The probiotic lactic acid bacterium (LAB) was reported in the gut of the honeybee Apis mellifera. Partial 16S rRNA gene sequences of the bacterial flora found in the gut of the honeybee workers, revealed the presence of seven novel strain of LAB. Five of the anaerobic LABs are closely related to five different strains of the lactic acid bacteria Lactobacillus kunkeei species, one is closely related to a strain of Lactobacillus plantarum species and the last one is identical to a strain of species Fructobacillus fructosus. We used inhibition assays on agar plates to evaluate the antagonistic effects of the newly identified honeybee LAB on Paenibacillus larvae larvae (P. l. larvae), American foulbrood (AFB) bacterial pathogen, growth in vitro. The individual LAB phylotypes showed different inhibition properties against P. l. larvae growth on agar plates, whereas a combination of all seven LAB phylotypes resulted in a total inhibition (no visible growth) of P.l.larvae. The results demonstrate that honeybee specific LAB possess beneficial properties for honeybee health. Enhancing growth of LAB or applying LAB to honeybee colonies should be further investigated.                 American foulbrood / Paenibacillus larvae / Lactic Acid Bacteria / Lactobacillus / inhibition