Simple analytical approach to determine the microbiological profile of bee pollen by MALDI-TOF

Abstract

Bee pollen is a natural food supplement collected by honeybees that has significant nutritional value and ther- apeutic qualities, including antioxidant, antibacterial, and anticarcinogenic properties. Due to its hygroscopic nature, bee pollen readily promotes microbial growth; however, there is no specific legislation for bee pollen in the European Union. Microbiological contamination therefore stands out as a crucial factor in determining its quality. To address this, the study focused on isolating bacterial and fungal colonies from bee pollen using various culture media, followed by a protein extraction based on a lysis process. Identification was achieved through the Biotyper-MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization coupled to Time-of-Flight de- tector) system. The greenness and blueness of the methods were assessed using different tools, which classified them as environmentally friendly and practical. Thirty-two samples were analyzed using proposed sample treatment approaches. Data were processed using chemometric techniques based on linear models. The results revealed the presence of both beneficial and potentially pathogenic species, including key representatives such as Bacillus licheniformis, Micrococcus luteus, and Aspergillus fumigatus. These findings highlight the importance of microbiota characterization in bee pollen and demonstrate that MALDI-TOF is a rapid and effective tool for profiling culturable microbial communities in this matrix.