In this study, three microdiets were tested on weaning of Florida pompano larvae: Otohime, Gemma and a reference diet LR803. The experimental system was stocked with 11-day-old larvae, which were co-fed micro-diets and live food from 11 dph to 17 dph then micro-diets only until 28 dph. Survival from 11 dph to 28 dph was similar for all treatments, with an average of 33%. At the end of the trial, the Gemma larvae were significantly longer and heavier than those larvae fed the other diets. Significant differences were observed in fatty acid composition of the diets and larvae between treatments. The Gemma larvae incorporated the lowest amount of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (ARA). However, they had the highest DHA/EPA and ARA/EPA ratios, which is in agreement with the concept that the proportions of polyunsaturated fatty acids could be of greater importance than their absolute amount. Results from the enzyme analysis suggest that fishmeal is suitable as the main protein source for Florida pompano larvae. This study gives new insights on Florida pompano early nutritional requirements and demonstrated the full functionality of the pancreas at 16 days post hatch, opening possibilities of an earlier weaning time.