Bee venom is a complex natural beekeeping product, traditionally used in apitherapy, with a wide spectrum of pharmacological properties. Research on the mineral content of bee venom is limited and challenging to compare across studies due to the varying regions where they are conducted. Our study aimed to assess the mineral content of bee venom and how supplementary feeding of bee colonies with probiotic products, essential oils, as well as rapeseed and acacia nectar and pollen, affects the mineral content and antioxidant activity of the venom. The parameters analyzed included moisture, pH, dry matter, ash, impurities, and levels of macro and micro elements and antioxidant activity. The moisture content of the samples was 10–22%, and pH ranged between 5.84 to 6.41. The macro element content of the venom showed that potassium was the most abundant macro element, followed by calcium, magnesium, and phosphorus. Pb was identified in samples collected indicating lead pollution in the area where the hives were located in the case of the three harvests. In all samples, the highest DPPH radical scavenging activity was observed at a concentration of 2.00 mg/mL, with sample V6 showing the maximum value of 87.05%.

Bee venom variability is driven by environmental and nutritional factors, yet their integrated effects remain poorly understood. This study provides a novel, comprehensive assessment combining dietary treatments with real-time environmental monitoring to evaluate their joint influence on the physico-chemical properties, total amino acid, mineral composition, and heavy metal content of Apis mellifera venom. A total of 32 samples collected between April and July 2025 were analyzed under both artificial feeding and natural foraging conditions. Moisture ranged from 11.2% to 19.2%, while pH remained stable (5.6–6.25). Total amino acids varied between 344.0 and 409.5 mg/g, with maximum values during the acacia period (>400 mg/g). Potassium was the dominant macroelement (3.19–11.37 mg/g), followed by Ca (0.80–3.68 mg/g) and P (0.31–1.84 mg/g). Microelements such as Fe (0.11–0.98 mg/g) and Mn (1.19–8.85 µg/g) showed pronounced seasonal variability. Lead reached up to 36.18 µg/g during natural foraging, while Cd (0.30–3.97 µg/g) was mainly associated with artificial feeding. By integrating nutritional and microclimatic determinants, this study demonstrates that floral origin and seasonal dynamics are the primary drivers of venom quality, while supplementation exerts secondary effects, and highlights the potential of bee venom as a sensitive bioindicator of environmental exposure.