Isolation and identification of lactic acid bacteria from stored Apis mellifera honey

This is the first assay that describes the isolation and identification of Lactic Acid Bacteria (LAB) from stored honey of the honey bee Apis mellifera meda. Honey samples were collected from stored honey of A. m. meda at the apiary of the Animal Science Research Institute of Iran, and were stored one year at room temperature, before LAB were isolated from stored honey using selective media. The isolates were Gram-stained and tested for catalase reaction. The 16S rRNA genes from extracted DNA of bacterial colonies were amplified with polymerase chain reaction (PCR) using LAB primers (27F and 1492R). All bacterial 16S rRNA genes were sequenced and deposited in GenBank. The 40 isolated strains yielded 8 distinct rRNA sequences of 8 different genera. Enterococcus in the stored honey of A. m. meda was dominantly comprised of Enterococcus hirae related-sequences (40%), Enterococcus sp. (20%), and Enterococcus faecium (10%) with other abundant sequences being related to Lactobacillus plantarum (25%) and Lactobacillus pentosus (5%).