FORAGE-BASED PARASITE CONTROL IN SHEEP AND GOATS IN THE NORTHEAST U.S.

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: OTHER GRANTS
• Reporting Frequency: Annual
• Accession No.: 0230779
• Grant No.: 2012-51300-20320
• Cumulative Award Amt.: $1,850,360.00
• Proposal No.: 2012-02290
• Project Start Date: Sep 1, 2012
• Project End Date: Aug 31, 2017
• Grant Year: 2012
• Program Code: [113.A]- Organic Agriculture Research & Extension Initiative

Recipient Organization
WEST VIRGINIA UNIVERSITY
886 CHESTNUT RIDGE RD RM 202
MORGANTOWN,WV 26505-2742

Performing Department
Plant & Soil Sciences

Non Technical Summary
This proposal establishes a multi-state, multi-disciplinary project to enhance productivity and profitability of organic livestock production in the Northeast US. Specifically, we plan to conduct coordinated research into the single most important limiting factor in organic sheep and goat production: gastro-intestinal nematode (GIN) parasites. We will evaluate high tannin forages and assess their efficacy for suppressing GIN parasites, identify the mechanism(s) of suppression, then demonstrate these improved pasture management practices on existing farms and extend these practices to organic and transitioning growers. A variety of intestinal roundworm species infect small ruminants in the Northeast and throughout the world. Haemonchus contortus (H. contortus, AKA barber pole worm) is generally considered the most important and has a life cycle typical of many other species. Symptoms of infection are associated with blood loss, and include anemia, pale color of mucous membrane, edema or bottle-jaw, severe loss of condition and poor growth. In severe infections, death may occur within a few days of the appearance of initial symptoms. Young lambs and kids tend to be most susceptible, as adults tend to develop resistance as they mature. Monitoring programs rely on clinical symptoms or on fecal egg counts (FEC), and growers must treat with anthelmintics, resulting in loss of organic certification. Low levels of infection may result in poor growth, sometimes without any other symptoms, thus making production unprofitable. One of the most promising findings in the last fifteen years has been the discovery that consumption of some forages containing condensed tannins (CT) can suppress GIN infection. CT-containing forages, by themselves, may have the potential to provide adequate control of GIN parasites. Tannins, in low to moderate concentration, increase nitrogen utilization in ruminants and subsequently have been shown to increase growth over and above the observed anthelmintic effect. Unfortunately, few studies have fully characterized the tannin profiles of the forages used. We will screen a range of cultivars, isolate the tannins involved, and screen for biological activity in vitro and in vivo. We will bring powerful analytical capability (MALDI-TOF MS) to characterize the molecular weight distribution and structural features of CT from the most promising BFT accessions based on agronomic performance, and in vitro and in vivo anthelmintic activity. Field and lab trials at URI and WVU will assess the BFT varieties for their effect on sheep by examining, anthelminthic effects, immune responses in sheep, and overall performance of organic lamb in the field. Additional grower trials will be coordinated through Cornell University to demonstrate grower adoption of these approaches.

Animal Health Component

40%

Research Effort Categories

Basic

30%

Applied

40%

Developmental

30%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
302	1610	1010	30%
307	1641	1110	50%
313	3610	1090	20%

Knowledge Area
302 - Nutrient Utilization in Animals; 313 - Internal Parasites in Animals; 307 - Animal Management Systems;

Subject Of Investigation
3610 - Sheep, live animal; 1610 - Pasture; 1641 - Trefoil;

Field Of Science
1090 - Immunology; 1010 - Nutrition and metabolism; 1110 - Parasitology;

Keywords

haemonchus

sheep

birdsfoot trefoil

condensed tannins

gastrointestinal nematodes

immunity

grazing management

small ruminants

Goals / Objectives
This proposal establishes a multi-state, multi-disciplinary project to enhance productivity and profitability of organic livestock production in the Northeast US. Specifically, we plan to conduct coordinated research into the single most important limiting factor in organic sheep and goat production: gastro-intestinal nematode (GIN) parasites. We will evaluate high tannin forages and assess their efficacy for suppressing GIN parasites, identify the mechanism(s) of suppression, then demonstrate these improved pasture management practices on existing farms and extend these practices to organic and transitioning growers. Research Objectives: 1) Evaluate birdsfoot trefoil cultivars to assess agronomic characteristics and tannin contents, identify which condensed tannins are critical for anthelminthic activity and characterize the structure/function relationship of the tannin profiles; 2) Assess the effects of high tannin forages on the larval and adult nematode response in vitro and in vivo, and measure key immune responses of the host animal; and 3) Evaluate herd health and economic outcomes of pasture mixes with high-tannin birdsfoot trefoil for suppression of intestinal parasites. Outreach and Extension Objectives: 4) Demonstrate the real-world potential to reduce parasite burden in small ruminants through on-farm comparisons of pastures with and without birdsfoot trefoil forages; 5) Promote grower adoption of organic parasite control practices through IPM workshops, field days, and presentations at grower organizations, and document changes in grower behavior; 6) Use traditional outreach tools such as short fact sheets, and articles in agriculture and livestock publications of interest to growers, as well as electronic media such as eXtension and eOrganic to develop Communities of Practice and provide results to growers, crop advisors, Extension agents, and others; and 7) Assist farmers to realize increased earnings by documenting four criteria among participating growers: a) reduced use of synthetic dewormers, b) increased animal productivity attributed to lower worm loads, c) reduced number of animals removed from organic marketing channels because of dewormer intervention and d) successful transition from conventional to organic farming due to increased confidence in coping with parasite issues.

Project Methods
Birdsfoot trefoil (BFT) accessions identified as having moderate to high levels of total condensed tannins (CTs) will be screened to assess tannin profiles, as well as plant survival, growth habit, flowering time, forage yield, regrowth after cutting, and any insect or disease damage. Hay will be used for profiling of condensed tannins and in vitro testing of anthelminic activity. CTs will be isolated from select BFT accessions. Purity of each CT fraction will be determined by MALDI-TOF MS, HPLC and elemental analysis and CTs will be quantified by the DMAC method and analyzed by mass spectrometry. Anthelmintic properties of BFT varieties will be evaluated using in vitro assays of H. contortus egg hatch, larval development and exsheathment and in vivo assays with: a) fistulated sheep fed selected varieties of BFT hay to assess larval exsheathment in the rumen, and b) lambs fed BFT hay and experimentally infected with H. contortus to determine the effect of BFT hay consumption on FEC, blood packed cell volume, weight gain and abomasal worm burden. Parasite-naive lambs (i.e., not previously exposed to GIN parasites), will be infected with 10,000 L3 H. contortus 4 weeks after weaning for 28 days and then treated with the anthelmintic drug Levamisole and rested for 4 weeks to test immune responses to BFT. Lymphocyte proliferation assays will be performed with 0, 10, 20, 40 μg/ml CT extract. The effects of feeding BFT on immunity will also be assessed using lambs with three infection statuses; naive, primary (first exposure to H. contortus) and challenge (second exposure to H. contortus). After feeding CT-rations for 2 weeks, lambs will then be experimentally infected with 10,000 L3 H. contortus larvae. Serum IgG, IgA and IgE antibody responses will be quantified on a total and antigen-specific basis. Fecal samples will be collected weekly and FEC enumerated. Four commercial cultivars will be established at the WVU Organic Research Farm in 2012 to evaluate productivity and phenology, and to produce adequate forage to assess palatability to sheep and to provide material as needed for feeding studies. A second multi-year field experiment will evaluate lamb performance and response to forages and grazing management in pastures interseeded with commercially available cultivars of a low-CT BFT, a high-CT BFT, or red clover as a control. Lambs will be exposed to either low or high levels of field infection achieved through grazing management. The experiment will consist of three grazing phases: 1) nursing lambs and ewes contaminate pastures; 2) nursing lambs and ewes re-graze contaminated pastures to achieve low or high infection rates, and 3) weaned lambs re-graze pastures to evaluate growth and maintenance of organic status with low or high CT BFTs. The experiment will test whether lambs exposed to a high level of infestation can be "rescued" by grazing high tannin forage pastures with an evasive management strategy and whether there is residual anthelmintic activity in lambs on BFT pastures.

Progress 09/01/12 to 08/31/17

Outputs
Target Audience:The Target Audiences were sheep and goat producers in the Northeast US. Specifically, we expanded field testing birdsfoot trefoil pastures with eight producers in 2016 to manage parasites in sheep or goats. Fiveparasite workshops and seven grower presentations were held from Oct. 2016 to November 2017, including one focused on veterinarians, with 15 veterinarian participants. Over 200 people participated in the workshops, and 167 people received FAMACHA training and certification.Seven field days and pasture walks focusing on organic management of sheep and goats and forage management to reduce parasite infection in sheep and goats were held during 2017. Workshops on alternative forages with potential to assist in parasite management were also taught by participating researchers and farmers. Changes/Problems:Sustainability of BFT stands and how to incorporate them into a grazing plan permitting weaned lambs and kids to have access to clean BFT fields for 4 weeks or more remains a major obstacle to wider adoption by growers. Two case study farms with average stands of 65% and 54% BFT during their grazing studies had the stands greatly reduced in later years with mugwort and/or conventional pasture plants outcompeting BFT. These losses may be attributed to grazing the stands too soon before the first killing frost in the fall (within 4 weeks) or to inability to control rhizome weeds. In contrast other farms completing BFT studies in 2015 or 2016 maintained stands of 37%, 54%, 57%, 86% in 2017. One experienced farm (57% BFT) was successful although they had been fighting an established stand of Canadian Thistle throughout the study. The farmer mowed the field short frequently to organically control this rhizome weed. In one case (37% BFT), the soil pH was 5.3 and never corrected thus Bruce BFT had little competition although more and more grasses came into the field each year possibly due to the BFT improving fertility. In the other cases, farmers were fairly aggressive about mowing the fields to set back conventional pasture plants and allow BFT to grow. Initially farmers and researchers were very concerned about mowing BFT low, as recommendations suggested that mowing below 5 inches slowed regrowth. Therefore farmers sometimes tried to mow above the BFT current height and below competition, for example at 8 inches. These higher mowings appeared to barely set back the weeds and did not provide as much advantage for BFT as mowing at 5 inches appeared to do at other experienced farms. However, more structured studies are needed to clearly determine what mowing heights are best for allowing a BFT stand to compete with weeds, clovers and cool season grasses and remain sustainable. Educational fact sheets for distribution have taken longer to develop than anticipated. One reason for delay is that statistical analyses of the on-farm grazing studies has just recently been completed. It is difficult to create factsheets advising people on the effect of BFT on worm loads and how best to use it and varieties to select when still in the midst of research with no clear answers to these questions. Continued research needs to be done to answer questions such as how to maintain high levels of BFT in stands, and the optimum way to manage pastures to maximize sheep health benefits. The BFT varieties used (Bruce and Pardee) do not act as highly efficient dewormers when grazed for 4 weeks at ≥25% of the biomass, or for longer periods if the proportion of BFT biomass in the forage is < 25 %. Results from the on-farm grazing studies suggest that farmers with a strong agronomy background or who hire out planting / establishment to consultants with such backgrounds can successfully establish stands of ≥25% BFT in the Northeast. Results also indicate that weight gains on BFT are improved for weaned lambs and kids being marketed as grass-fed as compared to same animals on typical forages especially when soil conditions are less optimal (acidic, low fertility, poorly drained) so that clovers and cool season grasses cannot easily compete with BFT. Worm loads and FAMACHA scores were improved over time in some trials for animals grazing BFT versus other forages. However, whether this was due to better nutrition or anthelmintic properties is not proven by these studies. What opportunities for training and professional development has the project provided?At least six graduate students and over 20 undergraduates received training on organic sheep production, nematode parasite management, and research methods as part of this project. Over 600 participants in our Integrated Parasite Control / FAMACHA© Training workshops from 2012 through 2017 received FAMACHA certification. One of the on-farm grazing trials took place at a farm learning center where ten farm interns were trained to take fecal samples and FAMACHA score goats. The List serv, [e-mail] was established in April 2014 to assist communication among the on-farm study farmers, extension staff and researchers with experience managing BFT test plots and grazing trials. Some of the farmers are organic inspectors or grazing consultants. Topics for 2016 have included frost seeding and assessing drought conditions in the region and its impacts on grower plots. How have the results been disseminated to communities of interest?Papers on the anthelmintic efficacy of birdsfoot trefoil and immunological responses of sheep were presented at professional meetings, and manuscripts for publication in peer-reviewed journals are in preparation. Results have been disseminated to grower and practitioner communities of interest primarily through presentations at various winter conferences and symposiums. Now that data from the on-farm planting and grazing studies has been statistically analyzed, factsheets and articles can be prepared for publication on eXtension and eOrganic, and in farmer magazines. Farmers conducting on-farm planting and grazing trials and researchers on the project met at Cornell Univ. in 2014 and again on June 26 - 27, 2017 to share their experiences. Representatives from eight of the farms and four researchers from WVU and URI were able to attend the 2017 meeting which also included Cornell extensionists, graduate students, parasitology lab staff and veterinarians participating in the project. Two other farmers attended online while 2 farmers were unable to attend for medical reasons. During the meeting, researchers and farmers toured two of the case study farms, nine farmers (one online) gave short presentations about their experiences and the 4 researchers provided in-depth presentations overviewing their research. The researcher presentations were videotaped and edited as Utube presentations. However, because the data has not yet been published they were not put on Utube but instead were only put on memory sticks for the four farmers unable to attend the meeting to review. Updated, preliminary results from on-farm grazing trials are available online to farmers through our project website at http://web.uri.edu/sheepngoat/orei-resources/ What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impacts achieved by this project include development of a repeatable protocol for in vitro assays of aqueous extracts of birdsfoot trefoil (BFT)for their effects on larval exsheathment within the rumen of fistulated ewes. The reduction in nematode infection and improved weight gain of lambs on BFT pastures was also observed on several grower farms. Growers who completed the workshops reported increased confidence in their ability to manage nematodes organically, and reported a decrease in costs of dewormer averaging $143 per grower. Objectives 1) Successful organic cultivation of BFT benefits from high seeding rates (e.g., 20 lb/Acre) to compensate for the inability to use herbicides at planting. All commercial cultivars tested appear to grow well in the Northeast US. Among accessions tested, high levels of condensed tannins do not appear to be correlated with poor agronomic performance. BFT hay was 50-60 % birdsfoot trefoil, and 40-50 % weeds. Perennial weed control remains a challenge for organic producers, necessitating new planting, cover crop, and management strategies to sustain good stands. Analysis of Proanthocyanidans suggests that these compounds may not be the only bioactive compounds in BFT. Variation in the ratio of catechins:gallocathecins in BFT accessions varying in anthelmintic efficacymay provide a focus for future work.. Objective 2) An in vitro assay evaluating exsheathment and L3 viability in an aqueous extract of BFT strains was completed using 13 BFT strains. Six of the 13 strains were commercially available in the US. Efficacy ranged from 0 to 75% exsheathment inhibition, and 17 to 92% L3 mortality at a concentration of 25 mg/mL (BFT extract); however efficacy of aqueous extracts was not correlated with proanthocyanidin content. In vivo exsheathment of Haemonchus contortus larvae within the rumen was determined using four fistulated ewes and 2000 H. contortus L3 contained within capsules suspended in the rumen for eight hours. FeedingBFT hay did not inhibit L3 exsheathment in the rumen. A feeding trial was conducted with hay from five varieties of BFT produced in summer of 2016. Fourteen lambs were experimentally infected with H. contortus and fed either BFT or control hay for 6 weeks. There was an apparent decrease in fecal egg count in lambs fed BFT hay. Objective 3) A replicated trial at WVU compared lambs grazing a low-tannin BFT cultivar (Norcen) and a moderately high-tannin cultivar (Pardee) versus a conventional pasture of orchard grass and red clover; using an avoidance grazing (3-day) versus a challenge grazing (7-day) rotation. Lambs grazed for approximately 120 days in 2014, 2015, and 2016, and for 60 days in 2017. Differences in lamb fecal egg count, body condition or weight gain between high and low tannin cultivars were variable, and were usually not statistically significant, however lambs tended to perform better on both trefoil cultivars than on the conventional orchard grass pastures. Lambs on the challenge grazing rotation generally did not differ significantly from those on the avoidance grazing rotation. The proportion of lambs that retained organic status at the end of the grazing trial was 33 % for orchard grass/red clover pastures, but 60 % for BFT. Organic status also was significantly greater for lambs on the avoidance grazing rotation than the challenge grazing rotation. Lambs from 2017 gained better on BFT pastures than on orchard grass, but showed no difference in % Organic lambs, Fecal Egg Counts, or FAMACHA scores. This may have been due to the shorter grazing period, or to a low BFT content of pastures (2-4 %) compared to previous years. Objective 4) Eight Northeast farmers who had established BFT demonstration pastures during 2015 or 2016 were able to conduct valid 8-wk. on-farm grazing trials during the 2017 growing season comparing lamb performance on BFT pastures versus other forages. Lamb weight gain was significantly better on BFT pastures than on conventional pastures for five of eight growers. Animals on BFT pastures were getting better nutrition than their cohorts on control (various other forage) pastures, so it is unclearwhether improvements are due to specific compounds in BFT that are antagonistic to worms, toimproved immunity, orto effects of better nutrition contributing to parasite tolerance. Objective 5) Five introductory goat and sheep parasite workshops, 41 workshops or field days, and 34 small ruminant integrated parasite control / FAMACHA Training workshops have been conducted as project outreach since Sept 1, 2013. Approximately 1523 people attended these events and 668 people became certified in FAMACHA. Participants in FAMACHA workshops showed improvement in before-and-afterquizzes to measure their knowledge of worm life cycles, evasive grazing practices and research on the possible role of high tannin forages and/or copper oxide wire particle oral dosing in worm control. A sampling of participants indicated that ~20% were interested in possibly planting BFT. A follow up survey to evaluate practices adopted or improved as well as economic and production impacts was distributed in March 2017. Past program participants (553)were invited to participate in this survey, and138 surveys were completed. The pasture management changes most adopted by respondents because of attendance at our workshops were increasing the pasture rest period and height of pasture during their pasture rotations. The pasture management practices most often cited as ones they planned to make were planting and grazing annual forages such as small grains and brassicas (58%) or planting and grazing a high tannin forage such as BFT (61%). Ninety four respondents indicated that they learned more about the potential of high tannin forages to suppress worm infections in small ruminants, 13 indicated that they had planted a high tannin forage as a result of the workshops. Objective 6)The primary form of outreach has been the 80 workshops conducted through the project that discusses the on-farm BFT grazing studies. The project resources developed during 2014 and 2015 continue to be distributed at workshops and field days and maintained on the project website at http://web.uri.edu/sheepngoat/orei-resources/. These resources include: Project Informational Flyer, BFT Establishment Fact Sheet and recordkeeping form, updated Case Studies on planting and establishing BFT organically. Objective 7) Fifty two respondents to the March 2017 survey indicated that they reduced their use of synthetic dewormers as a result of practices they adopted or improved upon because of our programs. Reported annual savings in dewormer costs ranged from $30 to $500, averaging $143. Although a third of the farmers reported improved animals performance, only a few provided monetary values for this improvement. When reported, earnings from improved animal productivity ranged from $100 to $1400 (average $557), savings in health costs (vet bills, labor to treat parasitized animals) ranged from $100 to $1000 (average $300), and savings from reduced animal mortality ranged from $400 to $2000 (average $1180). Three farms indicated that as a result of adoption of these practices they have reduced the number of animals they have to remove from their organic marketing channels while 52 farms reported that they have seen a reduction in the number of animals they have to deworm or the number of times a year they have to deworm. About 71% of respondents planning to raise goats or sheep reported increased confidence in their ability to cope with GIN parasites in their future flock because of our programs while 71% of veterinarians and educators also responded that they felt more confident advising goat and sheep farmers on GIN parasite management.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Stanton, tatiana. L. and M.L. Thonney (2016). Integrated control of internal parasites in small ruminants In: Proceedings of the 22nd Annual Dairy Sheep Assoc. of North America Symposium, Ithaca, NY (2-4 Dec, 2016). Pages 81-89.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Barone, C., Ferguson, S., Zajac, A., Brown, R., Reed, J., Krueger, C., Petersson, K. In vitro screening of the anthelmintic efficacy of birdsfoot trefoil commercial varieties and cultivars against ovine Haemonchus contortus. American Society of Animal Science Joint Annual Meeting (ASAS JAM), Salt Lake City, Utah, July 18-22, 2016. Abstract No 699. Oral Presentation.
• Type: Websites Status: Published Year Published: 2016 Citation: website, Northeast Small Ruminant Parasite Control at http://web.uri.edu/sheepngoat/orei/.
• Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Garza, J.J., S. P. Greiner, S.A. Bowdridge. 2017. Ovine neutrophils bind and impair H. contortus third-stage larvae in a breed and infection status-dependent manner requiring vital NET formation. Parasit. Immunol. Submitted
• Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Mata-Padrino, D.J., D.P. Belesky, C.D. Crawford, B. Walsh, S.A. Bowdridge. 2017. Effects of grazing birdsfoot trefoil-enriched pasture on managing Haemonchus contortus infection in Suffolk crossbred lambs. J. Anim. Sci. Submitted
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Garza, J.J., S.P. Greiner, S.A. Bowdridge. 2017. Serum-mediated H. contortus larval aggregation differs by larval stage and is enhanced by complement. Parasit. Immunol. 39:e12409.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Shepherd, E.A., J.J. Garza, S.P Greiner, S.A. Bowdridge. 2017. Effects of ovine peripheral blood mononuclear cells on Haemonchus contortus larval death in vitro. Parasit. Immunol. 39:e12424

Progress 09/01/15 to 08/31/16

Outputs
Target Audience:The Target Audiences in Year 4 were sheep and goat producers in the Northeast US. Specifically, we continued field testing trefoil pastures with four producers in 2016 to manage parasites in sheep or goats. Nine parasite workshops and four grower presentations were held from Oct. 2015 to September 2016. Over 200 people participated in the workshops. Seventy three people received FAMACHA certification. Two field days focusing on organic management of sheep and goats and forage management to reduce parasite infection in sheep and goats were held during 2016. Workshops on alternative forages with potential to assist in parasite management were also taught by participating researchers and farmers. Changes/Problems:URIlost the midsummer harvest of the six commercial varieties of BFT grown under organic management due to unexpectedly heavy rainfall on the freshly cut BFT.No further harvesting of BFT was possible in 2016 due to drought. We missed the June flowering on the seed production plots due to delay in obtaining bumblebees. Plants were clipped, and flowered again in August, but seed set was low due to heavy leafhopper presence and drought stress. Although the BFT stands were productive, Canada thistles were a problem on one farm resulting in the stand having to be mowed 3 days before the grazing study commenced and again during the study to prevent the thistles from going to seed. Another farm also opted to mow during their grazing trial the BFT stand was getting too mature. However, regrowth was rapid at both farms. Worms were absent at another farm. A major problem occurred at a farm where yearling goats were pulled from a good sized herd traditionally grazing 4 acres of diverse forage every 4 days for multiflora rose and barberry control in cattle pastures. The yearlings were not content to intensively graze small paddocks of BFT for 4 day periods. Instead two jumpers eventually led the entire treatment group to escape from the paddocks regularly and the study had to be halted.Drought caused most of the participatingfarmers to spend more time on their flocks during the summer and therefore did notconduct afield day. These farmers have committed to making presentations at winter conferences and/or producer meetings. Clear evidence for, and analysis of, the specific bioactive compounds in BFT has not been elucidated.Previously reported assays for nematode control efficacy were found to be impractical, however new assays have beendeveloped. Work in this area will continue in 2017. What opportunities for training and professional development has the project provided?Graduate and undergraduate students at WVU, URI and Cornell have obtained hands-on training in organic sheep management, FAMACHA and Body Condition Scoring, and analysis of Fecal Egg Counts. Seventy three participants in our Integrated Parasite Control / FAMACHA© Training workshops from January through July 2016 in NY and RI received FAMACHA certification. One of the on-farm grazing trials took place at a farm learning center where four farm interns were trained to take fecal samples and FAMACHA score goats. The List serv, [e-mail] was established in April 2014 to assist communication among the on-farm study farmers, extension staff and researchers with experience managing BFT test plots and grazing trials. Some of the farmers are organic inspectors or grazing consultants. Topics for 2016 have included frost seeding and assessing drought conditions in the region and its impacts on grower plots. How have the results been disseminated to communities of interest?Papers on the anthelmintic efficacy of birdsfoot trefoil and immunological responses of sheep were presented at the American Society of Animal Science Joint Annual Meeting, July 18-22, Salt Lake City, Utah. Current project activities and/or preliminary results were disseminated to Northeast sheep and goat farmers and educators at 11 parasite workshops and 2 field days throughout the project year. Updated, preliminary results from on-farm grazing trials are available online to farmers through our project website at http://web.uri.edu/sheepngoat/orei-resources/. What do you plan to do during the next reporting period to accomplish the goals?What do you plan to do during the next reporting period to accomplish the goals? URI will harvest BFT for hay during the summer of 2017. Based upon the feeding trial results with the fistulated ewes, it is anticipated that lambs infected with gastrointestinal nematodes, will be fed either control hay or the most promising commercial varieties of BFT to test for anthelmintic efficacy in vivo. Test an organic extract of the 45 BFT accessions and 6 commercial varieties using in vitro anthelmintic assays. Expand the seed production blocks using greenhouse-grown plants from seed harvested in 2016, and are planning to harvest seed again in 2017 Wisconsin will continue analysis of condensed tanninsd in birdsfoot trefoil accessions. WVU will repeat the replicatedlamb grazing trial in BFT pastures.. Cornell will statistically analyze the data from ten on-farm grazing trials that took place in 2015 and 2016. Conduct 2017 grazing trials with 2 farms frost seeding BFT. Present preliminary results from the BFT on-farm grazing trials at the Dairy Sheep Assoc. of North America 22nd Symposium on Dec 2-4, 2016, Cornell Univ., Ithaca, NY. IPM workshops, field days, and other conference presentations are planned for 2017. Continue to maintain and update project website with new resources and events including detailed progress bulletin and more conclusive results from the on-farm case studies. Publish preliminary results in farmer newsletters and magazines. Summarize results from the follow up survey sent out in spring/summer 2016. Administer the follow-up survey during spring/early summer 2017 to the 2016 program participants to document changes in producer behavior and project impacts and outcomes, and summarize the results in late summer. Project Diorectors will organize a final meeting with the farmers participating in our grazing trials to share research results from the research institutes (URI, WVU, Univ. of Wisconsin - Madison) Develop an eXtension fact sheet summarizing the results from the BFT on-farm grazing studies. Publish results from these studies.

Impacts
What was accomplished under these goals? Objectives1) Evaluate birdsfoot trefoil cultivars to assess agronomic characteristics and tannin contents, identify which condensed tannins are critical for anthelminthic activity and characterize the structure/function relationship of the tannin profiles. Winter survival data were collected on the BFT nursery. The late winter/early spring of 2016 was extremely harsh for plants, with extreme freeze events in February and April, and most accessions suffered extensive losses. Seed production blocks were established for PI 193725, PI 325379, and a mixed population of accessions that had shown high efficacy and good agronomic performance. Seed was harvested from all blocks, but yields were low due to drought. Analysis of Proanthocyanidans suggests that these compounds may not be the only bioactive compounds in birdsfoot trefoil.Suppression of gastrointestinal nematodes appears to be correlated with reactivity with proteins.Assays based on the 8-terminal flavan unit may not be well correlated with bioactivity.For example, the cultivar Bruse has high levels of proanthocyanidans (condensed tannins), but performed poorly on nematode assays. 2) Assess the effects of high tannin forages on the larval and adult nematode response in vitro and in vivo, and measure key immune responses of the host animal; The protocol for in vivo larval exsheathment assay using the fistulated ewes has been completed.A feeding trail of 3 commercial varieties of BFT plus a control is underway.It is expected to be completed during the 2016/2017 winter. Expression of IL-4 was detected in resistant hair sheep, but not in susceptible wool sheep, indicating that upregulation of associated genes may be associated with mechanisms that reduce establishment of nematode parasites. 3) Evaluate herd health and economic outcomes of pasture mixes with high-tannin birdsfoot trefoil for suppression of intestinal parasites. Outreach and Extension Objectives: A replicated trial at WVU compared lambs fed a low-tannin cultivar (Norcen) and a moderately high-tannin cultivar (Pardee) to a conventional pasture of orchard grass and red clover; using an avoidance grazing (3-day) versus a challenge grazing (7-day) rotation. Differences in lamb fecal egg count, body condition or weight gain between high and low tannin cultivars were not statistically significant in 2014 or 2015, however the challenge grazing rotation, as expected, resulted in lambs with lower body condition and lower body weight, compared to the avoidance grazing rotation. In 2016, Body condition and weight were significantly greater, and Famacha scores were lower, for lambs on birdsfoot trefoil cultivars than on orchard grass/red clover at most dates. Fecal Egg Counts in 2016 usually did not differ significantly, and there were few differences between lambs on the 3-day versus 7-day grazing rotations. After 3 years of trials, the proportion of lambs that retained organic status at the end of the grazing trial was 33 % for orchard grass/red clover pastures, but 60 % for birdsfoot trefoil. Trials at grower farms in 2015 showed lamb weight gain was equal to or better than on conventional pastures, however fecal egg counts and FAMACHA scores did not differ significantly. Data from the 2016 trials are being analyzed. 4) Demonstrate the real-world potential to reduce parasite burden in small ruminants through on-farm comparisons of pastures with and without birdsfoot trefoil forages; Four Northeast farmers (NY) who had established BFT demonstration pastures during 2015 conducted 8 wk. on-farm grazing trials during the 2016 growing season comparing performance on BFT pastures versus other forages. Project staff collected fecal egg counts, live weight changes, FAMACHA scores and any health interventions required (deworming etc.) in addition to evaluating the forages. One farm that frost seeded BFT in 2014 and 2015 delayed their grazing trial one year due to severe drought conditions. 5) Promote grower adoption of organic parasite control practices through IPM workshops, field days, and presentations at grower organizations, and document changes in grower behavior; Four Integrated Parasite Control / FAMACHA© Training workshops were conducted from January through July 2016 in NY and RI. Approximately 79 farmers, youth, livestock extension educators and veterinarians attended these workshops; Seventy three participants received FAMACHA certification. A pre and post test was administered to workshop participants to track changes in knowledge. Average scores increased from 51 % to 76%. Forty four percentage of respondents at the FAMACHA training in RI indicated that they planned to implement at least one new practice besides FAMACHA scoring as a result of the training. Seven parasite presentations or workshops targeting goat and sheep farmers were conducted in NY, RI and VT. Both URI and WVU organized summer field days in 2016 for farmers to tour the BFT plots/pastures and learn about ongoing research . 6) Use traditional outreach tools such as short fact sheets, and articles in agriculture and livestock publications of interest to growers, as well as electronic media such as eXtension and eOrganic to develop Communities of Practice and provide results to growers, crop advisors, Extension agents, and others; The project resources developed during 2014 and 2015 continue to be distributed at workshops and field days and maintained on the project website at http://web.uri.edu/sheepngoat/orei-resources/. These resources include: Project Informational Flyer, BFT Establishment Fact Sheet and recordkeeping form, updated Case Studies on planting and establishing BFT organically. 7) Assist farmers to realize increased earnings by documenting four criteria among participating growers: a) reduced use of synthetic dewormers, b) increased animal productivity attributed to lower worm loads, c) reduced number of animals removed from organic marketing channels because of dewormer intervention and d) successful transition from conventional to organic farming due to increased confidence in coping with parasite issues. A follow up survey of 41 questions to evaluate practices adopted or improved as well as economic and production impacts was developed and finalized by Cornell and URI during the winter 2016. Cornell issued an email invitation in March 2016 to 553 program participants throughout the Northeast who had provided contact information and had had sufficient time to begin implementing new parasite control practices and/or document improvements as a result of their education.

Publications

• Type: Journal Articles Status: Submitted Year Published: 2016 Citation: Shepherd, E.A., J.J. Garza, S.P Greiner, S.A. Bowdridge. 2016. Effects of ovine peripheral blood mononuclear cells on Haemonchus contortus larval death in vitro. Parasit. Immunol. In review
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Garza, J.J., S.P. Greiner, S.A. Bowdridge. 2016. Serum-mediated H. contortus larval aggregation differs by larval stage and is enhanced by complement. Parasit. Immunol. In revision.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Jacobs, J.R., K.N. Sommers, A.M. Zajac, D.R. Notter, S.A. Bowdridge. 2016. Early IL-4 gene expression in abomasum is a critical difference in resistance to H. contortus. Parasit. Immunol. 38:333-339.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Barone, C., Ferguson, S., Zajac, A., Brown, R., Reed, J., Krueger, C., Petersson, K. In vitro screening of the anthelmintic efficacy of birdsfoot trefoil commercial varieties and cultivars against ovine Haemonchus contortus. American Society of Animal Science Joint Annual Meeting (ASAS JAM), Salt Lake City, Utah, July 18-22, 2016. Abstract No 699. Oral Presentation.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Garza, J., S.A. Bowdridge. 2016. Serum induces H. contortus larval aggregation via complement and antibody complexes that differ by larval stage. 61st Annual Meeting AAVP San Antonio, TX
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Garza, J., S.A. Bowdridge. 2016. Haemonchus contortus induced neutrophil extracellular trap formation differs between resistant and susceptible breeds of sheep. 61st Annual Meeting AAVP San Antonio, TX
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Garza, J., S.A. Bowdridge. 2016. Haemonchus contortus fourth stage larval excretory/secretory protein inhibits serum-mediated larval aggregation. 61st Annual Meeting AAVP San Antonio, TX
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Jacobs, J., S.A. Bowdridge. 2016. Interleukin-4 and its downstream targets are rapidly upregulated in immune cells of St. Croix sheepexposed to H. contortus larval antigen in vitro. 61st Annual Meeting AAVP San Antonio, TX
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2016 Citation: Russ, B., J. Garza, S.A. Bowdridge. 2016. Isolation of third stage H. contortus cuticle and cuticle protein. 61st Annual Meeting AAVP San Antonio, TX

Progress 09/01/14 to 08/31/15

Outputs
Target Audience:The Target Audiences in Year3 were sheep and goat producers in the Northeast US. Specifically, we started field testing trefoil pastures with sixproducers in 2015 to manage parasites in sheep or goats. Sixteen parasite workshops were held from Dec 2014 to October 20145 Over 404 people participated in the workshops. Fifty three people received FAMACHA certification. Five field days focusing on organic management of sheep and goats and forage management to reduce parasite infection in sheep and goats were held during 2015. Three workshops on alternative forages with potential to assist in parasite management were also taught by participating researchers and farmers. Changes/Problems:We were only able to take one cutting of BFT for hay last year due to unavailability of haying equipment in early August. Designing a container that can be suspended in the rumen to securely contain the ensheathed larva but still allow for free flow of rumen "juices" has been problematic.Previously reported methods in the literature result in a high percentage of escaped larvae and ultimately result in an infected fistulated ewe.We are close to solving this issue. Grfazing trials at WVU have been dleayed by poor stand establishment in 2014. BFT stands in 2015 were substantially improved, and we expect a full complement of replicates in 2016. As anticipated, there have been some challenges and concerns for the 11 demonstration farms. Problems have included animal loss from predators, extremely low worm populations during the grazing study, difficulty providing a similar group of control animals, low BFT yields because of adverse weather challenges and/or compulsions to mow the BFT stands in the midst of the grazing study. Six farms conducted grazing trials in 2015. However, the amount of BFT available for animals to consume was limiting on two farms and worms were absent on another farm, leaving 3 farms with good parameters for comparison. What opportunities for training and professional development has the project provided?Eight Integrated Parasite Control / FAMACHA Training workshops provided training on integrated parasite management and FAMACHA certification to 149 people from November 2014 to October 2015. Approximately 200 farmers, youth and extension livestock educators attended introductory parasite management workshops (4), field days (6) or symposium presentations (1) providing information on innovations in sheep and goat parasite management and showcasing project activities. Many of the workshops included training in fecal worm egg identification and counting. The 2015 grazing trials provided opportunities for 3 Cornell undergraduates to become prolific at quantitatively analyzing and reporting small ruminant fecal samples including how to do highly specialized PNA assays using a lectin binding stain. One Cornell undergraduate obtained an extension internship to learn agronomy field evaluation techniques while working side by side with extension field staff at two of the demonstration farms. However, funding for these students was provided by cosponsors of the project rather than OREI. The List-serv, [e-mail] was established in April 2014 to assist communication among the on-farm study farmers, extension staff and researchers with experience managing BFT test plots and grazing trials. Some of the farmers are organic inspectors or grazing consultants. Thus the interchange between farmers and staff has been very educational. Example topics for 2015 have included: fertility management, weed and nurse crop management, and effective seeding methods for the on-farm demonstration BFT pastures. How have the results been disseminated to communities of interest?Dr. tatiana Stanton, Cornell University is a member of the eXtension Goat Industry team and attended a meeting in February 2015 at Tuskegee University, Alabama. Dr. Stanton updated the resources section with the OREI project website and will expand sections on the role of forage management in small ruminant parasite control, incorporating resources and knowledge gained by this grant project with assistance from project partners at URI and Virginia Tech. URI presented a poster on the anthelmintic efficacy of commercial varieties and cultivars of birdsfoot trefoil against Haemonchus contortus at the 2015 Annual Meeting of the American Association of Veterinary Parasitologists, July 2015, Boston, MA. Dr. Stanton presented a research report on parasite management in sheep and goats that included information on planting and establishing Birdsfoot trefoil organically on two of the demonstration farms at the Eastern Northern NY Ag. Dev. Program Annual Meeting, Miner Institute, Feb 27, 2015, Chazy, NY. 26 participants, mostly extension agronomy educators and farmers.Fifteen Cornell Cooperative Extension livestock educators participated in a November 2014 In-Service Training that included a 1 ½ hr. session on "innovations in sheep goat parasite management" where the rationale for this BFT project and problems encountered in establishing BFT organically were discussed in depth. Cornell Univ. presented preliminary res results from one on-farm study comparing effects of grazing BFT and/or dosing with COWP on worm counts and lamb performance during the Northeastern IPM Center's First Annual Online Conference , Oct 20, 2015. 20 speakers, 100+ online participants. Current project activities and/or preliminary results were disseminated to sheep and goat farmers at 12 parasite workshops and 7 field days throughout the project year. What do you plan to do during the next reporting period to accomplish the goals?It is anticipated that BFT feeding trials using the fistulated ewes will begin in January 2016. Based upon the feeding trial results with the fistulated ewes, it is anticipated that lambs, experimentally infected with Haemonchus contortus, will be feed either control hay or the most promising commercial varieties of BFT to test for anthelmintic efficacy in vivo. An organic extract of the 45 BFT accessions and 6 commercial varieties will be testing using in vitro anthelmintic assays. The grazing trials at WVU will be repeated in 2016, and immunological assays will be conducted at WVU using condensed tannin extracts. We will analyze and summarize 2015 data for the 6 on-farm grazing trials and the 5 new BFT seedings. and conduct 2016 grazing trials with the 5 farms that seeded BFT during 2015. Sustainability of BFT plantings from the 6 farms where BFT was established in 2014 and grazed in 2015 will be assessed. We hope to repeat grazing trials on two farms that participated in grazing trials in 2015. IPM workshops, field days, and other conference presentations are planned for 2016. Preliminary results will be publishedin farmer newsletters and magazines.A follow-up survey to program participants will be administeredto document changes in producer behavior and project impacts and outcomes.

Impacts
What was accomplished under these goals? Objective 1. Field evaluation of accessions at URI is continuing, and approximately 2000 pounds of Birdfsfoot trefoil (BFT) hay from six commercial varieties was harvested in 2015 for lambfeeding trials. UWisconsin has isolated 3.5 g of tannin extract from the variety Bruce for use in immune and in vitro assays. Objective 2, URI hasplaced rumenfistulas in four ewes and is continuing development of in vivo assay methods to be usedfor feeding trials in 2016. In vitro assays assessin anthelminthic efficacy, egg hatch and L1/L2 mortality were conduicted on an aqueous extract of forty-five BFT accessions and six commercial varieties. Anthelminthis effocacay differed among varieties at 3 mg/ml. Using aqueous extracts of BFT powder, egg hatch inhibition at 3 mg powder/mlranged from zero to 100 %, and one BFT variety induced 100 % inhibition at 0.75 mg/ml/butthere was no correlation between inhibition of egg hatch and the DMAC concentration of condensed tannins in the freeze-dried BFT powder. Objective 3. A replicated trial at WVU compared lambs fed a low-tannin cultivar (Norcen) and a moderately high-tanin cultivar (Pardee) to a conventional pasture of orchard grass and red clover; using an avoidance grazing (3-day_ versus a challenge grazing (7-day) rotation. Differences in lamb fecal egg count, body condition or weight gain between high and low tannin cultivars were not statistically significant in 2014 or 2015, however the challenge grazing rotation, as expected, resulted in lambs with lower body condition and lower body weigth, compared to the avoidance grazing rotation. Trials at three grower farms showed lamb weight gain was equal to or better than on conventional pastures, however fecal egg counts and FAMACHA scores did not differ significantly. Objective 4. Six Northeast farmers established BFT demonstration pastures in 2014 and conducted on-farm grazing trials in 2015. Five additional growers established BFT demonstration pastures in Spring 2015 and all will compare lamb or goat performance on BFT pastures versus conventional pastures. Lower FAMACHA scores and improved weight gain occurredwith BFT pastures, but data have not yet been analyzed statistically (http://web.uri.edu/sheepngoat/files/NYdemofarms_BFTstudy_2014-2015.pdf). Objective 5.Eight Integrated Parasite Control / FAMACHA Training workshops were conducted from January through October 2015 in PA, NY, VT and RI. Approximately 150 people participated in these workshops.Pre and post quizzes to measure changes in knowledge about integrated parasite management were completed by participants at five of the workshops. Average before and after results for the percentage of questions answered correctly changed from 55.2% to 77.5%. Participants completed post quiz evaluations in three VT, one RI and one NY FAMACHA workshops. Approximately 41%, 42%, 55%, 59% and 60% of the participants, respectively, plan to adopt parasite management practices as a result of attending the workshops including implementing or improving FAMACHA scoring, fecal egg counting, selective deworming and smart drug use, as well as several pasture management practices including improved rotation, lower animal density, mixed species grazing, and use of condensed tannin forages. Seven field days or case study presentations were conducted in NY, RI and WV focusing on current project research and preliminary results from grazing trials at demonstration farms. Objective 6. The project resources developed during 2014 continue to be distributed at workshops and field days and maintained on the project website at http://web.uri.edu/sheepngoat/orei-resources/. These resources include: Project Informational Flyer, BFT Establishment Fact Sheet and recordkeeping form, Case Studies on planting and establishing BFT organically showcasing 4 project demonstration farms. The project website is continually updated and maintained with resources and events. A detailed progress bulletin summarizing all project research and extension activities to date has been drafted and is awaiting completion of the 2015 growing season research activities for final development. It will be posted on the project website and circulated to demonstration farms and producer list servs and email networks. Objective 7. A follow up survey to evaluate practices adopted or improved as well as economic and production impacts has been drafted by Extension staff. It will be completed and administered during Winter 2016 to those program participants who have agreed to provide contact information (total to date ~ 452 participants - will be fewer once we edit for duplicates and members of the same farm), and who have had sufficient time to begin implementing new parasite control practices and/or documenting improvements as a result of their education. The survey can be administered online, or in hard copy format or by phone if preferred by the producer.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Barone, C., Ferguson, S., Brown, R., Zajac, A., Reed, J., Petersson, K. Anthelmintic efficacy of commercial varieties and cultivars of birdsfoot trefoil against Haemonchus contortus. Association of Veterinary Parasitologists (AAVP), Boston, MA, July 11-14, 2015. Abstract No. 94. Poster presentation.

Progress 09/01/13 to 08/31/14

Outputs
Target Audience: The Target Audiences in Year 2 were sheep and goat producers in the Northeast US. Specifically, we surveyed12 sheep producers to serve as participants in field testing in 2015, and hosted a range of workshops. Eighteen parasite workshops were held from Dec 2013 to Nov 2014. Approximately 379 people participated in the workshops. One hundred people received FAMACHA certification. Two field days focusing on organic management of sheep and goats and forage management to reduce parasite infection in sheep and goats were held during 2014. Three workshops on alternative forages with potential to assist in parasite management were also taught by participating researchers and farmers. The presentations are available on the web at http://www.ansci.cornell.edu/goats/CSGSymposium/index.html . Changes/Problems: Because of poor stands in some plots at WVU, only two replicates were used for the grazing study in 2014, and the proportion of BFT in some plots was relatively low. This may have limited any treatment effect. Plots were replanted in fall, 2014, and we expect to have all three replicates in 2015. As a result, an additional greenhouse experiment was developed to better compare condensed tannin concentration among commercial cultivars. Delays in isolating adequate amounts of condensed tannins have delayed immune response studies at WVU, but we anticipate having these underway in 2015. Planting of BFT for on-farm studies was delayed a year into the study because of problems with research test plots. Originally we planned to conduct 16 on-farm grazing trials. We have four additional farmers interested in joining the on farm studies and planting BFT in 2015. However, we need to decide whether to involve them or instead plan on having presently participating farms conduct repeat grazing trials in 2016. What opportunities for training and professional development has the project provided? Two graduate students traveled to the University of Wisconsin for a week-long hands-on training in the preparation and analysis of condensed tannin extracts in the laboratory of Dr. Jess Reed, a collaborator on this project. Dr. Katherine Petersson, also traveled to UW for several days to observe the extraction procedures and meet with Drs. Reed and Krueger on coordinating the BFT extraction phase of these studies. Four researchers, two graduate students, and two staff attended the Cornell Sheep and Goat Symposium, participating in workshops and professional development in pasture management, and sheep health workshops. Grower IPM workshops provided training on intestinal parasite management to approximately 379 people from Dec 2013 to Nov 2014. One hundred participants received FAMACHA certification. The List serv, [e-mail] was established in April 2014 to assist communication among the on-farm study farmers, extension staff and researchers with experience managing BFT test plots and grazing trials. Some of the farmers are organic inspectors or grazing consultants. Thus the interchange between farmers and staff has been very educational. Example topics have included: historic distribution of BFT Fusariun wilt in the Northeast US; addressing low soil P availability issues organically; using prilled lime, millet or killed (roasted) seed of similar size to solve seed volume problems when broadcasting BFT seed; sourcing organically approved lime applicators; on line lime calculators to determine actual ENV rates applied, etc. How have the results been disseminated to communities of interest? A preliminary research report was presented at the Crop Science Society of America annual meeting. Research results were presented in oral presentations by Dr. Jim Kotcon, Dr. Katherine Petersson, Dr. Tatiana Stanton, and Dr. Anne Zajac at the WVU Organic Farm Field Day, and at the Cornell Sheep and Goat Symposium. Results of previous work have been disseminated in grower workshops and field days as noted previously. The eXtension Goat Industry meeting will take place Feb. 17-19, 2015 in Alabama at Tuskegee University. Dr. tatiana Stanton will be attending. In the past she helped develop and edit the Goat Industry's parasite management resources. During 2015, she plans to expand sections on the role of forage management in small ruminant parasite control incorporating the resources this grant project has developed thus far. She will also be expanding sections on the role and considerations of copper oxide wire particles in barber pole worm management programs. What do you plan to do during the next reporting period to accomplish the goals? In 2015, we will collect a third year of agronomic performance data from the URI nursery, and produce seed from the most promising accessions. URI will also evaluate grazing tolerance and possibly feeding preference for sheep using the grazing trial pasture. In addition, sufficient hay will be produced to conduct in vivo analysis of anthelminthic efficacy on fistulated sheep. Due to the extensive amount of time required for freeze drying the various accessions of BFT prior to the preparation of an extract, BFT whole leaf and whole leaf powder will be tested for anthelmintic efficacy using the egg hatch assay, artificial exsheathment and larval development via coproculture. This will serve as a screening method to help direct the selection of candidates for seed propagation as well as prioritization of candidates for preparation of condensed tannin extracts. Four sheep will be fistulated over the winter 2015 and preliminary feeding trials on the BFT hay collected during 2014 will be conducted. Due to the small volume of hay collected, varieties will be combined and tested as a group versus control hay to test the effect of BFT on the exsheathment of Haemonchus contortus infective larvae (L3). After the preliminary feeding trials with BFT in the fistulated sheep using composites of the hay produced in 2014, commercial hay varieties grown at URI and harvested during the 2015 season will be individually fed to the fistulated sheep to test the effect of BFT consumption on larval exsheathment in the rumen. At WVU, the large grazing trial will be repeated and expanded, with the goal of three complete replicates of each treatment and grazing interval. Data to be collected will include lamb weight gain, Fecal Egg Counts, FAMACHA, Body Condition Index, and proportion of lambs that reach market weight with organic status. Forages will again be analyzed for condensed tannin content to assess the role of season and grazing treatment on tannin concentrations. A large pasture of Bruce BFT has been established to produce BFT forage to assess the role of condensed tannins in immune responses of naïve lambs. In addition, a portion of these lambs will be assessed for intestinal worm burden at Virginia Tech. Identification of the bioactive fractions of condensed tannins will continue at UW. On-farm comparisons of internal parasite infection in lambs or kids grazing pastures with and without birdsfoot trefoil forages are planned for Jul/Aug 2015 at 6 participating farms in New York and Maine that established BFT pastures in 2014. Another 5 farms will plant BFT during 2015. We have several invitations to conduct workshops on integrated parasite management in small ruminants throughout the Northeast US.

Impacts
What was accomplished under these goals? Impacts: This project is in early phases, impacts have been associated with preliminary results at research sites. Growers interested in organic sheep and goat production have demonstrated sustained interest in use of high-tannin forages to manage parasites, and 11 growers are now planting birdsfoot trefoil (BFT) pastures for on-farm grazing evaluations in 2015. Because BFT is slow to establish, research results are still preliminary. Progress and activities: Objective 1) Evaluate birdsfoot trefoil cultivars to assess agronomic characteristics and tannin contents, identify which condensed tannins are critical for anthelminthic activity and characterize the structure/function relationship of the tannin profiles We collected a second year of agronomic performance data at University of Rhode Island (URI) for accessions in the nursery. We harvested and froze sufficient leaf tissue of all of the accessions and commercial varieties to produce 1 kg freeze-dried tissue of each accession/variety. Freeze drying is in progress. A second hay field with 6 commercial varieties was established at URI, bringing the total hay acreage to ~ 1 acre per variety. We harvested 2 cuttings of hay from the original field established in September 2012. We established a grazing trial using the 20 best accessions from the nursery and all of the commercial varieties. This trial will be used to evaluate grazing tolerance of the accessions. We repeated a grazing trial at West Virginia University (WVU) to evaluate management practices to control weeds and enhance stands of four commercial cultivars. Birdsfoot trefoil growth and production of condensed tannins were determined before and after grazing at WVU. Grazing and mowing both improved BFT stand density compared to unmanaged plots. Foliage samples from a WVU greenhouse trial were assayed for condensed tannins before and 7 days after an initial clipping using Near InfraRed Spectrometry to evaluate condensed tannin development in response to simulated grazing. We have isolated between 859.8 to 3,472.7 mg of proanthocyanidins (PAC) from Bruce at the University of Wisconsin (UW). We have not yet validated a BFT standard so we are using PAC A2, a dimer that is available from chemical suppliers, and a PAC standard that we isolated and validated from cranberries. We have also carried out MALDI-TOF MS on the isolated BFT PAC from Bruce and these spectra are very informative. In addition to the hetero-oligomers that contain both catechin and gallocatechin monomers, there is another series that appears to be glycosylated with a pentose unit. We are ready to distribute the isolated PAC for use in vitro studies. Objective 2) Assess the effects of high tannin forages on the larval and adult nematode response in vitro and in vivo, and measure key immune responses of the host animal; We conducted a small feeding trial at URI with 4 sheep using the BFT hay harvested in 2014. Although, the sheep readily consumed the BFT hay, there was a negative correlation between the amount consumed and the percentage of BFT in the hay. A larval development assay using copro-culture was established, an in vitro larval development assay is under development at URI. Objective 3) Evaluate herd health and economic outcomes of pasture mixes with high-tannin birdsfoot trefoil for suppression of intestinal parasites. We conducted a large replicated grazing trial at WVU. Two ewes and their 4 lambs were grazed on a low-tannin BFT cultivar, Norcen; a moderately high-tannin cultivar, Pardee; or tannin-free pastures of orchard grass and red clover (OG/RC). Flocks grazed 2 replicates of each pasture for 3 or 7-day periods with 60 or 30-day return frequencies respectively. Forage mass was insufficient for the originally planned grazing duration, so sheep were moved to other pastures in 27 days instead of 60 until weaning, at which time lambs only were returned to the pasture for a second round of grazing. Lamb weight gain was greatest on pastures with the best BFT stands. Although Fecal Egg Counts and weight gain did not differ among pasture treatments or grazing duration, all 8 lambs on Pardee retained organic status at the end of the experiment, compared to only 6 of 8 lambs on Norcen and 4 of 8 on OG/RC. 4) Demonstrate the real-world potential to reduce parasite burden in small ruminants through on-farm comparisons of pastures with and without birdsfoot trefoil forages; Eleven Northeast growers from Maine and New York, interested in participating in the on-farm trials were surveyed. Extension and farmer involvement in the studies were reviewed. Growers have completed soil sampling and most have plantings underway, or will plant BFT in spring 2015. Ten of the participating farms attended a meeting for the OREI project on Oct. 3, 2014 in Ithaca, NY with extension staff and researchers. Researchers gave progress reports and all participants shared experiences planting BFT or preparing land. Requirements for the 2015 grazing trials were reviewed. 5) Promote grower adoption of organic parasite control practices through IPM workshops, field days, and presentations at grower organizations, and document changes in grower behavior; Approximately 379 people participated in parasite workshops held from Dec 2013 to Nov 2014, and 100 people received FAMACHA certification. Pre and post quizzes to measure changes in knowledge about innovations in integrated parasite management were completed by participants at some workshops. Before and after results for the percentage of questions answered correctly increased by an average of 15 % for 8 workshops. Participant evaluations were available from 4 other workshops, scores averaged 4.5 out of 5 possible points where 5 indicated "very useful". Two field days focusing on organic management of sheep and goats and forage management to reduce parasite infection in sheep and goats were held. Three workshops on alternative forages with potential to assist in parasite management were also taught by participating researchers and farmers. 6) Use traditional outreach tools such as short fact sheets, and articles in agriculture and livestock publications of interest to growers, as well as electronic media such as eXtension and eOrganic to develop Communities of Practice and provide results to growers, crop advisors, Extension agents, and others; and OREI Flyer - Overview of the research project and rationale for the potential of BFT as a parasite management tool. Distributed to participants at all succeeding workshops including 200 attendees of the 2014 Cornell Sheep and Goat Symposium. BFT Establishment Fact Sheet and record form - 6-page fact sheet outlining the potential use of high tannin forages to reduce parasite infections, challenges to trying to establish and maintain relatively pure stands of BFT, overview of organic methods of growing and management and educational resources. Case Studies on planting and establishing BFT organically at 4 study farms - Portions of these case studies were presented at workshops during the 2014 Cornell Sheep & Goat Symposium. These resources are available at http://web.uri.edu/sheepngoat/orei-resources/ 7) Assist farmers to realize increased earnings by documenting four criteria among participating growers: a) reduced use of synthetic dewormers, b) increased animal productivity attributed to lower worm loads, c) reduced number of animals removed from organic marketing channels because of dewormer intervention and d) successful transition from conventional to organic farming due to increased confidence in coping with parasite issues. Documentation will begin for growers for whom we have obtained contact information to follow up on farmer changes in behavior. Follow up contact info was obtained from 329 workshop participants in 2013 and 2014.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Uphold, R., J. Kotcon, T. Griggs, and E. Rayburn. 2014. Tannin content in birdsfoot trefoil in response to herbivory and season. Abstract, Crop Science Society of America Annual Meeting. available at: https://scisoc.confex.com/scisoc/2014am/webprogram/Paper89371.html
• Type: Websites Status: Published Year Published: 2014 Citation: Northeast Small Ruminant Parasite Control, at http://web.uri.edu/sheepngoat/orei/ .

Progress 09/01/12 to 08/31/13

Outputs
Target Audience: Target audiences in Year 1 were primarily sheep and goat producers in the Northeast US. We conducted 10 integrated parasite management workshops including FAMACHA certification with NE goat and sheep farmers and veterinary staff and technicians from October 2012 through November 2013. The workshops covered parasite life cycles, dewormer options and methods, parasite monitoring techniques, and the principles of IPM, forage-based parasite control, dewormer resistance, and refugia maintenance. Recent innovations in parasite management such as evasive grazing techniques, the use of high tannin forages and copper oxide wire particles were also discussed. Most workshops also included hands-on fecal egg counting. Twoon-farm field days were also conducted, one in New York, one at West Virginia University, focusing on new methods of parasite control. Participants at most of these workshopswere asked to provide contact information if they were willing to participate further in the project, including monitoring parasite management pratices, and participatng in grower trials. Approximately 240 people particpated in workshops at Cornell, URI, and WVU, of which approximately 150 provided their willingness to collaborate, an indication of the intense interest this project evokes among organic and transitionalsheep producers. List of Small Ruminant Integrated Parasite Management Workshops included FAMACHA certification, hands-on fecal egg counting and discussion of innovative parasite management techniques, etc. Cornell Sheep and Goat Symposium 2012, Cornell Univ. Ithaca, NY Date: October 27, 2012 Time: 8:30 am to 2:15 pm Workshop leaders: Dr. Dwight Bowman, Dr. Steve Hart, Dr. tatiana Stanton (organizer) Participants:40 (primarily farmers) NOFA-NY Winter Conference, Saratoga Springs, NY Date: January 25, 2013 Time: 9 am to 4:30 pm Workshop leaders: Dr. Katherine Petersson, Dr. tatiana Stanton, Dr. Anne Zajac DVM Participants: 25 (3 veterinarians, remainder sheep and goat farmers) Katherine and Anne in New England Date: January 26, 2013Time: Workshop leaders: Dr. Katherine Petersson, Dr. Anne Zajac DVM Participants: Glynwood Farm Center, Cold Springs, NY Date: May 8, 2013 Time: Workshop leaders: Dr. tatiana Stanton Participants: 21 ( 1 veterinarian, remainder were farmers or farm interns from several farms) State University of New York Delhi, Delhi, NY Date: May 18, 2013 Time: 10 am to 2:30 pm Workshop leaders: Dr. tatiana Stanton Participants: 26 (9 farmers, 2 extension educators, 15 vet. tech students) Dighton, MA Date: June 22, 2013Time: Workshop leaders: Dr. Katherine Petersson and Holly Burdett Participants: 6 farmers Cornell Goat & Sheep Health Day, Cornell Univ., Ithaca, NY Date: Sept 21, 2013 Time: 9:15 am to 3 pm Workshop leaders: Dr. Dwight Bowman, Dr. tatiana Stanton, Dr. Michael Thonney Participants: 38 farmers Cornell Cooperative Extension (CCE) of Oneida County, Waterville, NY – did not include hands-on fecal egg counting Date: Oct 12, 2013 Time: 10 am to 3 pm Workshop leaders: Dr. tatiana Stanton Participants: 11 (1 agricultural college student, 1 extension educator, 9 farmers) CCE of Otsego & Rensselaer Counties & the Leatherstocking Shepherds Assoc., New Berlin, NY Date: Oct 26, 2013 Time: 9:30 am to 3:30 pm Workshop leaders: Dr. Rachel Frazer, DVM and Dr. tatiana Stanton Participants: 17 (1 veterinarian, 1 extension educator, 15 farmers) WVU Sheep parasite Workshop, Morgantown, WV Date: nov. 2, 2013Time: 9:00 am to 4 pm Workshop leaders:Dr. Scott Bowdridge and Dr. David Belesky Participants:Approximately 30 Cornell University Veterinary College, Ithaca, NY - did not include hands-on fecal egg counting Date: Nov 10, 2013 Time: 1 pm to 4 pm Workshop leaders: Dr. tatiana Stanton Participants: 7 veterinary students focusing on small ruminants Pasture Field Days on Parasite Management in Sheep and Goats no FAMACHA certification, focused on evasive grazing techniques, condensed tannin forages and methods to incorporate copper wire oxide particles use into an integrated program WVU Organic Research Farm Field day – Morgantown, WV Date:Aug. 8, 2012Time 1 pm to 7 pm Workshop and tour leaders: Dr. James Kotcon, Dr. David Belesky, Dr. Domingo Matapadrino Participants:Approximately 120 total, of which the sheep workshps and tours attracted approximately 40 people. Northeast Organic Farming Assoc. of NY (NOFA-NY) – Field Day at Asgaard Goat Dairy Date: Aug 12, 2013 Time: 1 pm to 4 pm Workshop leaders: Dr. Dwight Bowman, Dr. tatiana Stanton, Rhonda Butler (farm owner), Betsy Hodge (extension educator) Participants: 16 farmers not including Asgaard farm interns and herdsperson, and NOFA extensionist Changes/Problems: Initial grazing trials at WVU scheduled for 2013 were delayed to 2014 due to poor pasture establishment. Additional work is underway to identify imporved agronomic management to encourage establishemnt using organic practices, as well as greenhouse trials to identify organic management approaches to optimize tannin concentrations in forages. What opportunities for training and professional development has the project provided? Two graduate students have begun Masters degree research as part of this project. Two additional students are planned to begin research in 2014. Grower IPM workshops provided training on intestinal parasite management to approximately 260 people in 2012-13. Seven presentations on "Innovations in Goat and Sheep Partasite Management" are planned for 2014. Interest in the project is reflected in the number of participants and the number of invitations for more presentations. How have the results been disseminated to communities of interest? Results of previous work have been disseminated in workshops and field days, as noted previously. Meetings are planned in 2014 to begin development of eOrganic outreach materials. What do you plan to do during the next reporting period to accomplish the goals? Initial grazing trials will commence at WVU in 2014, and pastures for on-farm trials will be seeded so that grazing may commence in 2015. Birdsfoot trefoil is slow to establish, and management practices to assure adequate stands in organic pastures will be evaluated. Extraction and identifcation of the active tannin components will be underway within the next few months, and adequate volumes of birdfoot trefoil hay for feeding trials will be collected in summer 2014. A planning meeting is scheduled for february 2014 to evaluate results and make decisions on preferred birdsfoot trefoil cultivars to recommend for grower trials. Seven presentations on “Innovations in Goat & Sheep Parasite Management” are scheduled this winter in NY and PA during the Northern NY Sheep and Goat Week, the Stone Barn Young Farmer Conference, the Catskills Regional Agriculture Conference, the NOFA-NY Winter Conference and the Northeast Pasture Consortium Conference. Interest in the topic is reflected in the number of invitations that resulted in these engagements.

Impacts
What was accomplished under these goals? Objective 1) Evaluate birdsfoot trefoil cultivars to assess agronomic characteristics and tannin contents, identify which condensed tannins are critical for anthelminthic activity and characterize the structure/function relationship of the tannin profiles; At URI, 51 BFT accessions with potential for high condensed tannin levels were obtained from germplasm repositories and breeders. Seedlings were grown up in the greenhouse and transplanted into a field nursery. Germination rates were sufficient to establish 60 plants per accession in the nursery for 36 accessions. Another 10 accessions were incompletely replicated with 40 plants, and 5 with only 20 plants. Three accessions had extremely poor germination and were not transplanted. Efforts continue to establish additional plants, with the goal of having all accessions fully replicated for 2014. Data were collected on growth habit, height, width, flowering and leaf color, and these nursery plots were rated for tolerance to leafhopper.Samples were collected for tannin analysis, and cuttings of each accession were rooted for greenhosue trials in 2014. Seven quarter-acre plots were seeded at URIin September 2012 at a rate of 8 lbs per acre, along with a nurse crop of oats at 80 lbs/acre. We seeded one plot of each of the following: Empire, Bull, Bruce, Norcen, Pardee, Leo, and a New York landrace. The field was mowed repeatedly with the blade set just above the trefoil to suppress weeds. At WVU, small plots (8 by100 feet) of cultivars Bull, Empire, Leo and Norcenestablished in 2012. In 2013 the plotswere subdivided to assess pasture management effects on stand establishment. Plotseither unmanaged or weregrazed or mowed to manage weeds. Sheep favored grazing birdsfoot tefoil over weeds in all varieties, however regrwoth after 30 days was better than in unmanaged controls. Mowing appeared to promote better stands and more regrowth than grazing. Greenhouse trials were established at WVU with cultivars Empire, Norcen, Pardee, and Grasslands Goldi, representing a range form low to high tannnin concentrations respectively. Plants will be clipped on 30 and 60-day frequencies and tannin concentration of regrowth will be compared to unclipped controls to determine whether BFT responds to age or mowing by altering tannin levels in shoot tissue. 2) Assess the effects of high tannin forages on the larval and adult nematode response in vitro and in vivo, and measure key immune responses of the host animal; At URI, anthelmintic assays required by this project are in varying stages of optimization. The in vitro egg hatch assay and larval exsheathment assay have been optimized and the in vitro larval development assay is in the process of being optimized. Donor lambs have been experimentally infected with Haemonchus contortus and are providing a ready supply of Haemonchus contortus eggs and infective larvae. The protocol for preparation of a crude condensed tannin extract of all BFT samples collected this summer is being optimized. At WVU, methods of cell separation and cell culture conditions were validated. Future experiments on immune responses will be conducted in 2014 as BFT hay and tannin extracts become available.. 3) Evaluate herd health and economic outcomes of pasture mixes with high-tannin birdsfoot trefoil for suppression of intestinal parasites. Field-scale plantings ofBirdsfoot trefoilwere established at WVU and URI. At WVU, four replicate 1-care plots each of Birdsfoot trefoil cultivarsNorcen (low tannin variety) andPardee (moderately high tannin levels) as well as orchard grass/red clover controls were planted in spring 2013. Due to excessive rainfall, weed pressure was high in some plots leading to poor stands. Two reps may need to be re-planted in 2014, however the remaining two reps averaged 20-30 % BFT stand composition and will be used for grazing trials in 2014.Due to slow establishment of BFT pastures, grazing trials were postponed until 2014. Outreach and Extension Objectives: 4) Demonstrate the real-world potential to reduce parasite burden in small ruminants through on-farm comparisons of pastures with and without birdsfoot trefoil forages; We conducted 11 integrated parasite management workshops including FAMACHA certification with NE goat and sheep farmers and veterinary staff and technicians from October 2012 through November 2013. These workshops covered parasite life cycles, dewormer options and methods, parasite monitoring techniques, and the principles of IPM, forage-based parasite control, dewormer resistance, and refugia maintenance. Recent innovations in parasite management such as evasive grazing techniques, the use of high tannin forages and copper oxide wire particles were also discussed. Most workshops also included hands-on fecal egg counting. . 5) Promote grower adoption of organic parasite control practices through IPM workshops, field days, and presentations at grower organizations, and document changes in grower behavior; We conducted2 pasture field days on new methods of parasite control. Pre and post tests were completed by most participants at seven of the IPM/FAMACHA workshops. Workshop participants were also asked to provide their contact information if willing to let us track whether their parasite management practices changed after participating in the workshops. Approximately 260 people participated in the workshops 6) Use traditional outreach tools such as short fact sheets, and articles in agriculture and livestock publications of interest to growers, as well as electronic media such as eXtension and eOrganic to develop Communities of Practice and provide results to growers, crop advisors, Extension agents, and others; and 7) Assist farmers to realize increased earnings by documenting four criteria among participating growers: a) reduced use of synthetic dewormers, b) increased animal productivity attributed to lower worm loads, c) reduced number of animals removed from organic marketing channels because of dewormer intervention and d) successful transition from conventional to organic farming due to increased confidence in coping with parasite issues.

Publications