Progress 09/01/18 to 08/31/22

Outputs
Target Audience:The principal target audience for this research project is small farmers in the Midwest looking to fill local niche markets for live or fresh shrimp products. AST provided design assistance to several major shrimp producers in the Midwest during this grant. Several have purchased small filters for their evaluation and several large filters have been sold to Canadian producers. In the early 2020-2021 time, two distinct designs for marine shrimp production facilities were developed. One design was proposed for placement in southern Florida and the second for an investment group in Michigan. Late in the 2020-2021 timeframe, almost all commercial development activities were halted; first by the COVID-19 impact on the restaurant industries and secondly by the Canadian import ban of post larvae shrimp. In 2022, after the COVID-19 induced hiatus, sales of large-scale units into the shrimp sector resumed. Changes/Problems:During the proposal stage, it was envisioned that the active nitrification would be maintained within the production tank by suspended solids while maintaining the total solids concentration below a targeted maximum (400-500 mg/L). The control systems required to control the suspended levels in the tank became prohibitively complex. So, it was decided to reverse the thinking and target accomplishing the nitrification in the external activated sludge basin. The change in thinking converges upon a fixed film and ASB approach described for the MDC series. Both system configurations consist of a PolyGeyser® and an ASB. In the ASB configuration, the PolyGeyser® is smaller and water is circulated though the ASB at a slow rate. The research team developed a unified design that changed the circulation pattern to maximize the benefits of pairing the PolyGeyser® and an ASB. What opportunities for training and professional development has the project provided?Training future engineers Over the course of this research project, AST has employed 23 undergraduate students from Louisiana State University from various engineering and marketing disciplines. The types of things the students have been taught are as follows: Fundamental lab protocol (emphasizing lab safety) Proper methods of taking, handling, and storing water samples Methods of measuring water quality parameters (total suspended solids, ammonia, nitrite, nitrate, dissolved oxygen, pH, alkalinity, temperature, salinity, sulfide, and turbidity) held to academic research standards as well as the meaning behind the measurements They were also shown how to utilize Standard Methods to figure out how to measure various water quality parameters How to compile and interpret data Strategies and techniques used to develop a targeted market. YouTube video marketing techniques High School Aquaponics Despite COVID, AST has continued to work to encourage the use of aquaponics in high school and middle-school STEM programs. In the last two years (2021/2022), the research group directly assisted in the setup and operation of three AST MDC aquaponics systems, two in Baton Rouge, Louisiana and 1 in Dallas, Texas. In addition, over a dozen MDC aquaponic systems we sold and installed in schools and colleges which were facilitated by our distributors. This appears to be an expanding trend. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Testing of the PolyGeyser® and biofloc combination lead the research team to the conclusion that the "in tank" biofloc component was too complex to operate by our intended user group. This complication stemmed from TSS limits (200-500 mg/L) placed on modern biofloc systems. Control of the TSS levels was inherently complicated as it required focused operator attention and process control. Additionally, the TSS limits inherently limited the nitrification capacity of the system. The decision was made to move the biofloc system to an external reactor. The external reactor chosen was the Aerated Sludge Basin (ASB), where TSS levels can exceed 10,000 mg/L and can be intuitively controlled. The relatively low shrimp densities (10 kg/m3 compared to normal fish densities of 60 kg/m3) and the need to maintain reasonable tank turnover rates lead to a PolyGeyser sizing that eliminated nitrification capacity as a concern. The ASB was relegated to the support role of a sludge digestor, which greatly extended water reuse while minimizing costly salt discharges. To this end, modeling and research was conducted on the design of a sludge degradation basin (SDB) that could denitrify and supplement nitrification depending on the duration of aeration and the availability of carbon. Sludge degradation basins were designed to match the sludge production for each of our rotomolded PolyGeyser® models. SDB management strategies for several RAS configurations (marine closed RAS, marine semi-closed, freshwater aquaponics, closed freshwater RAS) were developed. New SDB management techniques enabled new styles of systems and product lines to be created such as the Modified Decoupled or "MDC" product line. These early airlifted PolyGeyser® systems still have external filters, but otherwise reflect the developing technology. The early market development in the freshwater aquaponic market has been particularly strong. Virtually all expansion of RAS marine shrimp production was halted during 2020 with the COVID closure of restaurants markets. The research team designed and evaluated the means of pneumatically connecting the PolyGeyser® bioclarifier to the SDB, techniques for recycling sludge digestion supernatants, the use of a hypolimnetic aerator (HLA) to create a vertically partitioned SDB, and the means of utilizing the SDB as a supplemental nitrification unit. These features are described in U.S. Patent #11,219,194 issued to Dr. Ron Malone on January 1, 2022 for application to both aquaponic and aquacultural applications. Units distributed under the "MDC" product line have been actively marketed and sold since late 2021. The HLA features have recently been introduced to the market (late 2022). These small-scale units are particularly popular with high school and college educational programs. Under this Phase II in 2019, at a scale of 0.25 m3, the research team successfully produced an average (across three replicates) of 10.2 kg/m3 in the fixed film format that seemed to be limited by nitrite. A production level at 10 kg/m3 density was also achieved in a 600 gallon fiberglass FIT system. The 10 kg/m3 density is approximately double that is routinely achievable in Biofloc systems, which is currently the most widely used form for tank based marine shrimp production. In the 2021-22, the research team successfully conducted 4.5 m3 tank system growout for stage B shrimp (1-19 gram) with final harvest densities ranging from 9-10 kg/m3. About one third of the attempts conducted in the 4.5 m3 scale were terminated early due system upsets. These upsets caused nitrite peaks leading to shrimp loss. System details, feed rates, backwash frequencies were being tested and evaluated which increased the probability of system upsets. Most of these failures stemmed from early system design errors in the airlift configuration and the pneumatic sludge exchange leading to sudden nitrite accumulations. The underlying cause was a reduction in recirculation flows stemming from poor backwash interval selection, backwash air delivery problems, or sludge movement to the ASB. The resulting drop in recirculation rates lead to inadequate oxygen levels which caused rapid nitrite increases. These failures were not related to shrimp density. The most of the successful 4.5 m3 growouts achieved or approached densities of 10 kg/m3. The research team determined that a production goal of 10 kg/m3 was a reasonable expectation. Limited commercial testing has verified that this is a reasonable commercial goal in a bare tank configured with an MDC configuration with an airlift, airlift compatible floating bead filter, and an SDB. The design of MDC inland shrimp culture is summarized in a document posted on the AST website. This document describes the design details and performance expectations for an airlifted PolyGeyser® in an MDC configuration for annual production goals of 5,000 to 20,500 lbs of 25 count shrimp. The proposed system consists of commercially available polymer components (tank, filter, sludge digestor) and fully driven by a mechanical air blower. No water pumps or electronic controls are required for the core RAS operation. The research team successfully designed, operated, and sold fiberglass FIT systems, but concluded that their cost was prohibitive for the intended market. Several prototype FIT systems were evaluated, but none of the designs demonstrated the ability to rectify a biofouled condition remotely. The FIT approach was abandoned for the more promising MDC approach. As part of the FIT development program the research team produced several scale models using 3-D printers. These FIT prototypes failed to meet the rigorous evaluation criteria set for FIT performance but were selected as a potential spin off product. The promising 8" (Nano 8000) model underwent detailed beta testing both in house and at five external locations. Sales of these "Nano" units began in July of 2021. In the first half of 2022, nearly fifty units were sold to upgrade existing R&D rack systems. In late 2022 AST began to fabricate the metal racks supporting the Danio Lab filters and R&D tanks. AST is now investigating the feasibility of injection molding for key components to bring the production cost down. This would allow a much broader market development for home aquaria. The Phase 1 economic modelling effort remains valid. These modelling efforts were based upon the use of fiberglass components and required a density of about 13 kg/m3 to produce a profitable outcome. To improve economic projections the "CaRASel" strategy, which is largely based upon the use of molded polyethylene components, has been developed and is actively being promoted for commercial production use at 10 kg/m3. An economic analysis was conducted on the MDC "CaRASel" system. Three system configurations were considered: 17,000 gallon system, 34,000 gallon system, and a 68,000 gallon system. The net profit of each system was calculated at various shrimp prices. Conservative assumptions were made for key parameters: harvest density 10 kg/m3, 6% interest on the startup loan, and a 15 year loan length. At a shrimp price of $16 per pound (based on the 2015 Purdue study) the 17,000 gallon system will generate a return on investment (ROI) of 5.5 years, the 34,000 gallon system will generate an ROI of 2 years, and the 68,000 gallon system will generate an ROI of 1.9 years. Return on investment is heavily dependent on the price of shrimp and harvest density. The price of shrimp is determined by the local market, but harvest density is managed by the skill of the farmer. We consider a harvest density of 10 kg/m3 to be conservative, and that a harvest density of 15 kg/m3 can be achieved by a skilled farmer. As with all aquaculture ventures, success is largely determined by the farmer's sales skills, husbandry, and ability, as well as the stability of the local market.

Publications

• Type: Other Status: Published Year Published: 2022 Citation: A white paper entitledModified decoupled (MDC) Model AST description & technical details by Ronald Malone and Connor Tiersch, dated September 2022, posted on the AST filters website: https://astfilters.com/technical-documents/
• Type: Other Status: Published Year Published: 2021 Citation: A white paper entitled Airlifted Polygeyser� RAS for the marine shrimp by Ronald Malone and Connor Tiersch, Dated Spring 2021, posted on the AST filters website: https://astfilters.com/technical-documents/
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: In Pursuit of Small-Scale Indoor RAS Production of Marine Shrimp by Ronald Malone, Matthew Louque, and Tim Pfeiffer was presented at Aquaculture 2019, a meeting of the World Aquaculture Society held in New Orleans, March 7-11, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Fixed film and suspended growth in small scale indoor production of marine shrimp by Marie Gutierrez-Wing, Matthew Louque, Timothy Pfeiffer, and Ronald Malone was presented at Aquaculture America 2020 in Honolulu Hawaii, February 9-12, 2020.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Clear-water fixed film carousel production of marine shrimp by R. F. Malone and Connor Tiersch was presented at the Aquaculture America 2021 in San Antonio, Texas, August 11-15.
• Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Failure testing the modified decoupled recirculating aquaculture system by Connor Tiersch and R. F. Malone was presented at the Aquaculture America 2021 in San Antonio, Texas, August 11-15.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Interim sizing guidelines for airlifted PolyGeyser� for the marine shrimp Liptopenaeus vannamei by Connor Tiersch, R.F. Malone, and Marie Gutierrez-Wing was presented at the Aquaculture 2022, a meeting of the World Aquaculture Society held in San Diego, California, February 28  March 4.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Use of localized sludge digestors to extend water reuse in closed inland marine shrimp RAS production by R. F. Malone, Connor Tiersch, and Marie Gutierrez-Wing at the 2022 RASTECH Conference and Tradeshow held on Hilton Head Island, South Carolina, March 30 & 31.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Linking localized sludge digesters to PolyGeyser� with pneumatic exchange to facilitate decoupled aquaponic operations. by R. F. Malone and Connor Tiersch, and Sam Tanner at the 2022 RASTECH Conference and Tradeshow held on Hilton Head Island, South Carolina, March 30 & 31.
• Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Use of localized sludge digesters in airlifted mineralized decoupled aquaponic systems by Ronald F. Malone, presented at the 11th Annual Aquaponics Association Conference in Oklahoma City, Oklahoma, September 30  October 2 2022.

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

Outputs
Target Audience:The principal target audience for this research project are small farmers in the Midwest looking to fill local niche markets for live or fresh shrimp products. AST provided design assistance to several major shrimp producers in the Midwest during this grant. Several have purchased small filters for their evaluation. After a COVID induced hiatus, sales of large scale unit into this sector are resuming. Changes/Problems:During the proposal stage it was envisioned that the active nitrification would be maintained within the production tank by suspended solids while maintaining the total solids concentration below a targeted maximum (400-500 mg/L). The control systems required to control the suspended levels in the tank became prohibitively complex. So, it was decided to reverse the thinking and target accomplishing the nitrification in the external activated sludge basin. The change in thinking converges upon fixed film and SDB approach described for the MDC series. Both system configuration consists of a PolyGeyser® and an SDB. In the SDB configuration, the PolyGeyser® is smaller and water is circulated though the SDB at a slow rate. The research team is looking at the development of a unified design that differs in the way it circulated allowing one design to serve distinctly two philosophical approaches. What opportunities for training and professional development has the project provided?Training future engineers Over the course of this research project, AST has employed 19 undergraduate students from Louisiana State University from various engineering and marketing disciplines. The types of things the students have been taught are as follows: Fundamental lab protocol (emphasizing lab safety) Proper methods of taking, handling, and storing water samples Methods of measuring water quality parameters (total suspended solids, ammonia, nitrite, nitrate, dissolved oxygen, pH, alkalinity, temperature, salinity, sulfide, and turbidity) held to academic research standards as well as the meaning behind the measurements They were also shown how to utilize Standard Methods to figure out how to measure various water quality parameters How to compile and interpret data Strategies and techniques used to develop a targeted market. YouTube video marketing 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?The research team is well positioned to develop a shrimp specific commercial PolyFit design that vertically integrates the SDB and PolyGeyser® for placement with the tank for the Geyser model that is linked to an external sludge digestion basin. The general configuration and engineering challenges are well known to the research and product development teams. This Endurance MDC precursor configurations will also be successful in other fresh and marine for the small-scale production of baitfish, ornamental fish, broodstock, fry production, and aquaponics. To improve the economic projections the Carousel strategy which is largely based upon the use of molded polyethylene components has been developed and is actively being promoted for commercial production use at 10 kg/m3. There are currently two AST customers that using the Carousel configuration as basis for their business plans. Further economic modelling will be undertaken after the carrying capacity of the MDC lines are established.

Impacts
What was accomplished under these goals? To determine the maximum sustainable nitrification capacity of the Activated Sludge Degradation Basin (ASB) component of a FIT system at the target temperature of 30°C and salinity of 25 ppt. Three replicated 280-liter tanks were configured with small custom PolyGeyser® for solids capture and a 10-liter ASB for aging the sludge. The systems were pneumatically configured with timers and solenoid valves to control the PolyGeyser® backwash frequency and to control dosing of sludge into the tank. The system proved difficult to manage, but, was successfully operated for two months with heavy feed loading (no shrimp). The research team concluded that knowledge base required to operate the system commercially would be prohibitive. The complexity resulted from the needed to maintain the tank suspended solids level between 400-500 mg/L while noting that the industry is trending towards even lower suspended solids levels. It was concluded that much simpler configuration would result from utilizing the ASB directly as a nitrification unit (external to the tank). To this end, modeling and research was conducted on the design of a sludge degradation basin (SDB) that could denitrify and supplement nitrification depending on the duration of aeration and availability of carbon. Sludge degradation basins were designed to match the sludge production for each of our rotomolded PolyGeyser models. Sludge basin management strategies for several RAS configurations (marine closed RAS, marine semi-closed, freshwater aquaponics, closed freshwater RAS) were developed and marketing of the "MDC" or "Modified DeCoupled" with direct sales, indirect sales through distributors, and online sales through Amazon. These early airlifted PolyGeyser Systems still have external filters, but otherwise reflect the developing technology. The early market development in the freshwater aquaponic market has been particularly strong. Virtually all expansion of RAS marine shrimp production was halted during 2020 with the COVID closure of restaurants markets. To determine the maximum volumetric shrimp density that can be supported by a bare tank unit and develop a suspended tier system to enhance the maximum volumetric shrimp density. Four 280-liter tanks were configured with a 0.7 ft3 Endurance 2000 PolyGeyser® filter, a 12.4-liter fluidized bed, and a foam fractionator. Ozonation and PHA capabilities were held in reserve. One system (#7) was dedicated to hold excess shrimp and three for replicated study of the fixed film strategy. Under this Phase II in 2019 effort the research team successfully produced an average (across three replicates) of 10.2 kg/m3 in the fixed film format that seemed to be limited by nitrite. A production level at 10 kg/3 density was also achieved in a 600 gallon fiberglass FIT system. The 10 kg/m3 is approximately double that routinely achievable in the biocfloc systems that form now widely used for tank based marine shrimp production. No further shrimp production studies were conducted in late conducted in late 2019, or, 2020 as AST centralized its R&D efforts closing its New Orleans Office and Innovation Park operations. The proposed greenhouse demonstration facility was delayed first by permitting problems, then COVID, and construction till the spring of 2021. In June of 2021, an experiment using three replicated 4.5 m3 systems (plus 1 RAS in reserve) with the intent of demonstrating production densities of 15-20 kg/m3 in a bare tank confirmation. To design and evaluate a prototype commercial size Hybrid Biofloc FIT unit for inland shrimp culture. The research team has designed an airlifted Endurance, SDB and Polytank system that can be used for fish or shrimp production under the marketing name MDC. The Endurance, SDB, and Airlift are external to the tank. This MDC configuration is considered a precursor to a true FIT configuration that would contain the filter and SDB internal to the tank itself. However, the unit functions identically to the in-tank configuration and may be hold a strong market position mimicking a FIT. The MDC systems are actively being marketed as a spinoff from this research effort. Proposed FIT precursors were printed using 3-D printers at about a 10 percent scale. The end caps, diffusers, screens, charge chambers, triggers, and pneumatic sludge components were printed in ABS plastic while the main hull was about an 18 inch clear 6" acrylic cylinder. Several versions of these PolyGeyser®s were fabricated and tested for biofouling resistance. These units proved highly effective as bioclarifiers under light to moderate loading regimes, but, tended to biofoul at higher loadings. This feature was problematic for a filter that would be hard to view when floating in a tank, so the filter was redesigned to eliminate the behavior. A 2 cubic foot (18 inches in diameter) unit is currently under fabrication and will be evaluated with shrimp in late summer of 2021. However, the performance of the 6 inch prototypes were so promising that the design team propagated an 8 inch design. This unit underwent detailed beta testing both in house and externally and is now being marketed by A Boston based company, Danio Lab, a division of South Boston Life Science Group (www.daniolab; www.sobolifescience.com) as a filtration upgrade for Zebrafish rack systems used widely for biomedical research. Sales of these "Nano "units began in July of 2021, and serious marketing thrust of this spinoff is anticipated for the fall of 2021. AST is investigating the feasibility of injection molding of key components to bring the production cost down, allowing a much broader market development for home aquaria. The research team is well positioned to develop a shrimp specific commercial PolyFit design that vertically integrates the SDB and PolyGeyser® for placement with the tank for the Geyser model that is linked to an external sludge digestion basin. The general configuration and engineering challenges are well known to the research and product development teams. This Endurance MDC precursor configurations will also be successful in other fresh and marine for the small-scale production of baitfish, ornamental fish, broodstock, fry production, and aquaponics. To determine the direct economic effects on the cost of shrimp production in a hybrid suspended growth system. The Phase 1 economic modelling effort remain valid. These modelling efforts were based upon the use of fiberglass components and required a density of about 13 kg/m3 to produce a profitable outcome. The research team was able to achieve a production level of over 14 kg/m3 in a single replicate of three and expects a sustainable production level in excess of 15 kg/m3 will be obtained. To improve the economic projections the Carousel strategy which is largely based upon the use of molded polyethylene components has been developed and is actively being promoted for commercial production use at 10 kg/m3. There are currently two AST customers that using the Carousel configuration as basis for their business plans. Further economic modelling will be undertaken after the carrying capacity of the MDC lines are established.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Fixed film and suspended growth in small scale indoor production of marine shrimp by Marie Gutierrez-Wing, Matthew Louque, Timothy Pfeiffer, and Ronald Malone was presented at Aquaculture America 2020 in Honolulu Hawaii, February 9-12, 2020.
• Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation:  Clear-Water Fixed Film Carousel Production of Marine Shrimp R. F. Malone and Connor Tiersch was presented at the Aquaculture America 2021 in San Antonio, Texas, August 11-15.

Progress 09/01/19 to 08/31/20

Outputs
Target Audience:Target Audience The principal target audience for this research project are small farmers in the Midwest looking to fill local niche markets for live or fresh shrimp products. The US remains a net shrimp importer, with annual shrimp imports of $7.5 billion in 2019 (https://www.ustradenumbers.com/import/shrimp-other-crustaceans/). AST provided design assistance to several major shrimp producers in the Midwest and may have had a major impact on the design approaches. These producers are already evaluating our filter approaches, but, have not committed to full-scale production, largely a Covid impact. AST has provided design assistance and filters to a major scale bait shrimp producer in Florida. A presentation entitled "Fixed film and suspended growth in small scale indoor production of marine shrimp" by Maria Teresa Gutierrez-Wing, Matthew Louque, Timothy Pfeiffer, and Ronald Malone was presented at Aquaculture America 2020 in Honolulu Hawaii, February 9-12, 2020. This talk reported on the results of growout studies on 6 replicated growout studies. Changes/Problems:During the proposal stage it was envisioned that the active nitrification would be maintained within the production tank by suspended solids while maintaining the total solids concentration below a targeted maximum (400-500 mg/L). The control systems required to control the suspended levels in the tank became prohibitively complex. So, it was decided to reverse the thinking and target accomplishing the nitrification in the external activated sludge basin. The change in thinking converges the fixed film and SDB approach described for the Endurance MDC series. Both system configuration consists of a PolyGeyser® and an SDB. In the SDB configuration, the PolyGeyser® is smaller and water is circulated though the SDB at a slow rate. The research team is looking at the development of a unified design that differs in the way it circulated allowing one design to serve distinctly two philosophical approaches. What opportunities for training and professional development has the project provided?Training future engineers Over the course of this research project, AST has employed 13 undergraduate students from Louisiana State University under various engineering disciplines. The types of things the students have been taught are as follows: Fundamental lab protocol (emphasizing lab safety) Proper methods of taking, handling, and storing water samples Methods of measuring water quality parameters (total suspended solids, ammonia, nitrite, nitrate, dissolved oxygen, pH, alkalinity, temperature, salinity, sulfide, and turbidity) held to academic research standards as well as the meaning behind the measurements They were also shown how to utilize Standard Methods to figure out how to measure various water quality parameters How to compile and interpret data Ins and outs of conducting a research project Istrouma High School Aquaponics The AST research and engineering staff has worked with the aquaponics program at Istrouma High School in Baton Rouge, Louisiana. This program focuses on teaching kids on a STEM track how to properly conduct scientific research. It also focuses on helping the students not on a STEM track on methods of entrepreneurship, like how to find and fulfill a need in the aquaponics market. AST is working directly with Istrouma high and we are planning on putting together a workshop for the teachers at the high school on the fundamentals of aquaponics and managing recirculating aquaculture systems. Istrouma High School will be the first location nationally to demonstrate the PolyGeyser®/aerated sludge configuration that will be used in the shrimp systems generated in this Phase II effort. Istrouma High purchased a small system in 2019, and, successfully operated it with students through three semesters and two summers. AST has refined the configuration and is now preparing to introduce a line formally under the "Modified DeCoupled (MDC)" system incorporates the new features that are effectively spin-offs from the ongoing shrimp research effort. The core MDC line contains a rearing tank, an Endurance®PolyGeyser®Bioclarifier, an airlift, and a sludge digestor. The unit is being marketed in three sizes with tank capacities ranging from 150-500 gallons. The unit is operated solely by airlift. Although the Covid Pandemic has effectively halted many in person school activities, AST continues development and continues to support the State's effort to make aquaponics one of the elements of the STEM program. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest. The current MDC models are currently being promoted to clients that call in for RAS filtration support. Costs have been finalized and the new MDC series will be posted on our home page (ASTfilters.com) and the units will likely be posted on Amazon Prime for sales. U-tube videos will be shot and uploaded to YouTube with links to our homepage and amazon. Products developed will be actively marketed by advertisement through major Aquacultural magazines and presentations will be made at a couple of major aquacultural conferences. A presentation entitled "Fixed film and suspended growth in small scale indoor production of marine shrimp" by Maria Teresa Gutierrez-Wing, Matthew Louque, Timothy Pfeiffer, and Ronald Malone was presented at Aquaculture America 2020 in Honolulu Hawaii, February 9-12, 2020. This talk reported on the results of growout studies on 6 replicated growout studies. What do you plan to do during the next reporting period to accomplish the goals?Our plan to accomplish our goals during the next reporting period. To determine the maximum sustainable nitrification capacity of the Aerated Sludge Basin (ASB) component of a Hybrid Biofloc FIT system at the target temperature of 25 oC and salinity of 25 ppt. The new MDC models configured with an endurance filter and an aerobic digestor will be loaded with marine shrimp to demonstrate the maximum carrying capacity. The units will be first tested with nitrification support from the bead filter only, then once maximum capacity is reached. Then the testing will be extended while utilizing the sludge digestor for supplemental nitrification until the loading maximum. The study will be replicated and results subject to statistical analysis. To determine the maximum volumetric shrimp density that can be supported by a bare tank unit and develop a suspended tier system to enhance the maximum volumetric shrimp density. Artificial substrate will be evaluated only if the density objectives (approximately 15 kg/m3) cannot be met under the MDC evaluations conducted under goal A. Maximum carrying capacity studies will be repeated if necessary, with artificial substrate. To design and evaluate a prototype commercial size Hybrid Biofloc FIT unit for inland shrimp culture. Testing and refinement of the Hybrid Biofloc FIT will continue until a suitable configuration is confirmed by inhouse testing. Units will then be distributed for testing at near bye universities and commercial operations. To determine the direct economic effects on the cost of shrimp production in a hybrid suspended growth system Density, growout times, and mortality data derived from the growout studies will be combined with the costs associated with the various filter configurations tested and input into the Phase I economic model to generate costs of shrimp production at different scales production.

Impacts
What was accomplished under these goals? What was accomplished under these goals? To determine the maximum sustainable nitrification capacity of the Activated Sludge Degradation Basin (ASB) component of a FIT system at the target temperature of 25°C and salinity of 25 ppt. Three replicated 280-liter tanks were configured with small custom PolyGeyser® for solids capture and a 10-liter ASB for aging the sludge. The systems were pneumatically configured with timers and solenoid valves to control the PolyGeyser® backwash frequency and to control dosing of sludge into the tank. The system proved difficult to manage, but, was successfully operated for two months with heavy feed loading (no shrimp). The system did demonstrate the ability to control nitrite levels below 1.5 mg-N/L (our original target) and was demonstrated capable of sustaining shrimp growth when a few test organisms (shrimp) were added. The research team concluded that knowledge base required to operate the system commercially would be prohibitive. The complexity resulted from the needed to maintain the tank suspended solids level between 400-500 mg/L while noting that the industry is trending towards even lower suspended solids levels. It was concluded that much simpler configuration would result from utilizing the ASB directly as a nitrification unit (external to the tank). To this end, modeling and research was conducted on the design of a sludge degradation basin (SDB) that could denitrify and supplement nitrification depending on the duration of aeration and availability of carbon. Initial research efforts were encouraging in system design and experiments to test out the theory will be conducted over the summer and fall of 2020. To determine the maximum volumetric shrimp density that can be supported by a bare tank unit and develop a suspended tier system to enhance the maximum volumetric shrimp density. Four 280-liter tanks were configured with a 0.7 ft3 Endurance 2000 PolyGeyser® filter, a 12.4-liter fluidized bed, and a foam fractionator. Ozonation and PHA capabilities were held in reserve. One system (#7) was dedicated to hold excess shrimp and three for replicated study of the fixed film strategy. The research team attempted to obtain PL10 larvae and to rear them to 1 gram in January and February and experienced heavy shipping losses. The research team managed to save about 10% of the PL10's and experienced a 100 percent loss of the second shipment, thus, severely limiting the amount shrimp available for testing. It was discovered that the PL10's that we were receiving had origin in Thailand and were in shipping the shipping process for nearly 3 days. A third shipment arrived in May 2019 (under warmer conditions) with minimal shipping losses. The lack of properly sized shrimp (1-2 gram) forced the research team to conduct the ASB experiments with feed only and severely limited the number of shrimp available for the fixed film grow out. In 2018, during the Phase I effort, the research team achieved 5 kg/m3 in a fixed film system, the industry standard for growout density for tank based biofloc systems. In 2019, the research team wanted to load the tanks so a 20 kg/m3 was achievable but were forced to limit the stocking density for a 15 kg/m3 target. Under this Phase II effort the research team successfully produced an average (across three replicates) of 10.2 kg/m3 in the fixed film format that seemed to be limited by nitrite. Another effort in 2019 to achieve a 15 kg/m3 target was unsuccessful despite good water quality (nitrite well below the targeted 1.0 mg N/L targeted for the experiment) and the research team is beginning to suspect issues with disease or the shrimp gene line are the main cause of growth issues. The team is also reaching out to other vendors who can supply PL's of a different gene line to ensure that the stock being used is robust to try to limit the untenable mortalities with shipping. There is no visual indication that the shrimp areal density is limiting tank density in our relatively shallow (0.61 meter) tanks. If it becomes apparent that areal density is an issue in the experiments, then false bottoms or curtains will be used to reduce the areal density and the experiment will be repeated again. To design and evaluate a prototype commercial size Hybrid Biofloc FIT unit for inland shrimp culture. A batch of 2,000 PL10's was loaded into a 2.3 m3 fiberglass FIT equipped with a 57-liter airlifted PolyGeyser® with about a 90-liter sludge basin in 2019.The unit was placed in a green house in the AST New Orleans production facility.At the end of the study, the system was revealed to be at a density of 10 kg/m3, despite the study ending early. The sludge basin was initially configured as an SDB, but to avoid turbidity issues the basin was operated statically, presumably, anaerobically. The unit was configured as PSE (Pneumatic Sludge Exchange) meaning the water between the filter and tank is partially exchanged between basin and the tank loop during each backwash event. In this configuration the basin would provide for denitrification (nitrate and nitrite removal), but not at a rate sufficient to control nitrate but not nitrite levels. The research team has generated cad drawings of a PolyFit configuration that will mimic the ratio's displayed by this successful fiberglass FIT configuration. The research team has designed an airlifted Endurance, SDB and Polytank system that can be used for fish or shrimp production. The Endurance, SDB, and Airlift are external to the tank. This configuration is considered a precursor to a true FIT configuration that would contain the filter and SDB internal to the tank itself. However, the unit functions identically to the in-tank configuration and may be hold a strong market position mimicking a FIT. This unit was informally evaluated inhouse by AST in several configurations on freshwater aquaponic systems and marine shrimp systems in the New Orleans Production Facility. A unit has been purchased and installed at local Baton Rouge high school where is being used for freshwater aquaponics. This configuration has just been added to the AST product line and a few units have been sold. This configuration was used to support PL growout to a 1-2 gram during 2019 and 2020 and showed promising water quality conditions. This configuration will be used in a formal inhouse growout study in the fall of 2020. The research team is well positioned to develop a shrimp specific commercial PolyFit design that vertically integrates the SDB and PolyGeyser® for placement with the tank. The general configuration and engineering challenges are well known to the research and product development teams. We await clarification on the achievable production densities to finalize the product line. This Endurance MDC precursor configurations will also be successful in other fresh and marine for the small-scale production of baitfish, ornamental fish, broodstock, fry production, and aquaponics. To determine the direct economic effects on the cost of shrimp production in a hybrid suspended growth system. The Phase 1 economic modelling effort remains valid and requires a density about 13 kg/m3 to produce a profitable outcome. Further economic modelling will be undertaken after the carrying capacity of the Endurance MDC lines are established and if any costs for additional carbon is needed.

Publications

• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: M. T. Gutierrez-Wing, M. Louque, T. Pfeiffer, and R. Malone. 2020. Fixed film and suspended growth in small scale indoor production of marine shrimp. Presented at Aquaculture America 2020, World Aquaculture Meeting held in Honolulu Hawaii, February 9-12, 2020.

Progress 09/01/18 to 08/31/19

Outputs
Target Audience:The principal target audience for this research project are small farmers in the Midwest looking to fill local niche markets for live or fresh shrimp products. Currently the US remains a net shrimp importer, with annual shrimp imports of 1.2 billion pounds (worth $6.7 billion). Shrimp is also the leading fresh or frozen product imported into the US, accounting for approximately 33% of all imports by weight (Sea Food Health Facts, 2014). AST provided design assistance to 2 major shrimp producers in the Midwest and may have had a major impact on the design approaches. These producers are already evaluating our filter approaches, but, have not committed to full-scale production. AST has proved design assistance and filters to a major scale bait shrimp producer in Florida. A presentation entitled "In Pursuit of Small-Scale Indoor RAS Production of Marine Shrimp" by Ronald Malone, Matthew Louque, and Tim Pfeiffer was presented at Aquaculture 2019, a meeting of the World Aquaculture Meeting held in New Orleans, March 7-11, 2019. This talk provided an update on the project's progress towards improving RAS designs. Changes/Problems:During the proposal stage it was envisioned that the active nitrification would be maintained within the production tank by suspended solids while maintaining the total solids concentration below a targeted maximum (400-500 mg/L). The control systems required to control the suspended levels in the tank became prohibitively complex. So, it was decided to reverse the thinking and target accomplishing the nitrification in the external activated sludge basin. The change in thinking converges the fixed film and ASB approach described for the Endurance MDC series.Both system configuration consists of a PolyGeyser and an ASB.In the ASB configuration, the PolyGeyser is smaller and water is circulated though the ASB at a slow rate.The research team is looking at the development of a unified design that differs in the way it circulated allowing one design to serve distinctly two philosophical approaches. What opportunities for training and professional development has the project provided?Training future engineers. Over the course of this research project, AST has employed several undergraduate students (5 so far) from Louisiana State University under various engineering disciplines. The types of things the students have been taught are as follows: Fundamental lab protocol (emphasizing lab safety) Proper methods of taking, handling, and storing water samples Methods of measuring water quality parameters (total suspended solids, ammonia, nitrite, nitrate, dissolved oxygen, pH, alkalinity, temperature, salinity, sulfide, and turbidity) held to academic research standards as well as the meaning behind the measurements. They were also shown how to utilize Standard Methods to figure out how to measure various water quality parameters How to compile and interpret data. Ins and outs of conducting a research project How have the results been disseminated to communities of interest?Istrouma High School Aquaponics The AST research and engineering staff has worked with the aquaponics program at Istrouma High School in Baton Rouge, Louisiana. This program focuses on teaching kids on a STEM track how to properly conduct scientific research. It also focuses on helping the students not on a STEM track on methods of entrepreneurship, like how to find and fulfill a need in the aquaponics market. AST is working directly with Istrouma high and we are planning on putting together a workshop for the teachers at the high school on the fundamentals of aquaponics and managing recirculating aquaculture systems. Istrouma High School will be the first location nationally to demonstrate the PolyGeyser/aerated sludge configuration that will be used in the shrimp systems generated in this Phase II effort. What do you plan to do during the next reporting period to accomplish the goals?The Phase 1 economic modelling effort remains valid and requires a density about 13 kg/m3 to produce a profitable outcome. Further economic modelling will be undertaken after the carrying capacity of the Endurance MDC lines are established. Product development and refinement will continue as per task descriptions.

Impacts
What was accomplished under these goals? To determine the maximum sustainable nitrification capacity of the Aerated Sludge Basin (ASB) component of a Hybrid Biofloc FIT system at the target temperature of 25 °C and salinity of 25 ppt. Three replicated 280-liter tanks were configured with small custom PolyGeyser for solids capture and a 10-liter ASB for aging the sludge. The systems was pneumatically configured with timers and solenoid valves to control the PolyGeyser backwash frequency and to control dosing of sludge into the tank. The system proved difficult to manage, but, was successfully operated for two months with heavy feed loading (No shrimp). The system did demonstrate the ability to control nitrite levels below 1.5 mg-N/L (our original target) and was demonstrated capable of sustaining shrimp growth when a few test organisms (shrimp) were added. The research team concluded that knowledge base required to operate the system commercially would be prohibitive. The complexity resulted from the needed to maintain the tank suspended solids level between 400-500 mg/L while noting that the industry is trending towards even lower suspended solids levels. It was concluded that much simpler configuration would result from utilizing the ASB directly as a nitrification unit (external to the tank). The original replicated system was disassembled and is currently being reconfigured. To determine the maximum volumetric shrimp density that can be supported by a bare tank unit and develop a suspended tier system to enhance the maximum volumetric shrimp density. Four 280-liter tanks were configured with a 0.7 ft3 Endurance 2000 PolyGeyser filter, a 12.4-liter fluidized bed, and a foam fractionator. Ozonation and PHA capabilities were held in reserve. One system (#7) was dedicated to hold excess shrimp and three for replicated study of the fixed film strategy. The research team attempted to obtain PL10 larvae and to rear them to 1 gram in January and February and experienced heavy shipping losses. The research team managed to save about 10% of the PL10's and experienced a 100 percent loss of the second shipment, thus, severely limiting the amount shrimp available for testing. If was discovered that the PL10's that we were receiving had origin in Thailand and were in shipping the shipping process for nearly 3 days. A third shipment arrived in our May (under warmer conditions) with minimal shipping losses. The lack of properly sized shrimp (1-2 gram) forced the research team to conduct the ASB experiments with feed only and severely limited the number of shrimp available for the fixed film growout. Last year, during the Phase I effort, the research team achieved 5 kg/m3 in a fixed film system, the industry standard for growout density for tank based biofloc systems. In the first year of this Phase II effort, the research team wanted to load the tanks so a 20 kg/m3 was achievable but were forced to limit the stocking density for a 15 kg/m3 target. Under this Phase II effort the research team successfully produced an average (across three replicates) of 10.2 kg/m3 in the fixed film format. The effort is supported by a comprehensive collection of water quality data. However, the research team felt it did not establish the maximum density achievable in the tank. Although the mean nitrite was well below the targeted 1.0 mg N/L targeted for the experiment, this nitrite level did not avoid molting mortalities as the animals approached a 20-gm weight. Thus, the assumption that the system would not be water quality limited was violated. The research team concluded that higher densities will be achievable. There was not indication of adverse interactions between animals. There was no indication that vibrio was an issue, they animals were much healthier in appearance than the Phase I animals. Considering that the research team was able to double the carrying capacity over the Phase 1 effort, this experiment was successful. Currently, the research team is conducting an interim study with the three replicated tank systems to 1) confirm that the mechanism of death is nitrite toxicity during the molt, and 2) to identify the mechanism causing the nitrite instability. Currently, we are also setting up for our next growout where we will rerun these 3 systems to target a higher stocking density (around 15 - 25 kg m-3). There is no visual indication that the shrimp areal density is limiting tank density in our relatively shallow (0.61 meter) tanks. If it becomes apparent that areal density is an issue in the experiments, then false bottoms or curtains will be used to reduce the areal density and the experiment will be repeated again. To design and evaluate a prototype commercial size Hybrid Biofloc FIT unit for inland shrimp culture. A batch of 2,000 PL10's were loaded into a 2.3 m3 fiberglass FIT equipped with a 57-liter airlifted PolyGeyser with about a 90-liter sludge basin. The unit was placed in a green house in the AST New Orleans production facility The sludge basin was initially configured as a ASB, but to avoid turbidity issues the basin was operated statically, presumably, anaerobically. The unit was configured as PSE (Pneumatic Sludge Exchange) meaning the water between the filter and tank is partially exchanged between basin and the tank loop during each backwash event. In this configuration the basin would provide for denitrification (nitrate and nitrite removal), but not at a rate sufficient to control nitrate but not nitrite levels. The unit is operating well and is believed to hold a shrimp density >5 kg/m3 as the shrimp approach 10 grams each (about half their harvest size). The research team is optimistic that this unit will produce more than 10 kg/m3 when it is harvested in early September. The research team has generated cad drawings of a PolyFit configuration that will mimic the ratio's displayed by this successful fiberglass FIT configuration. The research team has designed an airlifted Endurance, ASB and PolyTANK system that can be used for fish or shrimp production. The Endurance, ASB, and Airlift are external to the tank. This configuration is considered a precursor to a true FIT configuration that would contain the filter and ASB internal to the tank itself. However, the unit functions identically to the in-tank configuration and may be hold a strong market position mimicking a FIT. This unit has (is being) informally evaluated inhouse by AST in several configurations on freshwater aquaponic systems and marine shrimp systems in the New Orleans Production Facility A unit has been purchased and installed at local Baton Rouge high school where is being used for freshwater aquaponics. This configuration has just been added to the AST product line and a few units have been sold. This configuration is currently being upgraded to comply with innovations described in a pending patent. This configuration will be used to support PL growout to a 1-2 gram during the next quarter. This configuration will be used in a formal inhouse growout study next year. The research team is well positioned to develop a commercial (shrimp specific) PolyFit design that vertically integrates the ASB and PolyGeyser for placement with the tank. The general configuration and engineering challenges are well know to the research and product development teams. We await clarification on the achievable production densities to finalize the product line. This Endurance MDC precursor configurations will also be successful in other fresh and marine for the small-scale production of baitfish, ornamental fish, broodstock, fry production, and aquaponics. To determine the direct economic effects on the cost of shrimp production in a hybrid suspended growth system. The Phase 1 economic modelling effort remains valid and requires a density about 13 kg/m3 to produce a profitable outcome. Further economic modelling will be undertaken after the carrying capacity of the Endurance MDC lines are established.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Malone,R., M. Louque, and T.J. Pfeiffer. 2019.In Pursuit of Small-Scale Indoor RAS Production of Marine Shrimp. Presented at Aquaculture 2019, World Aquaculture Meeting held in New Orleans, March 7-11, 2019.