Project Summary The over-arching goal of this Phase II SBIR proposal is to optimize and implement high quality standard reference diets for the Zebrafish (Danio rerio), an animal model of critical importance to the understanding of human health and development of vertebrate organisms. A key problem in the industry is that high quality standardized reference diets, with appropriate physical properties and adequate feed management strategies, have not been developed for D. rerio. Our solution for the industry, driven by a synergistic collaboration between Meridian Biotech and the laboratory of Dr. Stephen Watts at the University of Alabama at Birmingham, is to develop several open formulation diets for larval, juvenile and adult Zebrafish, which will achieve a desired level of control and whereby use of this modern and effective biomedical animal model is substantially improved and more standardized. This commercial-academic collaboration is a significant value add to this applied science program. Key ingredients in this formulated diet will include protein packages that contain defined and effective sources, including a microbial based Single Cell Protein (SCP), an enriched protein source obtained from bacterial biomass. Diets with these protein sources have been developed by Meridian Biotech, several of which have improved shrimp and fish farming feed technologies markedly; thus, we wish to apply this technology to D. rerio and, potentially, other relevant animal models. Our mutual Phase II Milestones and Specific Experimental Aims are many across both years of the program. Specific Aims for Milestone 1 (Year 1 work) include optimizing the protein content and amino acid profile of our most promising ingredient and testing it against the existing SCP-fortified diet used in pilot Phase I work, comparing the best diet to the competitor diets currently used in academic cores across the US biomedical research enterprise, and characterizing deeply at the phenotypic and genetic levels the benefits of our diet. Milestone 1 will seek to identify and progress the best diet and drive its market penetration, initially in academia. Specific Aims for Milestone 2 (Year 2 work) include further optimizing the amino acid, fatty acid, neutral fat, and energy of the SCP ingredient and scaling up manufacturing to meet demands at Meridian’s new US-based facilities, sharing the best diet formulation with academic Zebrafish cores to drive adoption of the diet, and centralizing the data gathering from the many academic cores for use by all and the NIH ORIP. Milestone 2 will seek to penetrate the academic Zebrafish husbandry marketplace by gaining Core adoption. In parallel with collaborative efforts within the academic marketplace, Meridian and commercial partners across both years will also pursue a Milestone 3 which seeks to expand the technologies and penetrate the industry fish aquaculture and farming marketplace, other production species, and potentially develop hum...