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The aim of this study was to evaluate antimicrobial activity of bee’s gastrointestinal tract (GUT) Lactobacillus kunkeei against pathogenic bacteria isolated from bees GUT. Content of the intestinal tracts microflora were cultured for isolation of different group of bacteria. Gut homogenates were plated on de Man, Rogosa and Sharpe agar plates and incubated for 48-72h at 30 °C anaerobically and McConkey agar and incubated for 24-48h at 37°C. Then, the identification of isolates with MALDI-TOF MS Biotyper was done. The bacterial strains Lactobacillus kunkeei, Bacillus cereus, Bacillus megatherium, Bacillus simplex, Bacillus thuringiensis, Delfia acidovorans, Escherichia coli, Enterococcus faecalis, Klebsiella aerogenes, Morganella morganii, Pseudomonas aeruginosa, Serratia marcescens and Sphingomonas parapaucimobilis, were isolated from gut content of bees. The disc diffusion method was used for the determination of antimicrobial activities of the Lactobacillus kunkeei supernatant against other isolated bacterial strains from GUT bees. The best antimicrobial activity of Lactobacillus kunkeei supernatant against Klebsiella aerogenes, Morganella morganii, Pseudomonas aeruginosa, Serratia marcescens. Lesser degree of antimicrobial activity against Bacillus cereus, Bacillus megatherium, Bacillus simplex, Bacillus thuringiensis were found in L. kunkeei supernatant. The moderate antibacterial activity against Delfia acidovorans, Escherichia coli, Enterococcus faecalis, Sphingomonas parapaucimobilis were found in L. kunkeei supernatant.

The aim of this study was to evaluate antimicrobial activity of bees gastrointestinal Lactobacillus spp. of against Paenibacillus larvae. Content of the intestinal tract was cultured for isolation of Lactobacillus spp. Gut homogenates were plated on de Man, Rogosa and Sharpe agar (MRS, Oxoid) plates and incubated for 48-72h at 30°C anaerobically. Then, the identification of isolates with MALDI-TOF MS Biotyper was done. The bacterial strains Lactobacillus gasseri, L. amylovorus, L. kunkeei, L. fructivorans, Paenibacillus larvae were isolated from gut content of bees. The disc diffusion method was used for the determination of antimicrobial activities of the Lactobacillus supernatant against two strains of Paenibacillus larvae. The best antimicrobial activity of Lactobacillus against Paenibacillus larvae from gut was found in L. gasseri supernatant. Lesser degree of antimicrobial activity against P. larvae was found in L. kunkeei supernatant. The strongest antibacterial activity against P. larvae CCM 4438 was found in L. gasseri and L. amylovorus and the least antibacterial activity was found in L. fructivorans.

Bacterial species of Enterobacteriaceae and the antimicrobial resistance of the isolates were detected in Apis mellifera L. bees gut. Gut content was cultivated on Meat peptone and McConkey agars at 30 and 37 °C, then, the isolates were identified with MALDI TOF MS Biotyper. Isolated strains were tested for antibiotic resistance to penicillins, cephalosporins, carbapenems, fluoroquinolones and aminoglycosides. Altogether, 12 species representing Enterobacteriaceae family were isolated. Firmicutes and Candida were represented by Bacillus megaterium and Issatchenkia orientalis . Isolated Enterobacteriaceae species were Enterobacter cloacae, Hafnia alvei, Klebsiella oxytoca, Morganella morganii, Serratia marcescens, Ser. liquefaciens, Raoultella ornithinolytica, R. planticola, R. terrigena, Pantoea ananatis, P. agglomerans, Rahnella aquatilis. Enterobacter cloacae, Hafnia alvei, Klebsiella oxytoca, Morganella morganii, Serratia marcescens, Ser. liquefaciens isolates exhibited the antimicrobial resistance more frequently than Raoultella ornithinolytica, R. planticola, R. terrigena, Pantoea ananatis, P. agglomerans, Rahnella aquatilis. Microflora of gut of bees could serve as a source of resistant microorganisms.

Carniolan honeybee (Apis mellifera carnica) is considered an endangered subspecies of Apis mellifera in Slovakia due to its illegal crossbreeding with allochthonous subspecies. Despite more than a century-long tradition of Slovak beekeeping, genetic research of Slovak honeybees has not yet been done. The aim of our study was to develop a molecular approach to characterize Carniolan honeybee population using polymorphic microsatellite markers. A total of 79 samples of Slovak Carniolan honeybee from 19 regions of Slovakia were analyzed together with 85 reference samples of Apis mellifera mellifera, Apis mellifera macedonica, Apis mellifera ligustica and Apis mellifera hybrid (Buckfast). Ten highly informative (PIC > 0.5) microsatellite markers running in two multiplex PCR reactions were used for molecular analysis of bees. The mean number of alleles per locus ranged from 4.6 (A. m. mellifera) to 11.5 (A. m. carnica). Bayesian and frequency-based methods implemented in software ‘GeneClass’ (v 2.0.h) have been used to assign individuals to populations. We were able to successfully assign 95% of individuals in actual sample size. After simulating 100,000 individuals using Bayesian Markov chain Monte-Carlo resampling, we correctly assigned 88% individuals to five tested populations. Population genetic structure was inferred through the Bayesian clustering method incorporated in software ‘Structure’ (v 2.3.4). Overall, 92% of the individuals were assigned to the same subspecies as originally declared. According to Structure analysis, Slovak population of A. m. carnica appears to be separated into two clusters, showing the relatively high level of gene flow from A. mellifera hybrid (Buckfast) and A. m. macedonica subspecies. It might be caused by rebuilding the Slovak population after colony losses using imported subspecies of the honeybee.

The honey bee, Apis mellifera, is critically important for the pollination of many economically important crops. Continued colony losses have called for a deeper understanding of both symbiotic and pathogenic microbial interactions, particularly as they relate to food storage and the pollination environment. Therefore, the aim of this study was to explore and characterize the bacteria colonizing the alimentary tract of the native honey bees using MALDI TOF MS Biotyper. Content of the intestinal tract was cultured for isolation of Gram-negative, Gram-positive microorganisms and yeasts. Then, the identification of isolates with MALDI-TOF MS Biotyper was done. Results showed that the most abundant genera in bees’ samples were Lactobacillus, Pseudomonas and Serratia. Altogether, 12 genera with 21 bacterial species and one yeast genus with two species were isolated. Bacteria were represented with Acidovorax facilis, Lactobacillus gasseri, L. amylovorus, L. kunkeei, L. fructivorans, Pseudomonas oryzihabitans, Ps. brenneri, Ps. indica, Micrococcus luteus, Serratia fonticola, Ser. marcescens, Ser. ureilytica, Hafnia alvei, Candida magnolia, Bacillus oleronius, B. horneckiae, Issatchenkia orientalis, Pantoea agglomerans, Enterobacter cloacae, Staphylococcus epidermidis, Staph. pasteuri, Shewanella profunda. The results of the study shows that the microflora of the bees gut is heterogenic and depend of locality and resources of environment for bees.