Patrycja Staniszewska, Anna Gryboś, Adam Staniszewski, Agata Kiec, Magdalena Podlodowska, Krzysztof Olszewski, Aneta Strachecka
In response to the growing problem of immune decline in honeybees (Apis mellifera), we investigated the potential of Ashwagandha (Withania somnifera) extract as a natural dietary supplement. The experiment was conducted under constant laboratory conditions, and worker bees were divided into three groups: a control group fed standard sugar syrup without additives, experimental group A1 receiving syrup supplemented with a low concentration of Ashwagandha extract, and experimental group A2 receiving a higher concentration of the extract. Hemolymph samples were analyzed at regular intervals for total protein concentration, protease and protease inhibitor activities, as well as non-enzymatic markers such as glucose and triglycerides. Ashwagandha supplementation increased total protein concentration and stimulated the proteolytic system in both experimental groups, with generally stronger effects in bees receiving the higher dose. At the same time, the balance between proteases and their inhibitors was maintained, indicating a coordinated and controlled physiological response. The supplemented bees also showed higher levels of glucose and triglycerides, suggesting improved energy status and metabolic support. Overall, the results indicate that Ashwagandha extract may act as a promising natural supplement supporting honeybee physiological resistance and immune homeostasis.