Characterization and antimicrobial susceptibility of spore-forming bacilli isolated from honeycomb

Aims of this study are isolation of effective microorganism, identification with conventional and molecular methods, antibiotic resistance, determination of sensitivity of the microorganism to the plant extracts in AFB disease suspected honeycomb. Muller Hinton, Yeast Exstract, potassium phosphate, glucose and sodium pyruvate medium (MYPG) was used for the production of microorganisms. Received from the 9 sample total of 21 gram-positive bacilli were isolated. For identification of isolates some physical and biochemical properties were tested. To confirm the diagnosis made in the traditional method, 16S rRNA ITS gene sequence analysis was performed using molecular diagnostic methods and family tree was created. The isolates 9 P.larvae, 7 Paenibacillus sp. and 3 Bacillus sp. were identified. Two strains (Pb2a, Pb3.2b2) were identified as Bacillus sp. with traditional identification, but they were identified as Paenibacillus sp. with molecular methods. Three strains (PB1b, PB3.3a1 and PB3.3b) were different in the genus level identification. Largely similarity between the molecular diagnosis and conventional diagnosis was observed. 4 of the 9 samples which AFB factors P.larvae have been identified. When we look at antibiotic resistance for two strains (PB5b and PB3.2b2) as sensitive to all antibiotics tested, it was observed that in all but three strains (PB5b, PB2a, PB3.2b2) carriying the multiple antibiotics resistance. Six of Paenibacillus larvae strains resistance to gentamicin, 5 strains modarate, 2 strains resistance to amikacin and one strain moderately resistant to trimethoprim/sulfamethoxazole were found.31 different aqueous and methanol extracts of plants (8 phanerogam, 23 moss) were tested against strains. Methanol extracts was determined to be more effective than the aqueous extracts. Thymus cilicicus, T. spicata ve Clinopodium umbrosum extracts was determined to be the most effective.