Anti-Bacterial Activity of Four Distinct Propolis Extracts against P. larvae and M. plutonius; Etiological Agent of American and European Foulbrood Disease of Honeybees.

Four different propolis samples obtained from different regions of Iran were evaluated for their antibacterial effects against the bacterial agents responsible for two important honeybee diseases. Paenibacillus larvae (P. larvae) and Melissococcus plutonius (M. plutonius), as the etiological agents of American foulbrood (AFB) and European foulbrood (EFB) diseases, were subjected to propolis ethanolic extracts in the agar well diffusion assay. The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) of the antibacterial effects of the samples against the two indicator organisms were determined by the microdilution technique using different concentrations of the propolis extracts. Finally, the synergistic antibacterial actions of the mixed propolis samples were determined, and their MIC and MBC values were recorded. A two-way analysis of variance was used to evaluate correlations among the diameters of the inhibition zones, the bacterial agents, and the propolis extracts. Based on our results, three of the propolis samples showed significant antibacterial effects against P. larvae and M. plutonius during the agar well diffusion assay. Furthermore, the antibacterial capacity of the propolis samples, when mixed in equal proportions, was significantly enhanced, as indicated by the obtained MIC and MBC values. Approximately, 0.02 mg/mL of mixed propolis samples was required for inhibiting the growth of both pathogens. A direct correlation was observed between propolis concentrations and their antibacterial activity. The results of the study are conclusive of the significant antibacterial actions of Iranian propolis samples against the etiological agents of the mentioned honeybee diseases, suggesting their probable use as a safe biological agent to control AFB and EFB diseases.

In vitro probiotic and safety attributes of Bacillus spp. isolated from beebread, honey samples and digestive tract of honeybees Apis mellifera.

Bacillus species isolated from honeybee Apis mellifera gut, honey and bee bread samples were characterized for their in vitro probiotic and safety attributes. Alpha and γ haemolytic cultures were tested for their antibiotic resistance, antibacterial spectrum, acid and bile tolerance, adhesion ability (auto-aggregation, co-aggregation and hydrophobicity) and phenol tolerance. Safety criteria included evaluation of virulence genes and cytotoxicity percentages. Bacillus isolates inhibited both Gram-positive and Gram-negative pathogens including Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and Streptococcus mutans, while none could inhibit Listeria monocytogenes. Among the isolates, Bacillus subtilis ZH05, ZB03 and ZG025 showed resistance to most of the tested antibiotics and were considered unsafe. B. subtilis (4) and B. licheniformis (1) tolerated acidic pH and bile conditions, never the less were more tolerant in simulated intestinal conditions vis-a-vis gastric conditions. In 0·5% phenol concentrations, B. licheniformis ZH02 showed highest growth, while, B. subtilis ZG029 demonstrated highest auto-aggregation (65 ± 4·6) and hydrophobicity (23 ± 3·6) percentages (P < 0·05). The isolates lacked virulence genes (hblA, hblC, hblD, nhe, cytK and ces), and their cytotoxic percentage on Caco-2 cell lines was ˂15%. Overall, honeybees appear to be a good source of Bacillus species exhibiting typical in vitro probiotic properties, which could be of commercial interest.

Role of Honey Bee Gut Microbiota in the Control of American Foulbrood and European Foulbrood Diseases.

American foulbrood (AFB) and European foulbrood (EFB) are the two most important honey bee brood diseases which impose heavy economic losses to the apiculture industry worldwide by reducing bee population and honey production. Treatment with antibiotics has led to the emergence of antibiotic-resistant strains, calling for alternative safe treatment procedures that could control these diseases. Honey bee gut microbiota is known to affect the overall health of honey bees by enhancing their resistance to a number of diseases via modulation of the immune response and production of different antimicrobial metabolites. The majority of these gut resident bacteria are identified as probiotic bacteria and secure the health of these tiny insects. In the present review, we highlighted the significance of the honey bee gut microbial community and their probiotic potency for the prevention of AFB and EFB diseases in honey bees.