The aim of the present study was to identify of bacterial species inhabiting the intestinal tract of Apis mellifera L. Content on intestinal tract was cultured for isolation of Gram-negative and Gram-positive bacteria. Content of gut was streaked onto MacConkey agar which was incubated for 24–48 h at 37 °C aerobically. For cultivation of Gram-positive and Gram-negative microflora, the inoculated Tryptone soya agar was incubated for 48–72 h at 30 °C. Then, the identification of isolates with MALDI-TOF MS Biotyper was done. After bacteria identification antibiotic resistance testing was done. Disc diffusion methods was applied and the following antibiotics discs were used: amikacin AMK (10 μg), gentamicin GEN (10 μg), tigecycline TGC (10 μg) and chloramphenicol CM (10 μg). Results show that the Gram-positive bacteria counts were from 2.17 ±0.04 to 3.21 ±0.02 log CFU/g and Gram-negative counts ranged from 2.36 ±0.14 to 4.05 ±0.04 log CFU/g in honey bee gut samples. Lactococcaceae, Micrococcaceae, Staphylococcaceae and Enterobacteriaceae were identified from Gram-positive and Gram-negative bacteria in bees gut. The Enterobacteriaceae were the most abundant and were represented with 8 bacterial species. The Gram-positive and Gram-negative bacterial species recovered exhibited the antimicrobial resistance to the antibiotics. Gram-positive and Gram-negative bacterial showed the more frequently resistance to tigecycline (9.5%) and amikacin (8.2%). The resistance to gentamycin (6.5%) and chloramphenicol (4.1%) was observed less frequently.