1. Medium weight laying hens were used for an assay to determine their isoleucine requirement between 26 and 36 weeks of age and again between 46 and 56 weeks of age. 2. Two isoleucine-limiting mixtures were formulated with similar amino acid profiles, one containing 198 g and the other 110 g crude protein per kg diet. These mixtures were blended to give a series of 11 diets with isoleucine contents ranging from 7.6 to 3.8 g/kg. The lowest protein diet was also fed with a supplement of L-isoleucine. Each of the 12 diets was given to 5 groups of 24 laying hens. 3. The daily isoleucine requirement of individual laying hens was estimated to be 9.48 mg/g egg output plus 44.47 mg/kg body weight per day for the 1st period and 12.11 mg/g egg output plus 6.86 mg/kg body weight per day for the 2nd period. Calculated optimum intakes of isoleucine for various ratios of cost of input to value of output are tabulated. For example, for a flock of medium weight hens producing an average of 50 g daily egg mass, the optimum isoleucine intake (mg/hen d) varied between 760 and 890 varying for ratios of costs to egg prices. 4. It is concluded that the isoleucine required per day does not decrease during the first laying year despite a decrease in rate of egg output.
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