Honeybees play a pivotal role in pollinating plants worldwide besides producing valuable products that are used commercially for several purposes. The main factor affecting the biological activities of honeybee colonies is the source of their food and components, especially the source of protein. Therefore, two experiments (in apiary and laboratory) were designed to use a novel protein source (Azolla pinnata) in an attempt to enhance the biological activities of honeybees. Nine colonies of honey bees were divided into three groups, each group received different protein source (azolla, maize pollen grains, sugar syrup only as control). We found that although the consumption rate of pollen patty exceeds the azolla patty, all estimated biological activities were higher in honeybee group fed on azolla patty than other groups. The estimated activities included the sealed brood area, Hypopharyngeal gland development (HPG) and its effect on royal jelly production, abdominal lipid content as well as the longevity of honey bee workers. Hence, the success of our idea of using Azolla as a promising source of protein in feed alternatives for honeybees has succeeded for the first time in the world, and it is considered as a new starting point for many researchers towards finding unconventional and environmentally safe feed alternatives.