Resume: Optimal protein content for pollen substitute is 30-35%. Lipid fraction of pollen act as phagostimulant. Vitamin suplement: provided per kilogram of diet: vitamin A, 5,000 IU; vitamin D3, 2,000 IU; vitamin E, 240 mg; vitamin C, 265.4 mg; thiamine, 5.4 mg; riboflavin, 6 mg; pyridoxine, 7 mg; niacin, 18 mg; folic acid, 20 mg; and inositol, 318 mg.

Abstract – The effects of different levels of dietary crude protein on the development, antioxidant enzymatic activity, and total midgut protease activity of honey bees were investigated in the study. A total of 30 colonies of bees with sister queens were used in the test. Dietary treatments were pure rape pollen (Control) and pollen substitutes (PS) with crude protein (CP) levels at 15%, 20%, 25%, 30%, and 35% (designated as PS15, PS20, PS25, PS30, and PS35), respectively. We compared the effects of these diets on honey bees by measuring diet consumption, bee development (egg hatch, pupation success, and pupal weight), and the protein content of emergent adult bees, their antioxidant status and the activity of their midgut digestive proteases. Bees consumed significantly more (P<0.001) natural pollen than any PS, and bees fed PS had similar diet consumption over the entire experimental period. However, the total protein intake was varied (P<0.05). PS with a protein level about 30% was recognized as excellent quality diet for maximum body weight, highest protein content and antioxidant enzymatic activity, and PS with a protein rate about 35% exerted the greatest effect on increasing percentage of hatch and percentage of pupation. All these results indicate that PS appeared to be a valuable proteinaceous food approximated to the pollen, and 30∼35% of dietary protein level was optimal to maintain the colony development.
    Provided per kilogram of diet: vitamin A, 5,000 IU; vitamin D3, 2,000 IU; vitamin E, 240 mg; vitamin C, 265.4 mg; thiamine, 5.4 mg; riboflavin, 6 mg; pyridoxine, 7 mg; niacin, 18 mg; folic acid, 20 mg; and inositol, 318 mg.
  Studies have shown that chemicals attractive to honey bees reside in the lipid fraction of the pollenkitt (Dobson 1988). Surprisingly, greater consumption of pollen did not enhance the development and growth performance of bees. Somerville (2005) revealed that what is probably more important for the growth and development of bees is the total protein intake of a colony but not simply food consumption. In this study, bees consumed significantly more pollen than PS30, but had lower development parameters, although the total protein intakes of these two groups were similar. This might have resulted from the different quantity of protein digested and absorbed since honey bees and bumblebees use proteolytic enzymes to digest dietary protein.
   In this experiment, PS diets were consumed in similar amounts even though their CP contents varied from 15% to 35%. This may partially be due to the fact that all PS diets had similar attractiveness.
   Malondialdehyde is formed as an end product of lipid peroxidation and therefore the extent of lipid peroxidation by reactive oxygen species can be monitored by MDA levels (Sumida et al. 1989). The lower MDA concentration of bees fed PS25, PS30 and PS35 than for bees fed PS15 indicated that bee antioxidant status was improved by increasing dietary protein content from 15% to 35% and therefore lipid peroxidation was reduced. The inferior antioxidant status of bees fed PS15 to those fed PS25, PS30, and PS35 also suggests that dietary protein at 15% by dry matter does not support the optimum physiology of bees. The observation that MDA concentration was reduced even though T-SOD activity was not affected by dietary protein concentration is probably explained by alterations in other antioxidant enzyme activities, such as glutathione S-transferase, glutathione peroxidase, and glutathione reductase, which also exist in insects.
  The higher midgut protease activity of bees fed the diet with 30% and 35% CP than those of bees fed other PS diets indicated that these groups of bees not only ate more protein but also digested and absorbed more protein than others.

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