Prirodna mikobiota i prisustvo aflatoksina B1 u polenu prikupljenom u Srbiji

Total fungal count, incidence of fungi and aflatoxin B1 (AFB1) concentration were studied in 33 samples of bee pollen randomly collected from beekeepers in Serbia. The total number of fungi was determined by dilution method whereas AFB1 was detected using the Enzyme-Linked Immuno-Sorbent Assay (ELISA). The mycological estimation showed the presence of nine genera of fungi as followed: Acremonium, Alternaría, Aspergillus, Cladosporium, Epiccocum, Fusarium, Mucor, Pénicillium and Rhizopus, with total number ranging from 1 x 103 to 1 x 105 CFU g-1. The results have shown the predominance of the fungi from the genera Aspergillus and Alternaria. Among Aspergillus species it was observed that the most frequent species was A. flavus with incidence of 27.27 %. Mycotoxin AFB1 was detected as 100% positive in all samples (100%) with an average concentration of 8.61 μg kg-1. The obtained results indicated that honey bee pollen must be strictly controlled during its manipulation in the harvesting and manufacturing. Therefore, the implementation of good manufacturing (beekeeping) practice to define procedures for honeybee products could be crucial to reduce the risk of possible contamination and provide natural and safety product without risk on the human health.Ukupan broj gljiva, učestalost (incidenca) gljiva i koncentracija aflatoksina B1 (AFB1) ispitivani su u 33 uzoraka polena sakupljenih od pčelara iz različitih regiona u Srbiji. Ukupan broj gljiva određen je primenom metode razređenja a AFB1 je određen primenom imunoadsorpcione enzimske metode (ELISA). Mikološkim ispitivanjima identifikovano je devet rodova gljiva: Acremonium, Alternaria, Aspergillus, Cladosporium, Epiccocum, Fusarium, Mucor, Penicillium i Rhizopus, sa ukupnim brojem od 1 x 103 to 1 x 105 CFU g-1. Najučestalije vrste gljiva su u rodovima Aspergillus i Alternaria. Među Aspergillus vrstama najučestalija je vrsta A. flavus sa incidencom od 27,27%. AFB1 je detektovan u svim uzorcima sa prosečnom koncentracijom od 8,61 μg kg-1. Dobijeni rezultati ukazuju da pčelarski polen mora biti strogo kontrolisan tokom prikupljanja i njegove dalje prerade. Zbog toga, sprovođenje dobre proizvođačke (pčelarske) prakse podrazumeva definisanje procedura za pčelarske proizvode što bi moglo biti presudno za smanjenje rizika od moguće kontaminacije i dobijanje prirodnih i bezbednih proizvoda bez rizika po zdravlje ljudi

Mold/aflatoxin contamination of honey bee collected pollen from different Serbian regions

Assessment of microbiological quality of bee collected pollen is very important, because of its use as a supplement in the human diet. In this study, 26 samples collected from different location in Serbia were tested for the presence of mold through mycologial analysis. The presence of aflatoxin B1, one of the most dangerous and the most widespread mycotoxin was also determined. It was established that 10 of the investigated samples were contaminated with some genera or species of mold, but all of the investigated samples were contaminated with aflatoxin B1. Considering that there is no unique and official procedure for mycological analysis of bee collected pollen, these findings suggest the need for their establishment. Mycological analysis should be followed by mycotoxicological analysis since the absence of mold does not confirm the absence of aflatoxin B1 in bee pollen

Clostridium botulinum spores in European honey bees from Serbia

A total of 61 honey bees from different regions of the Republic of Serbia were analyzed for Clostridium botulinum (C. botulinum) spores. The microbiological methods and molecular methods (multiplex PCR/mPCR and PCR method) were utilized to examine multiple subunits of each honey bees samples. The C. botulinum spores in PCR-positive samples were estimated by the most probable number method (MPN). The presence of C. botulinum spores, by applying mPCR and PCR methods, was detected in 1 of the 61 honey bees (1.64%). Using MPN method, the number of spores in positive sample was 110/kg. Detection of C. botulinum spores directly from untreated honey bees, without prior enrichment, is impossible by applying PCR. Using conventional microbiological methods, detection of C. botulinum spores in dead honey bees is not possible without preenrichment. Therefore, conventional, microbiological methods are not suitable for the detection of C. botulinum spores in honey bees. In order to detect C. botulinum spores in honey bees using PCR methods, due to the small and/or unequal distribution of spores in the samples, it is desirable to use multiple subunits/replicates for each sample examined