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National Institute of Food and Agriculture Small Business Innovation Research Phase I - grant proposal. Comparative replacement of conventional fish feed components with Algal Biomass in Florida Pompano. By Gaiergy corp.
National Institute of Food and Agriculture Small Business Innovation Research Phase I - grant proposal. Comparative replacement of conventional fish feed components with Algal Biomass in Florida Pompano. By Gaiergy corp.
National Institute of Food and Agriculture Small Business Innovation Research Phase I - grant proposal. Comparative replacement of conventional fish feed components with Algal Biomass in Florida Pompano. By Gaiergy corp.
Small Business Innovation Research Phase I Grant Proposal
Comparative Replacement of Comparative Replacement of Comparative Replacement of Comparative Replacement of Conventional Fish Feed Components Conventional Fish Feed Components Conventional Fish Feed Components Conventional Fish Feed Components with Algal Biomass in Florida Pompano with Algal Biomass in Florida Pompano with Algal Biomass in Florida Pompano with Algal Biomass in Florida Pompano
By
Gaiergy Corp.
Project Investigators
Jason Masters Nathan Brennan, PhD Kevan Main, PhD President Staff Scientist Senior Scientist Gaiergy Corp. Mote Marine Lab World Aquaculture Society 1610 North Ocean Blvd., #1205 1600 Ken Thompson Pkwy President Pompano Beach, FL 33062 Sarasota, FL 34236 Mote Marine Lab DUNS: 808480581 1600 Ken Thompson Pkwy Sarasota, FL 34236 1
Table of Contents Abstract ................................................................................................................................................... 2 Project Narrative ..................................................................................................................................... 3 1. Response to Previous Review ....................................................................................................... 3 2. Responsiveness to USDA SBIR Program Priorities ......................................................................... 3 Energy Efficiency & Renewable Energy Connection .......................................................................... 3 NIFA Societal Challenges .................................................................................................................. 3 3. Identification & Significance of the Problem or Opportunity ........................................................ 4 4. Background and Rationale ........................................................................................................... 5 Previous Feed Trials ......................................................................................................................... 7 Algae Feed Formulation ................................................................................................................... 7 5. Relationship with Research or R&D .............................................................................................. 8 6. Technical Objectives .................................................................................................................... 9 7. Work Plan .................................................................................................................................... 9 Algae Production System ............................................................................................................... 10 8. Related Research or R&D ........................................................................................................... 13 9. Potential Post Application .......................................................................................................... 14 10. Satisfying the Public Interest .................................................................................................. 15 Strategic Goal 1 ............................................................................................................................. 15 Strategic Goal 2 ............................................................................................................................. 15 Strategic Goal 3 ............................................................................................................................. 16 Strategic goal 4 .............................................................................................................................. 16 Bibliography ......................................................................................................................................... 17 Equipment ............................................................................................................................................. 20 Letters of Collaboration ......................................................................................................................... 24
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Abstract
The opportunity exists to create better fish feed formulations at lower costs while reducing the amount of fish meal incorporated into a formulation by producing, harvesting and pelletizing high proteinaceous algae into species specific fish feeds. Total fish produced by aquaculture is now surpassing the amount of wild caught fish. The feeds that are used for aquaculture though are still comprised of 30% to 50% fishmeal. This means that as aquaculture continues to grow, it will produce ever-increasing pressure on wild caught fish to be used for fish feed if no other alternative was set in motion.
One of the current alternatives that have been set in motion is to replace fishmeal in fish feeds with a higher percentage of terrestrial vegetative sources because it costs less and would relieve overfishing pressure. There are two problems with this. The first is that the chemical compound structures in fishmeal differ significantly from terrestrial vegetative sources, such that the resulting feed fed to fish are not converted into fish biomass at the same per unit rate as a fishmeal diet. Second, the fish meat resulting from a diet increasingly comprised from terrestrial vegetative sources is increasingly lacking in essential fish oils and protein structures that make wild caught fish a more beneficial food source versus an aquacultured fish.
Currently, algae production for biofuels has gained significant investment from the federal government and private industry. One of the significant problems with the algae-to-biofuel industry is what to do with the biomass that remains after the bioenergy is extracted from the algae cell. Our research proposal can also address this since that residual biomass can be used for fish feeds instead of fish meal or terrestrial vegetation.
Our proposal intends to research the production of specific algae species to be harvested and impounded into a Pompano-specific fish feed and then test that fish feed against commercially available fish feeds and previously researched unique Pompano feeds. The intent is to utilize the algae's ability to produce high-quality proteins and essential marine oils and compounds to provide a cheaper and more nutritious biomass replacement than either fishmeal or terrestrial vegetative sources.
We conduct this research from the perspective of a small scale, rural aquaculture farmer. In doing this we can establish the viability of small scale aquaculture to produce their own fish feed at least as economical as a large-scale feed producer. We will grow enough algae to be harvested on-site and processed into our Pompano specific formula and then fed to juvenile Pompano that will be raised to a market size of approximately 1.5 pounds. The market-ready Pompano will then be tested for their meat quality, lipid quality and marketability in the form of taste tests.
This research will provide conclusive evidence necessary for a proposed commercial Pompano production facility to utilize algal-enhanced species specific fish feeds that project a 20% to 25% reduction in input costs through the use of these algal-enhanced feeds.
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Project Narrative
1. Response to Previous Review This is the first submission for this application so we have nothing to add to a response to a previous review.
2. Responsiveness to USDA SBIR Program Priorities
Energy Efficiency & Renewable Energy Connection
There is significant work ongoing between the USDA, Department of Energy and the Department of Defense regarding algal-based biofuels. These initiatives are making progress towards a future where liquid drop-in fuels derived from algae could become a reality. Whenever that future is actually realized, there'll still be an immense amount of biomass that needs viable outlets. The one inescapable reality right now is that the algae industry mimics the oilseed industry in that there is greater value in the biomass than in the oil. To that extent, the renewable energy connection must identify with the fact that there is greater profitability on the biomass side than on the transportation fuel side. This research has direct applicability towards bridging the gap between the reality of today and the potential reality of tomorrow or that future when algal biofuels become economically viable. For algal-based biofuels to become energy-efficient and sustainable they must address the residual biomass remaining after oil extraction. This work will help to show that the biomass composition used in this research is either an appropriate use of biomass for aquaculture or indicate a direction where a compositional shift might prove to be even more effective or that algal biomass does not improve nor hinder growth characteristics of Florida Pompano given today's resources. NIFA Societal Challenges
In response to the application requirements regarding NIFA societal challenge areas, we will address how our proposal addresses the four challenge areas listed below. Global Food Security & Hunger As our population increases combined with globalization and increasing climate change, the price volatility of primary food products along with their availability is also increasing. To combat the availability and security of food supply, the world population must begin to diversify their food product origins even more than they are today. In the United States and most industrialized countries, we have optimized our agricultural industries so that they are centralized specific areas. But this leaves us vulnerable to specific climactic actions and terrestrial limitations on soil, nutrients and water availability. The concept of building species specific fish feeds means that aquaculture operations that cultivate indigenous fish populations would be better served by a locally derived feedstock partially comprised of algae that the fish have 4
genetically optimize themselves to utilize. This research is one specific step towards building a specific feed for a specific fish. The methods used here can be used in other aquaculture operations in first, second and third world countries to increase fish production while decreasing input costs. Climate Change With regard to climate change, since algae are the largest group of primary producers on the planet contributing over 70% of the world's oxygen, this proposal has a potential direct effect, albeit very minor, towards curbing carbon dioxide emissions due to anthropogenic effects. For large scale algae companies to be able to produce the algae in massive quantities over short periods of time, they will need access to large concentrated sources of carbon dioxide. While the algae could grow on atmospheric carbon dioxide concentrations, the timeframe in which they would be able to utilize the atmospheric carbon dioxide would cause the initial profitability of an endeavor such as this to be in doubt. Therefore, initial algae operations are being positioned adjacent to CO 2 emitters such as coal-fired power plants and other industrial manufacturing companies. The carbon dioxide that they emit is being sequestered, scrubbed for heavy metals and then dissolved into an aqueous phase for photosynthetic conversion by the algae into fats, proteins and carbohydrates. The research conducted here could prove to be a marketable outlet for sequestered carbon dioxide potentially shipped offsite for algae-farms or this research could be used as adjacent business development for high CO 2 emitters. Both potentials become a positive influence toward mitigating climate change. Sustainable Bioenergy With regard to sustainable bioenergy, the research conducted here complements the value added to a bioenergy enterprise by creating a co-product that is currently a higher value than the bioenergy commoditized product. While we are neither creating nor researching bioenergy, algal-based biofuels must contend with the fact that approximately 50% of the algal cell will be made up of proteins, fats and carbohydrates that cannot be used for biofuel. So for the bioenergy production to be truly sustainable, it must have defined outlets for the specific types of residual fats, proteins and carbohydrates that are left after the bioenergy portion has been removed from the algal cell. Childhood Obesity This proposal does not have any direct relation to curbing childhood obesity other than potentially providing more non-processed protein fish into the markets and restaurants.
3. Identification & Significance of the Problem or Opportunity
As aquaculture production surpasses wild-caught fish, there is an increasing need to reduce fish- feed costs while increasing fish-feed quality. Fish-feed quality primarily addresses palatability, 5
digestibility, feed-to-biomass conversion and overall health of the fish. Current aquaculture feed production efforts are pushing for more terrestrial vegetation sources to comprise the majority of the meal, but this introduces new problems. The difference in the chemical structure of the proteins and lipids in oilseed crops fed to fish result in slower growth rates, low biomass conversion ratios, and a decrease in the beneficial lipids such as DHA, EPA and other carotenoids (Rossi, 2011) (Rossi, Davis, & Rhodes, 2010) (Rhodes, et. al, 2012). Furthermore, as oilseed crops prices continue to rise, a fish feed dependent upon oilseed crop prices does not solve the need to lower fish-feed prices in the long-term. Between 2005 and 2009, the fish feed input costs of soybean meal, fish meal, corn, wheat and rice rose by 67%, 55%, 284%, 225% and 180%, respectively (Rana, Siriwardena, & Hasan, 2009). The cost projections in 2012 for the same basket of products are expected to double again due to weather related issues during the 2012 growing season.
A secondary problem exists in the bioenergy field, specifically with regard to algal-based biofuels. This huge investment, including both federal and private funding into the algal-based biofuel arena has been almost neglecting the fact that on average 50% or greater of the total biomass of a non-genetically modified algal cell is non-bioenergy material. Most algae companies have rushed to the assumption that the biomass will take care of itself. This is an unfortunately ignorant position. Our proposal is but one of many necessary steps to provide a diverse secondary market for the residual biomass created by algal-based biofuels.
A viable long-term solution would be to create species specific fish feeds based upon algal biomass. Gaiergy Corporations SBIR proposal intends to conduct a comparative study using conventional fish feeds versus algae-enhanced fish feeds fed to juvenile Florida Pompano, Trachinotus carolinus, until they reach a market size of approximately 1.5 pounds. We will choose at least two commercially available fish feeds and create at least 2 unique fish feeds containing algae. From the various previous research using open fish-feed formulations and documented conventional feed performance trials that did not utilize algae, we will create our own unique feed by replacing those ingredients derived from terrestrial vegetation, meat & poultry by-products and fishmeal with algal biomass that we will grow ourselves. (Groat, 2002) (Hauville, Main, & Barrows, 2012) (Rhodes, Zhou, Davis, Chappell, & Hanson) (Riche, 2009) (Rossi, 2011).
4. Background and Rationale
The Florida Pompano is chosen because there is a direct commercial profitability aspect related to developing a species specific fish-feed for this species that is sustainable and cost effective. Currently, the South Florida area market prices for Florida Pompano can reach anywhere between $14 and $18 per pound. Considerable research has already been performed by Harbor Branch Oceanographic Institution, Mote Marine Laboratory, the USDA and other established aquaculture research institutions to prove that Pompano can be grown in shore-based recirculating aquaculture systems (RAS) (Groat, 2002) (Hauville, Main, & Barrows, 2012) (Riche, Weirich, Pfeiffer, Wills, & Davis, 2009). Since Pompano are considered to be continuous spawners (Watanabe, 1995), Pompano are a good choice for aquaculture. When compared to 6
growing tilapia, which receives anywhere between $3 and $5 per pound, a strict business choice can be made to direct new commercial efforts towards growing and selling Florida Pompano. The problem resides with rising feed costs making this type of aquaculture unsustainable.
When considering just the revenue case, raising Pompano for sale is an easy choice versus another fish such as tilapia. But when you consider the costs involved in raising Pompano including a salt water recirculation system and carnivorous dietary requirements, it becomes clearly evident that lowering the current cost structure for raising this species is the limiting factor why Pompano aquaculture has not taken off (Main K. , Resley, Rhody, Nystrom, Stevens, & Adams, 2010). One of the problematic considerations towards growing Florida Pompano revolves around the salinity concentration that the Pompano require to reach a healthy market weight within a reasonable timeframe. (Riche & Williams, 2010) (Wills, Pfeiffer, & Riche, 2010). The other consideration is lowering feed costs while improving nutritive value.
While other researchers are continuing on researching water chemistry and physical containment characteristics, including the two co-PIs working with Gaiergy Corporation on this proposal, Gaiergy Corp. needs to focus on lowering feed costs while increasing nutritive value, because we know feedstock costs are going to be continually increasing and a defined algal-based fish feed has not been successfully developed for Florida Pompano when considering the future overwhelming supply of algal biomass that will be coming to market as algal-based biofuels become a reality.
Further rationale to support this type of research involves understanding the market pricing dynamics involved with a fish meat that is highly desired, therefore highly priced versus a fish whose labor is not as highly valued and its supply has to compete against cheap Chinese imports. For our example we will again use tilapia. While there is wide market acceptance of this fish and commercial aquaculture farms raising this fish do thrive, they have to compete against lower production costs from Chinese-raised tilapia. This international competitive market pressure drives the price per pound down to the import cost of the Chinese tilapia. While this market price pressure is good for the consumer, is a barrier to entry for new aquaculturists.
To continue with this market pricing rationale discussion, it is important to understand how the market will change regarding price and availability and how new higher-valued products incentivize or lower the barrier to entry for new aquaculturists. When you first introduce your product to the market, since it is in limited supply you can charge a relatively higher price, such is currently seen with the price of Florida Pompano. This allows the new businessman to reap a higher profit with a lower volume of product offerings for the first few years. And if the businessman a smart, he/she either introduces new high dollar value items or increases the production of its current product offering while lowering his cost of production through increasing economies of scale. Through time, as the availability of the fish product increases due to increasing production and other competitors entering the marketplace, the price that you can charge now has to decrease since the supply is increasing. So this type of research proposal lays the groundwork for small-scale, high-dollar value fish production that incentivizes new investment into aquaculture due to the market pricing dynamics detailed above.
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The technical approach for our research is to utilize the previous research that has been conducted on Florida Pompano using conventional feeds and unique feeds (Gatlin, et al., 2007) (Cavalin & Weirich, 2009) (Groat, 2002) (Lazo, Davis, & Arnold, 1998) (Rossi, 2011), and then adding a specific percentage of algal biomass to both feeds through grinding, mixing and then re- extruding modified fish pellets. We will be utilizing previous research to establish reasonable and defensible protein, lipid, carbohydrate and vitamin/mineral compositional percentages. This will be the feed-starting point for our research. Previous Feed Trials It has been shown that juvenile Pompano are fairly indiscriminate feeders. Wild caught juvenile gut content analyses have shown crustaceans, mollusks, amphipods and crab larvae (Armitage & Alevizon, 1980). Feed conversion ratios, the amount of dry feed fed to a fish versus the wet weight body mass gained, for Pompano have been indicated in the range of 3.5 for conventional dry feeds (Williams T. , 2008). The effect of fish oil, specifically menhaden oil on weight gain was shown to have the most beneficial results when the percentage of oil to total unit mass of fish feed was between 4% to 8% lipid (Williams, Lovell, & Hawke, 1985). Diets that had oil percentages higher than 8% showed decreasing digestible energy to digestible protein ratios. This should not be confused with the total lipid content though. Protein composition in fish-feed is typically between 40% and 55% depending on the manufacturer, life-stage requirements and the species (www.skretting.us ,www.zeiglerfeed.com or www.rangen.com ). The type of protein historically used in fish-feeds is fishmeal from sources such as menhaden, anchovy and herring. Current research is turning more toward terrestrial vegetative protein sources (Rhodes, Zhou, Davis, Chappell, & Hanson) (Riche & Williams, 2010) as a lower cost alternative. The majority of Pompano specific fish feeds created by researchers over the past 15 years have protein concentrations ranging between 40% and the 55% protein, between 8% and 16% lipid, about 2% or 3% vitamin and mineral mix while the rest can be variably assigned to ash, carbohydrate and other binders (Lazo, Davis, & Arnold, 1998)(Riche & Williams, 2010) (Rossi, Davis, & Rhodes, 2010) (Gonzales-Felix, Davis, Rossi, & Perez-Velazquez, 2010) (Main K. , Resley, Rhody, Nystrom, Stevens, & Adams, 2010). Algae Feed Formulation
Since there are thousands of strains of algae, we needed to select just a few species of algae that would be appropriate for our research. The basic criteria for our algae needed to be: 1. Simple to grow, 2. High protein content, 3. Established previous research, 4. Wide tolerance to temperature and salinity, 5. Similarity of lipids to wild stock composition, and 6. Enriched carotenoid complexes. In Becker's book, Microalgae (1994), he constructed a simple table comparing the chemical composition normal human food sources to a few algal species.
By comparing the protein percentages in the Table 1 to the protein percentages in the feed trials and conventional feeds, the algae that we select for this research needs to have a protein percentage greater than 50% of its dry weight biomass. It has to be higher than 50% because on addition into our unique Pompano-specific formulation, the resultant protein percentage in the 8
formulation needs to be maintained at the same or greater level to ensure that we are delivering at least the same nutritional value to our Pompano subjects.
Table 1. Gross comparison of protein percentages from terrestrial and algal sources
The algae species that we will be using for our proposal include: Spirulina maxima, Dunaliella salina, Chlorella vulgaris, Chlorella pyrenoidosa and Haematococcus pluvialis. We are choosing a widespread of algae to ensure compositional diversity and increase the likelihood of community competition dynamics that will mitigate potential community crashes or contamination events usually seen in monoculture systems. While the D. salina species is marine tolerant, the H. pluvialis is grown mostly in freshwater. We will not be growing these two species in the same containment vessel at the same time. But we need the option, if we so choose, to grow our algae in marine or freshwater. The H. pluvialis has been shown to be easy to grow in small containment systems and is a excellent food source for the compound astaxanthin, which is a beneficial aquaculture compound (Dominguez, Ferreira, Coutinho, Fabregas, & Otero, 2005).
5. Relationship with Research or R&D
The significance of this research has at least three direct effects: 1. the results of this research will be directly applied to creating a commercial aquaculture firm in South Florida that provides Pompano grown on algal-enriched feeds. 2. The results of this research will be used to approach other current algal-based biofuel companies as a potential joint venture or outright technology sale to grow the required algal biomass that needs to be incorporated into species specific fish feeds. 3. Follow-on phase 2 proposals will analyze and hopefully normalize a wide diversity of unique feeds previously used in feed trials on Florida Pompano so that future researchers may utilize all previous research against a normalized platform. 4. Phase 2 proposals will also include research to build upon the current phase 1 results to develop small, diversified, just-in-time (JIT) aqua feed production platforms so that this Commodity/Algae Protein % Commodity/Algae Protein % Baker's Yeast 39 Spirogyra sp. 6-20 Rice 8 Dunaliella bioculata 49 Egg 47 Dunaliella salina 57 Milk 26 Euglena gracilis 39-61 Meat Muscle 43 Prymnesium parvum 28-45 Soya 37 Tetraselmis maculata 52 Scenedesmus obliquus 50-56 Porphyridium cruentum 28-39 Scenedesmus quadricauda 47 Spirulina platensis 46-63 Scenedesmus dimorphus 8-18 Spirulina maxima 60-71 Chlamydomonas rheinhardii 48 Synechococcus sp. 63 Chlorella vulgaris 51-58 Anabaena cylindrica 43-56 Chlorella pyrenoidosa 57 9
technology does not need to reside in large-scale feed producers. Rather, it can be utilized by coastal agrarian populations for their aquaculture feed needs.
6. Technical Objectives
The goal of this project is to develop and test an algae-based high protein diet to improve the growth performance of a high-value marine finfish, Florida pompano Trachinotus carolinus. Our Phase I project will address this goal through the following technical objectives: 1. Produce suitable algae strains to be used in the marine finfish diet. 2. Develop an algae-based high protein finfish pelletized diet. 3. Compare growth performance of juvenile pompano fed algae based and traditional high- protein diets of similar composition and physical properties. 4. Compare product quality and taste characteristics of pompano reared on the above diets. 5. Distribute market sized pompano reared on algae based and traditional diets to the commercial market. During Phase I, algae production will occur at The marine feed will be developed byPhase I will allow us to evaluate the application of the algae infused diets in pilot commercial-scale production trials based at Mote Marine Laboratory, Mote Aquaculture Park, located in Sarasota, Florida. The animals will be reared in a zero-discharge integrated recirculating aquaculture system which co-produces marine plant species as they process the finfish wastewater.
7. Work Plan
Objective 1: Order Necessary Feed Mixes, Equipment & Consumables
The equipment that is required is laid out of the following sections. Assuming that this project is funded, Gaiergy Corporation will initiate purchasing all the required components, consumables and perishables that are required for this project. The capital equipment that will be purchased will be purchased only from vendors in the United States except for the small-scale fish feed extruder. The longest leadtime for any specific piece of equipment is 2 months. The remainder of the equipment can be purchased from local vendors, online retailers and mail-order catalogs. All individual purchases will be below the 10% of total project cost threshold established by the federal government.
Objective 2: Replicate Gaiergys Previously Deployed Algal Production System 10
Algae Production System
The algae production system has already been designed and placed in-use by Gaiergy Corporation for another private organization. The technology for the system is fairly simple and has been used in various parts by multiple organizations for many years. For this particular research though, a simple downsized modification will be used to produce the required amount of algal biomass that needs to be incorporated into the fish feed.
For this purpose, the algae will be produced in simple translucent cylinders that are commercially available. The number of cylinders that will be deployed in this study will range between 6 and 15 x 80 gallon cylinders. The cylinders will be exposed to normal day light and LED enhanced light. Gaiergy Corporation is done extensive testing on multiple algal species and has found that an augmented light cycle produces high-density cultures in medium to large-scale vessels in the shortest amount of time. With photon flux densities approaching 2000 mol per square meter and algal densities increasing beyond 1 million cells per milliliter, the high photon flux density does not negatively affect the algal community so long as sufficient circulation exists in the containment field. With sufficient circulation, combined with variable shading effects from other algae in direct line of sight to the light source, high algal densities (>15 million cells/ml) can be maintained in a confined environment under economical conditions.
Objective 3: Start Continuous Production of Small Scale Algal Batches
We will begin algal production within two weeks of gaining project approval. Water for the system will be provided by a reverse osmosis. The algae strains will be obtained from any number of commercially available sources such as the UTEX Culture Collection, The National Center for Marine Algae and Microbiota, or Carolina Biosciences. Nutrients will be provided by hand using a modification of F/2 Guillards medium. The LED light source will be focused in the red and blue light spectrum emitted by LEDs with focal dispersions no greater than 45. Algal densities will be monitored on a daily basis via a bright field/florescent algae counter that Gaiergy Corporation has previously used for its algal studies. Temperature, pH and oxygen will be monitored via a Hachs HQd bench top meter. Light intensity will be monitored by Li-Cors LI-193a spherical underwater light sensor.
Objective 4: Harvest & Spray Dry Algae
Algae maturation can take anywhere between three and 16 days depending upon various factors. Our algae will mature ready for harvest within 7 to 10 days. Harvesting/Gross De-watering of the algae will be conducted through a simple solid bowl centrifuge. Gaiergy Corporation has already conducted multiple harvest experiments using multiple technologies including dissolved air flotation, agitated filtration, stacked disk centrifugation, electro coagulation and new centrifugal methods developed by Evodos. For this research, the most economical choice will be a small solid bowl centrifuge. Considering the realistic maximum potential biomass of algae as a function of cylinder volume, a small conventional solid bowl centrifuge will provide the lowest harvest cost per wet biomass unit.
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The algae will then be stored in nitrogen blanketed containment vessels at a temperature below 45F. The wet algal biomass will then be transported to a pharmaceutical, pilot size spray drying system. There are four known commercial entities that have pilot spray drying capabilities. All spray dried providers work on and as-needed basis. Therefore, when Gaiergy Corporation has the wet biomass available, we will package that biomass immediately and ship it to the spray drying contractor. Once the spray drying contractor has spray dried the algae into a powder, the contractor will then either vacuum seal the algae into Mylar bags or place the dried algae into rigid containers that are blanketed by nitrogen. The contractor will then ship the spray dried algae back to Gaiergy Corporation.
Objective 5: Pelletize Algal-Enhanced, Pompano Specific Fish on As-Needed Basis Utilizing the Meadow Stone Burr Grain Mill, we will create a flower from the conventional feed that we purchased for our study. The grain mill is a commercially available appliance that produces extremely fine-grained flour and will have little problem performing this task. The spray dried algae will then be added to the pulverized commercial fish feed into different ratios. We will make a 5% and a 20% algal addition. Since the feed rate for the Pompano will be dependent upon body mass we will start by creating 2 x 100 pound batches of Pompano specific fish feed.
The first 100 pound batch will have a 5% mix of algal-based biomass. Therefore, 5 pounds of algae will be added to 95 pounds of pulverized fish feed. Since the average fish feed moisture content is still between 10% and 15%, and the spray dried algae will have a residual moisture content of approximately 3% to 6%, we will add about 20 mL of water to every liter of powder so that the extruding process will work better, thus creating a pellet that floats. We assume approximately 20 mL of water because we cannot specifically say how much moisture will be entrained in the commercial feed at the time of pulverization nor can we specifically know what the humidity will be like on the day of processing. It will have to be adjusted when the process begins to create the best feed that we need.
The same process will be used for the second 100 pound batch except 20% of the biomass will come from the spray dried algae. We do not anticipate a fourfold increase in the amount of water necessary to create the proper expansion during pelletization, but we would expect it to probably double due to the lower water content and higher additive percentage to the whole premix. For the 20% algal-enhanced Pompano fish feed, we will experiment with a wet-harvest algae incorporated directly into the pulverized commercial feed as a way to eliminate spray drying and then re-adding water for the pelletization process. Since the pellet mill that we are using cannot accept 100 pounds of product all at once, we can experiment with the wet-harvest production in 10 pound samples. In other words, we will take 2.2 pounds of wet algae and add that to 8 pounds of pulverized conventional feed. We will mix that together and then send that through the pellet mill. On average you can expect anywhere between 10% and 20% moisture loss through the extruding process and subsequent drying as the temperature of the newly extruded feed cools down.
One of the goals of this proposal is also to show that species specific fish feeds are high-quality feeds that have a perishability attached to them. Since there is a perishablity associated with producing fresh, high-quality feeds, we will not be producing all of the feed that we will need all 12
at one specific time. Rather, we will be continuously producing small batches of our Pompano specific fish feed mimicking what a shore-based Pompano aquaculture facility would also be doing. In this manner we show that small-scale algae production can complement small scale finfish aquaculture production.
This is not meant to eliminate large-scale fish feed producers, but it will provide an alternative for aquaculture farmers to choose whether they want to produce their own feeds or rely upon a commercial provider. This research may also be used by those same commercial providers to improve their current feed offerings for those customers that may request Pompano-specific fish feeds.
As a precaution against equipment failure, we have enlisted Dr. Sajid Alavi from the Kansas State University, Department of Grain Science and Industry to collaborate with us on extrusion technical efficiency and if needed, we can send our product to Kansas States extruder for processing.
Objective 6: Compare growth performance of juvenile pompano fed algae based and traditional high-protein diets of similar composition and physical properties.
Feed trials will be conducted in FH1 (Figure 1). This system is equipped with one 35.65 m 3
rearing tank (tank 7 or T7) and six 8.9 m 3 rearing tanks (tank 1 or T1, tank 2 or T2, etc.) connected to a common water filtration system (Figure 1). System water continually recirculates through the fish tanks then flows through a series of filtration systems that includes solids filtration, biofiltration, UV and ozone disinfection, foam fractionation, and liquid oxygen injection. Because the system is a zero-discharge recirculating water system, wastewater will be processed by three concurrent wastewater treatment systems (WT-1, GH-1, and GH-2, Figure 1) as part of a separate study.
In this study, experimental pompano will be reared in T1 T6, and each tank will be fitted with a separately-controlled automatic feeder used to dispense the respective feed treatments. Rearing salinity will be maintained at 11-15 ppt and water temperatures will be primarily controlled by ambient weather, but maintained several degrees warmer than the outside temperature because the system is situated both indoors and within a greenhouse. Water quality will be monitored daily for dissolved oxygen (D.O., mg/l), Temperature ( 0 C), salinity, and pH using a hand-held meter. Dissolved oxygen and temperature will also be continuously monitored in each tank using a pre-existing computer-controlled monitoring device that makes adjustments to the supply of liquid oxygen to oxygen cones and/or oxygen concentrators. The system is also connected to emergency oxygen supply lines and an automatic phone alert system if variables (D.O., Temperature, and water level) fall outside the desired levels. Ammonia and nitrite will also be monitored twice every week to insure that these variables are within desired levels (<0.5 ppm ammonia; <0.4 ppm nitrite).
Pilot work from USDA fellowship research conducted at MAP, has shown that growth juvenile performance was better in the MAP commercial-scale rearing system compared with smaller systems with fewer fish per experimental trial combination (Brennan et al., unpublished data). Therefore, two weeks prior to the study commencement, each experimental tank (T1 T6) will 13
be stocked with 120 pompano, age 45 days post-hatch (dph). These fish will originate from on- site larval rearing tanks and be acclimated from sea water to the study salinity prior to stocking. Because of high variability in genetic growth performance with this non-domesticated strain, experimental trial fish will include the fastest growing individuals of the cohort (e.g. largest 10% of the cohort). Three replicates of each feed treatment will be conducted (i.e. T1-T6 will be stocked accordingly). Additional pompano from the same larval rearing cohort will be stocked into the larger 35.65 m 3 tank (tank 7 or T7) and will be used as additional source fish if necessary.
Using the best available information to select a comparable commercial diet to the algae-based experimental diet, pompano juveniles (60 dph, approximately 20g) in each experimental tank will be fed one of the two above diets as extruded pellets at a rate of 3% mean body weight per day. Adjustments to the daily feed rate will occur according to typical growth projections obtained from previous pompano rearing experience. Optimal stage-specific pellet sizes and feed delivery schedules, and evaluation schedules will be used appropriately throughout the study based on findings from the USDA fellowship study. Growth parameters will be measured at the beginning of the study, then at three week intervals throughout the study. During each evaluation, measured variables will include body wet weight (g), total length (mm), fork length (mm), standard length (mm), Fultons Condition Factor (K), size distribution (growth uniformity), Specific Growth Rate (SGR) and Food Conversion Efficiency (FCE), where:
SGR = (ln(final mean fish weight) ln(initial mean fish weight))/days of study duration
FCE = Weight of feed delivered/(initial mean fish weight final mean fish weight)
Response variables (BW, fish length, K, SGR, and FCE) will be statistically compared using a one-way analysis of variance in SAS software (SAS Institute, Inc., Cary, North Carolina) using feed brand as a class variable and a least-squares means procedure to compare specific treatment means. Statistical differences will be considered significant at the = 0.05 level. Size frequencies of initial and final lengths and weights will be compared by length frequency plots. All pompano in T1-T6 will be harvested upon completion of the study.
Objective 7: Conduct Taste Trials with Local Restaurants and General Public
Once all of the data is collected regarding objective six, then we will look towards arranging taste trials at local restaurants. The fish will be donated to the restaurant or chef for simple preparation for taste testers. Our one stipulation for accepting the donation is that we be granted access to survey the patrons eating the fish.
8. Related Research or R&D
Significant work is being conducted on floor pompano by a multitude of different researchers. The primary interest of the majority of these researchers though is in the improvement of recirculating aquaculture systems (RAS), determining low salinity production parameters and incorporating more terrestrial vegetative sources into fish feeds. The impetus behind researching 14
higher amounts of terrestrial vegetation feed sources comes from the need to find a replacement for fishmeal and fish oil impounded into current fish feed formulations. As the oceans wild catch continues to dwindle while the human population and its protein consumption continues to increase, aquaculture feeds will not be able to depend upon wild caught fishmeal sources as an affordable input cost into fish feeds.
Mote Marine Laboratory is one of the institutions that is at the forefront of pompano research. The two co-PIs, Dr. Main and Dr. Brennan are both working on pompano research. The facility at mote Marine laboratory is one of the highest quality research facilities for pompano.
Directly on the other side of state from mote Marine laboratory is Harbor Branch Oceanographic Institute (HBOI). Before the USDA, agricultural research station closed down at HBOI,
9. Potential Post Application
Gaiergy was founded in 2006. It has operated as a boutique consultancy in the field of renewable energy and oceanographic science. Mr. Masters has worked for The National Oceanic and Atmospheric Demonstration (NOAA), the Department of Energy and various oceanographic universities and private algae companies. Gaiergy has a developed business plan for commercial pompano production with defined outlets for the sale of the fish into the South Florida market. Using the research of local scientists, such as the co-PIs involved in this proposal, USDA aquaculture scientists and HBOI scientists, Gaiergy developed a business plan that while it shows significant profitability potential, there is also volatility risk that is too high in the current economy's assessment to warrant funding. So, research must be conducted to provide answers that will be able to be used in the business plan to lower the internal volatility risk associated with fluctuating feed cost prices.
The results of this SBIR funding will be used to modify a business plan regarding whether or not to pursue on-site fish feed production for a commercial pompano operation. The results are anticipated to have a considerable cost-cutting benefit on the order of 20% to 25% cost reduction over a 3 to 5 year. While current commercial products are offered by producers such as Cargill, Rangan, Skretting, Zeigler and Star Milling, the products that they offer are not focused directly on pompano. Some of the products are geared towards Marine finfish and even within that product offering there are subset product offerings that vary protein, lipid, nutrient and carotenoid percentages. But the reality is that those products are still to generalistic, thus resulting in non-optimized conversion of feed into body mass.
There is a definite potential for this research to be used by the federal government. Currently, the USDA, DOE and DOD are engaged in intense public/private funding for algal-based biofuels. While there has been some success in creating algal-based biofuels, they have come at a cost that cannot currently compete with petroleum-based fuels. There will be more success if the focus is pushed more towards high dollar-value products from algae than commoditized fuels. This research and subsequent commercialization will be one specific example of that. 15
This research is not to suggest to the federal government that they should abandon algal-based biofuels, but it is a more prudent and profitable path for the federal government to pursue those products that can reap a higher dollar value per unit sold in the short term while the industry total volume output is allowed to grow over the next 5 to 10 years. If the volume output was allowed to focus on these higher dollar value items begin with, then as volume output increases along with time, the price for these food items will come down, thus allowing the algal oil to be pushed into the biofuel market at a lower per unit production cost than if the federal government continue down its current investment path where algal oil production costs are greater than $26 per gallon with no outlet for the algal biomass.
What advantages will the proposed research have over existing technology and application, performance, technique, efficiency or cost?
10. Satisfying the Public Interest
The Florida Pompano is chosen because there is a direct commercial profitability aspect related to developing a species specific fish-feed for this species that is sustainable and cost effective. Currently, the South Florida area market prices for Florida Pompano can reach anywhere between $14 and $18 per pound. Gaiergy Corporation already has significant interest from local restaurants, fishmongers and grocery stores in the South Florida area that would carry Pompano if there was a steady supply.
Strategic Goal 1
This research can satisfy the need to assist rural communities to create prosperity so there self- sustaining and economically thriving, because if this research is funded, then the results can be directly used for small, rural, coastal aquaculture interests. One of the specific goals of this research is to developing means for on-site, species specific fish feed production. When over 50% of the total cost of production for aquaculture resides in the fish feed (Hasan & Halwart, 2009), and that fish feed is created from one of five production facilities, as transportation cost increase combined with increasing feed input costs, small scale, rural aquaculture startups need to be able to produce their own feeds when the economics justify it. Strategic Goal 2
Our research addresses the strategic goal of enhancing our water resources, restoring and conserving our private working lands by providing a business case to grow algae off of wastewater or reclaimed water for the purpose of fish feed feedstock. One of the larger problems in municipal wastewater treatment plants is the tertiary clarification of wastewater before it is released back into the environment. At the point of tertiary treatment, the nutrient levels in the wastewater clarifiers are appropriate for these types of algae growing operations. A secondary 16
point of research for a phase 2 SBIR will be to investigate using tertiary clarifier water as the feedstock for algae growth. Strategic Goal 3
This research will definitely be promoting agricultural bio production through aquaculture old production of a much desired finfish. As our nation progresses into more globalized society that is influenced more and more by climactic changes, we need to be able to diversify our product offerings and methods to produce those product offerings. This research speaks directly to increasing that agricultural/aquacultural production, by improving the quality of the aquacultural product and diversifying the methods in which that product is grown.
Strategic goal 4
This research can lead to increasing safe, nutritious and balanced meals for children because the product in which intend to grow, Florida Pompano, is a Marine finfish that not only provides protein but also a healthy portion of omega-3 fatty acids including DHA and EPA. All of these compounds, the USDA has promoted as part of their heart healthy lifestyle.
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Bibliography Armitage, T., & Alevizon, W. (1980). the diet of Florida Pompano (Trachinotus carolinus) along the East Coast of Central Florida. Florida Science, 19-26. Cassiano, E., Ohs, C., Weirich, C., & Rhyne, A. (2010). Improvements in Growth, Survival, and Stress Resistance of Florida Pompano Trachinotus carolinus Larvae Fed the Calanoid Copepod Pseudodiaptomus pelagicus. Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AQ2010/AQ2010_0755.pdf Cavalin, F., & Weirich, C. (2009). Larval performance of aquacultured Florida Pompano (Trachinotus carolinus) fed rotifers (Brachionus plicatilis) enriched with selected commercial diets. Aquaculture, 67-73. Gatlin, D. M., Barrows, F. T., Dabrowski, K., Gaylord, T. G., Hardy, R., Herman, E., et al. (2007). Expanding the Utilization of Sustainable Plant Products in Aquafeeds: A Review. Aquatic Resources, 551- 579. Glencross, B., Booth, M., & Allan, G. (2007). A feed is only as good as its ingredients - a review of ingredient evaluation strategies for aquaculture feeds. Aquaculture Nutrition, 17-34. Gonzales-Felix, M., Davis, A., Rossi, W., & Perez-Velazquez, M. (2010). Evaluation of apparent digestibility coefficient of energy of various vegetable feed ingredients in Florida Pompano, Trachinotus carolinus. Aquaculture, 240-243. Groat, D. (2002). Effects of Feeding Strategies on Growth of Florida Pompano Trachinotus carolinus in Closed Recirculating Systems. Baton Rouge: Lousiana State University. Halwart, M., Soto, D., & Arthur, R. (2007). Cage Aquaculture: Regional Reviews and Global Overview. Rome: Food and Agriculture Organization of the United Nations: Aquaculture Management & Conversation Service. Hauville, M., Main, K., & Barrows, F. (2012). Effect of Different Micro-Diets on Florida Pompano Trachinotus carolinus - Weaning Success, Growth and Fatty Acid Incorporation. Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AA2012/AA2012_0392.pdf Heilman, M., & Spieler, R. (1999). The daily feeding rhythm to demand feeders and the effects of timed meal-feeding on the growth of juvenile Florida Pompano, Trachinotus carolinus. Aquaculture, 53-64. Lazo, J., Davis, A., & Arnold, C. (1998). The Effects of Dietary Protein Level on Growth, Feed Efficiency and Survival of Juvenile Florida Pompano (Trachinotus carolinus). Aquaculture, 225-232. 18
Main, K., Resley, M., Rhody, N., Brennan, N., & Nystrom, M. (2011). Developing Florida Pompano Hatchery Technology for Offshore Aquaculture, Land-Based Systems and Stock Enhancement in the Gulf of Mexico. New Orleans: Aquaculture America. Main, K., Resley, M., Rhody, N., Nystrom, M., Stevens, T., & Adams, C. (2010). An Overview of Florida Pompano Trachinotus carolinus Research at Mote Aquaculture Research Park. Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AQ2010/AQ2010_1003.pdf Rana, K., Siriwardena, S., & Hasan, M. (2009). Impact of Rising Feed Ingredient Prices on Aqua Feeds and Aquaculture Production. Rome: United Nations, food and agricultural organization, fisheries and aquaculture technical paper. Rhodes, M., Zhou, Y.-G., Davis, A., Chappell, J., & Hanson, T. (n.d.). Growth response of Florida Pompano (Trachinotus carolinus) fed taurine supplemented high soy diets compared to fish meal based diets. 2012. Riche, M. (2009). Evaluation of Digestible Energy and Protein for Growth and Nitrogen Retention in Juvenile Florida Pompano, Trachinotus carolinus. Journal of the World Aquaculture Society, 45- 57. Riche, M., & Williams, T. (2010). Apparent digestibile protein, energy and amino acid availability of three plant proteins in Florida Pompano, Trachinotus carolinus in seawater and low-salinity water. Aquaculture Nutrition, 223-230. Riche, M., & Williams, T. (2010). Apparent digestibility protein, energy and amino acid availability of three plant proteins in Florida Pompano, Trachinotus carolinus in seawater and low-salinity water. Aquaculture Nutrition, 223-230. Riche, M., & Williams, T. N. (2010). Apparent Digestible Protein, Energy and Amino Acid Availability of Three Plant Proteins in Florida Pompano, Trachinotus carolinus in Seawater and Low Salinity Water. Aquaculture Nutrition, 223-230. Riche, M., Weirich, C., Pfeiffer, T., Wills, P., & Davis, M. (2009, February). Trials Advance Low-Salinity Culture of Cobia, Pompano & Other Species. Retrieved from Global Aquaculture Advocate: http://naldc.nal.usda.gov/download/42079/PDF Rodrigues, R., Schwarz, M., Delbos, B., & Sampaio, L. (2007). Acute tocicity and sublethal effects of ammonia and nitrite for juvenile cobia Rachycentron canadum. Aquaculture, 553-557. Rossi, W. (2011). Development of Alternative Protein-based Diets for the Intensive Production of Florida Pompano Trachinotus carolinus. Auburn, Georgia: University of Auburn. 19
Rossi, W., Davis, A., & Rhodes, M. (2010). Replacement of Fish Meal by Meat and Bone Meal in Practical Diets for Florida Pompano (Trachinotus carolinus). Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AQ2010/AQ2010_0505.pdf Rossi, W., Davis, A., & Rhodes, M. (2010). Replacement of Fish Meal By Poultry-by-Product Meal in Practical Diets for Florida Pompano (Trachinotus carolinus). Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AQ2010/AQ2010_0508.pdf Watanabe, W. (1995, pp185-205). aquaculture of the Florida Pompano and other jacks (Family Carangidae) in the Western Atlantic, Gulf of Mexico and Caribbean basin: status and potential. Culture of High-Value Marine Fishes. Waimanalo, Hawaii: Oceanic Institute. Weirich, C., & Riche, M. (2006). Acute tolerance of juvenile Florida Pompano, Trachinotus carolinus, to ammonia and nitrate at various salinities. Aquaculture Research, 855-861. Weirich, C., & Riche, M. (2008, April). Pompano Reared to Market Size in RAS Low Salinity Culture Successful. Retrieved from Global Aquaculture Advocate: http://naldc.nal.usda.gov/download/42075/PDF Weirich, C., Riley, K., & Davis, M. (2005). Captive Reproduction, Larviculture of Florida Pompano. Retrieved 2005, from Global Aquaculture Alliance: http://pdf.gaalliance.org/pdf/GAA-Weirich- Jun05.pdf Weirich, C., Wills, P., Baptiste, R., Woodward, P., & Riche, M. (2009). Production Characteristics and Body Composition of Florida Pompano Reared to Market Size at Two Different Densities in Low- Salinity Recirculating Aquaculture Systems. North American Journal of Aquaculture, 165-173. Williams, S., Lovell, R., & Hawke, J. (1985). Value of menhaden oil in diets of Florida pompano. Prog Fish Culture, 159-165. Williams, T. (2008). An Assessment of Alternative Feed Ingredients in Practical Diets for Florida Pompano (Trachinotus carolinus) Held in Low Salinity Recirculating Systems. University of Maine. Wills, P., Pfeiffer, T., & Riche, M. (2010). Production of Florida Pompano Trachinotus carolinus in Low Salinity Systems. Retrieved from World Aquaculture Society: https://www.was.org/documents/MeetingPresentations/AQ2010/AQ2010_0471.pdf Woods, M. (2009, February). Growing Premium Seafood - Inland. Agricultural Research, pp. 14-16.