Source: ICHTHUS UNLIMITED, LLC submitted to NRP
FROM WASTE/POLLUTION TO PRIME AQUAFEED INGREDIENT: USE OF TUNA PROCESSING WASTE SOY AND COTTON SEED MEAL TO PRODUCE HIGHLY NUTRITIOUS MEALS FOR AQUACULTURE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1022868
Grant No.
2020-33610-31980
Cumulative Award Amt.
$100,000.00
Proposal No.
2020-00985
Multistate No.
(N/A)
Project Start Date
Sep 1, 2020
Project End Date
Apr 30, 2022
Grant Year
2020
Program Code
[8.7]- Aquaculture
Recipient Organization
ICHTHUS UNLIMITED, LLC
109 S 32ND ST
WEST DES MOINES,IA 50265
Performing Department
Fish Nutrition
Non Technical Summary
Growth of alternative protein sources is poised to accelerate, potentially claiming up to a third ofthe protein market by 2054, profoundly affecting agriculture, food technology, end products, andthe animal feed market. Here, interdisciplinary research is proposed to increase the efficiency andefficacy of US-based aquaculture through development of nutritious, cost-effective alternatives totraditional marine protein feedstuffs. This will entail development of protein concentrates andcomplete diets from tuna processing wastes and co-products such as low antinutrient soybean mealand gossypol-free cottonseed meal by applying novel extrusion technology. The most promisingproducts will be evaluated for nutrient digestibility and production potential based on assessment ofgrowth performance and physiological capacity via trials conducted under controlled laboratoryconditions with red drum at Texas A&M University and with yellowfin tuna at the AchotinesLaboratory using well-established, standardized protocols. The developed technologies(manufacturing techniques and feedstuff products) will be subjected to economic analysis andextended to various user groups including seafood processors, feed manufacturers andaquaculturists through established networks. Thus, the developed products from this project willmake more efficient use of tuna processing wastes and other co-products to replace fishmeal andother costly components of aquaculture diets, increasing the cost-effectiveness of aquacultureproduction while limiting the expense and logistical constraints associated with disposal of seafoodprocessing wastes.
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
30252301010100%
Goals / Objectives
Characterize the chemical and nutritional composition of tuna processing byproducts and specialty cotton and soybean mealAdapt novel extrusion technology to produce nutritional replacements for fishmealEvaluate the nutritional and digestibility values of the developed productsin diets for red drum (Sciaenops ocellatus) and yellowfin tuna (Thunnus albacares), as representatives of high-valued marine carnivorous fish speciesConduct preliminary economic and market analysis of developed technologies and extend to various user groups including seafood processors, feed manufacturers and aquaculturists, with the ultimate benefit being realized by the human consumer
Project Methods
Chemical Characterization. Tuna processing byproducts (TPBs) will be obtained from industry collaborators and subjected to compositional analyses. Standard assay procedureswill be used. Similarly, specialty soybean meal (SBM) and low-gossypol cottonseed meal (CSM) will be obtained from collaborators the Ohio Soybean Promotion Council and Cotton Inc., respectively, and previous evaluation of their chemical profile will be used in the processing of the tuna wastes.II. Manufacturing new ingredients. From earlier nutritional studies and field assessments, tuna by-products have been identified as the most promising and are available from collaborator Ahi+ Inc. (Pier 36, Honolulu, IA 96817). A significant amount of TPBs is currently discarded representing a liability to the processing plants. Novel extrusion technology (Twin Thermal Extrusion, TTE) offers the advantage of making use ingredients with high moisture and/or oil such as TPBs. Extrusion processing also subjects materials to high temperatures and pressure, resulting in cooking and pasteurization, thus eliminating any threat of potential pathogens in TPBs. Wenger's Extrusion Technology Center (WETC, Sabetha, KS) has TTEs of various capacities, along with drying, grinding and other related equipment to conduct pilot-scale manufacturing associated with this project.TPBs will be ground into a homogenous mush using a pilot-scale shredder and analyzed for proximate composition as outlined above.The wet processing waste will be mixed with a suitable amount of dry SBM and CSM to obtain an appropriate physical consistency.Each mixture then will be extruded at 145-155 C (320-340 F), with much of the moisture driven off as steam.The resultant blended products will be milled with a hammer mill to a US standard sieve #80 (180 µ) and used as one of the major ingredients for experimental red drum and yellowfin tuna diet formulations.Nutritional evaluations: Red drum will serve as the screening species and only the best performing treatments will be evaluated in yellowfin tuna. The resulting blended products and complete diets containing such products will be evaluated first in a series of feeding and digestibility trials with red drum. These trials will be conducted with juvenile red drum at the Texas A&M University Aquacultural Research and Teaching Facility (TAMU-ARTF) outside of College Station, TX. Selected treatments will be assessed with yellowfin tuna at the Achotines Laboratory of the Inter American Tropical Tuna Commission (AL-IATTC) in Panama. Both procedures at ARTF and AL-IATTC will be approved by the University Laboratory Animal Care Committee of TAMU.Red drum trials. Initially, we will screen the following ratios dry meal (a 50:50 SBM and CSM blend, SBM-CSM) to wet tuna byproduct: 50:50, 40:60, 30:70 (a). The best combination (compositionally and functionally) will be used at the following inclusion levels as replacement of menhaden fishmeal (MFM): 70, 85 and 100% replacement in a fishmeal-based reference diet.Red drum feeding trials will be conducted with juvenile (5-10 g) fish, and subsequently with sub-adult (100-150 g) fish.Procedures for conducting such feeding trials are well established at the ARTF. Experimental diets containing the various meals derived from fish processing wastes will be substituted at three levels for MFM in a control diet.The experimental diets will be fed to juvenile red drum in 110-L glass aquaria containing brackish water (7 ppt) and connected as recirculating systems.Salinity will be maintained using synthetic sea salt concentrate and sodium chloride.Temperature will be maintained at 28 ± 1 C.Flow rate through each system will be about 1 L/min and sufficient to maintain optimal water quality.A sand filtration system also will be used to remove particulate material, and nitrogenous waste will be removed by a biofilter.Supplemental aeration will be used to maintain dissolved oxygen levels close to saturation, and other water quality parameters will be monitored routinely to keep them at acceptable levels.A 12:12 light: dark cycle will be maintained with fluorescent lights controlled by timers.Each diet will be fed to fish in three randomly assigned units for approximately 8 weeks.Fish will be fed at a rate approaching apparent satiation and collectively weighed each week.Responses to be measured will include weight gain, feed intake, feed efficiency ratio and protein efficiency ratio according to well established procedures.Yellowfin tuna trials. Based on screenings conducted on red drum the two, most promising dietary treatments will be evaluated with species-specific modifications for yellowfin tuna. Tuna will be initially maintained in 7,000 L (4.6 m diameter) quarantine tanks for 2 weeks and treated with formalin (200 ppm) against external parasites. Prior to transferring the fish into the trial tanks (8.5 m diameter x 3 m deep; 170 m3 capacity [170,000 L], semi-closed recirculating system, ~ 3 water exchanges/day) fish will be treated with a broad-spectrum antibiotic (nitrofurazone, 100 ppm) which will be applied for approximately 3 min of transit time between quarantine and trial tanks. Husbandry conditions will be maintained throughout the rearing period at 27 - 28 C for water temperature, 5.5 - 6.3 ppm dissolved oxygen, < 0.05 ppm un-ionized ammonia and 28 - 29 ppt salinity. Performance of YFT fed each of the two experimental diets will be evaluated. Each experimental diet will be fed over a 3-month period and performance indicators (weight gain, instantaneous growth rate, feed intake, feed efficiency ratio and protein efficiency ratio) will be evaluated at the beginning and end of each period.Representative samples of red drum and tuna fed each diet will be obtained at the beginning and end of each feeding trial and analyzed for proximate and mineral composition.Digestibility determination: An indirect method will be used for digestibility determinations using sub?adult red drum. Yttrium oxide (Y2O3) will be the non?digestible marker included in a reference diet at 0.1% of dry weight (Gaylord et al. 2008). Reference diets will be formulated as previously described for red drum. Test diets containing 30% test ingredient and 70% reference diet will be used to determine the digestibility coefficients of TPBs. The reference and test diets will be processed and stored following published procedures.Fecal samples will be collected 6 h post?feeding at each collection period. If needed, red drum can tolerate subsequent fecal collection efforts which will occur at weekly intervals and pooled until adequate sample is obtained for each diet/tank assignment.Crude protein, lipid, dry matter, ash, gross energy and phosphorus will be analyzed as previously described and Y2O3 will be determined via inductively coupled plasma optical emission spectroscopy after wet decomposition with concentrated nitric acid. The apparent digestibility coefficients of the test ingredients will be calculated from a comparison of the apparent nutrient digestibility of the reference diet alone and in combination with the test ingredients using testablished equations.Apparent digestibility coefficients (ADCN) of nutrients and energy of the testdiets and reference diet will be determined also using established equations.?: Data from the digestibility determination and feeding trial will be subjected to analysis of variance and Duncan's multiple?range test using the Statistical Analysis System. Differences in treatment means will be considered significant at P < 0.05.

Progress 09/01/20 to 04/02/22

Outputs
Target Audience:We will recast tuna by products + biodiesel by products as novel, value added, sustainable fishmeal replacement ingredients (FMR) for animal feeds. These ingredients will have entry in pet food, high value marine fish, swine, and poultry. Based on preliminary market & economic analyses we will develop market intelligence and will produce a very compelling message for customer (global aquafeed manufacturers, GAM) interactions. The target audience for this project are such GAMs. Our message may include FMR attributes such as sustainability, superior nutrition, enhanced digestibility, scalability, etc. GAMs and other feed consolidators will consider alternative feedstuffs only if savings in production can be realized with similar performance. Novel ingredients resulting from this project have ecological appeal and may also have important price advantages over fishmeal. Benefits may include cost savings, protein concentration, flowability, marine lipid capturing, functional benefits at the extruder, advantages in animal performance and environmental benefits. Changes/Problems:For this project, cottonseed kernels with no gossypol were selected as a special ingredient to be blended with cold dried tuna to produce a nutritious and sustainable feed ingredient. Because the high percentage of oil content in cottonseed kernels (~34%), a batch of 1,300 lbs was sent to InstaPro - a R&D company located in Grimes, IA, with the objective to reduce the oil content to 10%. Unfortunately, InstaPro failed to reduce the oil content and the cottonseed meal had to be sent to another institution - the Northern Crops Institute at North Dakota State University for further processing. The processing at NDSU was more difficult and time consuming because the cottonseed kernels were ground to a meal at InstaPro. Otherwise, flaked kernels could have been used. Nevertheless, the engineers at NDSU successfully decreased the oil content on the cottonseed meal from 33.8% to 9.81% after performing two consecutive hexane extraction followed by two consecutives ethanol washes (Figure 13). Although the final oil content was satisfactory, the oil extraction process was not practical nor cost-effective. We have identified a better option for the cottonseed oil extraction process through Pure Green Extraction, located in Reedsport - OR. Processing protocols will be updated on Phase II of this project, including pre-processing steps such as de-linting and flaking. What opportunities for training and professional development has the project provided?One master student, a Ph.D. student and a post doctoral afiliate have been trained in ingredient processing, diet formulation, diet manufacturing and design of aquaculture recirculation systems How have the results been disseminated to communities of interest?In 2022 outreach programs, workshops and publications will be targeted to end users ensuring an efficient information distribution process. A competitive advantage of this project is that the generated technology will use established dissemination channels which have been proven effective. Specialized meetings including World Aquaculture Society Meetings, European Aquaculture Society Meetings, Fish Nutrition & Feeding Symposium, Fish Nutrition Workshop, etc., will be used as ideal venues to deliver invited presentations. Together with peer-reviewed publications these meetings have significant marketing importance because global aquafeed manufacturers send affiliated personnel to these specialized meetings to maintain a competitive edge on applied aquatic animal nutrition. As more information is generated on terrestrial species a similar strategy will be used to ensure efficient communication with other stakeholders. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? All goals were accomplished. 1.After manufacturing the tuna by product meal (TBPm) and consolidating the amendment ingredients, an aliquot of each was sent to a third part laboratory for characterization of its full chemical composition including amino acid and fatty acid profiles. These results were contrasted between those of IU's TBM with similar products of marine origin. 2. A better processing option to dissipate moisture was evaluated and used to develop the TBPm. The low heat sublimation process renders a low moisture product with no heat denaturation. The nutritional integrity is preserved and the product is stabilized with very low water activity(~0.3). 3. Digestibility trials were conducted with hybrid striped bass because collaborator Texasw A&M University did not have suitable red drum at the time of testing. Hybrid striped bass is a rapidly growing fish and a suitable replacement for red drum. Yellowfin tuna was also not available from collaborator the Inter American Tropical tuna Commission but the fishmeal replacement prototypes were tested on long fin amberjack, a metabolicaly demanding fish with super fast growth. Results indicate that the prototype product may work well with tuna and this will be tested under SBIR Phase II on late summer 2022 with bluefin tuna. 4.Preliminary economic and market analysis of prototype fishmeal replacement product was completed and it would be viably priced below fishmeal while delivering similar performance.

Publications

  • Type: Conference Papers and Presentations Status: Under Review Year Published: 2022 Citation: Book of Abstracts Aquaculture 2021


Progress 09/01/20 to 08/31/21

Outputs
Target Audience:Seafood processors in Honolulu, HI and San Diego, CA where byproducts originate Feed mills (petfood) interested in fishmeal replacement products Equipment manufacturers: freeze drying, extrusion, grinding, pulverizing Aqua feed manufacturersinterested in fishmeal replacement products Changes/Problems:Problems encountered - 2020 has been an extremely year challenging, and many problems have emerged which relate to the COVID-19 pandemic. Safety restrictions applied to all "non-essential" business in CA from March to May 2020, which resulted in unexpected delays. The safety restrictions also affected our collaborators at Texas A&M University (re-scheduling the digestibility trial) and both NY and TX freeze drying facilities. Because growth trials require digestibility data and are planned downstream from digestibility determinations, growth trials involving red drum, rainbow, trout, and tuna have been delayed also. Difficulties in identifying suitable freeze-drying collaborators relate to the fact that most operate with products oriented for human consumption and, although the tuna byproducts have rigorously kept the cold chain and may be considered as human grade, the moment it was disclosed that this was intended as ingredients for fish diets, several freeze-drying options were closed. In addition, COVID-19 also brought bankruptcy to one of the largest freeze-dry companies - Parker Freeze Dry, which had already agreed to process the tuna byproduct. What opportunities for training and professional development has the project provided?Technical training on processing methods RAS system design and assembly Interns: Alejandro Molina Beatriz Oliva Greg Albrecht John Lopez Jesse Lopez Stephenie Jones 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?Produce experimental blends Conduct digestibility studies Formulate test diets Feeding trials with red drum, rainbow trout and tuna

Impacts
What was accomplished under these goals? The first months of this project were dedicated to securing the raw materials necessary to prepare the experimental ingredients. This has been challenging during the COVID-19 pandemic, but we are making reasonable progress. In addition, Ichthus Unlimited (IU) solidified agreements with the Fish Technology Center in Bozeman, MT (for coextrusion and potential additional testing with rainbow trout), Texas A&M University (for digestibility studies with red drum) and the Muenster Milling Company for pre-processing. The main ingredient - tuna by-product is in the final processing phase (freeze-dry [FD] process). Presently the IU team has conducted pilot trials in collaboration with FD companies in NY and TX and already conducted the chemical characterization of this novelty ingredient, together with that of specialty Ag-products with no antinutrients. For growth trials with marine species a new culture system has been designed and it will be constructed in our facility in San Diego in Q2 2021, where performance trials will be conducted. The IU team has also invested considerable time/effort in securing the balance of the budget necessary to accomplish/expand experimental objectives for this project. Competitive initiatives have been targeted with compelling proposals. IU personnel enrolled in Larta's commercialization assistance program to develop and strengthen a commercialization plan and the Phase II project proposal. COVID-19-related delays have made it necessary to apply for a no-cost extension which was granted extending the period of performance through 4.30.2022. Therefore, the Phase II proposal will be submitted on Feb 2022. Experimental Ingredients - All the dry feed ingredients necessary to manufacture the test ingredients were secured during 2020. The main raw ingredient - 2,000 lbs of tuna scraps was obtained and processed (ground) July through November. The material is ready the next phase of the processing and more details will be described in the next sections. Processing Tuna Scraps - Developing a new ingredient can be challenging, especially those based on animal ingredients containing high moisture and lipid such as tuna processing byproducts. This is one of the major obstacles in producing high quality meals from tuna scraps; therefore, exploring freeze-drying as a suitable as dehydration process would allow for the preservation of the nutritive value and organoleptic properties of tuna byproducts. To optimize the freeze-drying process a reduction in size is necessary so the initial process was divided in 3 phases: 1) grinding, 2) freeze-drying and 3) fine grinding. Chemical Composition of Test Ingredients The chemical characterization of the feed ingredients is an important step in optimizing formulation of fish diets. Samples of fishmeal, NutrivanceTM, Enzo MealTM, cottonseed meal (ProFloTM) and tuna the byproduct meal were sent to a third-party laboratory to determine the chemical composition of the test ingredients prior to formulate the experimental blends. To assure safety and freshness of the tuna byproduct meal, biogenic amines were quantified. Levels of tyramine (≤ 5 ppm), putrescine (38.5 ppm), cadaverine (49 ppm), histamine (31 ppm), agmatine (≤ 5 ppm), spermidine (41.5 ppm) and spermine (≤ 5 ppm) were lower than those found in high quality (fresh) fishmeal by Opstvedt et al (2000). High quality fishmeal must be low in biogenic amines because this leads to a high protein digestibility (Aksnes Mundheim et al. 2004). In contrast, fishmeal produced from stale raw materials with a high content of biogenic amines has previously been shown to reduce the specific growth rate of Atlantic salmon (Salmo salar) and survival, feed consumption and final biomass of blue shrimp (Litopenaeus stylirostris) compared to fishmeal produced from fresh raw materials (Opstvedt et al. 2000, Tapia-Salazar et al. 2004).

Publications