated that human proinsulin mRNA expression level was undetectable at non-pregnant stage, significantly induced at pregnant and lactation stages, and reduced to basal line at involution stage. The similar profile was observed for human proinsulin protein expression level in mammary gland tissue at different stages. These results demonstrated that our transgene construct is successful to induce human insulin expression in milk. The results have been presented in this year's Gordon Mammary Gland Biology Conference by a graduate student.

Publications:
Xi Qian, and Feng-Qi Zhao. (2013). Interactions of the ubiquitous octamer-binding transcription factor-1 with both the signal transducer and activator of transcription 5 and the glucocorticoid receptor mediate prolactin and glucocorticoid-induced beta-casein gene expression in mammary epithelial cells. International Journal of Biochemistry and Cell Biology 45, 724-735

Feng-Qi Zhao. (2013). Octamer-binding transcription factors: genomics and functions (invited review). Frontiers in Bioscience 18, 1051-1071

1a

Hatch
Single Nucleotide Polymorphisms of Glucose Transporters as Breed Selection Markers for Milk Productivity in Dairy Cattle .

Principal Investigator: F. Zhao

Accomplishments & Outputs:
The specific aim of this DCE project is to analyze the associations of polymorphisms of glucose transporters (mainly GLUT1, the predominant GLUT isoform expressed in bovine mammary gland) with milk production traits in Holstein cows. We have carried out substantial work on the project in the last one and half years, specifically: a) We have collected blood samples from 1500 cows on 5 local farms (Nordic, Fairmont, Foster Brothers, Knoxland, and Newmont Farms) and obtained the production data and breed values from the DHIA for each individual cows. b) We have isolated genomic DNA from each blood sample. c) At first, DNA of 5 cows with highest breeding value and 5 cows with lowest breeding value was selected from each farm and these 50 DNA samples were mixed together. We amplified and sequenced 2kb promoter and the individual exon sequences (Exon 1 to Exon 10) and their flanking regions of bovine GLUT1 using the DNA pool. Only one SNP on exon 2 was found in our samples, which is a C/T synonymous SNP and is consistent with the SNP rs208845183 in the NCBI SNP database. This SNP does not change the amino acid sequence of GLUT1 protein, thus, it may not have much physiological significance and we did not genotype this SNP in our cows. We were not able to detect any other SNPs, including three expected missense SNPs found in NCBI SNP database: rs133236740, rs134711046 and rs135388257 on exon 6. These SNPs are of most interest because they change consecutive 2 amino acids of GLUT1 protein. d) We thought the mixing 50 cows' DNA might dilute the DNA samples with SNPs, so next we mixed DNA samples of 5 cows with highest breeding value and 5 cows with lowest breeding value from each individual farm and sequenced each exon sequence from the DNA pool of each of 5 farms. Similarly, only SNP rs208845183 was detected from these smaller DNA pools. e) To further check the possibility that our DNA pools may miss low frequency SNPs, we next decided to focus on SNPs of rs133236740, rs134711046 and rs135388257 on exon 6 in big number of cows. Primers were designed for each SNP and 500 cows from different farms were genotyped for these 3 SNPs by Tm-shift method. For each SNP, 2 forward allele-specific primers and 1 common reverse primer were designed. Each of the 2 forward primers contains a 3'-terminal base that corresponds to one of the two SNP allelic variants. GC rich tails with different length were attached to the 5' of the 2 forward primers. PCR products from these 2 forward primers can be distinguished by a melting curve on a real-time PCR machine because they have distinct Tm. 500 cows were genotyped for 3 SNPs but they all have the same genotype. f) To investigate whether these 3 SNPs are breed-specific, we sequenced exon 6 of 8 Jerseys DNA samples. We did not find any cow carrying the SNP. Unfortunately, based on the tremendous work we have done, no missense SNP could be found in bovine GLUT1 gene. In next year, we will look for the SNPs in other genes.

Publications:
Shun Kitaoka, Anthony D. Morielli, and Feng-Qi Zhao. (2013). FGT-1 is a mammalian GLUT2-like facilitative glucose transporter in Caenorhabditis elegans and its malfunction induces fat accumulation in intestinal cells. PLoS One 8(6), e68475

Huawei Zeng, Jun Liu, Matthew I. Jackson, Feng-Qi Zhao, Lin Yan, and Gerald F. Combs, Jr. 2013. Fatty liver accompanies an increase of Lactobacillus acidophilus in the hind gut of C57/BL mice fed a high-fat diet. Journal of Nutrition 143(5), 627-631.

Yong Shao, Emma H. Wall, Thomas B. McFadden, Yong Misra, Xi Qian, Ruth Blauwiekel, David Kerr, and Feng-Qi Zhao. (2012). Lactogenic hormones stimulate expression of lipogenic genes but not glucose transporters in bovine mammary gland. Domestic Animal Endocrinology 44, 57-69.

1b

Hatch/Multistate
Biological Control of Arthropod Pests and Weeds .

Principal Investigator: B. Parker

Accomplishments & Outputs:
This 5 yr project is evaluating growth factors of Acer mono in different coldhardiness zones to assess its invasiveness and develop methods for safe and ecological use for early detection of Asian longhorned beetle (ALB). Results will document the coldhardiness of A. mono and define the range for its use as a sentinel tree. It will provide guidelines for how to grow the tree to facilitate its use as a sentinel tree and provide evidence regarding its invasive threat. In spring 2013 sites in each targeted USDA plant coldhardiness zone (5,4,3) were visited and nets to prevent deer browse were removed. In August data on crown condition, dieback, pest damage, the presence or absence of root sprouts and their status were taken. In the fall, each site was revisited, and nets re-applied. The following data were recorded as in the past: caliper diam. at 5.08 cm from the ground, length of new growth, crown condition, dieback, tree height, pest damage and flowering or seed production. Climatic conditions over the past two years were extreme for this region, which impacted the health of the trees. For example, in spring 2012, record breaking warmth, triggering early budbreak, followed by cold that caused frost damage to the foliage. In addition, unusually hot and dry conditions occurred in summer 2012, further stressing the trees. These conditions were some of the most severe that trees would be expected to experience in these zones. After two growing seasons, no substantial differences in survival of the small trees grown above or in-ground were observed, though differences were noticeable among zones. About 20 percent of the small trees died in all zones, but 75 percent sent out new sprouts indicating the roots survived. Survival of the large in-ground trees was generally higher in all zones. Tree mortality during the summer of 2013 was not observed. None of the trees flowered or produced samaras in 2013. Data on shoot growth were separated into two categories, trees that survived (twig growth), and those for which the main stem died but sprouted from the ground (root sprout). Sprouts on test trees were retained as this may provide a suitable trap tree form, based on observations that ALB was particularly attracted to short bushy trees. A clear treatment effect was not observed for tree growth. In zone 3, the in-ground trees grew more than the above-ground trees, whereas differences were not substantial for zones 4 and 5. Little or no deer browse or rodent damage occurred in the 2012-2013 winter. Results to date show the above-ground growing method had no adverse impact on tree survival, and could be suitable for sentinel trees. At some sites, it was difficult to dig holes for the in-ground trees, due to rocky soil. The above-ground method facilitates deployment of these trees where planting a tree in the ground is difficult. It appears A. mono, though not completely killed by low temperatures, is not sufficiently coldhardy to thrive in zone 3, but can survive in zones 4 and 5. Seeds from several specimen trees at two arboretums were collected to test germination at sites within the coldhardiness zones in 2013.

Publications:
none

1b

Hatch
New approaches towards maximizing maple sap yields .

Principal Investigator: T. Perkins

Accomplishments & Outputs:
The goals of this project were to investigate methods to increase sap yields through both improved sanitation and better vacuum transfer in maple tubing systems. Tubing systems typically become less productive as they age, primarily due to increased levels of microorganisms within the system. Such contamination hastens "drying" of tapholes, reducing sap yields. Several experiments were conducted to examine approaches to reduce such losses. In general, spout and tubing cleaning, the use of new spouts annually, the use of check-valve spouts, or replacement of droplines increased yields by varying amounts. We constructed an economic model for producers to use as a tool to estimate the net effects of various strategies on sap yields and net profit. In several other studies we examined possible approaches aimed at increasing vacuum at the taphole, including: reduced taps per lateral, elimination of lateral lines (1/2"), larger-diameter lateral lines, and dual-lateral 5/16" lines. In general, any efforts made to reduce tubing-based restrictions in liquid and air flow are helpful in achieving better vacuum levels at the taphole. The use of dual-lateral lines produced the largest increase in sap yields compared to conventional systems, with an average of 17% increase. Results have been disseminated to the maple industry via a series of presentations to producer groups.

Publications:
Perkins, T.D. and A.K. van den Berg. 2012. Relationships Between Tubing System Age and Sap Yield. Maple Syrup Digest 24A(1): 11-16.

Perkins, T.D. and A.K. van den Berg. 2012. Tubing Cleaning - Methods Used in the U.S.. Maple Syrup Digest 24A(3): 11-18.

1b

North American Maple Syrup Council
Identification of Effective Maple Tubing Cleaning Practices .

Principal Investigator: A. van den Berg

Accomplishments & Outputs:
Microbial contamination of the tubing collection system reduces annual yields of sap from maple trees by an average of 10% annually. Annual cleaning of tubing systems is a method by which maple producers attempt to mitigate these yield reductions. A wide variety of cleaning techniques are currently used for this process, including rinsing the system with pressurized air and water, or with solutions such as peroxide or bleach. However, the effectiveness of these cleaning techniques at increasing annual sap yield is currently unknown. In fact, it is possible that many of these practices have limited effects on yield, and thus by employing them, producers could be wasting substantial amounts of money, time, and other resources each year. Since it is estimated that over 87% of producers currently use some method of tubing cleaning, it is imperative to determine which of these methods is most effective at preventing significant annual reductions in sap yield. This will enable producers to increase their annual revenues by employing a cleaning method with an established ability to improve annual sap yield, and/or by eliminating expenditures on the use of ineffective cleaning practices. Thus, the overall objective of this study is to determine which tubing cleaning practice, if any, results in the greatest increase in sap yield relative to the yield obtained when no cleaning treatment is used. To accomplish this objective, generally, a field experiment will be conducted to quantify the volume of sap collected from tubing systems treated with three cleaning techniques identified as most effective at reducing microbial populations through preliminary screening experiments, and the volume of sap collected will be compared to that of control systems which are not cleaned. Preliminary experiments have been completed and identified bleach, peroxide, and isopropyl alcohol as potentially effective treatments. Research plots have been established and initial cleaning treatment of the tubing in field plots is ongoing. Sap yield data from treatment plots will be collected in the spring 2014 maple production season. As research is ongoing, no dissemination of results has yet occurred. It is possible that many or all of the methods maple producers currently use to clean tubing are ineffective at preventing yield losses from microbial contamination (do not result in greater sap yields than not cleaning at all), and thus result in a substantial waste of producers' money and time each year. As producers report spending an average of 36 hours each year on tubing cleaning, the value of this potential loss is substantial (between $300-900 in labor costs alone). This research will identify a tubing cleaning method that does result in significant sap yield increases, or, likewise, establish that no method results in sap yields significantly greater than not cleaning at all. This information will enable producers to immediately increase their revenues by ceasing use of ineffective cleaning methods, and, if a significantly effective cleaning method is identified, by employing a practice that results in significant annual sap yield increases.

Publications:
none

1b

Hatch
Investigation of a new paradigm in maple syrup production .

Principal Investigator: T. Vogelmann

Accomplishments & Outputs:
Vermont's maple producers face increasing challenges to maintaining economically sustainable operations. Rising fuel prices and costs to implement the practices necessary to meet consumer demands for heightened assurance of product safety and quality have rapidly increased the cost of producing maple syrup. Without corresponding increases in productivity, these costs must be passed on to consumers in order to avoid significant reductions in annual income. It is thus imperative to find ecologically sustainable mechanisms to increase the productivity of maple production operations to enable producers to continue to thrive and provide an affordable, high quality product to consumers. The objective of this project is to investigate whether novel strategies of maple crop management can provide mechanisms to increase the productivity of maple production operations, and thus help maintain the long-term economic sustainability of maple syrup production operations in Vermont. We completed the second year of a three-year experiment to determine the quantity of sugar that can be extracted annually using alternative systems of maple crop management. During the 2013 maple production season, sap was collected and sugar extraction quantified from trees tapped with three alternative systems of maple crop management. The experiment will be repeated during the 2014 production season, and the data from all 3 (2012-2014) years will be used to calculate the average quantity of sugar extracted annually from trees of each treatment, and to determine which treatment yielded the highest production of sugar. As the experiment is ongoing, there are no data to report at this time. When available, the results and new information generated in this project will be communicated to maple producers using a variety of mechanisms, including a technical bulletin posted on Extension websites and published in industry journals, and presentations delivered at maple industry conferences and meetings.

Publications:
none

1d

National Institute of Food and Agriculture/Department of Agriculture
Using ""New"" Alternatives to Enhance Adoption of Organic Apple Production Through Integrated Research and Extension .

Principal Investigator: L. Berkett

Accomplishments & Outputs:
Apples are an important component of New England's diversified agriculture. Although there is significant interest in organic production, there are very few organic apple orchards in New England, in part, because of the challenges associated with the traditional apple cultivar grown (McIntosh). However, because of recent shifts in consumer preference for newer cultivars, growers are planting different apple cultivars. Growers want to know what the potential is for sustainable and profitable organic production with the newer apple cultivars that are being planted in the region. This project holistically examines the opportunities and challenges of organic production within the two major orchard systems growers are using to change to new cultivars and with five of the top apple cultivars that growers identified as important to the future of the industry. Specific project goals and the accomplishments during the past year were: 1. Continue to evaluate new apple cultivars and incorporate research-generated knowledge of apple ecosystem dynamics into organic production systems to determine sustainability and profitability. -- Field data on tree growth, fruit yield and quality, and disease and pest incidence on fruit and foliage were collected on all cultivars in two OrganicA research orchards. These data were included into the overall dataset for the project to continue to assess cultivar performance under organic management. Cumulative, long-term yield, fruit quality, and pest incidence data will be used to determine overall profitability of the cultivars under the two systems used in the project. 2. Field test commonly recommended organic foliar nutrient sources and evaluate their impacts on fruit yield, quality, tree nutrition and health including impact on disease and arthropod pests. -- Norwegian kelp extracts commonly used in organic production systems were assessed in an organic orchard. In 2012, complete results from this study were presented locally to Vermont apple growers, and internationally, at the 2nd International Organic Fruit Symposium. 3. Evaluate the benefits of different ground cover strategies in promoting tree health, plant and soil water status, and yield and fruit quality. -- Research on organic ground cover strategies for apple trees was completed and an on-farm organic research project to improve zinc nutrition in apple trees was continued in Maine. 4. Continue to collaboratively develop and implement with stakeholders a multi-dimensional extension program that addresses their priorities and needs, enables whole farm planning, improves competitiveness, and enhances the ability of growers to grow and market high quality organic apples. --A multi-dimensional extension program continued to provide stakeholders with organic apple information that included on-site visits, orchard tours, newsletter articles, websites, and grower meetings. Organic apple production information and research summaries were distributed to diverse audiences which included commercial apple growers, scientists, extension personnel, industry personnel, undergraduate and graduate students and the general public in the region.

Publications:
Berkett, L.P., Bradshaw, T.L., Griffith, M.C., Kingsley-Richards, S.L., Darby, H.M., Parsons, R.L., Moran, R.E. and Garcia, M.E. 2013. Disease and Arthropod Evaluation of Five Apple Cultivars under Organic Management in Vermont, USA. Acta Hort. (ISHS) 1001:235-248

Bradshaw, T.L., Berkett, L.P., Griffith, M.C., Kingsley-Richards, S.L., Darby, H.M., Parsons, R.L., Moran, R.E. and Garcia, M.E. 2013. Assessment of Kelp Extract Biostimulants on Arthropod Incidence and Damage in a Certified Organic Apple Orchard. Acta Hort. (ISHS) 1001:139-145

Bradshaw, T.L., Berkett, L.P., Griffith, M.C., Kingsley-Richards, S.L., Darby, H.M., Parsons, R.L., Moran, R.E. and Garcia, M.E. 2013. Assessment of Kelp Extract Biostimulants on Disease Incidence and Damage in a Certified Organic Apple Orchard. Acta Hort. (ISHS) 1001:265-271

Bradshaw, T.L., Berkett, L.P., Griffith, M.C., Kingsley-Richards, S.L., Darby, H.M., Parsons, R.L., Moran, R.E. and Garcia, M.E. 2013. Assessment of Kelp Extract Biostimulants on Tree Growth, Yield, and Fruit Quality in a Certified Organic Apple Orchard. Acta Hort. (ISHS) 1001:191-198

Garcia, M.E., Moran, R.E., Berkett, L.P., Bradshaw, T.L., Darby , H.M., Kingsley-Richards, S.L. and Parsons, R.L. 2013. Performance in the Early Production Years of Two Organic Orchards Established by Different Methods: Newly-Planted and Top-Grafted. Acta Hort. (ISHS) 1001:161-165

1d

Cornell University
Northern Grapes: Integrating Viticulture, Winemaking, and Marketing of New Cold-Hardy Cultivars Supporting New and Growing Wineries .

Principal Investigator: L. Berkett

Accomplishments & Outputs:
The Northern Grapes Project is an interdisciplinary research and extension project involving scientists, extension specialists, enologists, and marketing experts across 12 states. The focus of the research are cold-climate wine grape varieties, new to both growers and consumers, which have spawned new small-winery industries in the Northeast and upper Midwest over the past decade. The challenges in these new agricultural endeavors include determining the best growing conditions, how to modify the higher acidity of the grapes and showcase their aroma, and how to build tourism networks to draw visitors to tasting rooms, where a majority of sales take place. Research was conducted in Vermont to evaluate performance of a selection of cold climate wine grape varieties that are important to the developing wine grape industry in the state. Research results and observations were distributed to growers through a newsletter and website. Over 250 people enrolled to receive project information via email and the total number of visits to the cold climate wine grape production website totaled over 6,459. In addition, research results were presented at scientific meetings in the United States and in Europe.

Publications:
Berkett, L.P. 2013. Cold Climate Grape IPM Updates. University of Vermont. 9 issues, 35 pp. http://pss.uvm.edu/grape/newsletters/

Berkett, L.P. 2013. The Cold Climate Grape Production website. University of Vermont. http://pss.uvm.edu/grape/

Berkett, L.P., Bradshaw, T.L., and S.L Kingsley-Richards. 2013. Cold Climate Wine Grape Cultivars: A New Crop in the Northeast and Upper Midwest Regions of the USA. Book of Abstracts of the IOBC-WPRS Working Group Meeting on Integrated Protection and Production in Viticulture. Ascona, Switzerland: 123.

Berkett, L.P., Bradshaw, T.L., and S.L Kingsley-Richards. 2013. Disease evaluation of selected cold climate wine grape cultivars in Vermont, USA. Book of Abstracts of the IOBC-WPRS Working Group Meeting on Integrated Protection and Production in Viticulture. Ascona, Switzerland: 132.

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Winter Bud Injury. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMwinterbudinjury.html

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Winegrape Field Testing for Harvest. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMfieldtesting.html

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Vine Phenology. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMphenology.html

Bradshaw, T.L., Berkett, L.P., and S.L. Kingsley-Richards. 2013. Horticultural Assessment of Eight Cold-Hardy Table Grape Cultivars in Vermont, 2009-2012. HortScience 48(9): 2-3

Bradshaw, T.L., Berkett, L.P., and S.L. Kingsley-Richards. 2013. Horticultural Assessment of Eight Cold-Hardy Wine Grape Cultivars in Vermont, 2009-2012. HortScience 48(9): 7

Bradshaw, T. L. and Berkett, L.P. 2013. Considerations for Starting a Commercial Winegrape Vineyard in Vermont. http://pss.uvm.edu/grape/ConsiderationsForStartingACommercial VineyardInVermont.pdf

Kingsley-Richards, S. and L.P. Berkett. 2013. Through the Grapevine: UVM Vineyard Blog 2013. http://pss.uvm.edu/grape/UVMvineyard/UVM2012Blog/UVM2012Blog.html

1d

Vermont Agency of Agriculture, Food, and Markets
Enhancing the Competitiveness and Sustainability of Organic Apple Production in Vermont .

Principal Investigator: L. Berkett

Accomplishments & Outputs:
Vermont apple growers need research-generated information on the feasibility of new apple varieties that are disease-resistant to enhance the competitiveness and sustainability of organic apple production in Vermont. Growers want to know what the potential is for sustainable and profitable organic apple production with the newer apple varieties since the older varieties, such as McIntosh, pose significant challenges to sustainable organic production because of their susceptibility to Apple Scab, a major fungal disease of apple. Current organic disease management requires multiple applications of organic fungicides to control this disease on McIntosh and other susceptible varieties that are planted in organic orchards. Previous research has shown that the organic fungicides (i.e., sulfur, lime sulfur) used for Apple Scab disease management have negative impacts on the trees (reduced photosynthesis) and beneficial organisms in the ecosystem. Incorporating disease-resistant apple varieties into organic production systems will allow for a significant reduction in organic fungicide use. Since the 1940s, numerous apple varieties resistant to Apple Scab have been bred and released but few have gained acceptance with U.S. apple growers or marketers. In the past, growers have cited uncertainty with the horticultural requirements of growing these varieties and consumer reluctance to buy new apple varieties as reasons for not adopting scab-resistant varieties on a large scale. However, in recent studies, trained taste panels have identified scab-resistant varieties that potentially are more acceptable to consumers, and consumers in general appear to be more willing to buy new varieties. While disease-resistant varieties offer the potential for organic apple production to be more sustainable and profitable, growers need information on which disease-resistant varieties are best suited for Vermont conditions. In addition, with the impending discontinuation of antibiotic use in organically-certified apple orchards, it is important to provide growers with information regarding the susceptibility of these new varieties to Fire Blight, a potentially devastating bacterial disease. This new information would be of significant value to growers since Fire Blight can potentially kill a tree in one growing season and can rapidly destroy a whole orchard if severe. Currently, there are no alternatives as effective as the standard antibiotic in managing this serious disease and growers would be significantly increasing their disease risk by planting apple varieties susceptible to Fire Blight. An organic orchard containing the most promising, new apple varieties that are disease resistant was established to research their potential for enhancing the sustainability and competitiveness of Vermont organic apple production. This is a long-term project and grant funds were used to support the second year of orchard establishment and research data collection.

Publications:
Berkett, L.P. 2012. Orchard Observations. April 11 issue. 5 pp. http://www.uvm.edu/~organica/ListservesBlogs/OrchardObservations04112012.pdf

Berkett, L.P. 2012. Orchard Observations. April 25 issue. 7 pp. http://www.uvm.edu/~organica/ListservesBlogs/OrchardObservations04252012.pdf

Berkett, L.P. 2012. Orchard Observations. May 9 issue. 3 pp. http://www.uvm.edu/~organicaListservesBlogsOrchardObservations 05092012.pdf

Berkett, L.P. 2012. Orchard Observations. May 24 issue. 4 pp. http://www.uvm.edu/~organica/ListservesBlogs/OrchardObservations05242012.pdf

Berkett, L.P. 2012. Orchard Observations. June 11 issue. 6 pp. http://www.uvm.edu/~organica/ListservesBlogs/OrchardObservations06112012.pdf

Berkett, L.P. 2012. Orchard Observations. June 26 issue. 3 pp. http://www.uvm.edu/~organica/ListservesBlogs/OrchardObservations06262012.pdf

1d

Hatch/Multistate
Multi-State Evaluation of Winegrape Cultivars and Clones .

Principal Investigator: L. Berkett

Accomplishments & Outputs:
Cold climate wine grape production is an emerging new food system in Vermont and the region offering exciting value-added and agri-tourism economic opportunities. A key challenge to this young industry is the selection of wine grape cultivars which will consistently produce high quality fruit under our variable environmental conditions. High quality fruit is the basis for quality wine production. This project is evaluating viticultural and Integrated Pest Management (IPM) characteristics of new cold climate wine grape cultivars and is part of a national research project (NE1020). This past growing season was the fifth year of grape production in the research vineyard containing six wine grape varieties which was planted at the University of Vermont Horticultural Research Center. Observations and data were collected in the vineyard throughout the growing season and at harvest to determine differences between the varieties. Standard research protocols including those established by the NE1020 technical committee were used. Results and observations were distributed to growers through a newsletter and website. Over 250 people enrolled to receive project information via email and the total number of visits to the cold climate wine grape production website totaled over 6,459. In addition, research results were presented at scientific meetings in the United States and in Europe.

Publications:
Berkett, L.P. 2013. Cold Climate Grape IPM Updates. University of Vermont. 9 issues, 35 pp. http://pss.uvm.edu/grape/newsletters/

Berkett, L.P. 2013. The Cold Climate Grape Production website. University of Vermont. http://pss.uvm.edu/grape/

Berkett, L.P., Bradshaw, T.L., and S.L Kingsley-Richards. 2013. Cold Climate Wine Grape Cultivars: A New Crop in the Northeast and Upper Midwest Regions of the USA. Book of Abstracts of the IOBC-WPRS Working Group Meeting on Integrated Protection and Production in Viticulture. Ascona, Switzerland: 123.

Berkett, L.P., Bradshaw, T.L., and S.L Kingsley-Richards. 2013. Disease evaluation of selected cold climate wine grape cultivars in Vermont, USA. Book of Abstracts of the IOBC-WPRS Working Group Meeting on Integrated Protection and Production in Viticulture. Ascona, Switzerland: 132.

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Winter Bud Injury. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMwinterbudinjury.html

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Winegrape Field Testing for Harvest. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMfieldtesting.html

Berkett, L.P., Kingsley-Richards, S.L., and T.L. Bradshaw. 2013. 2013 Vine Phenology. University of Vermont. http://pss.uvm.edu/grape/UVMvineyard/2013UVMphenology.html

Bradshaw, T.L., Berkett, L.P., and S.L. Kingsley-Richards. 2013. Horticultural Assessment of Eight Cold-Hardy Table Grape Cultivars in Vermont, 2009-2012. HortScience 48(9): 2-3.

Bradshaw, T.L., Berkett, L.P., and S.L. Kingsley-Richards. 2013. Horticultural Assessment of Eight Cold-Hardy Wine Grape Cultivars in Vermont, 2009-2012. HortScience 48(9): 7

Bradshaw, T. L. and Berkett, L.P. 2013. Considerations for Starting a Commercial Winegrape Vineyard in Vermont. http://pss.uvm.edu/grape/ConsiderationsForStartingACommercial VineyardInVermont.pdf

Kingsley-Richards, S. and L.P. Berkett. 2013. Through the Grapevine: UVM Vineyard Blog 2013. http://pss.uvm.edu/grape/UVMvineyard/UVM2012Blog/UVM2012Blog.html

1e

New England Floriculture Inc
Hardiness of Herbaceous Perennials - Reacclimation after Midwinter Thawing .

Principal Investigator: L. Perry

Accomplishments & Outputs:
Many retail and wholesale nurseries grow herbaceous perennials, and many such growers in northern climates need to successfully overwinter plants either in production, or held from the previous year, in overwintering structures such as greenhouses. They need to know what low temperatures plants will tolerate and survive, in order to adjust their heating and culture accordingly. In many cases perennials may be overwintered at lower temperatures, with lower heat inputs, thus saving not only fuel but lessening the impact of burning excess fuel on global warming. Such results will result in significant economic savings in both fuel and plant losses, and are being shared in presentations, grower articles, and online (perrysperennials.info). Based on these results, future studies will evaluate the effect of longer deacclimation periods up to 10 days. In January when plants have been shown in previous studies to have maximum hardiness, two perennial species (Becky Shasta daisy and Route 66 coreopsis) were exposed to various periods of deacclimation including 1, 3 or 5 days of 16C for 8 hours during the day, returning to 4C at night; or 3 days at 16C. There were insufficient plants of coreopsis for the 1 day treatment. Plants were then immediately placed at -2C for 2 days to freeze solid, then after 6 replicate plants were removed, remaining plants were subjected to controlled freezing temperatures of -5, -8, -11, and -14C for a half hour once they reached each, and then returned to a 3-5C greenhouse. In April, plant regrowth was assessed using a visual rating scale of 1-5 (1 = dead, 3-5 = increasing salable quality, specific traits assessed varying by cultivar). Such ratings have been shown in previous studies to be reflective of dry weights. For coreopsis, there were no significant difference among freezing temperatures after 3 days at 16C or 5 days of fluctuating temperatures. The only significant differences among temperature treatments from either the control (no prior deacclimation to freezing) or 3 days of fluctuating temperatures with non-saleable (rated less than 3) plants at -14C. This relatively new and popular coreopsis, from this study, appears both hardy to low temperatures and possible affected by several days of deacclimation only at the lowest (-14C) temperature exposure. Becky Shasta daisy had plants rated significantly lower after -8 and lower exposures after all deacclimation treatments, and from the control, with most plants dead or mostly so. Plants exposed to -5 rated lower in all cases (3 range) than those after -2C (4 to 5 range). Those at two treatments were significantly different between these two temperatures, however, this may be an artifact due to higher standard errors of the mean (0.2 compared to 0.1 for other treatments). This popular daisy, from this study, appears hardy to only -5C, with no differences among these deacclimation treatments at either -2 or -5C.

Publications:
none

1e

Hatch
Herbaceous Perennial Hardiness .

Principal Investigator: L. Perry

Accomplishments & Outputs:
Many retail and wholesale nurseries grow herbaceous perennials, and many such growers in northern climates need to successfully overwinter plants either in production, or held from the previous year, in overwintering structures such as greenhouses. They need to know what low temperatures plants will tolerate and survive, in order to adjust their heating and culture accordingly. In many cases perennials may be overwintered at lower temperatures, with lower heat inputs, thus saving not only fuel but lessening the impact of burning excess fuel on climate change. Such results will result in significant economic savings in both fuel and plant losses, and are being shared in presentations, grower articles, and online (perrysperennials.info). Based on these results, other perennial species important to the trade will be similarly frozen and evaluated, including another catmint, coreopsis, gaillardia, and Russian sage to compare, along with additional new species not previously evaluated. In early January and late February, 6 perennial species were exposed to controlled freezing temperatures of -2, -5, -8, -11, and -14C and then returned to a 3-5C greenhouse. Two other perennials were similarly frozen in January only, due to insufficient numbers from prior plant losses. In April, plant regrowth was assessed using a visual rating scale of 1-5 (1 = dead, 3-5 = increasing salable quality, specific traits assessed varying by cultivar). Such ratings have been shown in previous studies to be reflective of dry weights. In January, four species (Joanna Reed catmint, Arizona Apricot Gaillardia, Filigran Russian sage, and Lucky Star coneflower showed no differences among temperatures, hardy to the lowest one (-14C). Plants at low temperatures, however, did not survive for these species after February freezing. Plants frozen to -14C for all were dead, and those from -11C exposure were significantly lower rated (unsaleable or below rating of 3) for catmint and the Russian sage. Arkansas bluestar or amsonia plants are both saleable after all freezing exposures, with no significant differences, on both dates except for those at -14C in January which were significantly lower rated than the -2 exposure plants. For the other three species in January (Pow Wow Wildberry coneflower, Crown of Gold evening primrose, Redshift coreopsis), plants from -11 or -14C exposure were dead or most plants died. Other plants of the coneflower rated low, below saleable; other plants of both the evening primrose and coreopsis (exposed to -2, -5, or -8C) rated saleable (above 3) with no significant differences; those of the coreopsis rating highly at 4.7 or above. For these species in February, only coneflower plants at the top two temperatures were barely saleable with low ratings (3.3 or less); coreopsis plants were similar to January results, those at the top three temperatures rating highly and those plants dead at the lowest two temperatures.

Publications:
none

1e

Environmental Protection Agency
Scouting to Reduce Chemical Pesticide Use in Greenhouse Ornamentals .

Principal Investigator: M. Skinner

Accomplishments & Outputs:
The goal of this project was to help growers of greenhouse ornamentals in Northern New England establish scouting programs by training seasonal workers, thereby reducing growers use of agrochemicals and increase their adoption of integrated pest management (IPM) and biological control. Two training programs were held attended by over 45 growers from ME, NH and VT. Over 60 percent of participants were staff, the primary target audience. The rest were owners or head growers. The 1-day training sessions included sections on establishing a scouting program, developing data sheets and entering data, insect and damage identification, disease diagnosis and soil and water testing methods and nutrient deficiency symptoms. Each participant developed an individualized scouting plan for their greenhouse operation, which they were to implement. Surveys were sent to participating growers before the workshop to gain baseline needs data. The surveys indicated that over 108 greenhouses were operated by workshop attendants totaling over 2.1 mil sq ft used for production of ornamentals and/or vegetables. Aphids, thrips and whiteflies were the most problematic insect pests for growers to manage. Powdery mildew, botrytis blight and bacterial leaf spots were the most challenging diseases. Environmental control and fertility issues were the top production issues that troubled attendants. Around 88 percent of attendants indicated that they used biological controls, and 94 percent said they planned to use chemical insecticides and/or fertilizers in the upcoming growing season. Over 88 percent of growers said they use sticky cards, disinfect areas, remove weeds and use new soil for pest management purposes. After the sessions, over 85 percent of the participants indicated they learned new skills they intend to use during the upcoming growing season. Almost 90 percent said they made new contacts they will draw on for information in the future. Growers indicated they liked the small size of the sessions, which allowed for more hands-on learning. One grower said she learned more in one hour of this training session than she learns in a whole day at most other workshops. Over 93 percent of the attendees used biological control in the last year, 51 percent used some form of a chemical pesticide. A listserv was established among growers to encourage interaction among the group. As a result of the session, growers that had fertilizer issues modified their type and rates and now monitor their water and fertilizer rates more frequently. Some purchased new meters. Many growers either designed new scouting routines or modified existing ones to monitor pests and diseases more effectively. Others indicated that they are more diligent in their pest management decisions choosing less toxic chemical insecticides if needed. Growers indicated that they would like to keep having hands on workshops and some indicated more IPM for vegetable production. It was clear from the outcome that small interactive workshops are more effective at disseminating information about IPM than large lecture-style conferences, though the smaller sessions are more labor intensive and expensive to organize.

Publications:
none

1e

American Floral Endowment
Granular Formulations of Insect-Killing Fungi with Plant-Medicated IPM Systems for Thrips .

Principal Investigator: M. Skinner

Accomplishments & Outputs:
This project evaluates a guardian plant system for thrips in spring bedding plants. It uses a flowering marigold on which predatory mites (Neoseiulus cucumeris) are released on the flowers and foliage; a millet-based granular formulation of Beauveria bassiana, an insect-killing fungus, mixed into the upper surface of the potting mix; and a thrips pheromone lure attached to a stake. This system is based on the concept that adult thrips are attracted from the crop to the marigolds, where they reproduce. Attractiveness may be enhanced by a pheromone lure. Thrips adults and immatures serve as prey for predatory mites on the flowers and foliage, sustaining mites and encouraging their dispersal throughout the crop. In the absence of thrips, mites feed on marigold pollen. Thrips that escape predation drop to the soil to pupate, where they become infected with the fungus. The granular formulation enables the fungus to colonize the potting mix thus, eliminating the need for reapplication. Trials were conducted at 5 operations in VT; 1 in NH. At 3 sites, trials were in single gutter-connected greenhouses. At the other sites, trials were in 3 separate, side-by-side, greenhouses. Each greenhouse contained 6 treatments: 1) marigold with a granular formulation of an experimental strain of B. bassiana incorporated into the upper layer of the potting mix, thrips pheromone lure and predatory mites; 2) marigold with a granular formulation of the B. bassiana GHA strain in the potting mix, thrips lure and predatory mites; 3) marigold with thrips lure only; 4) marigold without thrips lure, fungus or mites; 5) yellow sticky card with thrips lure; and 6) yellow sticky card without lure. For 12 wks marigolds and sticky cards and 3 plants located within 1 m of each marigold were monitored bi-weekly for thrips and other arthropods and damage. At the start and end of the experiment, soil samples were taken to determine the level of fungal inoculum. In both years, over the 12 wk, higher numbers of thrips were detected on marigolds than on random crop plants near the marigolds. At all sites, there was more damage on the marigolds than on the crop. On many occasions, marigolds without the mite/fungus treatment had over 50 percent foliar damage; whereas nearby crop plants had less than 10 percent damage. There were more thrips on marigolds with no mite/fungus treatment than on those treated with mites and fungi. Over 95 percent of the thrips were on the marigolds. Both fungal treatments provided similar thrips suppression. Differences in thrips numbers on plants with and without lures were not significant. Low numbers of mites were detected on the marigolds, but not on adjacent crop plants. Results demonstrate the effectiveness of the system for reducing thrips numbers and damage. Predatory mites and both fungi persisted for over 12 wks, providing a sustained source for biocontrols. Though mites were found in marigolds, they were not found on adjacent crop plants, suggesting that mite dispersal from the marigolds is minimal, or they escape detection. The lures did not enhance thrips attractiveness of the marigolds or sticky cards.

Publications:
Skinner, M., C.E. Frank Sullivan, S. Gouli and B.L. Parker. 2013. Granular formulations of insect-killing fungi in combination with plant-mediated IPM systems for thrips. American Floral Endowment Special Research Report #216. http://www.endowment.org/images/stories/research/Research_Insect_Management/216srr.pdf

1e

New Hampshire Plant Growers Association
Novel Methods for Energy Conservation in Northern New England Greenhouses .

Principal Investigator: M. Skinner

Accomplishments & Outputs:
Three major problems with the bubble system were identified that needed redesigning to improve operation and to enable it to run cost-effectively over time: 1) Leakage of soap solution along the gutter; 2) Attachment of the foam generators; 3) Support for the greenhouse plastic over the foam generators. A specially formulated biodegradable and non-phytotoxic soap solution is used in the system, to which a small amount of biodegradable antifreeze is added to prevent freezing. Aluminum gutters run the length of the greenhouse to collect the solution as bubbles degrade, and return it to the tanks. Nonetheless, there have been recurrent problems with leaks due to expansion and contraction of the gutter sections. To eliminate this problem the aluminum gutters had to be welded, requiring removal of the gutters. The welded joints were filed down to facilitate installation of the splines which hold the plastic in place. The bubble system relies on a novel foam generating device into which the soap solution is injected. Air is forced through the generator which is capped with a fine mesh, producing a thick stream of bubbles that flows into the void between the two layers of plastic. The foam generators (14 per side) were attached with rubber grommets along a 4-inch diam. aluminum pipe at the peak of the greenhouse. The grommets were a weak link. The greenhouse plastic pressed down on the generators and sheared off the grommets, detaching them. A method of attaching the generators was designed so they were screwed into the pipe with a threaded coupling which was inserted into the pipe, after which the generators were reattached. The foam generators are positioned between the two layers of plastic covering the greenhouse. Squirrel fans are used to keep the space between the plastic inflated. To reduce the risk of puncturing the plastic, it is critical to keep it from resting on the foam generators. It was necessary to devise a system to hold the plastic above the generators. Special risers were designed to raise the black pipe above the generator, fabricated from PVC. They were designed to fit within the trough that held the black pipe, and are held in place with zip ties. As a result of these refinements, the bubble system operated from Dec. 2013 to Apr. 2013; the longest period of operation since it was installed three years ago. A significant reduction in gas consumption was observed in the bubble house compared to the one with the curtain or the unimproved greenhouse. Information on this project was presented at two events. In July, we hosted the Board of Directors of the American Floral Endowment, a leading national floriculture industry association, and gave a tour of the research site. This group of 20 leaders in the industry was fascinated with the project and impressed by the grower involvement. In addition we gave a presentation to over 45 greenhouse growers from New Hampshire for the annual meeting of the New Hampshire Horticulture Association. The growers were interested in the technology, and look forward to an on-site demonstration in the coming year.

Publications:
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1e

Natural Resources Conservation Service/Department of Agriculture
Innovative Technologies for Energy Conservation in Vermont Greenhouses .

Principal Investigator: M. Skinner

Accomplishments & Outputs:
The goal of this project is to improve the design of the bubble greenhouse insulation system and implement improvements in a demonstration greenhouse. The bubble system insulates hoop houses by injecting soap bubbles between the two plastic layers. The project compares energy savings among greenhouses with a bubble system, an energy curtain and an unimproved house. It also includes demonstration and outreach activities, such as production of a video to show the value of energy conservation to growers. The three objectives are: 1) install and demonstrate a control mechanism to limit the quantity of bubbles injected into the plastic layers; 2) install light sensitive controller on the curtain; and 3) produce and disseminate a video on the bubble and energy curtain insulation systems. The bubble generation mechanism is controlled by a digital timer. We wanted to design and install a device to automatically shut off the system when it was full. Because bubble quality changes depending on the bubble solution concentration and the weather conditions (temperature and humidity), it was difficult to set the timer to maximize on the bubbles but not over fill the system, which results in bubbles escaping out the damper. We have not been able to design a suitable electronic sensor system but continue to consider options. We investigated several approaches and discussed the matter with engineers. We tested a system designed by an engineer from Israel, who maintains the rights to the bubble system. Despite these efforts, to date none have worked effectively. We continue to consider options to solve this problem. A light-sensitive curtain controller was installed and is in operation. This assured that the energy curtain was closing at the same time as the bubble system started up which made energy consumption comparisons between the two houses more comparable. This system is much more dependable than relying on a greenhouse worker remembering to close the curtain. The grower has used the controller in the summer months as well, by making the curtain close at very high solar radiation levels, so it provides shading and cooling. Work is underway to produce a short video for growers describing the systems being tested, and how they reduce energy consumption. Initial filming of the greenhouse in operation was shot and digitized. A 3D animated drawing of the bubble greenhouse and the curtain system was produced. These drawings show the mechanical components and will be included in the video. There were two opportunities for us to present information about energy conservation, and specifically these systems during the spring and summer. We hosted the Board of Directors of the American Floral Endowment, a leading national floriculture industry association, and toured of the research site. This group of 20 leaders in the industry was fascinated with the project and impressed by the grower involvement. We also gave a presentation to over 45 greenhouse growers from New Hampshire for the annual meeting of the New Hampshire Horticulture Association. The growers were interested in the technology, and look forward to an on-site demonstration in the coming year.

Publications:
none

1e

Hatch/Multistate
The Working Group on Improving Microbial Control of Arthropod Pests .

Principal Investigator: M. Skinner

Accomplishments & Outputs:
This is a multi-faceted project to enhance the marigold guardian plant system. It assesses the comparative attractiveness of yellow marigolds to western flower thrips (WFT) in non-flowering and flowering stages of various types of bedding plants of different colors; evaluates the compatibility of disease-suppressing treatments with liquid and granular formulations of experimental and registered entomopathogenic fungi; and tests persistence of granular formulations of experimental isolates and commercial fungal products in potting media. In Yr 1 greenhouse cage trials were conducted in the spring with flowering and non-flowering blue and yellow calibrachoas; and in the late summer with white and red petunias. In the spring trial, yellow calibrachoas had more WFT than the blue one; and averaged over the entire trial period, the flowering marigolds had more WFT than both the flowering and non-flowering calibrachoa treatments. When foliar damage was averaged over the experiment, marigolds had greater foliar damage than the non-flowering calibrachoas. Less damage was observed on the marigolds than on the flowering calibrachoas. In the summer trial, there were more WFT on the red petunias than on the white ones; and when averaged over the experiment, there were more WFT on marigolds than on the petunias. When foliar damage was averaged over the experiment, marigolds had slightly greater foliar damage than the non-flowering petunias, and slightly less damage than the flowering petunias. Data from both trials indicate that marigolds are highly attractive to thrips in both flowering and non-flowering calibrachoas and petunias, but tend to be more attractive when the crop plants are in non-flowering stages. In addition, by using marigolds in non-flowering calibrachoas and petunias, damage on plants could be less than if used when the calibrachoas are flowering. Lab experiments were done to test the compatibility of select chemical fungicides (Subdue MAXX and Banrot) and the antagonist Trichoderma harzianum to insect-killing fungi (Beauveria bassiana (GHA, a commercial strain, and an experimental strain), Metarhizium anisopliae). The entomopathogenic fungi were assessed for their survival and growth rate following a combined application with the fungicides. T. harzianum was combined with either B. bassiana or M. anisopliae and the rate of growth of the fungal colonies and conidial germination was determined. For the first 2 d both colonies grew independently without a significant impact on each other. After that colonies of the entomopathogens did not increase in size, whereas the T. harzianum colonies increased by 22-27 mm. After four days colonies of the entomopathogenic fungi were completely surrounded by T. harzianum, though they remained viable for 1 mo. Studies are underway to assess conidial germination, testing different timing and combinations of the entomopathogenic fungi, the chemical herbicides and T. harzianum. Germination is being evaluated after 18, 20, 22, 24 and 48 hr. Similar trials will be done to test the growth rate of the entomopathogenic fungi in the future.

Publications:
none