Progress 08/01/18 to 12/31/19
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Reviewers of our proposal expressed concerns that feeding hop-rich feeds could cause negative effects due to phytoestrogen exposure. Experiments are on-going to assess exposure levels in THYM fed animals to XN and 8PN in rumen fluid samples and in blood. Studies were conducted on serum samples from THYM fed and control ration animals, taken 2 hr after feeding. These were sent to the laboratory of Professor Jan F. Stevens, Oregon State Univ, Corvalis, OR for xanthohumol metabolite analysis by LC/MS/MS. Levels of xanthohumol (XN) and XN metabolites in the THYM and Control serum samples. At this point, some caution is warranted regarding use of THYM supplements in female cattle pending more study. These substances may also have beneficial impacts that warrant further study. This work was not originally considered in the Phase 1 application, and therefore no money was budgeted for this work. This work, totaling $3,937 wasfunded with money reallocated from the infrastructure budget for the project. Unanticipated project management needs necessitated payment for some additional outside support. Payment for these services was pulled from underages on the shipping, materials and supplies, facilities upfit and equipment / facility rentalbudgets. Even with the adjustments, we managed to finish a complex and multi-faceted study while staying under budget. What opportunities for training and professional development has the project provided?This project provided an opportunity for undergraduate and graduate students at Warren Wilson College and NC State University to participate in meaningful scientific research. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Project Impact As US and world population continues to grow, and in the face of a changing global climate, demand for products that improve feed efficiency, reduce disease, reduce greenhouse gas emissions and reuse waste by-products will skyrocket. We have the potential to develop a feed supplement material comprised of natural, plant-based materials that are currently disposed of as waste in the craft brewing process. This brewing waste stream consists of spent trub, hops and yeast. In this report, we refer to the product as THYM (Trub, Hops, Yeast Mixture). This supplement material provides a valuable protein source, has natural antibiotic properties, reduces methane production in cattle, and can improve the rate of weight gain in animals. Our work has provided positive indications in every area of research performed in Phase 1 supporting the benefits of this potential product and the feasibility of producing this at a reasonable cost. Our 9-week feeding trial has been completed showing a positive trend of increased average daily weight gain and feed efficiency with THYM compared to a control. We have developed provisional infrastructure and processes to harvest and mix THYM from a variety of craft beer styles and have determined cost effective means to dry the material. Finally, we have conducted vitro studies, confirming the methane inhibiting properties of the hop acids found in THYM. The results of the work accomplished in Phase I strengthens our belief in the fundamental premise and value, both commercially and in terms of societal good, of this project. Namely, this product has the potential to translate a costly waste product into a protein rich food source for animals, humans and other living organisms and has significant potential co-benefits, such as methane reduction, disease management and positive impacts on overall organism health. Progress toward achieving objectives: The following provides a brief discussion of each of the 6 objectives identified above in terms of the progress made during the reporting period toward achieving these objectives: 1) Develop infrastructure at Highland Brewing Company for harvesting THYM. Work has been completed to develop the infrastructure required to harvest THYM, and to mix and store THYM temporarily prior to dewatering. This includes the purchase of a lobe pump, equipment to support large dewatering bags, and minor infrastructure improvements. Each batch was carefully collected,and the overall makeup was noted for review during the feeding study and the various dewatering studies. 2) Evaluate cost and labor intensity of several small-scale drying methods to reduce the moisture content to 15% or less. Significant work to evaluate and test low technology/ small volume processes was completed early in the project. Subsequent work focused on the evaluation of several high tech, industrial scale solutions. The interim and final technical reports contain details on data collected on filtration/ processing rates, moisture content, hop acid retention, detailed nutritional analysis and other data points. Higher technology, larger scale equipment and processes have the potential to produce a very low moisture, shelf stable product, potentially in a single step process. These studies have also proven a high rate of retention of valuable proteins and hop acids. Preliminary cost/benefit analyses of the technologies indicate that these higher tech solutions can produce the dried product at a reasonable cost, supporting the potential to commercialize this product. 3) Measure total nutritional parameters of THYM as produced and during experimental drying steps. For each batch of THYM data was collected on mix pH, hop acid content, protein content and percent dry matter. See final technical report for details on data collected on retention of proteins and hop acids and other information on the results of dewatering/ drying. In addition to this data, a full nutritional analysis was performed by Dairy One in Syracuse, NY on the raw THYM (pre-dewatering) as well as various samples of dried product from different processing runs with results showing a high rate of retention of valuable proteins and hop acids. The outcomes support the premise that THYM can be dewatered without losing the beneficial proteins, yeast and hop acids. 4) Evaluate THYM as a feed additive in growing cattle diets. A nine-week feeding trial, was performed at NC State University's Mountain Research Station in Waynesville, NC. Feed ration mix data for the experiment, average daily gain, feed efficiency, serum urea nitrogen (SUN), short chain fatty acid (SCFA) concentrations are all reported in detail in the final technical report. The analysis of the feed trial data indicates positive trends for average daily gain and feed efficiency for THYM versus the control group. Overall, THYM had no negative effects on animal health or performance. These results provide favorable evidence that THYM may enhance animal growth and feed efficiency. 5) Extend knowledge of the effects of hop- and protein- rich co-products on bovine rumen metabolism using batch anaerobic cultures with bovine rumen microbe samples obtained at slaughter. Samples of mixed bovine rumen microbes were collected at Wells Jenkins Wells Abattoir, Forrest City, NC. These samples were tested in the lab at Warren Wilson College with various feed substrates and with varying levels of THYM added to certain samples. We have extended earlier findings to positively confirm in 6 separate experiments that the hop-acid-rich craft brewing coproduct THYM has significant rumen microbial activity as judged by strong inhibition of methane inhibition with moderate additions of THYM. These studies also included further evaluation of individual hop acids inhibition of methane production, dose responsiveness, and a detailed analysis of spent yeast from different beer styles. 6) Consolidate the results of experimentation noted above to develop a final process and product that can be sold or franchised around the world anywhere there are concentrations of breweries. We achieved all of the objectives laid out in our Phase I application. As a result of the research conducted in Phase I, we have learned the following: Dewatering THYM using multiple potential technologies andscalable processes can be accomplished at a reasonable cost, with minimal impacts to brewing operations and in a manner that produces a shelf stable, easy to handle, dry product that retains valuable nutrients and hop acids. THYM has the potential to enhance weight gain and feed efficiency in beef cattle. The nutritional characteristics of this product indicate a broad range of potential applications/ markets including additional animal species, and even humans. THYM has the potential to support meaningful reductions in bovine methane production. Repurposing THYM for this purpose reduces (or potentially eliminates) an existing waste stream that is both costly to the brewery owner and negatively impactful to the municipal wastewater treatment system. We intend to expand on the successes of our Phase I work in Phase II to include at least two additional feed trials at major US universities focused on assessing proper dosage levels and expanding the size of the trial to provide more statistical evidence of the potential benefits of THYM as an animal feed product, as well as an in vivo methane study, using rumen fistulated Holstein steers to provide further and stronger evidence of the methane inhibiting properties of THYM. Further research on the dewatering process including downstream processes to package, enhance shelf stability and provide alternatives for the physical property of the finished product (i.e. powders, pellets, blocks, etc.) will be pursued as will research on potential applications beyond beef cattle including, but not limited to dairy, swine, poultry, aquaculture, and humans.
Publications
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Progress 08/01/18 to 07/31/19
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Reviewers of our proposal expressed concerns that feeding hop-rich feeds could cause negative effectsdue to phytoestrogen exposure.Experiments are on-going to assess exposure levels in THYM fedanimals to XN and 8PN in rumen fluid samples and in blood. Blood samples from TMR and TMR+THYManimals have been sent off to Professor Jan Fred Stevens, Oregon State, Corvalis, OR for LC/MS/MSmeasurements (Stevens and Maier, 2016). This work was not originally considered in the Phase 1application, and therefore no money was budgeted for this work. It is expected to cost $4-5000and will be funded with money reallocated from the infrastructure budget for the project. Additional scope and unanticipated project managment needs necessitatedpayment for some additional consulting help. Payment for these services was pulled from underages on the brewery labor and facilities upfit budgets. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?As outlined in our interim technical report, submitted in early Augustand summarized above, we have several major areas of work remaining to complete the Phase 1 objectives. All of these will be completed by December 31, 2019 which is the end of our Phase 1 grant period. These major work areas are as follows: 1) Complete reviews of high-tech, larger scale dewatering technologies and processes. We have work ongoing and planned with major manufacturers and suppliers of dewatering equipment. This includes a variety of technologies including filter presses, drum dryers, and spray dryers. The goal of this work is to prove viability of these technologies, develop an understanding of potential process flows for production scale application, develop high level cost estimates and identify the appropriate next steps for further evaluation and research in subsequent phases. This work is being completed at Highland Brewing Company as well as at various manufacturers/ suppliers facilities. 2) Complete the in vitro studies using bovine rumin microbe samples as outlined in our application. This work is currently ongoing at Warren Wilson College and will be completed by the end of the grant period. 3) Finalize remaining data analysis from the feed trials that were completed, notably to conduct final analysis of feed efficiency for this study. This work is being conducted by NC State University and will becompleted by the end of the grant period. 4) The next section of this report (Changes / Problems) containsa detailed discussion of work that has been added to the scope of this project to address concerns that feeding hop-rich feeds could cause negative effects due to phytoestrogen exposure. This work is being conducted at Oregon State University and will be completed by the end of the grant period.
Impacts
What was accomplished under these goals?
Project Impact As US and world population continues to grow, and in the face of a changing global climate, demand for products that improve feed efficiency, reduce disease, reduce greenhouse gas emissions and reuse waste by-products will skyrocket. As outlined in our original Phase 1 application, we have the potential to develop a feed supplement material comprised of natural, plant-based materials that are currently disposed of as waste in the craft brewing process. This feed supplement material provides a valuable protein source, has natural antibiotic properties, reduces methane production in cattle, and can improve the rate of weight gain in animals. The process we propose also has the potential to provide significant positive benefits to municipal wastewater treatment systems from reduced suspended solids loading and reduced Biological Oxygen Demand (BOD), both of which are significant issues in these systems. Our work thus far has provided positive indications in every area of research that support the benefits of this potential product and the feasibility of producing this at a reasonable cost. In summary, our 9-week feeding trial has been completed showing a positive trend of increased average daily weight gain with Rumensin® (7%) and THYM compared to a control. In addition, we have developed provisional infrastructure and processes to create THYM batches from a variety of craft beer styles. We have also conducted research on multiple approaches to dewatering and stabilizing THYM and have concluded that simple, low technology manual processes can cost effectively dewater THYM with high retention of valuable proteins and hop acids. Finally, we are conducting in vitro studies, which will help bolster the data proving the methane inhibiting properties of the hop acids found in THYM. The results of the work accomplished thus far strengthen our belief in the fundamental premise and value of this project, both commercially and in terms of societal good. Progress toward achieving objectives: Significant work has been completed toward accomplishment of the project objectives above. The Interim Technical Report, submitted on August 6, 2019 provides extensive detail on the work completed, including documentation of data, statistical analysis and preliminary conclusions. 1) Develop infrastructure at Highland Brewing Company for harvesting THYM. Major activities completed - work has been completed to develop the infrastructure required to harvest THYM, and to mix and store THYM temporarily prior to dewatering. This includes the purchase of a lobe pump, equipment to support large dewatering bags, and minor pipe infrastructure improvements. Data Collected - Data was collected for each batch on the input streams (e.g. the beer styles and batches collected for each batch of mixed THYM).Because craft brewing involves many different styles, every batch of THYM was unique in makeup. Each batch was carefully collected and the overall makeup was noted for review during the feeding study. Subsequent collections were small for specific drying technologies, but the overall makeup of the THYM followed the same similar procedures. This work has been completed in full. 2) Evaluate cost and labor intensity of several small-scale drying methods to reduce the moisture content to 15% or less. Major activities completed - Significant work has been completed on evaluation and testing of low technology/ small volume processes. Work is in progress on the evaluation of several high tech, industrial scale solutions. Data Collected - See interim technical report for data collected on filtration rates, moisture content, retention of proteins and hop acids and other key statistics. The end result of this work will be the cost/benefit analyses of all the technologies that will help to guide technology used, test the usability of the product, and identify technologies that are appropriate for individual end users. Summary statistics and discussion of results - See interim technical report Key outcomes or other accomplishments realized - The results of the work to date has proven that small scale, low-tech solutions can effectively reduce moisture content to a range of levels, including as low as 10% when coupled with air drying. These studies have also proven a high rate of retention of valuable proteins and hop acids. This work is approximately 50% complete, with ongoing work to explore larger scale technologies and processes. 3) Measure total nutritional parameters of THYM as produced and during experimental drying steps. Major activities completed - For each batch of THYM data was collected on mix pH, hop acid content, protein content and percent dry matter. Data Collected - See interim technical report for data collected on retention of proteins and hop acids and other information on the results of dewatering/ drying. Summary statistics and discussion of results - The results of the work to date has proven that small scale, low-tech solutions to dewater THYM result in a high rate of retention of valuable proteins and hop acids. Key outcomes or other accomplishments realized - The outcomes thus far support the premise that THYM can be dewatered without losing the beneficial proteins, yeast and hop acids. Ongoing work is underway to evaluate the shelf stability of the dried THYM. This work is ongoing. 4) Evaluate THYM as a feed additive in growing cattle diets. Major activities completed - Completion of nine week feeding trial, described in detail in the Technical Report. Data Collected - Feed ration mix data for the experiment, average daily gain, feed efficiency, serum urea nitrogen (SUN), short chain fatty acid (SCFA) concentrations. Summary statistics and discussion of results - Work is still underway to fully analyze the feed trial results, however average daily gain did show a positive trend for THYM versus the control group. Key outcomes or other accomplishments realized - Preliminary results provide favorable evidence that THYM may enhance animal growth and feed efficiency. As such, THYM may provide protein supplement benefits that allow for a reduction in other, potentially more expensive protein in the feed ration mix. This work is largely complete, with the exception of a few remaining data analysis tasks. 5) Extend knowledge of the effects of hop- and protein- rich co-products on bovine rumen metabolism using batch anaerobic cultures with bovine rumen microbe samples obtained at slaughter. Major activities completed - Experiments are ongoing to strengthen our hypothesis that alpha and beta hop acid are responsible for the methane and SCFA modulations seen when spent craft brewers yeast is incubated with rumen microbes. This work is ongoing and will be concluded by the end of the Phase 1 Grant. 6) Consolidate the results of experimentation noted above to develop a final process and product that can be sold or franchised around the world anywhere there are concentrations of breweries. At the conclusion of the Phase 1 grant period we anticipate completing all of the objectives identified in the application as well as additional objectives added to the work subsequent to the application and described later in this progress report. At that point we expect to have developed a roadmap to further evaluate and perfect the production process including batch mixing, dewatering, stabilizing and finished product production.In subsequent phases of work we anticipate the need to expand feeding trials to include larger populations, different segments of the feeding cycle and varying dosage in the feed mix. We may also expand these feeding trials to other animals and organisms. We expect to develop a work scope document detailing the recommended additional feed trials. Finally, we will have a full set of in vitro studies on the effects of THYM on bovine rumen metabolism, which will likely inform recommendations for further studies, including potential in vivo studies.
Publications
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