Pine honey is a honeydew honey produced in the East Mediterranean region (Greece and Turkey) from the secretions of the plant sucking insect Marchalina hellenica (Gennadius) (Coccoidea: Marchalini-dae) feeding on living parts of Pinus species. Nowadays, honeydew honey has attracted great attention due to its biological activities. The aim of this study was to study unifloral pine honey samples produced in Greece regarding their physicochemical parameters and antioxidant and antibacterial activity against five nosocomial and foodborne pathogens. These honeys showed physicochemical and microscopic characteristics within the legal limits, except for diastase activity, a parameter known to be highly variable, depending on various factors. Substantially higher levels of H₂O₂ were estimated compared to other types of honeydew honey, whereas protein content was similar. The total phenolic content was 451.38 ± 120.38 mg GAE/kg and antiradical activity ranged from 42.43 to 79.33%, while FRAP values (1.87 to 9.43 mmol Fe⁺²/kg) were in general higher than those reported in the literature. Various correlations could be identified among these parameters. This is the first attempt to investigate in depth the antibacterial activity of pine honey from Greece and correlate it with honey quality parameters. All tested honeys exerted variable but significant antibacterial activity, expressed as MIC and MBC values, comparable or even superior to manuka honey for some tested samples. Although honey antibacterial activity is mainly attributed to hydrogen peroxide and proteins in some cases (demonstrated by elevated MICs after catalase and Proteinase K treatment, respectively), no strong correlation between the antibacterial activity and hydrogen peroxide concentration or total protein content was demonstrated in this study. However, there was a statistically significant correlation of moisture, antioxidant and antibacterial activity against Klebsiella pneuomoniae, as well as antioxidant and antibacterial activity against Salmonella ser. Typhimurium. Interestingly, a statistically significant negative correlation has been observed between diastase activity and Staphylococcus aureus antibacterial activity. Overall, our data indicate multiple mechanisms of antibacterial activity exerted by pine honey.

Bees are important pollinators worldwide, promoting sustainability in agriculture and natural ecosystems. Moreover, honey bees produce a variety of honey bee products (beehive products). Honey is the main edible bee product. The consumption of pollen, bee bread, royal jelly, and propolis is becoming more popular nowadays. All these products are characterized by high nutritional value and/or bioactivity. A high microbial diversity has been reported in bees and beehive products, forming distinct microbial communities. The honey bee gut microbiome actively promotes good health and nutrient availability for the host. Furthermore, it prevents food spoilage and contributes to the maintenance of good hygiene conditions in the hive. Pseudomonads are often reported in investigations on bee and bee product microbiomes. Diverse Pseudomonas species demonstrate high metabolic adaptability, producing a wide range of bioactive enzymes and secondary metabolites. Several studies have provided evidence that Pseudomonads might play a role in bee well-being and the bioactivity exerted by honey bee products, though further research is warranted to fully understand the effects and mechanisms. The aim of this narrative review is to highlight the importance of Pseudomonads in the context of up-to-date knowledge regarding the bee and bee product microbiomes.

It has been suggested that microorganisms present in honey are a potential source of antimicrobial compounds. This study aimed to isolate and characterize bacteria from 46 Greek honey samples of diverse botanical and geographical origin and to determine whether these bacteria demonstrate antibacterial activity against five important nosocomial and foodborne pathogens. In total, 2014 bacterial isolates were obtained and screened for antibacterial activity. Overall, 16% of the isolates inhibited the growth of Staphylococcus aureus, 11.2% inhibited the growth of Pseudomonas aeruginosa and Acinetobacter baumannii, 10.2% inhibited the growth of Salmonella Typhimurium and 12.4% of the isolates affected the growth of Citrobacter freundii. In total, 316 isolates that inhibited the growth of more than two of the tested pathogens were grouped by restriction fragment length polymorphisms (RFLP) analysis of the 16S rRNA gene amplicon. Fifty of them were identified by 16S rRNA gene sequencing. The majority, 62% of the isolates, belonged to the genus Bacillus. Only 10% of the isolates were identified as Gram-negative bacteria. Furthermore, in several bacterial isolates, genes encoding polyketide synthases and nonribosomal peptide synthetases that catalyze the biosynthesis of secondary metabolites which might contribute to the exerted antimicrobial activity, were detected. This study demonstrates that honey microbiota exerts antimicrobial activity and is a putative source of secondary metabolites against important nosocomial and food pathogens that warrants further investigation.

Pine honey is a unique type of honeydew honey produced exclusively in Eastern Mediterranean countries like Greece and Turkey. Although the antioxidant and anti-inflammatory properties of pine honey are well documented, few studies have investigated so far its antibacterial activity. This study investigates the antibacterial effects of pine honey against P. aeruginosa PA14 at the molecular level using a global transcriptome approach via RNA-sequencing. Pine honey treatment was applied at sub-inhibitory concentration and short exposure time (0.5× of minimum inhibitory concentration –MIC- for 45 min). Pine honey induced the differential expression (>two-fold change and p ≤ 0.05) of 463 genes, with 274 of them being down-regulated and 189 being up-regulated. Gene ontology (GO) analysis revealed that pine honey affected a wide range of biological processes (BP). The most affected down-regulated BP GO terms were oxidation-reduction process, transmembrane transport, proteolysis, signal transduction, biosynthetic process, phenazine biosynthetic process, bacterial chemotaxis, and antibiotic biosynthetic process. The up-regulated BP terms, affected by pine honey treatment, were those related to the regulation of DNA-templated transcription, siderophore transport, and phosphorylation. Pathway analysis revealed that pine honey treatment significantly affected two-component regulatory systems, ABC transporter systems, quorum sensing, bacterial chemotaxis, biofilm formation and SOS response. These data collectively indicate that multiple mechanisms of action are implicated in antibacterial activity exerted by pine honey against P. aeruginosa.

In this study, the chemical composition of essential oils (EOs) extracted from Origanum vulgare ssp. hirtum Lamiaceae, (oregano), Salvia officinalis Lamiaceae (sage), Mentha pulegium Lamiaceae (pennyroyal), and respective hydrosols (HSs) has been investigated by Gas Chromatography–Mass Spectrometry (GC-MS). The antimicrobial activity was assessed against two oral pathogens: Gram-positive bacterium Streptococcus mutans and the fungus Candida albicans by determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal/Fungicidal concentration (MBC/MFC). Three-fold diluted solutions were dispensed into each well of a 96-well microtiter plate and, after incubation, MIC was determined by visual monitoring. The MBC/MFC was determined by transferring a small quantity of sample contained in each replicate well of the microtiter plates to appropriate culture media using a microplate replicator. The EOs of the tested herbs showed antimicrobial properties, especially the EO oil of O. vulgare, which exerted the highest antimicrobial activity. HSs of S. officinalis and M. pulegium exerted no antimicrobial activity, in contrast to oregano HS, which displayed strong antimicrobial activity. In all cases, a higher number of compounds were detected in EOs than in the corresponding HSs. The major compounds of sage EO were detected to be α-thujone (25.1%), 1,8-cineole (15.8%) and β-pinene (10.0%), while the HS was characterized by the presence of 1,8-cineole (32.6%), borneol (22.6%) and α-thujone (22.4%). Pennyroyal EO and HS consists mainly of pulegone (62.1 and 50.6%, respectively). Carvacrol was the major component present in EO (63%) and HS (97.3%) of oregano, probably contributing to the antimicrobial activity. Further research is needed in order to elucidate the antimicrobial mechanisms of specific compounds present in essential oils and hydrosols of Lamiaceae grown in Greece against oral pathogens.

Satureja pilosa Velen. (Lamiaceae) is a perennial and melliferous aromatic–medicinal subshrub which is range-restricted in adjacent parts of Greece and Bulgaria and locally in Italy, known in Northern Greece as wild oregano (“agriorigani”) and traditionally collected from the wild for culinary purposes. Since the ethnopharmacological data and modern biological activities of Satureja spp. suggest promising applications in skin conditions, the present study aimed to investigate the hitherto unknown phenolic content of cultivated S. pilosa and its potential biological activities, focusing mainly on wound-healing and anti-inflammatory effects. An HPLC-PDA-MS-targeted phytochemical investigation, along with NMR, allowed for the isolation and characterization of the main constituents, resulting in 18 compounds. Representative extracts and purified compounds were tested for wound-healing activity on NIH/3T3 fibroblasts. The butanol extract exhibited a significantly higher cell migration rate (73.4%) compared to aqueous (50.6%) and methanolic (49.6%) ones, enhancing the cell migration more rapidly at both concentration levels, whilst rosmarinic acid was the most potent among the isolated compounds, with a migration rate of 64.0% at the concentration level of 10⁻⁵ mg/mL, followed by 3,4-dihydrophenyllactic acid (54.7%). Moreover, potential effects on endothelial activation processes were explored, including the leukocyte–endothelial cell interaction during inflammatory processes and the migratory capacity during angiogenic actions, since these processes are commonly associated with skin diseases. Finally, extracts and purified compounds demonstrated weak antibacterial potential against two important pathogens (Staphylococcus aureus and Pseudomonas aeruginosa), suggesting that further investigation is warrented.