Progress 08/01/16 to 09/30/17

Outputs
Target Audience: Aquaculture Feed Companies and the USDA Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?These exciting findings will be presented at the upcoming Aquaculture America in February 2018. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impact Statement: According to the FAO, demand for fish will triple by 2050. Feed is the largest and most important component to ensuring safe, abundant and affordable fish supply. Most fish feeds contain a minimum level of fishmeal for adequate protein, amino acid profile, and omega-3 oils to attain good fish growth and meat quality. Global capture fisheries have been level for several decades, so fishmeal prices have risen dramatically, and alternatives to fishmeal are needed. Microalgae is part of the natural food chain for fish, and algae cultivation for a feed ingredient offers an attractive alternative to fishmeal. Global Algae Innovations has developed economical, scalable, algae production technology that will enable cost competitive production of algae meal for aquaculture feed. During this Phase 1 project, two types of algae were cultivated using Global Algae Innovations low-cost production technology, and the resulting algae meal was evaluated via compositional analysis and digestibility trials at the USDA's Agricultural Research Service, Bozeman, Montana. The first algae species was grown for high protein content and a favorable amino acid profile. The compositional analysis confirmed that the amino acid profile is favorable for aquaculture, and the digestibility trial found that the digestible protein content of the algae meal was greater than fishmeal, so this algae meal has the potential to be a very high quality aquaculture feed ingredient for protein. The second algae species was grown for omega-3 oil content. The compositional analysis found that the 22% of the fat was omega-3 oil, and the digestibility trial found that the fat digestibility coefficient is 89%, so this high omega-3 algae meal has the potential to be a replacement for the omega-3 oil normally supplied by fishmeal in aquaculture feeds. Objective #1: The key technical objective of the Phase 1 research is to develop and provide initial validation of a high value new algal feed ingredient for aquaculture. Growth of both the high protein algae, GAI-220, and the high omega-3 oil algae, GAI-230 was conducted using Global Algae's outdoor cultivation facility in Hawaii (see objective 2 for specifics). After growth and harvest of both algae species, samples were submitted for several analytical measurements that will further determine if these two species are suitable for aquaculture feed. Samples were sent for toxin detection, amino acid and macronutrient composition, fatty acid profiling, and fish feeding trials, the last in collaboration with the USDA Agricultural Research Service. Toxin analysis: • For the GAI-220 high-protein algae strain, a blue-green algae, we tested for the following toxins: Anatoxin-a, Cylindrospermopsin, Lyngbyatoxin-a, Microcystin or Nodularin, and Paralytic Shellfish Toxins (Saxitoxins). Results indicated no toxins present in our species. • For the GAI-230 high omega-3 algae strain, a pennate diatom, we tested for domoic acid, a toxin known to induce shellfish poisoning. Results indicated no toxin present in our species. Amino acid and lipid composition: • GAI-220 had a high crude protein content, 71% by dry weight. This data further reinforces the findings initially documented and described in the proposal. • GAI-230, 18.8% of the dry weight algae material consisted of oil. Fatty acid analysis determined 22.6% this oil in the algae is omega-3. Fish feeding trials: • The digestibility coefficient is a measurement used determine the amount of macronutrient and amino acids that were digested in the fish, rainbow trout for our feed trial. Compared to the reference diet, a formulated diet considered to have the ideal nutritional elements for fish growth, both algae strains performed well and exhibited digestibility coefficients on par with the reference diet and fish meal. The measured protein digestibility coefficient for the GAI-220 algae meal was 81%, for GAI- 230 was 76%, and for fishmeal was 79%. The ARS indicated that these findings, particularly with the high protein algae strain GAI-220, were some of the best results reported from these studies especially for an algae feedstock. • GAI-230 was cultivated primarily for the omega-3 content, so the fat digestibility coefficient is important. Again the digestibility coefficients were on par with fishmeal. The fat digestibility coefficient for GAI-230 was 89% and for fishmeal was 92%. Objective #2: Global Algae will produce, harvest and dry algae strain GAI-220 as well as at least one high omega-3 oil diatom or green algae. These materials will be combined to create a new algal feed ingredient. Initial progress: Using Global Algae's novel, low cost, high efficiency open-raceway bioreactor, CO2 supply, and harvest systems, a total of 7 batches of GAI-220, the high protein algae species, were grown to large-scale at our outdoor cultivation facility in Kauai. Data fundamental to characterizing growth productivity, such as pigment levels, optical density, screening for contaminants, was collected and deposited in our online database. Additionally, growth data was correlated with detailed reporting on weather and solar radiation. These datasets will be essential to further optimize GAI-220 for higher growth efficiency. Initial growth productivity data suggested that the fertilizer components which provide the nutrients for the algae are not quite optimized for this specific species. Media optimization experiments were performed at the laboratory scale using our state-of-the-art robotics system and results determined the ideal nutrient concentration suited to increase growth. This data will be used in continued production of GAI-220 and future work proposed in Phase II of the project. Conclusions: Overall, data collected during the scale up and harvest of the algae, samples processed for analytical measurements, and the fish feeding trials with the USDA Agricultural Research Service all exemplify the use of both the high protein and high omega-3 algae strains to be well suited for development as a fish meal replacement for aquaculture.

Publications

Progress 08/01/16 to 07/31/17

Outputs
Target Audience:Aquaculture Feed Companies and the USDA Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?As per our collaboration with the USDA Agricultural Research Service, Nutrient Composition and digestibility data of two Global Algae Innovations algae strains was collected. This data will help determine if either of these algae species are suitable for an aquaculture fish feed product. The data will be presented to fish feed formulators and scientists within the aquaculture community in an effort to commercialize an algae based aquaculture fish feed product. What do you plan to do during the next reporting period to accomplish the goals?Nutrient and fertilizer tests are currently underway, which will provide additional data to further increase the growth productivity of the high protein algae strain.

Impacts
What was accomplished under these goals? Impact Statement: According to the FAO, demand for fish will triple by 2050. Feed is the largest and most important component to ensuring safe, abundant and affordable fish supply. Most fish feeds contain a minimum level of fishmeal for adequate protein, amino acid profile, and omega-3 oils to attain good fish growth and meat quality. Global capture fisheries have been level for several decades, so fishmeal prices have risen dramatically, and alternatives to fishmeal are needed. Microalgae is part of the natural food chain for fish, and algae cultivation for a feed ingredient offers an attractive alternative to fishmeal. Global Algae Innovations has developed economical, scalable, algae production technology that will enable cost competitive production of algae meal for aquaculture feed. During this Phase 1 project, two types of algae were cultivated using Global Algae Innovations low-cost production technology, and the resulting algae meal was evaluated via compositional analysis and digestibility trials at the USDA's Agricultural Research Service, Bozeman, Montana. The first algae species was grown for high protein content and a favorable amino acid profile. The compositional analysis confirmed that the amino acid profile is favorable for aquaculture, and the digestibility trial found that the digestible protein content of the algae meal was greater than fishmeal, so this algae meal has the potential to be a very high quality aquaculture feed ingredient for protein. The second algae species was grown for omega-3 oil content. The compositional analysis found that the 22% of the fat was omega-3 oil, and the digestibility trial found that the fat digestibility coefficient is 89%, so this high omega-3 algae meal has the potential to be a replacement for the omega-3 oil normally supplied by fishmeal in aquaculture feeds. Objective #1: The key technical objective of the Phase 1 research is to develop and provide initial validation of a high value new algal feed ingredient for aquaculture. Growth of both the high protein algae, GAI-220, and the high omega-3 oil algae, GAI-230 was conducted using Global Algae's outdoor cultivation facility in Hawaii (see objective 2 for specifics). After growth and harvest of both algae species, samples were submitted for several analytical measurements that will further determine if these two species are suitable for aquaculture feed. Samples were sent for toxin detection, amino acid and macronutrient composition, fatty acid profiling, and fish feeding trials, the last in collaboration with the USDA Agricultural Research Service. Toxin analysis: • For the GAI-220 high-protein algae strain, a blue-green algae, we tested for the following toxins: Anatoxin-a, Cylindrospermopsin, Lyngbyatoxin-a, Microcystin or Nodularin, and Paralytic Shellfish Toxins (Saxitoxins). Results indicated no toxins present in our species. • For the GAI-230 high omega-3 algae strain, a pennate diatom, we tested for domoic acid, a toxin known to induce shellfish poisoning. Results indicated no toxin present in our species. Amino acid and lipid composition: • GAI-220 had a high crude protein content, 71% by dry weight. This data further reinforces the findings initially documented and described in the proposal. • GAI-230, 18.8% of the dry weight algae material consisted of oil. Fatty acid analysis determined 22.6% this oil in the algae is omega-3. Fish feeding trials: • The digestibility coefficient is a measurement used determine the amount of macronutrient and amino acids that were digested in the fish, rainbow trout for our feed trial. Compared to the reference diet, a formulated diet considered to have the ideal nutritional elements for fish growth, both algae strains performed well and exhibited digestibility coefficients on par with the reference diet and fish meal. The measured protein digestibility coefficient for the GAI-220 algae meal was 81%, for GAI-230 was 76%, and for fishmeal was 79%. The ARS indicated that these findings, particularly with the high protein algae strain GAI-220, were some of the best results reported from these studies especially for an algae feedstock. • GAI-230 was cultivated primarily for the omega-3 content, so the fat digestibility coefficient is important. Again the digestibility coefficients were on par with fishmeal. The fat digestibility coefficient for GAI-230 was 89% and for fishmeal was 92%. Objective #2: Global Algae will produce, harvest and dry algae strain GAI-220 as well as at least one high omega-3 oil diatom or green algae. These materials will be combined to create a new algal feed ingredient. Initial progress: Using Global Algae's novel, low cost, high efficiency open-raceway bioreactor, CO2 supply, and harvest systems, a total of 7 batches of GAI-220, the high protein algae species, were grown to large-scale at our outdoor cultivation facility in Kauai. Data fundamental to characterizing growth productivity, such as pigment levels, optical density, screening for contaminants, was collected and deposited in our online database. Additionally, growth data was correlated with detailed reporting on weather and solar radiation. These datasets will be essential to further optimize GAI-220 for higher growth efficiency. Initial growth productivity data suggest that the fertilizer components which provide the nutrients for the algae are not quite optimized for this specific species. Going forward, we will perform optimization tests to identify the best concentration of specific nutrients, such as phosphorus and nitrogen, in order to further increase growth. Additional testing will also be done with our high omega-3 algae species. Conclusions: Overall, data collected during the scale up and harvest of the algae, samples processed for analytical measurements, and the fish feeding trials with the USDA Agricultural Research Service all exemplify the use of both the high protein and high omega-3 algae strains to be well suited for development as a fish meal replacement for aquaculture.

Publications