Progress 09/01/19 to 02/28/23

Outputs
Target Audience:Training had the largest outcome that will impact the trainees to give them the opportunity to continue in the field of aquaculture. The trainees have learned in the time with the project valuable lessons on fish husbandry and with live animals. The trainees also gain experience on work ethics and what is means to be on a team. The experienced and information gained from the project on the observations on feeding and fish behavior to improve in the results of grow-out trials that will be used in future trials. The project achieved the following positve outputs for support for targeted audiences. The project has tested its newly developed diets (semi-moist and dry feed); The project has established a baseline for and optimal stocking density for commercial grow-out; Provided hands-on training on feed manufacturing and fish farming, train a local aquaculture workforce; Possible new grow-out feeding regime to lower cost with the incorporation of raw fish incorporated with pelleted feeds; Transfer the technology locally and throughout the Pacific region, Marshall Islands and Micronesia; 40% average survival in the hatchery, 97% survival of juveniles from the nursery, and 6-9 month grow-out survival of 75%; Produced local feeds with the production capacity of 135 kg/hour; Since project began in 2013, have trained over 70 local Marshallese in feeds and fish technologies. 1. There have been many inquiries on the availability of resources and opportunities to do cage grow-out in the outer atolls in the RMI. We have engaged in informal meetings with other atoll Mayors. Local government agencies have shown great support with the project. 2. Majuro Local Government, general public, Marshall Island Marine Resource Authority, College of the Marshall Islands, high schools, local vendors, other local governments, general public and other institutions in the region can benefit from the project to be used as a model. 3. We have continued discussions on the impacts to the livelihood and employment opportunities for local fisherman and farmers in Majuro. 4. Important is the potential for possible stock enhancement for the depleting fishery in Majuro. 5. The positive financial impacts from project employment in aquaculture for the sustainability for families and the local/regional communities. Changes/Problems: Moi Broodstock: In 2020-2021, some of our broodstock matured and produced enough viable eggs to complete Objective 1. Broodstock have not produced any viable eggs since May 2021, and waiting for maturity of remaining broodstock. We have enough maturing female/male broodstock in two primary brood tanks, 47 and 44 fish. Upcoming broodstock in another tanks, 35 fish (1 year old) and newly collected juveniles, 85 fish. Algae Paste for Live Feed Production: Due to the lack of a consistent supply chain of air freight and mail due to the current COVID pandemic, we are not able to receive consistent algae paste shipments for live feed production for the maintenance and mass production of rotifers. The project has incurred the cost of live algae production to fill the gaps in shipments. PI re-allocated funds for equipment to purchase local equipment and supplies for the mass production of live algae: tanks, plumbing, blower for the outside production, and supplies to equip a small live algae room for stock cultures. Microalgae is essential for the production of rotifers for the feeding of moi larvae. Currently, have live algae cultures in production and a minimum of one flight a month for cargo and freight. On Objective 1c. Intermediate Cage Trial: Feeds: SM 2016 and Dry 2018.experienced higher than normal initial stocking mortality of the juveniles (3%). One cage was fed the control feed, Semi-Moist 2016 and other Dry 2018. The 65 dph (March 15, 2021) juveniles was stocked in two cages at 5,000 fish each.The loading densities are very low for the 30 m³ cages. Continued to have high mortality for the next 30 days (1 - 5 % daily). Majority of the mortalities occurred overnight, suspecting predators, jellyfish, something stinging or stressing the fish at night. Daily dissolved oxygen and temperature recorded twice daily. Days of high/low tides, monitored dissolved oxygen levels throughout the day and night where normal, 5.2 - 6.9 mg/L.After the initial 30 days (95 dph) in the stocked cages, mortality decreased, but daily mortality was still higher than the normal (2-20 fish). At 112 dph, the two cages experienced more than 50% mortality. A combined total of 4,921 fish was divided into eight cages, each 600 fish.The experimental design of the feed cage trial was changeddue to contamination of fishmeal. New feeding commercial diet, EWOS Pacific and a combination of EWOS Pacific with local by-catch/reject skipjack tuna from purse seine vessels. Theory of High Mortalities There were no noticeable signs of internal/external parasites. Dissolved oxygen was normal. The higher than normal mortalities were also observed in the juveniles in tanks at the hatchery fed the same pellets. During every cohort of juveniles stocked into cages, 200-400 fish in the nursery tanks at the hatchery for observation and comparison. Both land based tanks and the cages experienced high mortalities and confirms mortality not due to environmental factors. With the same cohort of fish, a small scale feeding trial in twelve 140 liter tanks was conducted to compare the SM 2016, Dry 2018, EWOS Pacific and grounded skipjack tuna. The observed mortalities in the tanks (fed SM 2016 and Dry 2018) experienced similar mortalities to the fish in cages. Important to note: the tanks that were fed commercial diets and grounded skipjack tuna did not have any mortalities. Feed Manufacturing During the Trial: All feeds (semi-moist and dry pellets) are made upon demand and placed in 20 ft. container for use and storage at 1-4°C. Batches of feeds are manufactured every 2-3 weeks. Vitamin/mineral premix and cassava flour are in dry storage. Due to Covid-19 and the border lockdown. The allocated fishmeal produced for the Moi project was moved and stored in a dry 20 ft. container at the fishmeal production facility. On March 13, 2021, while picking up fishmeal bags and talking with the General Manager, I noticed that the technician was spraying flying insect spray in the container with fishmeal bags and closes the door. I found out that, he does this to kill the flies. He has done this after each time we pick up fishmeal bags. 110 - 50 kg bags of fishmeal remained. Flying insect spray was discontinued. All exposed fishmeal bags exposed to the air in the container was discarded and fishmeal bags buried was used to produce feeds. Daily mortality decreased. Due to delays from Covid-19 lockdown of borders and lack of their labor force, have confirmed fishmeal will be produced in September 2021. Conclusion The fishmeal was contaminated with flying insect spray. Consulted with Dr. Warren Dominy and Dr. Dong-Fang, they both advised to discontinue with fishmeal, insecticide will kill the fish.Changed the feeds used until new batches of fishmeal are produced in September 2021. Four cages are feed commercial pellets (EWOS Pacifica) and modified commercial pellet/raw grounded skipjack tuna.? Density trial: With excess fish for stocking and increase volume of cages, the stocking densities where changed from 33to 37fish/m³ (1500), 66 to 69fish/m³ (2750) and additional density of 25 fish/m³ (1000 fish).The trial was terminated at 270 dph, the fish are 9 months old. Ourprevious growth data suggest that the fish should have an average weight of 454 g or one pound at 9 months, but the collected weight of 330 g.was recorded at termination. The fishmeal analysis is very important to ensure the quality of feeds produced. The crude protien of the fishmeal used in the experiment was 43.06%. This might explain, why the growth rate was slower than before, but further investigation is required. During the previous 8 years, the crude protien has been consistently in the 54% range. What opportunities for training and professional development has the project provided?The hands on trainings showed a clearimpact on the knowleegded gained bytrainees and gave them anopportunity to continue in the field of aquaculture. The trainees have learned in the time with the project valuable lessons on fish husbandry and with live animals. The trainees also gain experience on work ethics and what is means to be on a team. The experienced and information gained from the project on the observations on feeding and fish behavior to improve in the results of grow-out trials that will be used in future trials. The staff and trainees has shown potential to lead in hatchery, feed manufacturing, and grow-out husbandry in the RMI. In the near future, the existing staff will be able to train other trainees for employment to other atolls in the RMI, were technology transfer of aquaculture can be a sustainable asset. Quality control of feed ingredients will need to be monitored, especially how it is stored. We had experienced contamination of fishmeal during the project, where flying insect spray was used in the sealed/closed storage container to eliminate flying insects. Continuous training on the project is vitalto achieve quality staff, trainings have been demanding and not all trainees are equipped to be staffed. The project also gave additional training to existing staff on the research and commercial production applications. 1. Trained local trainees to become Hatchery, Feed mill and Cage Managers. 2. Increase in knowledge of fish husbandry and feed manufacturing. 3. Increased skills set of the local workforce. 4. Improved survivalof Moi. 5. Establishment of a workforce pipeline. 7. Increased production of fish for local and commercial consumption. 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?Commercialization Analysis The survival, growth, and weight gain of the fish indicated successful achievement of the project targets. ATMI has a good grasp on the baseline approach to the feeds and grow-out (density, feeding, length of grow-out, timing, etc.) of the fish. However, the current data provides limited information for the full commercialization analysis. However, based on the limited data below, we can make a simple calculation: 40% average survival in the hatchery, 97% survival of juveniles from the nursery, and 6-9 month grow-out survival of 75%; and 17,961 lbs of fish were produced during the 3-month grow-out. Based on the production output and estimated market value, $4.15 a pound, the grow-out trial can bring $76,332.35 in revenue. However, the cost of the project was $100,000+. Therefore, the project is not profitable at this rate of survival and growth. In addition, the project was designed and submitted in 2017, with the assumption that the distribution will remain the same. However, the COVID pandemic officially shut down ocean and air transportation for months, which severally impacted access to raw resources for feeds and distribution channels for the final product. In combination with the fact that there was only one grow-out trial in the commercial cages, the project experienced a complex situation, some elements of which were outside of the team?s control. Based on the available data, below are the recommendations for further research and development: Conduct several more grow-out runs, with consistent results, over a 2-year span. The optimal scenario would be achieving 7-8 grow-out runs with consistent results for 2 years, which will show that ATMI can have a consistent supply of fish for the commercial demand, Increase the survival rate in the hatchery (higher than 40%) in order to achieve better economics for each grow-out, Invest at least $350,000 annually over 2 years, Wait for the market conditions (air and ocean distribution channels) to stabilize to the pre-COVID or similar conditions. Overall, the project is not ready to move into the commercialization phase. It requires further research and development.

Impacts
What was accomplished under these goals? Objective 1: Objective 1: 8-week trial comparing three feeds:SM 2016, SM 2018 and Dry pellet 2018. a. Feeds Trial #1: Start: 11.30.20. Completed: 1.25.21 b.Feeds Trial #2 Start: 2.03.21. Completed: 4.31.21 Proximal Analysis of Feeds. Sample Name DM (%) Ash (%) CL (%) CP (%) Semi-moist Phase I 99.41 13.81 15.34 50.12 Semi-Moist Phase II 98.76 19.97 6.41 55.62 Dry Pellet Phase II 98.48 18.74 6.76 59.85 Results Trial #1 and #2 Survival Feed Type Trial #1 Trial #2 SM 2016 86.7 ± 6.7 72.2 ± 3.8 SM 2018 85.6 ± 3.8 73.3 ± 3.3 Dry 2018 88.9 ± 1.9 77.7 ± 10.7 Final Growth Trial #1 Trial #2 Feed Type Total Length (cm) Weight (g) Total Length (cm) Weight (g) SM 2016 16.9 ± 2.2 56.9 ± 9.5 16.6 ± 4.3 49.8 ± 6.5 SM 2018 16.5 ± 3.0 57.6 ± 10.1 16.5 ± 2.2 46.9 ± 8.2 Dry 2018 18.6 ± 1.6 69.2 ± 7.8 17.2 ± 1.2 58.6 ± 8.6 FCR and SGR Trial #1 Trial #2 Feed Type FCR SGR % FCR SGR % SM 2016 1.92 3.99 2.10 4.22 SM 2018 2.02 4.11 2.30 4.15 Dry 2018 1.36 4.41 1.45 4.51 c. Intermediate Cage Trial: Feeds: SM 2016 and Dry 2018. Selected SM2016 with best survival, growth and FCR to compare with Dry 2018. Due to contamination of fishmeal. New feeding regime was implemented. Feed Trial Continued: (fed twice daily to satiation) Control: Commercial Diet (EWOS Pacific) - everyday Treatment: Commercial Diet (EWOS Pacific) - fed 2 days/week Skipjack tuna (ground) - fed 4 or 5 days/week The experiment ended, October 5, 2021, 270 dph. Water Quality DO (mg/L) Temperature (C°) AM PM AM PM Control 5.9 ± 0.2 6.3 ± 0.2 28.5 ± 0.4 29.4 ± 0.7 Treatment 5.8 ± 0.1 6.4 ± 0.2 28.6 ± 0.5 29.5 ± 0.6 Survival (%) Feeds 1 2 3 4 Average Control EWOS Pacific 86.8 91.8 85.3 89.8 88.5 ± 2.9 Treatment Combination Raw By-Catch/EWOS 90.0 91.8 89.2 91.5 90.6 ± 1.3 Survival: No significant difference. Average Final Weight (g) Feeds Fed 1 2 3 4 Average Control EWOS Pacific 365.8 382.2 355.4 372.4 369.0 ± 11.3 Treatment Combination Raw By-Catch/EWOS 451.3 438.2 433.2 418.4 435.3 ± 13.6 Average weight at harvest. Significant (P < 0.05) Feeds Fed Control 1 2 3 4 EWOS Total Feed (kg) 256.6 242.2 254.8 250.7 Treatment 1 2 3 4 Skipjack By-Catch (kg) 1085.2 1025.6 1002.6 1100.5 80% water loss (dehydrator) 217.1 205.1 200.5 220.1 EWOS Fed per cage (kg) 88.9 91.3 79.0 82.6 Total Treatment Dehydrated + EWOS (kg) 306.0 296.4 279.5 302.7 Grounded skipjack was dried in a dehydrator at 130 F° for 2 days. The 2-day dried fish was grounded with coffee grinder and dehydrated for another 3 days. 80% of moisture from the skipjack was removed from dehydrator. This calculations was used for feed conversion ratio. Feed Conversion Ratio (FCR) Cage 1 2 3 4 Control (EWOS) 1.7 1.6 1.8 1.6 Treatment (EWOS + Skipjack Tuna) 1.6 1.6 1.5 1.7 No significant Difference between feeds. Cost of Feeds Skipjack Tuna (whole) = $50 per (1 MT) 1000 kg = $0.05/kg EWOS Pacific 3mm/4mm = $2.57/kg The combination of commercial diet with grounded skipjack tuna (by-catch), the fish wassignificantly bigger than fish only fed commercial diet. To add to the commercial viability and reduce the cost of production of aquaculture sustainability in the Marshall Islands, the use of by-catch fish can prove to increase profitability and sustainability of fish farming. Results with Significant Differences for Future Evaluation Feeds Fed Total Feeds Fed (kg) Final Weight (g) Cost of Feed ($US) EWOS 250.7 ± 6.5 369.0 ± 11.3 $644.29 ± 16.62 EWOS + Skipjack Tuna 296.1 ± 11.8 435.3 ± 13.6 $272.25 ± 15.12 Objective 2: Conduct density trials to establish the most optimal density for Moi March 2, 2022, 60 dph juveniles were transported to cages and fed commercial diet (EWOS) until the new batch of fishmeal was produced on March 24, 2022 for the production of SM 2018 to be the feed used in the remainder of the trials. Fish Density Stocked in Cages No. Fish Density (fish/m³) No. Cages 1000 25 3 1500 37 4 2750 69 4 Density of 25 fish/m³ is a standard that we used to stocked in the past with favorable results. The grant objective was to use 33 and 66 fish/m³ andstockedfour additional cages at 25fish/m³as controls. The experiment began April 1, 2022 with fish at 90 dph and was terminated,October 30, 2022, 302 dph.? Stocked Harvest Harvest wt Feed Fed FCR No.Fish Surv % lbs g (kg) 1000 745 74.5 0.75 340.9 866 3.4 1000 701 70.1 0.81 368.2 832 3.2 1000 722 72.2 0.74 336.4 915 3.8 1500 1023 68.2 0.69 313.6 1286 4.0 1500 1123 74.9 0.75 340.9 1198 3.1 1500 982 65.5 0.82 372.7 1206 3.3 1500 1022 68.1 0.73 331.8 1145 3.4 2750 1745 63.5 0.65 295.5 1755 3.4 2750 1596 58.0 0.69 313.6 1728 3.5 2750 1622 59.0 0.71 322.7 1897 3.6 2750 1645 59.8 0.69 313.6 1740 3.4 Stock Density Survival (%) Final Wt (g) FCR 25 fish/m³ 72.3 ± 2.2 348.5 ± 17.2 3.5 ± 0.3 33 fish/m³ 69.2 ± 4.0 339.8 ± 24.7 3.5 ± 0.4 66 fish/m³ 60.1 ± 2.4 311.4 ± 11.4 3.5 ± 0.1 Survival: (P < 0.01 ) 25 and 33 fish/m³ to 66 fish/m³ Final Wt: (P < 0.05 ) 25 and 33 fish/m³ to 66 fish/m³ No significant difference between 25 and 33 fish/m³ Objective 3: Conduct commercial-scale trials to establish the most optimal and economical open ocean cage grow-out requirements for Moi. Stocked juveniles into nurse net for grow-out trial, November 10, 2022. Have reportedthe resultsof the growout trial to the end of the grant, February 28, 2023. The fish where continued to be fed SM 2018 formulation. Results for 4 Months Grow Out Age (dph) DO (mg/L) Temp (C°) Total Feed (kg) Number Fish Survival (%) Weight (g) FCR Stocked 65 5,500 5.1 ± 0.3 3.28.23 175 5.76 ± 0.68 28.0 ± 0.8 1,655 5,123 93.1 145.7 ± 16.4 2.3 At termination of trial, the local staff was able to complete daily maintenance and husbandry of the growout of moi. The fish was sightly behind on growth during the four month growth cycle and anticipated that the fish could catch up later to more agressive feeding. Objective 4: Refine the commercialization plan with the data from Phase II Collecting data for cost analysis. The data collected will support and evaluate commercialization/business plan. Due to the limited space to complete the final report, he information for the commercialization plan will be in the section: "What do you plan to do during the next reporting period to accomplish the goals?" Objective 5: Train the local workforce on the feeds and fish production. See next section: "What opportunities for training and professional development has the project provided?".

Publications

Progress 09/01/21 to 08/31/22

Outputs
Target Audience:1. There have been many inquiries on the availability of resources and opportunities to do cage grow-out in the outer atolls in the RMI. We have engaged in informal meetings with other atoll Mayors. Local government agencies have shown great support with the project. 2. Majuro Local Government, general public, Marshall Island Marine Resource Authority, College of the Marshall Islands, high schools, local vendors, other local governments, general public and other institutions in the region can benefit from the project to be used as a model. 3. We have continued discussions on the impacts to the livelihood and employment opportunities for local fisherman and farmers in Majuro. 4. Important is the potential for possible stock enhancement for the depleting fishery in Majuro. 5. The positive financial impacts from project employment in aquaculture for the sustainability for families and the local/regional communities. Changes/Problems:Objective 1: Conducting feeding trials to establish the most optimal feeds for Moi. (c): Change in feeds used for Intermediate Cage Feed Trial, due to suspected contamination of fishmeal. Fishmeal is produced by a local vendor in Majuro, Marshall Islands from by-catch and reject skipjack tuna from purse seine fleet that off-loads at Majuro Port.The experimental design of the feed cage trial was changed due to possible contamination of fishmeal, experienced by high mortalities. Feeding regime change with commercial diet, EWOS Pacific and a combination of EWOS Pacific with local by-catch/reject skipjack tuna from purse seine vessels. EWOS Pacific is the commercial diet that we keep in stock. We collect frozen skipjack from a local vendor Pan Pacific Foods (PPF) in Majuro, transport to our grinder at feedmill and process enough feed for 5-7 days (stored in freezer). Each day with feeding of skipjack, containers of feed are defrosted and fed to the designated cages. Feed Trial Continued: (fed twice daily to satiation) Control: Commercial Diet (EWOS Pacific) - everyday Treatment: Commercial Diet (EWOS Pacific) - fed 2 days/week Skipjack tuna (ground) - fed 4 or 5 days/week Objective 2: Conduct density trials to establish the most optimal density for Moi. The density was adjusted to the amout available fish for the trial. The adjustment should not effect the results. An additional treatment was added, 25 fish/m³, is standard density from previous preliminary trials. The grant objective densities were 33 and 66fish/m³. The experiment will have the following densities, 25, 37, 69fish/m³. What opportunities for training and professional development has the project provided?1. Trained Cage Manager for grow-out of fish. 2. Trained locally, one (1) for Team Lead position and eight (8) Fish Technicians. Training had the largest outcome that will impact the trainees to give them the opportunity to continue in the field of aquaculture. The trainees have learned in the time with the project valuable lessons on fish husbandry and with live animals. The trainees also gain experience on work ethics and what is means to be on a team. The experienced and information gained from the project on the observations on feeding and fish behavior to improve in the results of grow-out trials that will be used in future trials. The staff and trainees has shown potential to lead in hatchery, feed manufacturing, and grow-out husbandry in the RMI. In the near future, the existing staff will be able to train other trainees for employment to other atolls in the RMI, were technology transfer of aquaculture can be a sustainable asset. Quality control of feed ingredients will need to be monitored, especially how it is stored. We had experienced contamination of fishmeal during the project, where flying insect spray was used in the sealed/closed storage container to eliminate flying insects. Training had the largest outcome that will impact the trainees to give them the opportunity to continue in the field of aquaculture. The trainees have learned in the time with the project valuable lessons on fish husbandry and with live animals. The trainees also gain experience on work ethics and what is means to be on a team. The experienced and information gained from the project on the observations on feeding and fish behavior to improve in the results of grow-out trials that will be used in future trials. How have the results been disseminated to communities of interest?The outputs from this project is trained staff and with the continuous training of trainees on the project. To achieve quality staff, trainings have been demanding and not all trainees are equipped to be staffed. The project also gave additional training to existing staff on the research and commercial production applications. 1.Increase in knowledge of fish husbandry and feed manufacturing. 2. Increased skills set of the local workforce. 3. Improved production of Moi. 4. Establishment of a workforce pipeline. 5. Increased production of fish for local and commercial consumption. 6. A locally trained workforce with an increased income What do you plan to do during the next reporting period to accomplish the goals?1. Objective 2: Complete analysis of Density trial. 2. Objective 3: Summarize data collected fromproduction scale of cage growout for commercial evaluation. 3. Objective 4: Data collected and evaluated from Objectives 1, 2, and 3 will be used to conduct cost analysis for baseline forcommerialization plan.

Impacts
What was accomplished under these goals? Objective 1: Objective 1: Conducting feeding trials to establish the most optimal feeds for Moi. Intermediate Cage Trial: Feeds: Thefeeds selected for the intermediate cage trial is SM 2016 and Dry 2018. The 65 dph (March 15, 2021) juveniles was stocked in two cages at 5,000 fish each. Experienced higher than normal initial stocking mortality of the juveniles (3%). One cage was fed the control feed, Semi-Moist 2016 and other Dry 2018. The loading densities are very low for the 30 m³ cages. Continued to have high mortality for the next 30 days (1 - 5 % daily). Majority of the mortalities occurred overnight, suspecting predators, jellyfish, something stinging or stressing the fish at night or maybe the feed. Daily dissolved oxygen and temperature recorded twice daily. On selected days of high/low tides, monitored dissolved oxygen levels throughout the day and night where normal, 5.2 - 6.9 mg/L. After the initial 30 days (95 dph) in the stocked cages, mortality decreased, but daily mortality was still higher than the normal (2-20 fish). At 112 dph, the two cages experienced more than 50% mortality from initial stocking. The surviving fish was combined and then divided into eight cages, each 600 fish. The experimental design was changed due to possible contamination of fishmeal, experienced by high mortalities. Feedschanged tocommercial diet, EWOS Pacific and a combination of EWOS Pacific with local by-catch/reject skipjack tuna. We collect frozen skipjack from a local vendor Pan Pacific Foods (PPF) in Majuro, transport to our grinder at feedmill and process enough feed for 5-7 days (stored in freezer). Feed Trial Continued: (fed twice daily to satiation) Control: Commercial Diet (EWOS Pacific) - everyday Treatment: Commercial Diet (EWOS Pacific) - fed 2 days/week Skipjack tuna (ground) - fed 4 or 5 days/week The remaining fish did very well on commercial diet (EWOS Pacific) and processed skipjack until termination of the experiment, October 5, 2021, 270 dph. Survival (%) Feeds 1 2 3 4 Average Control EWOS Pacific 86.8 91.8 85.3 89.8 88.5 ± 2.9 Treatment Combination Raw By-Catch/EWOS 90.0 91.8 89.2 91.5 90.6 ± 1.3 Survival: No significant difference. Average Final Weight (g) Feeds Fed 1 2 3 4 Average Control EWOS Pacific 365.8 382.2 355.4 372.4 369.0 ± 11.3 Treatment Combination Raw By-Catch/EWOS 451.3 438.2 433.2 418.4 435.3 ± 13.6 Average weight at harvest. Significant (P < 0.05) Explanation of the Feeds Fed Table: EWOS Pacific Plus pellet sizes 2 mm, 3 mm and 4 mm was fed in combination through the experiment period. The ratio changed through the trial. To make a comparison between pellets and raw skipjack for feed conversion ratio, the moisture will need to be calculated out of the grounded skipjack. Grounded skipjack was dried in a dehydrator at 130 F° for 2 days. The 2-day dried fish was grounded with coffee grinder and dehydrated for another 3 days. 80% of moisture from the skipjack was removed from dehydrator. Feed Conversion Ratio (FCR) Cage 1 2 3 4 Control (EWOS) 1.7 1.6 1.8 1.6 Treatment (EWOS + Skipjack Tuna) 1.6 1.6 1.5 1.7 No significant Difference. Control FCR: 1.7 ± 0.1, Treatment FCR: 1.6 ± 0.1 Cost of Feed Skipjack Tuna (Whole) = $50 per (1 MT) 1000 kg = $0.05/kg EWOS Pacific 3 mm/4mm = $2.57 / kg It was very unfortunate that the fishmeal was contaminated. We were able to continue with the remaining fish with a separate feeding regime, to determine if using raw fish as a supplement/replacement for commercial dry pellets and its effect on survival, growth and cost of production to grow fish to market size. No significant differences between the two feeding regimes, cages fed commercial diet (EWOS) and combination of EWOS and grounded skipjack tuna: water quality, survival and feed conversion ratio. There have been some promising results from the misfortune of the fishmeal. Further research will have to be investigated based on the results of the use of grounded skipjack tuna as supplemental feed. With the combination of commercial diet with grounded skipjack tuna (by-catch), the fish fed the combination was significantly bigger than fish only fed commercial diet. To add to the commercial viability and reduce the cost of production of aquaculture sustainability in the Marshall Islands, the use of by-catch fish can prove to increase profitability and sustainability of fish farming. Results with Significant Differences for Future Evaluation Feeds Fed Total Feeds Fed (kg) Final Weight (g) Cost of Feed ($US) EWOS 250.7 ± 6.5 369.0 ± 11.3 $644.29 ± 16.62 EWOS + Skipjack Tuna 296.1 ± 11.8 435.3 ± 13.6 $272.25 ± 15.12 ? Graph 1. Daily mortalities from stocking of cages at 65 dph, March 15, 2021, through changing of feeds at 112 dph, May 1, 2021 and termination of trial at 270 dph, October 5, 2021. A change in feeds used due to evidence that the fishmeal was contaminated with flying insect spray at the fishmeal production facility. Until new batches of fishmeal are produced (estimated December2021)the feeds were changed in the experiment. Four cages are feed commercial pellets (EWOS Pacifica) and modified commercial pellet/raw grounded skipjack tuna Figure below. New feeding regime began May 1, 2021 and will continue until tentative termination date of October 5, 2021. Based on data and observation, concluded that the fishmeal was contaminated. Consulted with Dr. Warren Dominy and Dr. Dong-Fang, they both advised to discontinue with fishmeal, insecticide will kill the fish. Objective 2: Conduct density trials to establish the most optimal density for Moi March 2, 2022, 60 dph juveniles were transported to cages and fed commercial diet (EWOS) until the new batch of fishmeal was produced on March 24, 2022. The new semi-moist diets from new fishmeal were tested on the remaining fish left in land based tanks. The transferred juveniles were then stocked into their respected cages for density trial.The grant objective was to use 33 and 66 fish/m³, but with the excess amount of fingerlings available, four additional cages were stocked as controls. Therefore, the final stocking density of the experiment are as followed: 25, 37 and 69 fish/m³. Density of 25 fish/m³ is a standard that we have stocked in the past with favorable results. Fish Density Stocked in Cages No. Fish Density (fish/m³) No. Cages 1000 25 4 1500 37 4 2750 69 4 The stocked experiment began April 1, 2022, juveniles 90 dph, with feeding of Semi-Moist Phase II pellet. Semi-Moist Phase II Pellet Composition Ingredients Composition (%) Fishmeal 68 Raw Fish 18 Cassava 12 Vit/Min Premix 2 Total 100 The density trial date of termination October 30, 2022. Objective 3: Conduct commercial-scale trials to establish the most optimal and economical open ocean cage grow-out requirements for Moi. Stocked juveniles into nurse net for grow-out trial, November 10, 2022. Targeted termination of grow-out trial, June 2023. Due to the end of the grant, will be reporting on the data collected up to the the end of the grant for the final report, February 28, 2023. Objective 4: Refine the commercialization plan with the data from Phase II Collecting data for cost analysis. The data collected will support and evaluate commercialization/business plan. Once the framework is finalized, ATMI and consultant can create a predictive business modeling for different stages of aquaculture development. After the project conducts its trials, the consultant will use the data from the previous years and current info to do actual calculations. Objective 5: Train the local workforce on the feeds and fish production. ATMI currently training Hatchery manager for cage operations. Have moved responsibilities of cage operation to trained broodstock husbandry technician, feedmill manager and live feeds technician. ?

Publications

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

Outputs
Target Audience: Nothing Reported Changes/Problems:Changes Objective 2:Conduct density trials to establish the most optimal density for Moi Would like to change the experimental perameters from density to another feed trial, since the first trial was inconclusive due to the contaminated fishmeal. Will stock two cages per feed, five feed comparisons. 1. Semi-Moist 2016 2. Dry Feed 2018 3. 100% Grounded Skipjack tuna 4. Four days skipjack / 2 days Commercial Pellet (EWOS) 5. Commercial Pellet (EWOS) The best outcome of the experiment can be utilized on Circular Cage Growout Trial (Objective 3) Problems Encountered Moi Broodstock: In 2020-2021, some of our broodstock matured and produced enough viable eggs to complete Objective 1. Broodstock have not produced any viable eggs since May 2021, and waiting for maturity of remaining broodstock. We have enough maturing female/male broodstock in two primary brood tanks, 47 and 44 fish. Upcoming broodstock in another tanks, 35 fish (1 year old) and newly collected juveniles, 85 fish. Algae Paste for Live Feed Production: Due to the lack of a consistent supply chain of air freight and mail due to the current COVID pandemic, we are not able to receive consistent algae paste shipments for live feed production for the maintenance and mass production of rotifers. The project has incurred the cost of live algae production to fill the gaps in shipments. PI re-allocated funds for equipment to purchase local equipment and supplies for the mass production of live algae: tanks, plumbing, blower for the outside production, and supplies to equip a small live algae room for stock cultures. Microalgae is essential for the production of rotifers for the feeding of moi larvae. Currently, have live algae cultures in production and a minimum of one flight a month for cargo and freight. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Complete the remaining objectives: Objective 2: Conduct density trialto establish the most optimal density for Moi with new fishmeal. 1. Intermediate Cage Trial: Fish Density, 33 fish/m³ and 66 fish/m³. 2. Tentative start date: November 2021, tentative completion date: May 2022 Objective 3: Conduct commercial-scale trials to establish the most optimal and economical open ocean cage grow-out requirements for Moi. Tentative stocking of 40 ft. diameter cage: March 2022 Tentative completion: August 2022. ?Objective 4: Refine the commercialization plan with the data from Phase II Collecting data for cost analysis. The data collected will support and evaluate commercialization/business plan. What's needed is an efficient method to understand the internal operations and costs, so selling fish wholesale can make financial sense. Since the project didn't have the actual numbers to work with during Year 1 due to the availability of eggs, the consultant focused on creating a framework for calculating the operational costs - identifying key factors, which result in different scenarios. Once the framework is finalized, ATMI and consultant can create a predictive business modeling for different stages of aquaculture development. After the project conducts its trials, the consultant will use the data from the previous years and current info to do actual calculations. Objective 5: Train the local workforce on the feeds and fish production. ATMI is currently training one person in broodstock husbandry, one for the cage manager position, and another person trained as a case site staff. November 2021, plan additional two trainees at cage growout site February 2022, planned trainings for one hatchery, one feedmill and two cage growout technicians. Due to the lack of eggs during the initial startup of the project and with the Covid-19 lockdown, ATMI is a year behind schedule. Objectives that where schedule to be competed in Year 1 will be completed in Year 2. Year 2 objectives will be completed in Year 3 (no-cost extension)

Impacts
What was accomplished under these goals? The overall goal of Phase II is to establish the most optimal and economical open ocean cage technology for Moi through R&D. Objective 1: Objective 1: Conducting feeding trials to establish the most optimal feeds for Moi. Viable eggs were collected in September 2020 and stocked in hatchery tank to produce juveniles for small tank trial. a.Feeds Trial #1 in small tanks: Start: 11.30.20. Completed: 1.25.21 8-week trial comparing three feeds, SM 2016, SM 2018 and Dry pellet 2018. Compiled data for: Survival, growth, feed conversion ratio and specific growth rate. b. Feeds Trial #2 in small tanks: Start: 2.03.21. Completed: 4.31.21 8-week trialcomparing three feeds, SM 2016, SM 2018 and Dry pellet 2018. Compiled data for: Survival, growth, feed conversion ratio and specific growth rate. c. Intermediate Cage Trial: Feeds: The results from the tanktrials indicated that there were no significant differences in survival, growth, SGR or FCR between the two semi-moist diets. For the intermediate cage trial, SM 2016 was selected primarily because of the total cost of the feed ingredients is lower than the SM 2018, $0.92/kg and $1.13/kg respectively. The crude protein is lower in SM 2016 (50.12%) compared to SM 2018 (55.62%) and it is at an acceptable level above 45% CP. The two feeds selected for the intermediate cage trial is SM 2016 and Dry 2018. Current reported results of 5-month square cage trial: Start: 3.15.21, with stocking of 65 dph fingerling, with tentative completion 9.10.21. The 65 dph (March 15, 2021) juveniles was stocked in two cages at 5,000 fish each. Experienced higher than normal initial stocking mortality of the juveniles (3%). One cage was fed the control feed, Semi-Moist 2016 and other Dry 2018. The loading densities are very low for the 30 m³ cages. Continued to have high mortality for the next 30 days (1 - 5 % daily). Majority of the mortalities occurred overnight, suspecting predators, jellyfish, something stinging or stressing the fish at night.Daily dissolved oxygen and temperature recorded twice daily. Days of high/low tides, monitored dissolved oxygen levels throughout the day and night where normal, 5.2 - 6.9 mg/L. After the initial 30 days (95 dph) in the stocked cages, mortality decreased, but daily mortality was still higher than the normal (2-20 fish). At 112 dph, the two cages experienced more than 50% mortality. A combined total of 4,921 fishwas divided into eight cages, each 600 fish. Theory of High Mortalities There were no noticeable signs of internal/external parasites. Dissolved oxygen was normal.The higher than normal mortalities were also observed in the juveniles in tanks at the hatchery fed the same pellets. During every cohort of juveniles stocked into cages, 200-400 fish in the nursery tanks at the hatchery for observation and comparison. Both land based tanks and the cages experienced high mortalities and confirms mortality not due to environmental factors. With the same cohort of fish, a small scale feeding trial in twelve 140 liter tanks was conducted to compare the SM 2016, Dry 2018, EWOS Pacific and grounded skipjack tuna. The observed mortalities in the tanks (fed SM 2016 and Dry 2018) experienced similar mortalities to the fish in cages. Important to note: the tanks that were fed commercial diets and grounded skipjack tuna did not have any mortalities. Feed Manufacturing During the Trial All feeds (semi-moist and dry pellets) are made upon demand and placed in 20 ft. container for use and storage at 1-4°C. Batches of feedsare manufactured every 2-3 weeks. Vitamin/mineral premix and cassava flour are in dry storage. Due to Covid-19 and the border lockdown. The allocated fishmeal produced for the Moi project was moved and stored in a dry 20 ft. container at the fishmeal production facility.On March 13, 2021, while picking up fishmeal bags and talking with the General Manager, I noticed that the technician was spraying flying insect spray in the container with fishmeal bags and closes the door. I found out that, he does this to kill the flies.He has donethis after each time we pick up fishmeal bags. 110 - 50 kg bags of fishmeal remained. Flying insect spray was discontinued. All exposed fishmeal bags exposed to the air in the container was discarded and fishmeal bags buried was used to produce feeds. Daily mortality decreased. Due to delays from Covid-19 lockdown of borders and lack of their labor force, ahve confirmed fishmeal will be produced in September 2021. Conclusion The fishmeal was contaminated with flying insect spray. Changed the feeds used until new batches of fishmeal are produced in September 2021.Four cages are feed commercial pellets (EWOS Pacifica) and modified commercial pellet/raw grounded skipjack tuna Figure below. New feeding regime began May 1, 2021 and will continue until tentative termination date of September 10, 2021. The remaining fish are doing well on commercial diet (EWOS Pacific) and processed skipjack. Current Data as of 7.10.21 Feed Average Survival Average Weight (g) EWOS Pacific 98.1% 192.4 Skipjack Tuna / EWOS Pacific 98.6% 249.6 Feed Trial Continued: (fed twice daily to satiation) Control: Commercial Diet (EWOS Pacific) - everyday Treatment: Commercial Diet (EWOS Pacific) - fed 2 days/week Skipjack tuna (ground) - fed 4 or 5 days/week Objective 4: Refine the commercialization plan with the data from Phase II Collecting data for cost analysis. The data collected will support and evaluate commercialization/business plan. What's needed is an efficient method to understand the internal operations and costs, so selling fish wholesale can make financial sense. Since the project didn't have the actual numbers to work with during Year 1 due to the availability of eggs, the consultant focused on creating a framework for calculating the operational costs - identifying key factors, which result in different scenarios. Once the framework is finalized, ATMI and consultant can create a predictive business modeling for different stages of aquaculture development. After the project conducts its trials, the consultant will use the data from the previous years and current info to do actual calculations. Objective 5: Train the local workforce on the feeds and fish production. ATMI is currently training one person in broodstock husbandry, one for the cage manager position, and another person trained as a cage site staff. November 2021, plan additional two trainees at cage growout site February 2022, planned trainings for one hatchery, one feedmill and two cage growout technicians. Due to the lack of eggs during the initial startup of the project and with the Covid-19 lockdown, ATMI is a year behind schedule. Objectives that where schedule to be competed in Year 1 will be completed in Year 2. Year 2 objectives will be completed in Year 3 (no-cost extension)

Publications

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

Outputs
Target Audience: Nothing Reported Changes/Problems:Describe major changes/problems in approach and reason(s) for these major changes. Moi Broodstock: August 2019, we lost six of our primary female broodstock.Since the loss of the broodstock, we have not had any spawns for the production of good viable eggs.There are 53 remaining broodstock that are in the system. Estimated 25 are females, and 28 are males. The brood is split into two tanks.In the next months, we have monitored the growth of the remaining broodstock.March 2020, we began to get sporadic spawns of eggs, not on the normal lunar cycle, but it is a positive sign that the females are maturing. The spawns started to be more on a cycle in April 2020, but the eggs are not viable.Eggs were deformed during development, which concluded that the fish might still be immature. April 2020, yearly collections, we collected 24 moi juveniles from nearby atoll for domestication and used for future broodstock.Moi are hermaphrodites. They are all males when juveniles, about 1.5 years of age, begin to transform into females.Juvenile collection every year, is necessary to maintain a productive brood. To date, the brood fish are currently spawning once a month, but the eggs are still not viable.We are on stand-by.I anticipate we will have viable eggs by November 2020. I amconfidentthat we will be able to complete objectives and goals with the addition of 1 year to 1.5 year no-cost extension. Algae Paste for Live Feed Production: Due to the lack of a consistent supply chain of air freight and mail due to the current COVIDpandemic, we are not able to receive consistent algae paste shipments for live feed production for the maintenance and mass production of rotifers.The projecthasincurred the cost of live algae production to fill the gaps in shipments (lack of consistency).PIre-allocated funds for equipment to purchase local equipment and supplies for the mass production of live algae: tanks, plumbing, blower for the outside production, and supplies to equip a small live algae room for stock cultures.Microalgae is essential for the production of rotifers for the feeding of moi larvae. Changes to the budget Due to the current situation, PI needed to be on the project more than expected. His time increased to 0.6 FTE. The project focused on troubleshooting and adjusting to the COVID conditions; therefore, we couldn?t take the expected number of inexperienced trainees/staff.Additional funds werere-allocated for supplies and equipment for the microalgae room and mass production of microalgae due to the inconsistency of air freight to the Marshall Islands. What opportunities for training and professional development has the project provided?The project is actively training interns to become staff. More training will be initiated when eggs/fish become available. How have the results been disseminated to communities of interest?The project hasn?t completed all necessary trials and experiments. The dissemination to the community will happen after they are completed. What do you plan to do during the next reporting period to accomplish the goals?The project plans to completethe two small scale feed trials and square cage feed trialin Year 2, pending the availability of the eggs.

Impacts
(N/A)

Publications