Progress 09/01/23 to 08/31/24
Outputs
Target Audience:Educational tours of the fish farm for undergraduate and graduate students. Changes/Problems:The serious illness and death of co-PI Shepherd in 2022 prevented the completion of research activities related to immunity impacted by alfalfa protein concentrate. His lab closed and no other facilities exist with the same capacity. What opportunities for training and professional development has the project provided?Through this study, we trained one postdoctoral researcher and a graduate student in research management and extension work with farmers at an aquaponic farm. How have the results been disseminated to communities of interest?Educational tours of the fish farm for undergraduate and graduate students. Trade journal article on research results. What do you plan to do during the next reporting period to accomplish the goals?This is the final year of the project. Additional journal articles are in preparation and will be submitted inthe coming year.
Impacts
What was accomplished under these goals?
Objective 1: Major activities completed / experiments conducted: Two 12-week feeding trials were conducted to assess the impact of alfalfa nutrient concentrate (ANC) and soybean concentrate (SBC) on yellow perch, focusing on growth, feed conversion ratio, and survival. Test diets containing 10% ANC or 10% SBC were prepared using protocols similar to those employed by commercial feed manufacturers. The first trial took place in a flow-through system under laboratory conditions, while the second trial was conducted on an aquaponic farm. Data collected: The two test diets were analyzed for proximate composition, amino acid composition, and mineral content. Both diets exhibited similar nutritional profiles. The results indicated that ANC and SBC had comparable effects on the growth and feed conversion ratio of yellow perch. No mortality was observed as a result of the different feed types. However, the overall growth of the fish was lower than in our previous studies on yellow perch. Feed efficiency was consistently above 2. Summary statistics and discussion of results: The preliminary results of this study suggest that ANC can replace SBC in feed for yellow perch. The observed poor growth of the fish may not be related to the nutritional quality of the test diets but rather to the physical quality of the feed pellets. The pellets were found to be floating due to their low density. Yellow perch prefer slow-sinking feed and showed reduced feeding efficiency with floating pellets. Therefore, future studies should focus on modifying the feed processing protocols to address this issue and validate the results. Key outcomes or other accomplishments realized: This study suggests that ANC can potentially replace SBC in yellow perch feed, and this finding may be applicable to other fish species as well. Further research is needed to confirm this conclusion. Additionally, research conducted on extension farms can assist in training both students and farmers This research provides valuable information for optimizing feed practices when using ANC.
Publications
- Type:
Other
Status:
Published
Year Published:
2024
Citation:
Deng, D.-F., and Samac, D. A. 2024. Alfalfa nutriente concentrate benefits for rainbow trout diets. Forage Focus (May 2024): 8-9.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2024
Citation:
Chen, H., Blaufuss, P. C., Deng, D.-F., Casu, F., Kraco, E. K., Shepherd, B., Sealey, W. M., Watson, A. M., Digman, M.F., and Samac, D. A. 2024. Impacts of alfalfa nutrient concentrate on pellet physical attributes, growth performance, metabolism and nutritional quality of rainbow trout, Oncorhynchus mykiss. Animal Nut. (in press)
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2023
Citation:
Sullivan, W., Deng, D.-F., Kraco, E., Blaufuss, P., Samac, D., Digman, M., and Sealey, W. 2023. Can alfalfa nutrient concentrate serve as a feed ingredient for feeding juvenile yellow perch (Perca flavescens)? National Animal Nutrition Program 2023 Summit. National Academy of Sciences, Washington DC, April 12, 2023.
|
Progress 09/01/19 to 08/12/24
Outputs
Target Audience:In 2021, a presentation on the project was given to stakeholder farmers and faculty, students, staff in the Minnesota Forever Green Initiative. This group conducts research to develop new crops and new products to protect soil and water through production of perennial crops. A presentation on the project was given at the Midwest Forage Association Symposium in the section on new uses of forages. The audience was composed of farmers, crop consultants, extension educators, industry leaders, and researchers. In 2022, presentations on the project were given to farmers, extension educators, industry representatives, and research scientists at the Midwest Forage Association annual symposium, the Forever Green project updates, at the AURI webinar on new uses for alfalfa, the North American Alfalfa Improvement Conference, and the World Alfalfa Congress. A poster presentation was made at the 2022 ASABE Annual International Meeting, Houston, TX. The presentation was to professional scientists involved in agricultural engineering. The presentations at Aquaculture America were given to international audiences including researchers, industry representatives, policy makers, funding agencies, educators and students. The National Animal Nutrition Program summit was mainly for researchers, industrial representatives, and federal funding agencies involved in animal feed ingredients and feed production research and production. Changes/Problems:Research was significantly delayed due to delays in obtaining funding from NIFA, establishing agreements with the cooperating universities, and no lab access for 24 months due to COVID-19 restrictions. Funds were not transferred from NIFA until April 13, 2020, seven months into the granting period. Establishing cooperative agreements with co-PI institutions took another month of processing. In addition, no sufficient production of phytase-APC was produced for feed testing due to the delay of the project. Instead, a commercial phytase was used as feed additive to APC, which was evaluated in a feeding trial. The serious illness and death of co-PI Shepherd in 2022 prevented the completion of research activities related to immunity impacted by alfalfa protein concentrate. His lab closed and no other facilities exist with the same capacity. What opportunities for training and professional development has the project provided?At the University of Wisconsin-Milwaukee, three undergraduate students, two graduate students, and one postdoctoral scientist were supported by this project. They were trained to develop feed formulations, conduct feed testing research and collaborate with farmers. This training included gaining experience in testing feed physical quality, evaluating the effects of feeds on fish growth, collecting feed fecal samples for nutrient digestion testing, and collecting samples for nutritional and physiological analysis of fish metabolic responses, and compiling data for dissemination. All students were involved in outreach, extension, and data dissemination. A MS graduate who worked on this project submitted his thesis in 2023 and presented the results at a national symposium. An undergraduate student at the University of Minnesota received training in protein isolation and enzymatic analysis and technical staff developed skills in protein extraction, concentration, and analysis. This project partially supported training an MS graduate and undergraduate student at the University of Wisconsin-Madison. The students gathered the TEA's capital cost and process information, developed the process unit operations, and performed a mass balance. The MS student presented our preliminary results at a national meeting. How have the results been disseminated to communities of interest?Educational tours of the fish farm for undergraduate and graduate students. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Under Objective 1, studies were done to evaluate alfalfa nutrient concentrate (ANC) as an ingredient in feed for rainbow trout. Five test diets contained varying ANC levels (0%, 5%, 10%, 15%, and 20%) to replace fishmeal (32% in the 0% ANC diet) to achieve equal digestible protein were formulated and processed using a cooking extrusion method. Analysis of feed pellets showed that pellet density increased with ANC levels, leading to sinking pellets at 20% ANC, accompanied by improved water stability, durability, and reduced oil leakage. Two feeding trials were conducted to test the diets in flow-through water systems with three replicates per diet. The first 10-week trial evaluated their impact on feeding, fecal physical quality, and the apparent digestibility coefficient (ADC) of dietary nutrients in rainbow trout (initial body weight, 18 ± 0.2 g). ANC inclusion did not impact the palatability and satiation feed intake of the fish (p > 0.050). However, the ADC of dry matter and phosphorus decreased in fish fed the 20% ANC diet, and alterations of fecal physical quality (p <0.010). The second 9-week trial investigated the growth performance, nutrition quality, and metabolism of rainbow trout (initial body weight, 19 ± 0.2 g). While all fish exhibited substantial growth, fish fed diets with 10% to 20% ANC displayed lower growth rates and higher feed conversion ratios compared to those fed with 0% or 5% ANC (P<0.001). The whole-body protein content was higher in fish fed 5% ANC compared to all other treatments (p=0.030). The biochemical parameters of plasma were similar across treatments except for decreased plasma phosphorus levels in the ANC-based diets (p=0.033). Significant changes were observed in liver metabolism including tricarboxylic acid cycle, amino acid and energy metabolism pathways in fish fed the 20% ANC diet versus the 0% ANC diet (p < 0.050). These results demonstrate that ANC inclusion improved pellet physical quality without impairing feeding behavior and nutritional quality of the fish but inclusion ≥ 10 % in the diet reduced fish growth. This study offers the first comprehensive assessment of the potential of ANC used in fish feed involving feed management, feeding evaluation, and the biological response. Moreover, our study underscores the necessity for additional research to explore the lasting impacts of ANC incorporation into fish feeds, particularly concerning nutrient utilization, fecal characteristics, and consequent water quality management in aquaculture settings. Future studies are needed to evaluate the potential of ANC in practical feed formulations with lower levels of fishmeal and compare its potential with other plant protein concentrates such as soy and corn protein. Another lab study was conducted to investigate the impact of the same test diets on yellow perch. Juvenile yellow perch (71 days post hatch) were raised in a flow through system held at 23±1°C for 9-weeks and fed a diet with fishmeal replaced with ANC (0%, 5%, 10%, 15%, 20%). Growth rate, feed conversion ratio, satiation feed intake, and protein retention were similar for fish fed different diets (P>0.05). Fish fed 20% ANC had lower ash, phosphorus, calcium, and manganese than those fed 0% ANC (P<0.05). A lower phosphorus apparent digestibility coefficient was determined in ANC than in menhaden fishmeal, which partially explains the lower phosphorus content in fish fed 20% ANC. This study suggests that ANC can be used as a partial protein source in perch feed, but more research is needed to address the concerns of low nutrient digestibility. These findings will benefit aquaculture producers as it demonstrates ANC can be utilized as an alternative to fishmeal without impacting the quality of the fish, and this would open a new avenue of income for alfalfa producers. A preliminary study (12 week feeding trial) was conducted to compare the utilization of ANC versus soy protein concentrate in an aquaponic farm and under laboratory conditions. The results showed that ANC had a similar effect on the growth performance of yellow perch compared to soy protein concentrate. Under Objective 2, a total of 42 alfalfa lines expressing the defensin gene were produced for analysis of peptide levels in plants and protein concentrates. Plant expressing the defensin peptide were increased by vegetative propagation. Methods were developed to increase plant vigor for testing defensin production and disease resistance. Plants were inoculated to test resistance to foliar disease (anthracnose), root rot (Aphanomyces root rot) and vascular wilt (Fusarium wilt). In contrast to in vitro assays in which defensin peptide inhibited fungal spore germination and growth, expression of defensin in plants did not affect disease resistance ratings and in some cases appeared to increase disease susceptibility. A total of 38 lines expressing phytase were established in field plots for quantification of phytase. Methods were optimized for juicing alfalfa plants, and for concentrating and drying protein precipitates. Phytase activity was measured with a commercial kit using fresh juice, fresh juice flash frozen and thawed, freeze dried juice, and dried plant herbage. The juice was also treated with 5 M HCl to reduce the pH to 4.0 to precipitate proteins or heated to 90°C for 10 minutes to precipitate protein. RNA was extracted from plants producing low and higher amounts of phytase and used for quantitative RT-PCR assays to measure phytase mRNA. Phytase activity was 0.20 to 0.8 Units/mg protein. On average, the fresh juice had highest activity of 0.43 U/mg protein, followed by frozen and thawed juice at 0.36 U/mg protein and dried plants at 0.24 U/mg protein. The amount of protein in the acid protein precipitate could not be measured. Activity in the precipitate ranged from 0.1 to 0.5 U/mg precipitate. The heat treatment resulted in inactivation of phytase activity. Ranking of individual plants for amount of phytase produced was similar for all extraction methods. Under Objective 3, we aimed to identify the most cost-efficient pathway to harvest and isolate alfalfa protein for aquafeed production. We compared four field harvest scenarios for collecting alfalfa for producing alfalfa protein concentrate - one included traditional whole-plant harvest and the other a novel system that harvests only the leaves. We developed a cost analysis of a screw press, membrane separation, and drying process for concentrating alfalfa protein. These costs can be combined with alfalfa production costs to determine the cost of alfalfa protein relative to alternative sources such as soybean meal. Leaf stripping with stem collection had a harvest cost of $33.23/kg, processing was $14.13/kg for a total of $47.36/kg feed produced. Leaf stripping without stem collection had a cost of harvesting of $20.26/kg feed, processing cost of $14.13/kg feed for a total of $34.39/kg feed produced. The direct cut was the least expensive method of producing alfalfa protein concentrate. For alfalfa directly cut in the field the harvesting the cost was $9.63/kg feed produced and processing was $5.71/kg feed for a total of $15.34/kg. The final cost is like the price for fish meal ($1500/ton) but higher than that of soybean meal ($350/ton). These results were presented at the 2023 American Society of Agricultural and Biological Engineering Annual International Meeting in Houston, TX.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Sullivan, W., Deng, D.-F., Kraco, E., Blaufuss, P., Samac, D., Digman, M., and Sealey, W. 2023. Can alfalfa nutrient concentrate serve as a feed ingredient for feeding juvenile yellow perch (Perca flavescens)? National Animal Nutrition Program 2023 Summit. National Academy of Sciences, Washington DC, April 12, 2023.
|
Progress 09/01/22 to 08/31/23
Outputs
Target Audience:Poster presentation at the 2022 ASABE Annual International Meeting, Houston, TX. The pesentation was to profession scientists involved in agricultural engineering. Changes/Problems:The metabolic analyses proposed to be done on fish receiving feed with different levels of alfalfa protein concentrate can not be done due to the death of co-PI Brian Shepherd. His lab has been closed and no other facilities exist with the same capacity. What opportunities for training and professional development has the project provided?This project partially supported training an MS graduate and undergraduate student at the University of Wisconsin. The students gathered the TEA's capital cost and process information, developed the process unit operations, and performed a mass balance. The MS student presented our preliminary results at a national meeting. 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?An on-farm feeding trial will be conducted. Production of defensin peptide in E. coli will be attempted and an ELISA assay developed for measruing the amout of defensin produced. Manuscripts describing results will be submitted for publication.
Impacts
What was accomplished under these goals?
Objective 1 Major activities completed / experiments conducted An 8-week trial was completed to test feeding response, fecal particle distribution, and digestibility of test diets with0, 5, 10, 15, 20% ANC to replace fishmeal. A 9-week experiment was completed to measure growth performance, nutrient utilization, and fish metabolism with the test diets. There were 15-30 fish/and with three replications/diet. Sufficient ANC was obtained for producing diets for an on-farm trial. Data collected Fillets of fish from the experiment were submitted for proximate analysis:moisture, crude protein, total fat, total carbohydrate, and dietary fiber. Summary statistics and discussion of results There was no significant difference in any of the factors measured based on percentage of ANC in the diet. Feed for the on-farm trial was produced. Objective 2 Major activities completed / experiments conducted Plant expressing the defensin peptide were increased by vegetative propagation. Methods were developed to increase plant vigor for testing defensin production and disease resistance. Plants were inoculated for testing resistance to foliar disease (anthracnose), root rot (Aphanomyces root rot) and vascular wilt (Fusarium wilt). Assays to produce the defensin peptide in E. coli are in progress. Data collected No difference in anthracnose resistance or Aphanomyces root rot resistance was observed for plants expressing the defensin peptide. The Fusarium wilt assay is in progress. Summary statistics and discussion of results In contrast to in vitro assays in which defensin peptide inhibits fungal spore germination and growth, expression of defensin in plants has not affected disease resistance ratings. Objective 3 Major activities completed / experiments conducted This project looks to identify the most cost-efficient pathway to harvest and isolate alfalfa protein for aquafeed production.We compared two field harvest scenarios for collecting alfalfa for producing alfalfa protein concentrate - one included traditional whole-plant harvest and the other a novel system that harvests only the leaves.We have developed a cost analysis of a screw press, membrane separation, and drying process for concentrating alfalfa protein. Data collected Costs to harvest and concentrate alfalfa protein were established. These costs can be combined with alfalfa production costs to determine the cost of alfalfa protein relative to alternative sources such as soybean meal. These values will be presented in a peer-reviewed publication that is in development.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Samac, D.A., Deng, D.-F., Shepherd, B.S., and Digman M.F. 2022. Alfalfa leaf protein concentrate in aquafeeds to enhance finfish production and reduce environmental impacts in aquaculture production. World Alfalfa Congress, San Diego, CA, Nov. 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
A.B. Hemmelgarn, D.F. Fares, D.A. Samac, D.F. Deng, B.S. Shepherd, M.F. Digman. Techno-economic assessment of protein concentrate derived from alfalfa leaves, Poster presentation at the 2022 ASABE Annual International Meeting, Houston, TX.
|
Progress 09/01/21 to 08/31/22
Outputs
Target Audience:Presentations to farmers, extension educators, industry representatives, and research scientists at the Midwest Forage Association annual symposium, January 2022; the Forever Green project update in May 2022, and at the AURI webinar on new uses for alfalfa, July 2022. Changes/Problems:The original plants to be used for defensin production were lost due to the COVID19 pandemic and had to be replaced. Production of phytase by the participants was found to be impractical and so a commercial source was used for addition to fish feed. What opportunities for training and professional development has the project provided?Graduate students received training in use of SuperPro Designer (v12). Undergraduates learned new biochemical assays and lab techniques. 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?Further metabolic analysis of fish will be carried out. Analysis of a feeding trial using feed containing phytase will be completed. Feed will be produced for an on-farm trial with the optimal concentration of alfalfa protein concentrate. Assays for defensins will be developed and used to measure the amount of peptides produced in alfalfa plants. Three manuscripts are in preparation on each objective.
Impacts
What was accomplished under these goals?
Objective 1 Major activities completed / experiments conducted An 8-week trial was completed to test feeding response, fecal particle distribution, and digestibility of test diets with 0, 5, 10, 15, 20% ANC to replace fishmeal. A 9-week experiment was completed to measure growth performance, nutrient utilization, and fish metabolism with the test diets. There were 15-30 fish/and with three replications/diet. Data collected In the 8-week trial, palatability and feed satiation, fecal particle distribution, total dry matter and phosphorus digestibility was recorded. In the 9-week trial, weight gain and feed conversion ratio were measured. From harvested fish, phosphorus, protein, and metabolic markers were analyzed. Summary statistics and discussion of results There was no significant difference in palatability or satiation feeding response as measured by feed intake % of body weight for the test diets. Fecal particle size distribution changed only in the diet with 20% ANC with the percentage of small and medium particles increasing, and large particles decreasing. Replacement of fishmeal with 10% and 20% ANC significantly decreased the digestibility of total dry matter and P, respectively. The ADC of protein was 89-90% for different test diets and was not impacted by ANC inclusion. Phosphorus as mmol/L plasma was slightly lower with 10% ANC but was not different from the control at other concentrations. Protein content of fillets was not affected by ANC in the diet. A change in branched chain amino acid degradation and energy metabolism occurred with inclusion of ANC in the diet. Key outcomes or other accomplishments realized Feeding was not impaired due to fishmeal replacement by ANC. When ANC was >10% in the feed, nutrient digestibility and growth were decreased. ANC-based feed could potentially alter some major metabolism pathways such as branch chain amino acid metabolism, TCA pathway, and ketone metabolism. Objective2 Major activities completed / experiments conducted Phytase activity was measured from transgenic plants with a commercial kit using fresh juice, fresh juice flash frozen and thawed, freeze dried juice, and dried plant herbage. RNA was extracted from plants producing low and higher amounts of phytase and used for quantitative RT-PCR assays to measure the mRNA accumulation for phytase. The juice was also treated with 5 M HCl to reduce the pH to 4.0 to precipitate proteins or heated to 90°C for 10 minutes to precipitate proteins. Phytase activity was measured in protein precipitates. Plants expressing MtDef5 (defensin) were developed and the plants expressing the highest amount of defensin mRNA were identified. Data collected, Summary statistics and discussion of results Phytase activity was 0.20 to 0.8 Units/mg protein. On average, the fresh juice had highest activity of 0.43 U/mg protein, followed by frozen and thawed juice at 0.36 U/mg protein and dried plants at 0.24 U/mg protein. The amount of protein in the acid protein precipitate could not be measured. Activity in the precipitate ranged from 0.1 to 0.5 U/mg precipitate. The heat treatment resulted in inactivation of phytase activity. Key outcomes or other accomplishments realized Active phytase enzyme could be recovered from fresh juice, thawed juice, acid precipitated protein and dried plant herbage. Ranking of individual plants for amount of phytase produced was similar for all extraction methods. Objective 3 Major activities completed / experiments conducted Four scenarios were outlined for field harvest of alfalfa. The costs of the equipment were obtained from the literature or vendor quotes. The processes for production of protein concentrate were outlined and costs of equipment obtained. Data collected. The equipment production costs were modeled including variable operating costs (components, materials, and labor), and fixed operating costs (general and administrative costs, overhead, maintenance, and insurance) per ASABE EP 496. Summary statistics and discussion of results. The team completed assessing the costs from the field edge to final product (e.g., concentration, drying). For alfalfa directly cut in the field the harvesting the cost was $9.63/kg feed produced and processing was $5.71/kg feed for a total of $15.34/kg feed produced. Leaf stripping with stem collection had a harvest cost of $33.23/kg, processing was $14.13/kg for a total of $47.36/kg feed produced. Leaf stripping without stem collection had a cost of harvesting of $20.26/kg feed, processing cost of $14.13/kg feed for a total of $34.39/kg feed produced. Key outcomes or other accomplishments realized. The direct cut was the least expensive method of producing alfalfa protein concentrate. The final cost is similar to the price for fish meal ($1500/ton) but higher than that of soybean meal ($0.43/dg).
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Deng, D.-F., Chen, H., Shepherd, B., Zohn, M., Casu, F., Digman, M., Samac, D. A., and Sealey, W. 2022. Utilization of alfalfa nutrient concentration in feed for rainbow trout, Oncorhynchus mykiss: Effects on pellet physical quality, nutrient utilization, and fish metabolism. World Aquaculture Society, San Diego, CA, March 4, 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Samac, D. A., Deng, D.-F., Shepherd, B. S., and Digman, M. F. 2022. Alfalfa leaf protein concentrate in aquafeeds to enhance finfish production and reduce environmental impacts in aquaculture production. North American Alfalfa Improvement Conference, Lansing, MI, June 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Hennelgarn, A. B., Samac, D. A., Deng, D.-F., Shepherd, B. S., and Digman, M. F. 2022. Techno-economic assessment of protein concentrate derived from alfalfa leaves. American Society of Agricultural and Biological Engineers, Houston, TX. July 17-20, 2022.
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Progress 09/01/20 to 08/31/21
Outputs
Target Audience:A presentation on the project was given to stakeholder farmers and faculty, students, staff in the Minnesota Forever Green Initiative. This group conducts research to develop new crops and new products to protect soil and water through production of perennial crops. A presentation on the project was given at the Midwest Forage Association 2021 Symposium in the section on new uses of forages. The audience was composed of farmers, crop consultants, extension educators, industry leaders, and researchers. Changes/Problems:Research was significantly delayed due to limited access to laboratory facilities and hiring personnel due to COVID-19 restrictions. What opportunities for training and professional development has the project provided?Two undergraduate students have been trained in the project to conduct a technoeconomic analysis of alfalfa protein concentrate production. 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?An on-farm feeding trial will be conducted to evaluate the optimized feed in a real-world setting. Plant exptracts, dried leaf material, and protein concentrate produced by several methods from transgenic plants expressing defensin peptide or phytase enzyme will be assayed for peptide and enzymatic activity. The model for estimating production costs for alfalf aprotein concentrate will be run under different scenarios to identify the most economical means of production.
Impacts
What was accomplished under these goals?
Aquaculture, the raising of farmed fish and shellfish, is the fastest growing food sector around the world and the most efficient means of providing animal protein to human populations. Bottlenecks to sustainable aquaculture include identifying acceptable plant proteins for fish feeds.Alfalfa foliage can be refined into a high protein concentrate (alfalfa protein concentrate; APC) that shows excellent promise for use in fish feeds as an alternative to fishmeal. As a result of the research, new products from refining alfalfa and processes for on-farm or near-farm refining of alfalfa can be promoted. Objective 1: Evaluate APC as an ingredient in feed for rainbow trout. Major activities completed / experiments conducted. Five test diets were formulated to include different levels of APC (0, 5, 10, 15, and 20%) to replace fishmeal. The physical properties of feed were determined. Palatability of feed by rainbow trout fingerlings was measured by determining the food intake and attempt to feed time during the second week of feeding of the assigned diets. Fecal samples were collected for the measurement of fecal particle distribution and nitrogen content after the fish were fed the experimental diet for 6 weeks. Satiation feeding was conducted 7 weeks after the fish had been fed with the assigned diets. Digestibility of test diets was determined at the end of 8 weeks feeding. A 9-week feeding trial was conducted to evaluate the effects of five test diets on the growth performance, nutrient utilization, fish health, and metabolism of juvenile rainbow trout. At the end of the trial, four fish from each replicate were used for whole-body proximate composition analysis and four fish per tank were used for measurement of individual body weight and were dissected to obtain the liver, intestine, carcass and gonads for calculations of hepatosomatic index, carcass index, and gonad index. Liver and intestines were collected for histology assessment or nutritional composition analysis using NMR spectroscopy. Data collected Data were collected on the effect of APC on physical quality of feed pellets, effect of APC on palatability and digestibility, and to determine the optimal APC based feed formulation for rainbow trout. Summary statistics and discussion of results The durability index of pellets was significantly higher in the feed with 10 to 20% APC than that in the control and the 5% APC diets. Bulk density of pellets was significantly increased by APC inclusion. The control and the pellets containing 5 to10% APC maintained neutral buoyancy while the pellets containing 15% APC were shown to sink slowly and 80% of the pellets from feed containing 20% APC sank within 10 min. Oil leaking from pellets was reduced by inclusion of APC. Water stability was significantly higher for feed pellets containing 10 to 20% APC than the control or 5% APC diet. Palatability was highest for fish fed 5% or 20% APC diets. Satiation feeding was not significantly different across different treatments. The apparent digestibility coefficient (ADC) of dietary protein was similar but the ADC of dry matter was significantly decreased in the 20% APC diet and inclusion of APC up to 10% resulted in a significantly lower ADC value compared to the control and 5% APC diets. The ADC values of other minerals except for Mg, Ca, and Mn tended to increase with the increasing levels of APC. The fecal protein content decreased with the increasing level of dietary APC. The test diets had no significant impacts on animal morphology (condition factor, carcass index, hepatosomatic index, visceral fat index). Satiation feeding was not changed due to inclusion of APC in the diets. Proximate compositions of whole fish and liver tissues were not significantly different among dietary treatments except that the protein content of fish fed the 5% APC diet was higher than that of fish on other diets. Biochemical parameters related to fish health were also similar among different treatments. Alanine and betaine were found to be significantly lower in fish fed the 20% APC diet compared with the control group; whereas leucine, valine, threonine, malate, and succinate were significantly higher in fish fed the APC diet compared with the control group. Altered levels of alanine and threonine and altered levels of succinate and malate may indicate modulation of energy metabolism. Higher levels of valine and leucine suggest TCA cycle inhibition since these metabolites can normally be converted to substrates of the TCA cycle. Key outcomes or other accomplishments realized. Replacement of fishmeal by APC changes the physically quality of feed pellets. The increased density of feed pellet due to inclusion of ANC may facilitate storage or shipping and make the feed easy to handle because of the increased durability. No adverse effect was found on fish health or end product nutrition related to use of ANC in the test diets. However, replacement of fishmeal by 10 to 20% significantly depressed growth rate, and the ADC of dry matter and phosphorus. The decreased digestibility of P is likely one of the reasons leading to the poor growth of fish fed the high ANC diets. Energy metabolism and osmoregulation were inhibited in the fish fed 20% APC. Inclusion of APC could alter the physical and nutrition quality of fecal samples such as residual particle sizes and nitrogen content. The results provide critical information for the management of culture systems and water quality. Objective 2: Evaluate APC with plant defensin and phytase for improving rainbow trout growth, well-being, and immunity. Major activities completed / experiments conducted. An additional 42 transgenic plants were generated expressing the defensin gene. These are currently being tested for defensin peptide concentration. Plants expressing phytase were established in field plots under regulations for field release by APHIS. Methods were optimized for juicing alfalfa plants, and for concentrating, and drying protein precipitates. The assay to quantify phytase in plants and protein concentrate was tested. Data collected. Transgenic plants were tested for presence of the marker gene nptII. Summary statistics and discussion of results. A total of 42 alfalfa lines expressing the defensin promoter were produced for further analysis of peptide levels in plants and protein concentrates. A total of 38 lines expressing phytase were established in field plots and quantification of phytase is underway. Key outcomes or other accomplishments realized. All notifications to APHIS were completed for establishing transgenic alfalfa expressing phytase in field conditions. The release was approved by the University of Minnesota Institutional Biosafety Committee. These results lay the ground work for determining the levels of peptide or enzyme that could be produced for use in aquaculture feeds. Objective 3: Carry out a technoeconomic and market analysis of APC production. Major activities completed / experiments conducted. Four scenarios have been outlined for field harvest of alfalfa. The costs of the equipment were obtained from the literature or vendor quotes. Data collected. The equipment production costs were modeled including variable operating costs (components, materials, and labor), and fixed operating costs (general and administrative costs, overhead, maintenance, and insurance) per ASABE EP 496. Summary statistics and discussion of results. Currently the team is assessing the costs from the field edge to final product (e.g., concentration, drying) and plans to compare the cost of the alfalfa protein concentrate to alternative protein sources. Key outcomes or other accomplishments realized. The needed background data has been obtained for producing the model and personnel trained in methods to assess models.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Coburn, J., Wells, M. S., Phelps, N., Gaylord, T. G., and Samac, D. A. 2021. Acceptance of a protein concentrate from alfalfa (Medicago sativa) by yellow perch (Perca flavescens) fed a formulated diet. Fishes 6(2):9. https://doi.org/10.3390/fishes6020009
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Coburn, J., Wells, M. S., Sheaffer, C. C., Ruan, R., and Samac, D. A. 2021. Comparison of feedstocks and methods to recover leaf proteins from wet fractionation of alfalfa for potential use in aquaculture, poultry, and livestock feeds. Agrosyst. Geosci. Environ. 4:e20184. https://doi.org/10.1002/agg2.20184.
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Progress 09/01/19 to 08/31/20
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Research was significantly delayed due to delays in obtaining funding from NIFA, establishing agreements with the cooperating universities, and no lab access due to COVID-19 restrictions. Funds were not transferred from NIFA until April 13, 2020, seven months into the granting period. Establishing cooperative agreements with co-PI institutions took another month of processing. All PIs have no access to laboratory facilities due to COVID-19 restrictions and have been teleworking since mid-March 2020. It has not been possible to hire technical, student or postdoctoral scientists due to the delay in fund transfer from NIFA and restrictions in lab access. 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?Research was significantly delayed due to delays in obtaining funding from NIFA and establishing agreements with the cooperating universities. We will now be back on track for conducting the research described in the proposal. During the year we will carry out all experiments to test the physical properties of feed prepared using alfalfa protein concentrate and establish the optimal concentration to use for rainbow trout based on digestibility studies. Assays will be developed to quantify defensin peptide and phytase in alfalfa plants and protein concentrate. Data will be collected for developing a technoeconomic and market analysis of APC production.
Impacts
(N/A)
Publications
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