Project description:Bee bread is one of the least studied bee products. In this study, ten bee bread samples were characterized using palynology and HS-SPME-GC-MS (headspace solid-phase microextraction gas chromatography-mass spectrometry). In total, over one hundred different volatile components were identified, belonging to different chemical groups. Only ten common components were detected in all the samples. These volatiles were ethanol, ethylene chloride, ethyl acetate, acetic acid, α-pinene, furfural, nonane, nonanal, n-hexane and isovaleric acid. Several other components were commonly shared among various bee bread samples. Over sixty detected compounds have not been previously reported in bee bread. The analysis required a mild extraction temperature of 40 °C, as higher temperatures resulted in the Maillard reaction, leading to the production of furfural. The profile of volatile compounds of the tested bee pollen samples was complex and varied. Some relationships have been shown between botanical origin and volatile organic compound profile.
Project description:The main goal of this study was the evaluation of the probiotic potential of 10 Bacillus spp. strains isolated from 5 bee bread and 3 bee pollen samples. The antagonistic interaction with Staphylococcus aureus and Escherichia coli was a primary criterion for the preliminary selection of the isolates. Three out of ten strains-PY2.3 (isolated from pollen), BP20.15 and BB10.1 (both isolated from bee bread)-were found to be possible probiotic strains. All these strains are safe for humans (exhibiting γ -hemolytic activity) and meet all essential requirements for probiotics in terms of viability in the presence of bile salts and acid conditions, hydrophobicity, auto-aggregation, and co-aggregation with the cells of important human pathogenic bacteria. They also assimilate more than 30% of cholesterol after 24 h of incubation. These three isolates are resistant to penicillin but sensitive (or exhibit moderate resistance) to the other nine antibiotics tested herein. On the basis of whole-genome sequencing, BP20.15 and BB10.1 were classified as B. subtilis and PY2.3 as B. velezensis. Moreover, genomic analyses revealed that all these isolates are potential producers of different antimicrobial compounds, including bacteriocins and secondary metabolites. The outcomes of this study have proven that some of the Bacillus strains isolated from bee pollen or bee bread are potential probiotics.
Project description:Bacteria are present in stingless bee nest products. However, detailed information on their characteristics is scarce. Thus, this study aims to investigate the characteristics of bacterial species isolated from Malaysian stingless bee, Heterotrigona itama, nest products. Honey, bee bread and propolis were collected aseptically from four geographical localities of Malaysia. Total plate count (TPC), bacterial identification, phenotypic profile and enzymatic and antibacterial activities were studied. The results indicated that the number of TPC varies from one location to another. A total of 41 different bacterial isolates from the phyla Firmicutes, Proteobacteria and Actinobacteria were identified. Bacillus species were the major bacteria found. Therein, Bacillus cereus was the most frequently isolated species followed by Bacillus aryabhattai, Bacillus oleronius, Bacillus stratosphericus, Bacillus altitudinis, Bacillus amyloliquefaciens, Bacillus nealsonii, Bacillus toyonensis, Bacillus subtilis, Bacillus safensis, Bacillus pseudomycoides, Enterobacter asburiae, Enterobacter cloacae, Pantoea dispersa and Streptomyces kunmingensis. Phenotypic profile of 15 bacterial isolates using GEN III MicroPlate™ system revealed most of the isolates as capable to utilise carbohydrates as well as amino acids and carboxylic acids and derivatives. Proteolytic, lipolytic and cellulolytic activities as determined by enzymatic assays were detected in Bacillus stratosphericus PD6, Bacillus amyloliquefaciens PD9, Bacillus subtilis BD3 and Bacillus safensis BD9. Bacillus amyloliquefaciens PD9 showed broad-spectrum of antimicrobial activity against Gram-positive and Gram-negative bacteria in vitro. The multienzymes and antimicrobial activities exhibited by the bacterial isolates from H. itama nest products could provide potential sources of enzymes and antimicrobial compounds for biotechnological applications.
Project description:A novel facultatively anaerobic, Gram-stain-positive, non-motile, non-spore-forming, catalase-negative bacterium of the genus Lactobacillus, designated strain Bb 2-3T, was isolated from bee bread of Apis cerana collected from a hive in Kunming, China. The strain was regular rod-shaped. Optimal growth occurred at 37 °C, pH 6.5 with 5.0 g l-1 NaCl. The predominant fatty acids were C18 : 1ω9c, C16 : 0 and C19 : 0 iso. Respiratory quinones were not detected. Seven glycolipids, three lipids, phosphatidylglycerol and diphosphatidylglycerol were detected. The peptidoglycan type A4α l-Lys-d-Asp was determined. Strain Bb 2-3T was closely related to Lactobacillus bombicola DSM 28793T, Lactobacillus apis LMG 26964T and Lactobacillus helsingborgensis DSM 26265T, with 97.8, 97.6 and 97.0 % 16S rRNA gene sequence similarity, respectively. A comparison of two housekeeping genes, rpoA and pheS, revealed that strain Bb 2-3T was well separated from the reference strains of species of the genus Lactobacillus. The average nucleotide identity between strain Bb 2-3T and the type strains of closely related species was lower than the 95-96 % threshold value for delineation of genomic prokaryotic species. The G+C content of the genomic DNA of strain Bb 2-3T was 37.4 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic analyses, strain Bb 2-3T is proposed to represent a novel species of the genus Lactobacillus, for which we propose the name Lactobacillus panisapium sp. nov. The type strain is Bb 2-3T (=DSM 102188T=ACCC 19955T).
Project description:Due to its high nutritional value and broad beneficial effects, the artichoke plant (Cynara cardunculus L.) is an excellent healthy food candidate. Additionally, the artichoke by-products are usually discarded even though they still contain a huge concentration of dietary fibers, phenolic acids, and other micronutrients. The present work aimed to characterize a laboratory-made gluten-free bread (B) using rice flour supplemented with a powdered extract from artichoke leaves (AEs). The AE, accounting for the 5% of titratable chlorogenic acid, was added to the experimental gluten-free bread. Accounting for different combinations, four different bread batches were prepared. To evaluate the differences, a gluten-free type-II sourdough (tII-SD) was added in two doughs (SB and SB-AE), while the related controls (YB and YB-AE) did not contain the tII-SD. Profiling the digested bread samples, SB showed the lowest glycemic index, while SB-AE showed the highest antioxidant properties. The digested samples were also fermented in fecal batches containing viable cells from fecal microbiota samples obtained from healthy donors. Based on plate counts, no clear tendencies emerged concerning the analyzed microbial patterns; by contrast, when profiling volatile organic compounds, significant differences were observed in SB-AE, exhibiting the highest scores of hydrocinnamic and cyclohexanecarboxylic acids. The fecal fermented supernatants were recovered and assayed for healthy properties on human keratinocyte cell lines against oxidative stress and for effectiveness in modulating the expression of proinflammatory cytokines in Caco-2 cells. While the first assay emphasized the contribution of AE to protect against stressor agents, the latter enlightened how the combination of SB with AE decreased the cellular TNF-α and IL1-β expression. In conclusion, this preliminary study suggests that the combination of AE with sourdough biotechnology could be a promising tool to increase the nutritional and healthy features of gluten-free bread.
Project description:Bee bread is the only fermented product of the beehive. It constitutes the main source of proteins, lipids, vitamins, and macro- and microelements in honeybee nutrition and it exerts antioxidant and antimicrobial properties, though research on these aspects has been limited so far. In this study 18 samples of Greek bee bread, two of which were monofloral, were collected during different seasons from diverse locations such as Crete and Mount Athos and were tested for their bioactivity. Samples were analyzed for their antibacterial properties, antioxidant activity, total phenolic content (TPC), and total flavonoid content (TFC). The antimicrobial activity of each sample was tested against Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmonella typhimurium. Our data demonstrate that all samples exert inhibitory and most of them bactericidal activity against at least two pathogens. Furthermore, all samples exert significant antioxidant activity, where the monofloral Castanea Sativa sample demonstrated superior antioxidant activity. Nevertheless, the antioxidant and antimicrobial activity were not strongly correlated. Furthermore, machine learning methods demonstrated that the palynological composition of the samples is a good predictor of their TPC and ABTS activity. This is the first study that focuses on the biological properties of Greek bee bread and demonstrates that bee bread can be considered a functional food and a possible source of novel antimicrobial compounds.
Project description:Pollen stored by bees undergoes a fermentation marked by the presence of lactic acid bacteria and yeasts. It results in bee bread. Past studies have singled out Starmerella (Candida) magnoliae as the most common yeast species in honey bee-stored bee bread. Starmerella species are ecological specialists with potential biotechnological value. The rarity of recent studies on yeasts in honey bees prompted us to generate new information on yeast diversity during the conversion of bee-collected pollen to bee bread. Bees and stored pollen from two apiaries in Belgium were sampled, a yeast isolation protocol was developed, yeast isolates were grouped according to their macro- and micromorphology, and representative isolates were identified using DNA sequences. Most of the 252 identified isolates belonged to the genera Starmerella, Metschnikowia, and Zygosaccharomyces. The high abundance of yeasts in fresh bee bread decreased rapidly with the storage duration. Starmerella species dominated fresh bee bread, while mostly Zygosaccharomyces members were isolated from aged bee bread. Starmerella (Candida) apis, a rarely isolated species, was the most frequent and abundant species in fresh bee bread. Yeasts from the bee's honey stomach and from pollen pellets obtained from bees hind legs were dominated by Metschnikowia species. The distinctive communities from pollen pellets over fresh bee bread to aged bee bread indicate a non-random distribution of these yeasts.
Project description:Bee pollen and bee bread have always been regarded as excellent natural resources for application in food and pharmaceutical fields due to their rich nutrient content and diversity of bioactive compounds with health-improving properties. Extensive studies on both bee products as ingredients for a healthy diet were reported, although the data concerning their metabolization on the gastrointestinal tract is quite limited. Here, we report, at each digestive stage, the bioactive profile for both bee products, their bioaccessibility levels and the antioxidant activity evaluation. The findings indicated that the average bioaccessibility level of total phenolic and total flavonoid content for bee pollen was 31% and 25%, respectively, while it was 38% and 35% for bee bread. This was reflected in a decrease of their antioxidant capacity at the end of in vitro gastrointestinal digestion, both in free radicals scavenging capacity and in reducing power. Moreover, within the 35 phytochemicals identified, the most affected by gastrointestinal digestion were phenylamides, with a complete digestibility at the end of the intestinal phase. Overall, our results highlight that bioactive compounds in both raw products do not reflect the real amount absorbed in the intestine, being bee bread more accessible in bioactive content than bee pollen.
Project description:Bacillus species isolated from Polish bee pollen (BP) and bee bread (BB) were characterized for in silico probiotic and safety attributes. A probiogenomics approach was used, and in-depth genomic analysis was performed using a wide array of bioinformatics tools to investigate the presence of virulence and antibiotic resistance properties, mobile genetic elements, and secondary metabolites. Functional annotation and Carbohydrate-Active enZYmes (CAZYme) profiling revealed the presence of genes and a repertoire of probiotics properties promoting enzymes. The isolates BB10.1, BP20.15 (isolated from bee bread), and PY2.3 (isolated from bee pollen) genome mining revealed the presence of several genes encoding acid, heat, cold, and other stress tolerance mechanisms, adhesion proteins required to survive and colonize harsh gastrointestinal environments, enzymes involved in the metabolism of dietary molecules, antioxidant activity, and genes associated with the synthesis of vitamins. In addition, genes responsible for the production of biogenic amines (BAs) and D-/L-lactate, hemolytic activity, and other toxic compounds were also analyzed. Pan-genome analyses were performed with 180 Bacillus subtilis and 204 Bacillus velezensis genomes to mine for any novel genes present in the genomes of our isolates. Moreover, all three isolates also consisted of gene clusters encoding secondary metabolites.