Royal Jelly Proteins as Markers for Authenticity and Quality of Honey

Major royal jelly proteins as markers of authenticity and quality of honey / Glavni proteini matične mliječi kao markeri izvornosti i kakvoće meda

Arh Hig Rada Toksikol. 2015 Dec 1;66(4):259-67

Until now, the properties of honey have been defined based exclusively on the content of plant components in the nectar of given plant. We showed that apalbumin1, the major royal jelly (RJ) protein, is an authentic and regular component of honey.

Apalbumin1 and other RJ proteins and peptides are responsible for the immunostimulatory properties and antibiotic activity of honey. For the quantification of apalbumin1, an enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal anti-apalbumin1 antibody. The method is suitable for honey authenticity determination; moreover it is useful for detection of the honey, honeybee pollen and RJ in products of medicine, pharmacy, cosmetics, and food industry, where presences of these honeybee products are declared.

Results from the analysis for presence and amount of apalbumin1 in honeys will be used for high-throughput screening of honey samples over the world. On the basis of our experiments which show that royal jelly proteins are regular and physiologically active components of honey we propose to change the definition of honey (according to the EU Honey Directive 2001/110/EC) as follows: Honey is a natural sweet substance produced by honey bees from nectar of plants or from secretions of plants, or excretions of plant sucking insects, which honey bees collect, transform by combining with major royal jelly proteins and other specific substances of their own, deposit, dehydrate, store and leave in the honey comb to ripen and mature.

Serbia Bee Pollen Rich in Potassium, Calcium, Magnesium, Iron and Zinc

Mineral content of bee pollen from Serbia / Sadržaj minerala u uzorcima pčelinjega peluda iz Srbije

Arh Hig Rada Toksikol. 2015 Dec 1;66(4):251-258

In this study we analysed mineral composition of bee pollen of different plant origin collected across Serbia using inductively coupled plasma - optical emission spectrometry. The most abundant elements were potassium, calcium, and magnesium.

The samples were also exceptionally rich in iron and zinc, which are very important as nutrients. Judging by our findings, mineral composition of bee pollen much more depends on the type of pollen-producing plant than on its geographical origin.

Bee Venom Shows Antibacterial Activity and Antibiotic-Enhancing Effects Against MRSA

Antibacterial Activity and Antibiotic-Enhancing Effects of Honeybee Venom against Methicillin-Resistant Staphylococcus aureus

Molecules 2016, 21(1), 79

Methicillin-resistant Staphylococcus aureus (MRSA), along with other antibiotic resistant bacteria, has become a significant social and clinical problem. There is thus an urgent need to develop naturally bioactive compounds as alternatives to the few antibiotics that remain effective.

Here we assessed the in vitro activities of bee venom (BV), alone or in combination with ampicillin, penicillin, gentamicin or vancomycin, on growth of MRSA strains. The antimicrobial activity of BV against MRSA strains was investigated using minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC) and a time-kill assay. Expression of atl which encodes murein hydrolase, a peptidoglycan-degrading enzyme involved in cell separation, was measured by reverse transcription-polymerase chain reaction.

The MICs of BV were 0.085 µg/mL and 0.11 µg/mL against MRSA CCARM 3366 and MRSA CCARM 3708, respectively. The MBC of BV against MRSA 3366 was 0.106 µg/mL and that against MRSA 3708 was 0.14 µg/mL. The bactericidal activity of BV corresponded to a decrease of at least 3 log CFU/g cells. The combination of BV with ampicillin or penicillin yielded an inhibitory concentration index ranging from 0.631 to 1.002, indicating a partial and indifferent synergistic effect. Compared to ampicillin or penicillin, both MRSA strains were more susceptible to the combination of BV with gentamicin or vancomycin. The expression of atl gene was increased in MRSA 3366 treated with BV.

These results suggest that BV exhibited antibacterial activity and antibiotic-enhancing effects against MRSA strains. The atl gene was increased in MRSA exposed to BV, suggesting that cell division was interrupted. BV warrants further investigation as a natural antimicrobial agent and synergist of antibiotic activity.

Neonicotinoid Concentrations Low in UK Honey

Neonicotinoid Concentrations in UK Honey from 2013

Pest Management Science
Accepted Article

Background

Concentrations of the neonicotinoid insecticides clothianidin, thiamethoxam and imidacloprid were determined in honey collected in Spring 2013 from a variety of locations in England. The honey was produced before the moratorium in the EU on the use of neonicotinoids in pollinator-attractive crops became effective.

Results

Neither imidacloprid, not its metabolites were detected in any honey samples. Concentrations of clothianidin ranged from < 0.02 µg/kg to 0.82 µg/kg and thiamethoxam concentrations were between < 0.01 µg/kg and 0.79 µg/kg.

Conclusion

Neonicotinoid concentrations were below those likely to cause any chronic mortality. The concentrations detected should provide a useful baseline against which the effectiveness of the moratorium in reducing exposure of honeybees can be measured.

Chemical Composition and Biological Activity of Brazilian Brown, Green and Red Propolis

Chemical Composition and Biological Activity of Extracts Obtained by Supercritical Extraction and Ethanolic Extraction of Brown, Green and Red Propolis Derived from Different Geographic Regions in Brazil

PLoS One. 2016 Jan 8;11(1):e0145954

The variations in the chemical composition, and consequently, on the biological activity of the propolis, are associated with its type and geographic origin. Considering this fact, this study evaluated propolis extracts obtained by supercritical extraction (SCO2) and ethanolic extraction (EtOH), in eight samples of different types of propolis (red, green and brown), collected from different regions in Brazil. The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH and ABTS), Artepillin C, p-coumaric acid and antimicrobial activity against two bacteria were determined for all extracts. For the EtOH extracts, the anti-proliferative activity regarding the cell lines of B16F10, were also evaluated. Amongst the samples evaluated, the red propolis from the Brazilian Northeast (states of Sergipe and Alagoas) showed the higher biological potential, as well as the larger content of antioxidant compounds. The best results were shown for the extracts obtained through the conventional extraction method (EtOH). However, the highest concentrations of Artepillin C and p-coumaric acid were identified in the extracts from SCO2, indicating a higher selectivity for the extraction of these compounds. It was verified that the composition and biological activity of the Brazilian propolis vary significantly, depending on the type of sample and geographical area of collection.

Composition, Biological Activity of Brazilian Propolis Vary Based on Type of Sample, Geographical Area of Collection

Chemical Composition and Biological Activity of Extracts Obtained by Supercritical Extraction and Ethanolic Extraction of Brown, Green and Red Propolis Derived from Different Geographic Regions in Brazil

PLoS One. 2016 Jan 8;11(1):e0145954

The variations in the chemical composition, and consequently, on the biological activity of the propolis, are associated with its type and geographic origin. Considering this fact, this study evaluated propolis extracts obtained by supercritical extraction (SCO2) and ethanolic extraction (EtOH), in eight samples of different types of propolis (red, green and brown), collected from different regions in Brazil.

The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH and ABTS), Artepillin C, p-coumaric acid and antimicrobial activity against two bacteria were determined for all extracts. For the EtOH extracts, the anti-proliferative activity regarding the cell lines of B16F10, were also evaluated.

Amongst the samples evaluated, the red propolis from the Brazilian Northeast (states of Sergipe and Alagoas) showed the higher biological potential, as well as the larger content of antioxidant compounds. The best results were shown for the extracts obtained through the conventional extraction method (EtOH). However, the highest concentrations of Artepillin C and p-coumaric acid were identified in the extracts from SCO2, indicating a higher selectivity for the extraction of these compounds.

It was verified that the composition and biological activity of the Brazilian propolis vary significantly, depending on the type of sample and geographical area of collection.

Antibacterial and Leishmanicidal Activity of Bolivian Propolis

Letters in Applied Microbiology

The antimicrobial activity of Bolivian propolis was assessed for the first time on a panel of bacteria and two endemic parasitic protozoa. Ten samples of Bolivian propolis and their main constituents were tested using the micro-broth dilution method against 11 bacterial pathogenic strains as well as against promastigotes of Leishmania amazonensis and L. braziliensis using the XTT-based colorimetric method. The methanolic extracts showed antibacterial effect ranging from inactive (MICs >1000 μg ml−1) to low (MICs 250-1000 μg ml−1), moderate (62∙5-125 μg ml−1) and high antibacterial activity (MIC 31.2 μg ml−1), according to the collection place and chemical composition. The most active samples towards Leishmania species were from Cochabamba and Tarija, with IC50 values of 12∙1 and 7∙8, 8∙0 and 10∙9 μg ml−1 against L. amazonensis and L. brasiliensis, respectively. The results show that the best antibacterial and antiprotozoal effect was observed for some phenolic rich propolis.

Bee Venom Component Can Mitigate Airway Inflammation

Bee venom phospholipase A2 suppresses allergic airway inflammation in an ovalbumin-induced asthma model through the induction of regulatory T cells

Immun Inflamm Dis. 2015 Aug 9;3(4):386-97

Bee venom (BV) is one of the alternative medicines that have been widely used in the treatment of chronic inflammatory diseases. We previously demonstrated that BV induces immune tolerance by increasing the population of regulatory T cells (Tregs) in immune disorders. However, the major component and how it regulates the immune response have not been elucidated.

We investigated whether bee venom phospholipase A2 (bvPLA2) exerts protective effects that are mediated via Tregs in OVA-induced asthma model. bvPLA2 was administered by intraperitoneal injection into control and OVA-challenged mice. The Treg population, total and differential bronchoalveolar lavage fluid (BALF) cell count, Th2 cytokines, and lung histological features were assessed. Treg depletion was used to determine the involvement of Treg migration and the reduction of asthmatic symptoms. The CD206-dependence of bvPLA2-treated suppression of airway inflammation was evaluated in OVA-challenged CD206(-/-) mice.

The bvPLA2 treatment induced the Tregs and reduced the infiltration of inflammatory cells into the lung in the OVA-challenged mice. Th2 cytokines in the bronchoalveolar lavage fluid (BALF) were reduced in bvPLA2-treated mice. Although bvPLA2 suppressed the number of inflammatory cells after OVA challenge, these effects were not observed in Treg-depleted mice. In addition, we investigated the involvement of CD206 in bvPLA2-mediated immune tolerance in OVA-induced asthma model. We observed a significant reduction in the levels of Th2 cytokines and inflammatory cells in the BALF of bvPLA2-treated OVA-induced mice but not in bvPLA2-treated OVA-induced CD206(-/-) mice.

These results demonstrated that bvPLA2 can mitigate airway inflammation by the induction of Tregs in an OVA-induced asthma model.

Honey Heals Skin Grafts Faster

Honey Dressing Accelerates Split-Thickness Skin Graft Donor Site Healing

Indian J Surg. 2015 Dec;77(Suppl 2):261-3

The management of the donor site after harvesting a skin graft is an important issue, as patients often report more discomfort at the donor site than at the recipient site. There is, however, a plethora of dressings available for the treatment and management of donor sites, yet, there is no widely accepted method established for these partial thickness wounds.

Honey has been found to be useful in the treatment of burns and other wounds, split-thickness skin graft donor sites are like partial thickness burn wounds and honey's healing effect on burn wound can also be expected on these types of wounds. Therefore, this study was undertaken to evaluate the effect of honey on skin graft donor sites.

From 2002 to 2004, 100 patients who have undergone skin grafting for various reasons formed the material of the randomized study divided into two groups of 50 each in honey-treated group and Vaseline gauze-treated group. Graft donor site area ranged from 30 to 48 cm(2), mean 32.6 cm(2). In the group treated with honey, 90 % of the patients had nil or only moderate pain, whereas in the group treated with Vaseline gauze,88 % had nil or mild pain (p > 0.001, not significant). There were no allergic reactions in any of the patients in either group. On opening of the dressing on the 7th day, epithelialization has occurred in 48 patients as compared to 39 in group 2, i.e., donor sites treated with Vaseline gauze (p < 0.05, statistically significant). By the 10th day, all the wounds healed in honey-treated group, whereas 76 % of wounds healed in Vaseline gauze-treated group (p < 0.05). At 1 month follow-up, the results were comparable in both groups, with regard to patient satisfaction.

In conclusion, honey-impregnated gauze causes less pain and heals donor sites wounds faster with good cosmetic result.

Oak and Chestnut Honeys Show Substantial Antioxidant Effects

Characterization of Anatolian honeys based on minerals, bioactive components and principal component analysis

LWT - Food Science and Technology
Volume 68, May 2016, Pages 273–279

Our aim is the characterization of Anatolian monofloral and honeydew honeys according to their mineral, vitamin B2, total phenolic contents and antioxidant activities. Five main elements (Ca, K, Fe, Cu, and Mn) were determined in 20 honey samples by inductively coupled plasma – optical emission spectrometry (ICP-OES). The vitamin B2 contents of honey samples were determined by the capillary electrophoresis method coupled with a sensitive laser induced fluorescence (LIF) detector. The total phenolic contents were analyzed with Folin–Ciocalteu's method. Two comparative antioxidant assays, namely cupric reducing antioxidant capacity assay (CUPRAC) and ABTS radical scavenging assay, were applied to detect the antioxidant power of honeys. Heather honeys were distinguished from others with significantly high vitamin B2 and iron contents. Considerably higher antioxidant capacities and Mn contents were observed for oak and chestnut honeys. Principal components analysis was applied to the analysis result in order to classify the honeys from different botanical origins.

Propolis Component Shows Anti-Cancer Properties

Caffeic acid phenethyl ester: Inhibition of metastatic cell behaviours via voltage-gated sodium channel in human breast cancer in vitro

Int J Biochem Cell Biol. 2015 Dec 24. pii: S1357-2725(15)30085-6

Caffeic acid phenethyl ester, derived from natural propolis, has been reported to have anti-cancer properties. Voltage-gated sodium channels are upregulated in many cancers where they promote metastatic cell behaviours, including invasiveness.

We found that micromolar concentrations of caffeic acid phenethyl ester blocked voltage-gated sodium channel activity in several invasive cell lines from different cancers, including breast (MDA-MB-231 and MDA-MB-468), colon (SW620) and non-small cell lung cancer (H460). In the MDA-MB-231 cell line, which was adopted as a 'model', long-term (48h) treatment with 18μM caffeic acid phenethyl ester reduced the peak current density by 91% and shifted steady-state inactivation to more hyperpolarized potentials and slowed recovery from inactivation. The effects of long-term treatment were also dose-dependent, 1μM caffeic acid phenethyl ester reducing current density by only 65%. The effects of caffeic acid phenethyl ester on metastatic cell behaviours were tested on the MDA-MB-231 cell line at a working concentration (1μM) that did not affect proliferative activity. Lateral motility and Matrigel invasion were reduced by up to 14% and 51%, respectively. Co-treatment of caffeic acid phenethyl ester with tetrodotoxin suggested that the voltage-gated sodium channel inhibition played a significant intermediary role in these effects.

We conclude, first, that caffeic acid phenethyl ester does possess anti-metastatic properties. Second, the voltage-gated sodium channels, commonly expressed in strongly metastatic cancers, are a novel target for caffeic acid phenethyl ester. Third, more generally, ion channel inhibition can be a significant mode of action of nutraceutical compounds.

Brazilian Propolis May Help Treat Mastitis in Cows

The effects of Brazilian propolis on etiological agents of mastitis and the viability of bovine mammary gland explants

J Dairy Sci. 2015 Dec 23. pii: S0022-0302(15)00941-8

The objective of this study was to evaluate in vitro the antimicrobial activity of Brazilian propolis from Urupema, São Joaquim, and Agua Doce (Santa Catarina State) and green propolis from Minas Gerais State, and the effects of propolis on bovine mammary gland explant viability.

The propolis samples differed in flavonoid content and antioxidant activity. Green propolis showed the highest content of flavonoids, followed by the sample from São Joaquim. The propolis from Urupema showed the lowest flavonoid content along with the lowest antioxidant activity. The total phenolics were similar across all studied samples. Despite phytochemical differences, the propolis samples from Minas Gerais, São Joaquim, and Urupema presented the same level of antimicrobial activity against Staphylococcus aureus strains. The reduction in S. aureus growth was, on average, 1.5 and 4 log10 times at 200 and 500 μg/mL, respectively. At concentrations of 1,000 μg/mL, all propolis reduced bacterial growth to zero.

On the other hand, when the propolis were tested against strains of Escherichia coli, the samples presented weak antimicrobial activity. Mammary explants were maintained in culture for 96 h without a loss in viability, demonstrating the applicability of the model in evaluating the toxicity of propolis. The origin and chemical composition of the propolis had an effect on mammary explant viability. We encountered inhibitory concentrations of 272.4, 171.8, 63.85, and 13.26 μg/mL for the propolis from Água Doce, Urupema, São Joaquim, and Mina Gerais, respectively. A clear association between greater antimicrobial activity and toxicity for mammary explants was observed. Of all propolis tested, the Urupema sample was noteworthy, as it showed antimicrobial activity at less toxic concentrations than the other samples, reducing bacterial growth to an average of 9.3 × 102 cfu/mL after 6 h of contact using 200 μg/mL of extract. The results demonstrate the potential for Brazilian propolis in the treatment of mastitis, although effectiveness is dependent on geographical origin and concentration.

The results from the mammary gland explant assays are promising for the investigation of other natural products with antimicrobial and anti-inflammatory properties that can be used in the intramammary treatment of subclinical mastitis and during dry cow therapy.

Royal Jelly May Help Treat Kidney Stones

Anti-inflammatory effects of royal jelly on ethylene glycol induced renal inflammation in rats

Int Braz J Urol. 2015 Sep-Oct;41(5):1008-13

OBJECTIVE:

In this study, anti-inflammatory effects of Royal Jelly were investigated by inducing renal inflammation in rats with the use of ethylene glycol. For this purpose, the calcium oxalate urolithiasis model was obtained by feeding rats with ethylene glycol in drinking water.

MATERIALS AND METHODS:

The rats were divided in five study groups. The 1st group was determined as the control group. The rats in the 2nd group received ethylene glycol (1%) in drinking water. The rats in the 3rd group were daily fed with Royal Jelly by using oral gavage. The 4th group was determined as the preventive group and the rats were fed with ethylene glycol (1%) in drinking water while receiving Royal Jelly via oral gavage. The 5th group was determined as the therapeutic group and received ethylene glycol in drinking water during the first 2 weeks of the study and Royal Jelly via oral gavage during the last 2 weeks of the study.

RESULTS:

At the end of the study, proinflammatory/anti-inflammatory cytokines, TNF-a, IL-1ß and IL-18 levels in blood and renal tissue samples from the rats used in the application were measured.

CONCLUSION:

The results have shown that ethylene glycol does induce inflammation and renal damage. This can cause the formation of reactive oxygen species. Royal Jelly is also considered to have anti-inflammatory effects due to its possible antiradical and antioxidative effects. It can have positive effects on both the prevention of urolithiasis and possible inflammation during the existing urolithiasis and support the medical treatment.

Italian Consumers Prefer Local Honey

Attitudes towards honey among Italian consumers: a choice experiment approach

Appetite. 2015 Dec 21. pii: S0195-6663(15)30127-6

Honey is becoming increasingly popular with consumers for its nutritional benefits as well as many other functions. The objective of this article is to determine which factors influence consumers' purchase intentions and to assess the importance of certain honey characteristics to enable identification of the constituents of an ideal honey profile. This information will lead to satisfaction of consumers' preferences and formulation of marketing strategies that support honey makers. We applied a choice experiment to the Italian honey market to define the preferences and the willingness to pay for key characteristics of the product. A face-to-face questionnaire survey was conducted in 2014 (January-July) among Italian consumers; it was completed by 427 respondents. A latent class model was estimated and four classes were identified, with different preferences, illustrating that respondents seem to be heterogeneous honey consumers Results suggest the "organic" attribute was more important than others factors, such as the place where the honey was produced (landscape), but less important than the country of origin; local Italian honey was preferred to foreign honey. Respondents showed a higher willingness to pay (WTP) for honey from their country of origin versus the production method used. Our results suggest that while organic beekeeping might be an important strategy for diversification, if suitable communication is not taken into consideration, the added value of the production method might not be perceived by consumers.

The Use of Propolis in Micro/Nanostructured Pharmaceutical Formulations

Recent Pat Drug Deliv Formul. 2015 Dec 29. [Epub ahead of print]

BACKGROUND:

Propolis is a resinous material with complex chemical structure, produced by bees using plant sources, displaying a wide spectrum of biological activities. Many studies have reported the use of this compound in pharmaceutical, medicinal, veterinary and dentistry areas, and the results have reported its pharmacological activities. Moreover, several propolis delivery systems have been proposed and their properties evaluated, indicating that they can be used. On the other hand, considering its chemical and physical characteristics, propolis could be used as a material to produce micro/nano-structured pharmaceutical formulations. This work reviews the recent studies of development of micro/nanostructured systems using propolis or its byproduct. In addition, patents were reviewed and categorized.

METHODS:

This review was based on a structured search of databases for peer-reviewed papers and patents using a focused review question and inclusion/exclusion criteria. The patent documents and the quality of retrieved articles were appraised using standard tools. The characteristics of screened patents and papers were described. In addition, a deductive qualitative content analysis methodology was applied to analyze the findings and interventions of included studies using a conceptual framework.

RESULTS:

Propolis identification, chemical composition and biological activity were discussed, and the information was structured and discussed. The literature shows the safety of this compound and the possibility to use it to prepare micro or nanostructured pharmaceutical systems. There are many strategies to modify the propolis delivery such as the use of micro/nanoemulsions, particles, thermoresponsive systems, films, hydrogels, lotions, creams, ointments, carbon nanotubes, liquid crystalline phases, and bio/mucoadhesive systems. Propolis can also be used as an excipient to produce drug-delivery systems.

CONCLUSION:

Many papers and submitted or granted patent applications were found, reporting from extraction method for propolis, which provides new possibilities for incorporation of propolis components in micro/nanostructured cosmetic products, foodstuffs and medicines.