Propolis is a material manufactured by bees and contains beeswax, bee salivary secretions and plant resins. Propolis preparations have been used for millennia by humans in food, cosmetics and medicines due to its antibacterial effects. Within the hive, propolis plays an important role in bees’ health, with much of its bioactivity largely dependent on the plant resins the bees select for its production. Few chemical studies are available on the chemistry of propolis produced by Australian honeybees (Apis mellifera, Apidae). This study aimed to determine the chemical composition as well as in vitro antimicrobial effects of propolis harvested from honeybees in subtropical eastern Australia. Honeybee propolis was produced using plastic frames and multiple beehives in two subtropical sites in eastern Australia. Methanolic extracts of propolis were analysed by liquid chromatography with ultraviolet detection and high-resolution mass spectrometry (ultra-high-pressure liquid chromatography (UHPLC)-UV-high-resolution tandem mass spectrometry (HR-MS/MS)) and by gas chromatography mass spectrometry (GC-MS). The resulting chemical data were dereplicated for compound characterisation. The two crude extracts in abs. ethanol were tested in vitro by the agar diffusion and broth dilution methods, using a phenol standard solution as the positive control and abs. ethanol as the negative control. Chemical constituents were identified to be pentacyclic triterpenoids and C-prenylated flavonoids, including Abyssinoflavanone VII, Propolin C and Nymphaeol C. The two propolis crude extracts showed bactericidal effects at the minimal inhibitory concentrations of 0.37–2.04 mg mL−1 against Staphylococcus aureus ATCC 25923. However, the extracts were inactive against Klebsiella pneumoniae ATCC 13883 and Candida albicans ATCC 10231. The antistaphylococcal potential of propolis was discussed, also in relation to honeybees’ health, as it warrants further investigations on the social and individual immunities of Australian honeybees.
This is a preview of subscription content, log in to check access.
The authors are thankful to Terry Braggins at Analytica Laboratories Ltd, NZ, for HR-MS analyses, to the Craig’s members of ‘Valley Bees’, Gympie, for beekeeping help and to Michael Howes at ‘Tyagarah Apiaries’ and Dieter Horstmann at ‘Eagle Farm’ Tyagarah, NSW, Australia, for supplying honeybee propolis from their beehives.
This study and CFM were financed through the University of the Sunshine Coast studentships.
Conceived and designed the experiments: CFM. Performed the experiments: CFM, JS and DP. Analysed the data: CFM. Contributed reagents/materials: PB. Wrote the manuscript: CFM, JS and PB.
Compliance with ethical standards
The authors declare that the experiments described in this article comply with the current laws for the conduct of scientific research in Australia.
Conflict of interest
The authors declare that they have no conflict of interest.
Berenbaum MR, Johnson RM (2015) Xenobiotic detoxification pathways in honey bees. Curr Opin Insect Sci 10:51–58CrossRefGoogle Scholar
Chen C-N, Wu C-L et al (2004) Propolin C from propolis induces apoptosis through activating caspases, Bid and cytochrome C release in human melanoma cells. Biochem Pharm 67(1):53–66PubMedCrossRefGoogle Scholar
Chen YW, Wu SW et al (2008) Characterisation of Taiwanese propolis collected from different locations and seasons. J Sci Food Agric 88(3):412–419CrossRefGoogle Scholar
Cui L, Ndinteh DT et al (2007) Isoprenylated Flavonoids from the Stem Bark of Erythrina abyssinica. J Nat Prod 70(6):1039–1042PubMedCrossRefGoogle Scholar
de Rijke E, Out P et al (2006) Analytical separation and detection methods for flavonoids. J Chromatogr A 1112(1):31–63PubMedCrossRefGoogle Scholar
Drescher N, Wallace HM et al (2014) Diversity matters: how bees benefit from different resin sources. Oecologia 176(4):943–953PubMedCrossRefGoogle Scholar
Ghisalberti E, Jefferies P et al (1978) Constituents of propolis. Cell Mol Life Sci 34(2):157–158CrossRefGoogle Scholar
Greco MK, Hoffmann D et al (2010) The alternative Pharaoh approach: stingless bees mummify beetle parasites alive. Naturwissenschaften 97(3):319–323PubMedCrossRefGoogle Scholar
Inui S, Hosoya T et al (2012a) Solophenols B–D and solomonin: new prenylated polyphenols isolated from propolis collected from the Solomon Islands and their antibacterial activity. J Agric Food Chem 60(47):11765–11770PubMedCrossRefGoogle Scholar
Inui S, Shimamura Y et al (2012b) A new prenylflavonoid isolated from propolis collected in the Solomon Islands. Biosci Biotechnol Biochem 76(5):1038–1040PubMedCrossRefGoogle Scholar
Kumazawa S, Hamasaka T et al (2004) Antioxidant activity of propolis of various geographic origins. Food Chem 84(3):329–339CrossRefGoogle Scholar
Kumazawa S, Ueda R et al (2007) Antioxidant prenylated flavonoids from propolis collected in Okinawa, Japan. J Agric Food Chem 55(19):7722–7725PubMedCrossRefGoogle Scholar
Kumazawa S, Murase M et al (2014) Analysis of antioxidant prenylflavonoids in different parts of Macaranga tanarius, the plant origin of Okinawan propolis. Asian Pac J Trop Med 7(1):16–20PubMedCrossRefGoogle Scholar
Marquez Hernandez I, Cuesta-Rubio O et al (2010) Studies on the constituents of yellow Cuban propolis: GC-MS determination of triterpenoids and flavonoids. J Agric Food Chem 58(8):4725–4730PubMedCrossRefGoogle Scholar
Massaro FC, Brooks PR et al (2011) Cerumen of Australian stingless bees (Tetragonula carbonaria): gas chromatography–mass spectrometry fingerprints and potential anti-inflammatory properties. Naturwissenschaften 98(4):329–337PubMedCrossRefGoogle Scholar
Massaro C, Katouli M et al (2014a) Anti-staphylococcal activity of C-methyl flavanones from propolis of Australian stingless bees (Tetragonula carbonaria) and fruit resins of Corymbia torelliana (Myrtaceae). Fitoterapia 95:247–257PubMedCrossRefGoogle Scholar
Massaro CF, Shelley D et al (2014b) In vitro antibacterial phenolic extracts from “sugarbag” pot-honeys of Australian stingless bees (Tetragonula carbonaria). J Agric Food Chem 62(50):12209–12217PubMedCrossRefGoogle Scholar
Massaro CF, Smyth TJ et al (2015) Phloroglucinols from anti‐microbial deposit‐resins of Australian stingless bees (Tetragonula carbonaria). Phytother Res 29(1):48–58PubMedCrossRefGoogle Scholar
Mathe C, Culioli G et al (2004) Characterization of archaeological frankincense by gas chromatography–mass spectrometry. J Chromatogr A 1023(2):277–285PubMedCrossRefGoogle Scholar
Melliou E, Stratis E et al (2007) Volatile constituents of propolis from various regions of Greece—antimicrobial activity. Food Chem 103(2):375–380CrossRefGoogle Scholar
Miorin P, Levy Junior N et al (2003) Antibacterial activity of honey and propolis from Apis mellifera and Tetragonisca angustula against Staphylococcus aureus. J Appl Micro 95(5):913–920CrossRefGoogle Scholar
Mutai C, Abatis D et al (2004) Cytotoxic lupane-type triterpenoids from Acacia mellifera. Phys Chem Chem Phys 65(8):1159–1164Google Scholar
Ponte F, Silva A et al (2008) Bee-honey, propolis and Eucalyptus globulus extract: preclinical toxicity study in rodents. Pharmacogn Mag 4(16):278Google Scholar