Development and Production of Standardized Reference Diets for Zebrafish Research

The over arching goal of this Phase I SBIR proposal is to begin to optimize and implement standard reference diets for the ZebrafishDanio rerioan animal model of critical importance to the understanding of human health and development of vertebrate organismsA key problem in the industry is that standardized diets or feed management strategies have not been developed for Danio rerioThese diets are essential to develop rigor and reproducibility in research where zebrafish is the appropriate modelOur solution for the industrydriven by a synergistic collaboration between Meridian Biotech and the laboratory of DrStephen Watts at the University of Alabama at Birminghamis to develop well defined open formulation diets for juvenile and adult Zebrafishthat will achieve a desired level of control and whereby use of this modern and effective biomedical animal model is substantially improvedThis commercial academic collaboration is a significant value add to this applied science programKey ingredients in this formulated diet will include protein packages that contain defined and effective sourcesincluding Single Cell ProteinSCPand enriched protein source obtained from bacteria or yeastDiets with these protein sources have been developed in collaboration with Meridian Biotech which have improved shrimp farming feed technologies markedlythuswe wish to apply this technology to Danio rerio and other relevant modelsThe over arching hypothesis of this SBIR proposal is that specific nutrientsprovided through selected ingredient compositionwill affect developmentgrowthreproductive capacityimmunocompetenceand disease pathology of Zebrafish and their offspring in a standard and defined mannerThis hypothesis is based on the fact that different commercial diets currently referenced in Zebrafish studies elicit different survival and growth rates within specific Danio rerio cohortsFeeding different commercial diets also causes differences in reproductive successindicating that one or more physiological functions are affected by the nutrient composition of the dietFurtherthese commercial diets are not an open formulation and thus may change without the knowledge of the scientist thereby his her research may become unverifiable and not repeatableAlsothe lack of control of diet influences other specific behavioralphysiologicalbiochemicalor molecular pathwaysleading to a loss of rigor and reproducibilityAs a consequenceundesired variability in the outcome of specific experimental trials designed to address developmentalpharmacologicaltoxicologicalor disease related questions have emergedconfounding progress in this fieldOur mutual Milestones and Specific Experimental Aims are straightforwardMilestoneseeks the development and optimization of candidate reference diets for Zebrafishbased largely on previous work in the lab at UABSpecific Aims for Milestonewillatest a small series of candidate diets that differ modestly in composition andbcontinue to develop specific outcome assessment tools to evaluate nutrient effects on organismalphysiologicaland molecular levels of biological organizationA lead candidate diet that is chemically defined and easy to manufacture will emerge from these initial studiesAdditional back up lead diets will also advance into Milestoneefforts belowEarly juvenile and adult Danio rerio will be evaluatedWe will compare our diet throughout the study to other diets used currently in Zebrafish husbandryMilestoneseeks to select of a Standard Reference Diet for ZebrafishSpecific Aims for Milestonewillatest coded and blinded lead and back up lead diets inindependent Zebrafish facilities across the US for rigorreproducibility and independent validation of the data andbdevelop assay metrics led by our laboratory among these Zebrafish facilities for quantitative and reproducible testing of dietsA standard reference diet is envisioned to emerge and be selected for further development in Phase II SBIR envisioned activities that will be spearheaded eventually by a new subsidiary of Meridian Biotech A key problem in the ZebrafishDanio reriohusbandry science and industry is that standardized diets or feed management strategies have not been developed for this important vertebrate animal modelOur solution for the industrydriven by a synergistic collaboration between Meridian Biotech and the laboratory of DrStephen Watts at the University of Alabama at Birminghamis to develop an open formulation diet for larvaljuvenile and adult Zebrafishthat will achieve a desired level of controlwhereby use as a modern and effective biomedical animal model is substantially improvedKey ingredients in this formulated diet will include protein packages that contain defined and effective sourcesincluding Single Cell ProteinSCPan enriched protein source developed from yeast or bacteriaGiven the utility of the Zebrafish as an animal model for human healthand coupled with the concomitant investment of research dollarsit is imperative that we develop openhigh quality standard reference diets to add to the confidencereproducibilityand exchange of results derived from this important animal modelMeridian also envisions that such efforts can be extended to other aquatic and laboratory models of human diseaseenvironmental animal modelsand to the agricultural industry