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Propolis in human and bee health conference / Sofia, Septembеr 28 – 29, 2018 3

Dear Colleagues and Friends,

I am honoured to welcome you on behalf of the Organizing Committee of the Second Conference Propolis in Human and Bee Health. The inspiring success of the First Propolis Conference in Glasgow, 2016, demonstrated a growing community of researchers round the world committed to understanding propolis and developing its potential. Propolis research is actually a research of complex and sophisticated relationships: the relationships between the society of honeybees, the plant world and the human society. We already know a lot about these relationships but there is still much to be discovered and understood. The Second Propolis Conference is a great opportunity to share research results, new approaches and ideas, views and visions for the development of propolis studies and applications for the benefit of society. It is also a celebration of human curiosity and endeavour to explore Nature.

The Conference is held in Sofia, the capital city of Bulgaria, place of ancient and contemporary culture. We hope that you will enjoy the Conference, meet old friends and make new ones!

Welcome to the Second ConferencePropolis in Human and Bee Health 2018!

Vassya Bankova, PhD, DScChair of the Organizing Committee

Propolis in human and bee health conference / Sofia, Septembеr 28 – 29, 20184

Scientific Organizing Committee

Chair:

Vassya BANKOVACorresponding member of Bulgarian Academy of Sciences (BAS)

President of the Bulgarian Phytochemical SocietyInstitute of Organic Chemistry with Centre of Phytochemistry (Bulgaria)

Scientific committee members:

Miguel VilAS-BOAS – University of Applied Sciences of Bragança, PortugalJanko BOziC – University of Ljubljana, Slovenia

Hugo Fearnly – The Apiceutical Researcjh Center, UKJames Fearnly – The Apiceutical Researcjh Center, UKShigenori KumAzAwA – University of Shizuoka, Japan

Hu Fu-liang – Zhejiang University, Hangzhou, ChinaMilena POPOVA – Institute of Organic Chemistry with Centre of Phytochemistry, BAS, Bulgaria

anant PraDakar – University of Bradford, UKalexandra SaWaya – State University of Campinas, SP, Brazil

Mauricio SfOrCiN – São Paulo State University, SP, Brazilluca raStrelli – University of Salerno, Italy

Marla SPivak – University of Minnesota, MN, USADavid wAtSON – University of Strathclyde, UK

technical committee:

Boryana truSHevakristina GeOrGieVA

tsvetinka GrOzdANOVA


SeCONd PrOPOliS CONfereNCe, SOfiA 2018Conference program

Friday, September 28, 2018

08:30 – 18:30 Registration

09:15 – 09:30 Official opening ceremony

Session I

09:30 – 10:10 Plenary Lecture 1. James Fearnly (UK) Exploring a new generation of medicines from the beehive.

10:10 – 10:50 Plenary Lecture 2. Marla Spivak (USA)What we know, and don‘t know, about the benefits of propolis to honey bee health

10:50 – 11:05 Short lecture 1. Ahmad Sulaeman (Indonesia)Hepatoprotective activity of Indonesian stingless bee propolis against toxicity of anti-tuberculosis drug on pulmonary Tb patients

11:05 – 11:30 Coffee break

11:30 – 12:10 Plenary Lecture 3. Stefan Stangaciu (Romania)Use of Propolis in Clinical Medicine. A review

12:10 – 12:25 Short lecture 2. Kai Wang (China)Emerging impact of bee propolis for gut health: evidence and mechanisms

12:25 – 12:40 Short lecture 3. Plamen Enchev (Bulgaria)Application of propolis – herbal ethanol extract and ointment for a leg with gangrene (clinical case)

12:40 – 12:55 Short lecture 4. Verica Milojkovic (Serbia)Propolis: a natural product – great potential as a medicine

12:55 – 13:10 Short lecture 5. Sevgi Kolayli (Turkey)Some apitherapeutic properties of chestnut propolis

13:10 – 14:20 Lunch


Session II

14:30 – 15:10 Plenary Lecture 4. Tim Regan (UK) An exploration into the relationship between propolis production and trypanosome burden

15:10 – 15:50 Plenary Lecture 5. Milena Popova (Bulgaria) Chemical profiling of tropical propolis: challenges and new data

15:50 – 16:05 Short lecture 6. Efrain Alday (Mexico)Plant origin authentication of Sonoran propolis and its antiproliferative effect on cancer cells: a bioactive poplar type propolis from Semi-arid zones

16:05 - 16:20 Short lecture 7. Alexandra C. H. F. Sawaya (Brazil)Chemical composition of extracts of geopropolis from Melipona quadrifasciata (Mandaҫaia) evaluation of its antioxidant activity

16:20 – 16:35 Short lecture 8. Vincenzo Zaccaria (Italy)Characterized propolis extracts, obtained with standardized extraction method, show similar chemical; profile (HPLC-ESI.MSN) and in vitro antibacterial, antioxidant and anti-inflammatory activity through evaluation of expression of miRNAS, mRNAS and proteins.


17:00 – 18:30 General discussion: Meeting of the International Propolis Research Group18:30 - 19:30 Poster session

20:30 – 23:00 Conference Dinner


Saturday, September 29, 2018

08:30 – 18:30 Registration

Session III

09:00 –9:40 Plenary Lecture 6. Shigenori Kumazawa (Japan)Chemical profile and botanical origin of stingless bee propolis from Thailand and Indonesia

9:40 – 10:20 Plenary Lecture 7. David Watson (UK)The immune modulatory and anti-protozoal effects of different propolis samples.

10:20 -10:35 Short lecture 9. Stephen Bloor (New Zealand)New antiproliferative acyl glycerols from New Zealand propolis and source poplar resin.

10:35 – 10:50 Short lecture 10. Risqa Rina Darwita (Indonesia)The efficacy of propolis fluoride in inhibiting dental caries activity on primay teeth

10:50 – 11:05 Short lecture 11. Konstantia Graikou (Greece)An overview of chemical studies and biological activities of Mediterranean propolis


11:45– 12:25 Plenary Lecture 8. Vassya Bankova (Bulgaria)Insights and pitfalls in propolis research

12:25– 13:05 Plenary Lecture 9. Badiaa Lyoussi (Morocco)Propolis: Pharmacological properties and medical applications of Propolis in modern medicine

13:05 – 13:20 Short lecture 12. Miguel Vilas-Boas (Portugal)A collaborative study for performance evaluation of analytical methods for propolis. An IHC trial

13:20 - 13:35 Short lecture 13. Katarína Bíliková (Slovakia)Molecular characterization of propolis and royal jelly components as tool for study of their multifunctional therapeutic effects.

13:35– 14:30 Lunch


Session IV

14:30 – 14:45 Short lecture 14. Michael Goblirsch (USA)Can propolis inhibit infection dynamics of honey bee (Apis mellifera L.) viruses in vitro?

14:45 – 15:00 Short lecture 15. Shankar Katekhaye (UK)Study of relationship between geographical location of collection, chemical composition and biological activity of propolis by multivariate data analysis

15:00 – 15:15 Short lecture 16. Amando Siuiti Ito (Brazil)Interaction of Artepillin C with model membranes: optical absorption and fluorescence spectroscopy studies

15:15 – 15:30 Short lecture 17. Şaban Keskin (Turkey)A new approach to propolis extraction

15:30 – 15:45 Short lecture 18. Cristina Almeida-Aguiar (Portugal)Portuguese propolis: a source of valuable bioactivities

15.45 – 16:15 Coffee break

16:15 – 16:30 Short lecture 19. Zbigniev Balion (Lithuania)Anticancer properties of aqueous and nonaqueous propolis extracts

16:30 – 16:45 Short lecture 20. Merve Keskin (Turkey)Antimicrobial effect of commercial propolis extract (BEEO©)

16:45 – 17:00 Short lecture 21. Boryana Trusheva (Bulgaria)New insights into tropical propolis: propolis from Pitcairn Island Short

17:00 - 17:15 Short lecture 22. Klemen Rihar (Slovenia)Toothpaste with propolis “Apident” shows antimicrobial activity in vitro

17:15 - 17:30 Short lecture 23. Hugo Fearanly (UK)ARC (Apiceutical Research Centre): Developing a global BeePharma network. Local kelp for local health

17:30– 17:45 Short lecture 24. Ashok K. Shakya (Jordan)Fatty acid analysis and biological activity of Jordanian Propolis

17:45– 18:00 Closing Ceremony


inviteD leCtureS


IL 1

arC (apiceutical research Centre): exploring a new generation of Medicines from the BeehiveJames Fearnly1

1Apicultural Research Centre Modern pharmaceutical medicine is in crisis. The search for the magic bullet is prov-ing ever more elusive and inadequate. Whilst the economically driven pharmaceutical jugger-naut trundle on, the definition and meaning of medicine is being redefined – from targeted, synthetic and single molecule to natural, synergistic and holistic. Science is turning back again to natural whole products with a new will to understand their complexity, intricacy and potency as medicines for man. Medicines from the beehive as with plant medicines have been used for millennia. In our short sightedness however, we have isolated the loudest instruments in the orchestra and synthesised them, but we have lost the symphony. We must thank the East Europeans for keeping alive a scientific understanding of bee medicines. Over 40 years ago the Kazan Veterinary Institute published research which showed that combining propolis with antibiotics increased the effectiveness of the antibiotics by up to 100 times. We had to wait till the 1990’s before the West began to publish scientific papers. Thanks to a brilliant combination of marketing and science, Manuka Honey has become known to millions of consumers worldwide as a natural antibiotic. It is a shame that this marketing drive has left those same consumers thinking only Manuka honey has antibiotic properties. Research into propolis, potentially the greatest medicinal jewel in the beehive treasure chest, has grown exponentially over the last ten years with Brazil, China, Turkey and India now major contributors. Research in UK has grown steadily over the last 15 years particularly at the university of Strathclyde in Scotland. They have developed a greater understanding of how the chemical compounds collected by the honey bee from plants and trees within their local environment is transformed by the honey bee into a product able to provide immune defence for the whole superorganism which is the bee colony. The relationship between climate and the antibacterial properties of propolis is now more clearly understood. The discovery of anti-trypanosome chemicals in propolis in areas where there is sleeping sickness has opened exciting new research possibility.ARC (Apiceutical Research Centre) founded in 2010 has grown out of these insights, in particu-lar the concept that the bee hive could be a source of a unique new generation of (Geographic) Medicines for man. Since 2011 ARC has been linking research activity round the world through a series of international conferences and more recently through the formation of IPRG (International Propolis Research Group). ARC’s Global BeePharma project further explores the concept of Geographic Medicine i.e. valuable therapeutics for man derived from the unique transformation by the honey bee of the plants defence mechanism (generated in response to its environment), into the honey bees immune support mechanism and the positive relationship of these immune defence products to the diseases effecting man in those same geographic areas. ARC has developed plans to build The BeeArc, a physical research centre and exhibition centre based in North Yorkshire UK and focussed on Apiceuticals – medicines from the beehive and sustain-able beekeeping.


IL 2

what we Know, and don‘t Know, about the Benefits of Propolis to Honey Bee Health

Marla Spivak1

1Department of Entomology, University of Minnesota, USA

When colonies of Apis mellifera nest in tree cavities, they line the nest interior with a propolis envelope. Colonies in standard beekeeping equipment made of smooth wood do not construct a propolis envelope. The benefits of an antimicrobial layer are both prophylactic and therapeutic; a propolis envelope can improve colony health by reducing stress on individual immunity and can directly inhibit some pathogens (e.g., American foulbrood) and parasites (e.g., small hive beetle)1, 2, 3. More research is needed on the costs and benefits of resin collection to individual bees, and the presence of a propolis envelope on colony health. The behavioral regulation of resin foraging at the individual bee and colony level is poorly understood: how do bees locate sources of plant resins; do they choose among resin sources; and how do they recruit nest mates to the resins? We do not understand the modes of action (contact and/or volatile emissions) of propolis on bee immune function, and against A. mellifera microbes and parasites. At the colony level, more studies are needed to explore propolis as a treatment against hive diseases, both alone and in combination with other mechanisms of social immunity (i.e. hygienic behavior). What modifications in manmade bee boxes are needed to stimulate colonies to construct a propolis envelope? And finally, unlike A. mellifera and stingless bees, the Asian honey bee, A. cerana, does not collect resin or incorporate propolis in its nest - why not?

Acknowledgements: National Science Foundation IOS 0717530 and IOS 1256992 to M Spivak

References:1 Simone-Finstrom M, Borba RS, Wilson M, Spivak M (2017) Propolis counteracts some threats to honey bee health. Review. Insects 8: 462 Simone-Finstrom M, Spivak M (2010) Propolis and bee health: the natural history and significance of resin use by honey bees. Apidologie 41: 295–3113 Borba RS, Spivak M (2017) Propolis envelope in Apis mellifera colonies supports honey bees against the pathogen, Paenibacillus larvae. Scientific Reports 7: 11429


IL 3

use of Propolis in Clinical Medicine. a review

Stefan Stangaciu1

1President of the Romanian and German Apitherapy Societies, Secretary general of the InternationalFederation of Apitherapy, Secretary general of the International Federation of Apitherapy, [email protected]

The aim of the work is to find common characteristics, rules and principles of the use of propolis in clinical medicine, at international level. Scientists from all over the world have found that propolis, due to its extremely rich composition in useful, natural, pharmacologically active compounds can offer many useful properties and uses.Methods. We have studied scientific and regular papers on propolis, papers written in over 30 countries in various languages (English, French, Spanish, German, Portuguese, Romanian). The main studied topics were propolis geobotanical origin, uses in the beehive, physico-chemical characteristics, composition, pharmacological effects, indications, contra-indications, administration, preparations and products. We have discussed directly, at various national and international conferences, with scientists and practitioners that studies/uses regularly propolis. We have afterwards corroborated everything with our own clinical experience and the clinical experience of many colleagues from all over the world.Main results. Propolis can offer over seventy useful properties and hundreds of uses in clinical medicine, both as a preventative and as curative agent in human and veterinary medicine. It is also an excellent product for Api-Cosmetology.Conclusions. Propolis is an extraordinary “tool” in the hands of a well informed medical doctor/health practitioner. It is by far the most important product to be kept in each home’s pharmacy and in each medical office or clinic, no matter the medical specialty/groups of diseases are prevented/treated. If propolis would be as nutritious as it is pharmacologically active, we would be able to name it, almost without hesitation, as a true panacea!

mailto:[email protected]


IL 4

an exploration into the relationship Between Propolis Production and trypanosome Burden

T. Regan1, M. Barnett1, T. Freeman1

1 The Roslin Institute, University of Edinburgh, UK

The European honey bee (Apis mellifera) plays a major role in pollination and food production, but is under threat from emerging pathogens and agro-environmental insults. As with other organisms, honey bee health is a complex product of environment, host genetics and associated microbes (commensal, opportunistic and pathogenic). Improved understanding of bee genetics and their molecular ecology can help manage modern challenges to bee health and production. Sampling bee and cobiont genomes, we characterised the metagenome of 19 honey bee colonies across Britain. Low heterozygosity was observed in bees from many Scottish colonies, sharing high similarity to the native dark bee, A. mellifera mellifera. Apiaries exhibited high diversity in the composition and relative abundance of individual microbiome taxa. Most non-bee sequences derived from known honey bee commensal bacteria or known pathogens, e.g. Lotmaria passim (Trypanosomatidae), and Nosema spp. (Microsporidia). However, DNA was also detected from numerous additional bacterial, plant (food source), protozoan and metazoan organisms. This leads to an anecdotal observation that colonies producing high levels of propolis carried lower trypanosome burden. We are currently investigating this potential anti-correlation at our apiary in Edinburgh.


IL 5

Chemical Profiling of tropical Propolis: Challenges and new Data

Milena Popova1, Boryana Trusheva1, Kristina Georgieva1, Vassya Bankova1

1 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev str., bl.9, 1113 Sofia, Bulgaria

Propolis is a beehive product valuable for both bees and humans. Its complex chemical composition and beneficial properties have attracted the scientific attention for many years. Extensive studies on propolis from different regions have revealed similar chemistry of propolis from temperate zones, its botanical origin is mainly Populus spp., and remarkably different for the tropical one. Due to diversity of tropical flora, several propolis types have been formulated most of them originating from Brazil, Cuba and Pacific Islands. However, in tropical climatic zone there are regions from where propolis is scantly or not investigated that may result in characterization of new types, and thus in isolation of valuable bioactive compounds. In this end, the study of tropical propolis entails challenges associated with diversity of geographical locations, plant sources and bee species. Propolis analysis is an added challenge due to need of reliable profiling, and identification, if possible, of majority of the compounds often being a number of structurally different, present in one sample. In relation to above-mentioned, our recent results on propolis collected by honey bees and/or indigenous bee species from Oman, Thailand, Fiji, Colombia and Vietnam will be summarized. Moreover, new data about isolated new natural constituent, and constituents found in propolis for the first time, such as tocotrienolic acid derivatives, prenylated benzophenons, flavanes, homoisoflavans, and prenylated xanthones, as well as their supposed plant sources will be also presented.

Acknowledgment: A partial support from a project between Bulgarian Academy of Sciences and Vietnamese Academy of Science and Technology is gratefully acknowledged.


IL 6

Chemical Profile and Botanical Origin of Stingless Bee Propolis from thailand and indonesia

Shigenori Kumazawa1

1 Department of Food and Nutritional Sciences, University of Shizuoka, Japan

Propolis is a resinous mixture of substances collected by honey bees from certain plants. We recently obtained stingless bee propolis from Thailand and Indonesia. To assess their potential utility, we analyzed the composition of these stingless bee propolis. Thailand propolis was from stingless bee species, Tetragonula pagdeni, from a mangosteen orchard in Chanthaburi. Nine prenylated xanthones were isolated from propolis and their structures were determined using high resolution electrospray mass spectrometry (HRESIMS) and NMR. These compounds were previously isolated from the pericarps of Garcinia mangostana. Therefore mangosteen fruit was paid attention. Comparative analysis of the 70% ethanol extracts of Thailand stingless bees propolis and the yellow resin from the fruit surface of Garcinia mangostana was performed using reversed-phase high-performance liquid chromatography coupled with HRESIMS. The extracts showed the similar chromatographic patterns. In conclusion, we suggest that the plant origin of Thailand stingless bee propolis is the yellow resin from fruit surface of G. mangostana1. Indonesian stingless bee (Tetragonula biroi) propolis was collected on South Sulawesi. We isolated four new compounds and nine known compounds from the propolis. Moreover, some of the isolated compounds were tested for xanthine oxidase inhibitory activity. As a result, one of the isolated compounds showed potent XO-inhibitory activity, with an IC50 value of 3.9 mM. This IC50 value is close to that allopurinol, which is used as anti-gout drug.

References: 1 Ishizu E, Honda S, Vongsak B, Kumazawa S (2018) Identification of plant origin of propolis from Thailand stingless bees by comparative analysis. Natural Products Communications, in press.


IL 7

the immune modulatory and Anti-protozoal effects of Different Propolis Samples

David G. Watson1, Harry de Koning2, Godwin Ebiloma2, John Igoli2, Weam Siheri1, Naif Alenzi1, Samyah Alanazi1, Sameah Alenezi1 William Harnett1.

1 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 1RE.2 Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK

Propolis almost always has moderate to strong activity against protozoa, with the strongest activity being against trypanosomatids. It has become evident that protozoan infection in bees is widespread with Lotmaria passim, a monoxenous parasite transmitted via infection of the rectum and a close relative of Trypansoma brucei, being widespread in European bee populations 1, 2. We have studied anti-protozoal activity in propolis from Libya, Saudi Arabia and Nigeria and most recently in samples from the UK. Samples from the UK have widely varying levels of activity against both T.brucei and Crithidia fasiculata, which is closely related to Crithidia mellificae which infects the honey bee. Using orthogonal partial least squares modelling it was possible to see that the highest activity in the UK samples seems to be associated with the abundance of pinobanksin esters. In a separate study the immunomodulatory effects of propolis were studied and it was found that propolis extracts from Brazil and the UK were strong inhibitors of nitric oxide formation by primary macrophages stimulated with lipopolysaccharide. In addition the propolis extracts lowered TNF-1β levels and IL-6 levels. Metabolomic profiling of the macrophages indicated that propolis samples had a direct effect on the conversion of arginine into citrulline and the recycling of citrulline back to arginine via argininosuccinate.

References:1 Ravoet, J.; Schwarz, R.S.; Descamps, T.; Yañez, O.; Tozkar, C.O.; Martin-Hernandez, R.; Bartolomé, C.; De Smet, L.; Higes, M.; Wenseleers, T. Differential diagnosis of the honey bee trypanosomatids Crithidia mellificae and Lotmaria passim. Journal of invertebrate pathology 2015, 130, 21-27.2 Schwarz, R.S.; Bauchan, G.R.; Murphy, C.A.; Ravoet, J.; Graaf, D.C.; Evans, J.D. Characterization of two species of trypanosomatidae from the honey bee Apis mellifera: Crithidia mellificae langridge and mcghee, and Lotmaria passim n. Gen., n. Sp. Journal of Eukaryotic Microbiology 2015, 62, 567-583.


IL 8

insights and Pitfalls in Propolis research

Vassya Bankova1, Milena Popova1, Boryana Trusheva1

1 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev str., bl.9, 1113 Sofia, Bulgaria

Some recent developments, and also some potential sources of erroneous interpretation of experimental results in propolis research are discussed. Latest advances in the knowledge about factors which influence the choice of propolis botanical sources by honeybees, as well as in the metabolomic approaches to propolis studies are considered. The problems and possible misinterpretations connected to the use of palinological data as an indication of the propolis source plant, to the evidence of changes in the chemistry of collected plant resin by bees and to the procedures for quantification of total phenolics in propolis are critically reviewed.


IL 9

Propolis: Pharmacological Properties and Medical applications of Propolis in Modern Medicine Badiaa Lyoussi1

1Laboratory of Physiology, Pharmacology and Environmental Health, University Sidi Mohamed Ben Abdallah, Fez, Morocco, [email protected]

Diabetes mellitus and its associated complications (cardiovascular, renal, hepatic, ophthalmic, neurological, and osteopathic-, endothelial-, and sexual-dysfunction, etc.) remain a burden worldwide in spite of the availability of a number of anti-diabetic drugs, many of which have adverse effects. There is a trend towards using natural products to control hyperglycemia and associated pathologies. Propolis, an apiproduct used by the honeybees as a sealant, has been rediscovered as a medicinal agent, reported to be of benefit in the treatment of allergies, bruises, burns, ulcers, sunburn, wounds, tumors, fatigue, sore throat, nasal congestion, respiratory ailments, flu, colds, acne, skin disorders, and shingles. Propolis consists of plant resins, balsams, wax, bee pollen and essential oils. The composition of propolis (of different colors) depends on the phytogeographical location, seasonal collection time, and botanical source. It is reported to contain more than 300 natural compounds such as polyphenols, phenolic aldehydes, sesquiterpene-quinones, coumarins, amino acids, steroids and inorganic compounds. Propolis is reported to have anti-angiogenic, antihypercholesterolemic, antihypertensive, anti-inflammatory, antimicrobial, antioxidant, anti-parasitic, anti-septic, anti-tumor, antiulcer, anti-viral, immune-stimulatory properties, and hepato-, cardio-, and neuro-protective actions. Diabetes and pathologies associated with it are mainly due to inflammation and oxidative stress, as a result of elevated levels of reactive oxygen species (ROS), which cause lipid peroxidation and protein oxidation. The anti-oxidant, oxygen radical scavenging activity of propolis (and its extracts) is mainly due to the presence of phenolics and flavonoids. The beneficial effects of propolis in diabetes have been confirmed by a number of studies in experimental animals. For example, propolis given orally to rats with streptozotocin (STZ)-induced diabetic rats significantly decreased plasma insulin and insulin resistance, reduced glycated hemoglobin, suppressed elevated hepatic enzymes, and increased hepato-renal glutathione peroxidase levels. An ethanolic extract of propolis administered to STZ-induced diabetic rats, reversed body and kidney weight loss, improved serum glucose and lipid profile, and renal function tests, as well as decreased oxidative damage [increased superoxide dismutase, glutathione, catalase and decreased malondialdehyde (MDA)] in the renal and pancreatic tissue. Similar results were obtained by us using Moroccan propolis in STZ-diabetic rats and diabetic 2 rats. The protective role of propolis against the ROS induced damages in diabetic rats and nephrotoxicity models gives hope that they may have similar protective action in humans. These studies in humans and in experimental model of diabetes in rats, provide a basis for therapeutic use of beehive products. Furthermore, several studies, including ours, have revealed that the observed beneficial effects of beehive products may be as a result of synergistic action of several constituents.



SHOrt leCtureS


SL 1

Hepatoprotective activity of indonesian Stingless Bee Propolis against toxicity of anti-tuberculosis Drug on Pulmonary tb Patients A. Sulaeman1, M. Mahani2, Hardinsyah1, E. Darma3, S. R. Mubarokah3, N. Nurjanah4

1Department Community Nutrition, Faculty of Human Ecology, Bogor Agricultural University, Indonesia, [email protected] 2Department Food Industrial Technology, Faculty of Agricultural Engineering, Padjadjaran University, Indonesia3Public Health Office, Bogor City, West Java, Indonesia.4Center of Research and Development for Biomedical and Basic Technology of Health, National Agency for Health Research and Development, Ministry of Health, Indonesia

The usage of rifampicin, isonoazid, pyrazinamide and ethambuthol to treat tuberculosis (Tb) leads to hepatotoxicity which affects patient’s body weight and nutritional status. Numerous studies have shown propolis to act as a hepatoprotector. This study aimed to clinically evaluate the ability of propolis to protect the liver from the toxic effects of anti tuberculosis drug (ATD) and its impact on body weight and nutritional status changes of patients with pulmonary Tb. Randomized clinical controlled trial, involving 50 subjects of lung Tb patients divided into three groups: control (P0), propolis 1 supplementation (P1) and propolis 2 supplementation (P2); each subject received intervention for 6 months. Biomarkers of liver toxicity; SGPT, SGOT, BT, SOD and GSH were measured three times and body weight and nutritional status were assessed weekly during treatment. Measurement of liver toxicity biomarkers showed control groups experienced high liver oxidative damage during the intensive treatment stage as characterized by bilirubin exceeding normal values. In contrast, the supplementation propolis groups showed no liver oxidative damage as indicated by normal bilirubin concentrations. P2 group showed excellent hepatoprotective activity as showed by the lowest bilirubin, the greatest decrease of SGPT and the best antioxidant status of GSH and SOD enzymes. This group also showed body weight and nutritional status increased from the beginning of the intervention, while the other group decreased first. Pursuant to all indicators, propolis 2 supplementation demonstrates liver protection from toxic effects of OAT and prevents decreased body weight and nutritional status of pulmonary Tb patients.

Acknowledgements: Ministry of Research and Higher Education of the Republik Indoensia for the research grants

References:1 Mahani, Sulaeman A, Anwar F, Damanik MRM, Hardinsyah, Ploeger A. 2018. Determination of Indonwsian native stingless bee propolis as complementary nutraceutical candidate of antituberculosis drug. Int J Pharm Pharm Sci 10 (4): 15-212 Mahani, Sulaeman A, Anwar F, Damanik MRM, Hardinsyah, Ploeger. 2018. Efficacy of Propolis Supplementation to Accelerate Healing Process and Body Weight Recovery of Pulmonary Tuberculosis Patients. J Gizi dan Pangan.



SL 2

emerging impact of Bee Propolis for gut Health: evidence and Mechanisms

Kai Wang1, Xiaolu Jin2, Qiangqiang Li1, Alexandra Christine Helena Frankland Sawaya3, Richard K. Le Leu4, Michael A. Conlon5, Liming Wu1,* and Fuliang Hu6,*

1 Institute of Apiculture Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China;2 Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China;3 Faculty of Pharmaceutical Science, Universidade Estadual de Campinas, UNICAMP, Campinas, Brazil;4 Central and Northern Adelaide Renal and Transplantation Service, Royal Adelaide Hospital, Adelaide 5000, Australia;5 CSIRO Health and Biosecurity, Adelaide, SA 5000, Australia6 College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.

Bee propolis is an important hive product and considered beneficial to health. However, evidence of its potential for improving gut health is still lacking. We hereby investigated the effects of dietary supplementation with propolis against dextran sulfate sodium (DSS)-induced acute colitis model in rats. Moreover, effects of propolis extracts on intestinal barrier function were explored in human intestinal epithelial cells. Rats were fed with control diet or supplemented with different amounts of Chinese propolis (0.1%, 0.2%, and 0.3%) to examine effects on acute colitis induced by 3% DSS in drinking water. Propolis at 0.3%, but not lower levels, significantly improved colitis symptoms compared with the control group, with a less pronounced disease activity index (DAI), a significant increase in colon length/weight ratio and an improved distal colon tissue structure as assessed by histology. Although short chain fatty acid levels in digesta were not altered by propolis supplementation, 16S rRNA phylogenetic sequencing revealed a significant increase in gut microbial diversity after 21 days of 0.3% propolis supplementation compared with controls including a significant increase in bacteria belonging to the Proteobacteria and Acidobacteria phyla. In human intestinal epithelial cells, propolis increased transepithelial electrical resistance with organized expressions in proteins related to TJ assembly, i.e., occludin and ZO-1, by activating AMPK and ERK signaling. Overall, these data provide new insights into understanding the beneficial effects of propolis on gut health and have important implications for human IBD prevention/treatment.

Acknowledgements: This work was supported by the National Natural Science Foundation of China under Grant (No. 31702287); the Agricultural Science and Technology Innovation Program (talent training programs, No. 20170815), and Modern Agroindustry Technology Research System from the Ministry of Agriculture of China under Grant (CARS-45).


SL 3

application of Propolis – Herbal ethanol extract and Ointment for a leg with Gangrene (Clinical Case)

Plamen Entchev1,2

1Hotel Hissar, Sana Spa Hotel, 2Apitherapy office Plovdiv

Propolis has been utilized in Bulgarian folk medicine for many centuries, mainly for slow healing wounds. This abstract will aim to describe the role of propolis in the case of a leg with gangrene. Propolis can affectively stimulate the granulation tissue, and speed up the recovery of slow healing wounds. In this case, I initially applied an ethanol–based propolis (after evaporation of the alcohol) on the wound on a daily basis. After, I applied herbal ethanol propolis extract and herbal propolis ointment. The wound was dressed and packed with propolis-herbal soaked (dry) gauze. The herbal propolis ointment was applied on the edges of the wound and around. During the treatment, regular surgical debridements were utialized when required. After almost six months of intense daily treatment using the methods described above, the wound finally closed and the leg was saved. This case showed that this type of dressing is very suitable for slow healing wounds containing a lot of necrotic tissue. Barely any antibiotics were required during the wound’s treatment. Even for a challenging case like this one, the propolis worked effectively. The final result is that this type of dressing is one of the most effective known to me for these types of wounds, which can be a field for further research.


SL 4

Propolis: a natural Product – great Potential as a Medicine

Verica Milojkovic

Family physician specialist, Licensed Apiphytotherapist, President of the Serbian Apitherapy Society, Executive member of the International Federation of ApitherapyE-mail: [email protected]; www.apiterapija.rs

Introduction. Propolis has been used in traditional medicine for centuries, while in recent years it has attracted great attention as a valuable or potential substance used in medicine1,2,3.Clinical experiences and scientific evidence point to its wide therapeutic potential in the treatment of respiratory tract diseases as well as the diseases of oral cavity, the digestive system and the skin. Propolis exhibits a broad spectrum of biological activities, such as anticancer, antioxidant, anti-inflammatory, antibiotic and antifungal. It also successfully resolves problems with Candid in the oral cavity and intestines. Propolis has shown interesting biomedical properties, such as improving wound healing, tissue regeneration stimulation and skin psoriasis treatment. Aim. The aim of this paper is to demonstrate the propolis effectiveness in treating bacterial infections of the throat and nose, the Candida infection of the tongue, as well as chronic wounds and psoriasis. Materials and Methods. Experiences in the treatment of psoriasis and chronic wounds with propolis.In this paper,the author has used propolis as tincture, nasal spray, oral spray, propolis cream for chronic wounds and cream for psoriasis.Results. The results confirm the propolis therapeutic efficacy. Propolis possesses antioxidant, anti-inflammatory, antibiotic, antifungal and regenerative properties. Conclusion. Therapies with propolis offer new possibilities for treatment. Propolis, a medical miracle from the hive, has given encouraging results in the treatment of bacterial and fungal infections, as well as in the treatment of psoriasis and chronic wounds.Propolis has been attracting scientific attention since it has many biological and pharmacological properties. Propolis has excellent chances to become an antibiotic of the future.References:1. Nunan, R., Harding, K. G., & Martin, P. (2014). Clinical challenges of chronic wounds: searching for an optimal animal model to recapitulate their complexity. Disease models & mechanisms, 7(11), 1205-1213.2 Kucharzewski, M., S. Kubacka, T. Urbanek, K. Wilemska-Kucharzewska, and T. Morawiec. (2013) Stan scheller: the forerunner of clinical studies on using propolis for poor and chronic nonhealing wounds. Evidence-based complementary and alternative medicine 2013. 456859.3 Kuropatnicki, A. K., Szliszka, E., & Krol, W. (2013). Historical aspects of propolis research in modern times. Evidence-Based Complementary and Alternative Medicine, 2013.


http://www.apiterapija.rs/


SL 5

Some apitherapeutic properties of chestnut propolis

Sevgi Kolayli1, Saliha Eksi2, Zihni Yazici2, Oktay Yildiz1, Hüseyin Şahin3, Zehra Can4, Şengül KaraoğluA2

1Karadeniz Technical University, Faculty of Sciences, Department of Chemistry, Trabzon, Turkey2RTU University, Faculty of Medicine, Department of Microbiology, Rize, Turkey3 Giresun University, Espiye MYO School, Giresun, Turkey4 Bayburt University, MYO School, Bayburt, Turkey

Composition and biological active properties of propolis is depends many different parameters, such as floral sources, collecting season, and beekeeper’s performance. In this study, phenolic composition and some apiterapeutic potentials of chestnut originated propolis samples were investigated. The raw propolis samples were obtained from Balıkesir region of Turkey. Antioxidant, anti-microbial, anti-inflammatory and anti-tumoral properties of the ethanolic samples were investigated. A wide screening antimicrobial tests againts some pathogenic microorganism of Escherichia coli, Yersinia pseudo tuberculosis, Helicobacter pylori, Klebsiella pneumonias subp. pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutants, Streptococcus agalactiae, Bacillus cereus, Lactobacillus acidophilus, Lactobacillus casei, Mycobacterium smegmatis, Candida albicans, Candida tropicalis and Saccharomyces cerevisiae were tested. Cytotoxic and anti-proliferative activities were measured in the four transformed human cell lines of cervical carcinoma (HeLa), lung adenocarcinoma (A-549), colon adenocarcinoma (HT-29) and endometrial adenocarcinoma (CRL-2923) and normal cell line of retina pigment (ARPE) were investigated. Anti-inflammatory properties were measured by their anti-human testicular hyaluronidase and urease of Helicobacter pylori. The results were showed that chestnut originated propolis samples were apitherapeutic potentials.

Acknowledgment: This study was supported by Tubitak 114Z370 project.


SL 6

Plant Origin Authentication of Sonoran Propolis and its antiproliferative effect on Cancer Cells: a Bioactive Poplar type Propolis from Semi-arid zones Efrain Alday1, Lucía Uribe1, Dora Valencia2, Adriana Garibay-Escobar1, Anna Lisa Piccinelli3, Luca Rastrelli3, Humberto Astiazarán-García4, Ramón Enrique Robles-Zepeda1, Javier Hernandez5,* and Carlos Velazquez1,*1 Department of Chemistry-Biology, University of Sonora, Blvd. Luis Encinas y Rosales s/n, 83000 Hermosillo, Son., Mexico.2 Department of Chemical Biological and Agropecuary Sciences, University of Sonora, Av. Universidad e Irigoyen, 83600 Caborca, Son., Mexico.3 Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, SA, Italy.4 Centro de Investigación en Alimentación y Desarrollo, CIAD, A.C., Carretera a La Victoria km 0.6, 83000, Hermosillo, Son., México.5 Unidad de Servicios de Apoyo en Resolución Analítica, Universidad Veracruzana, 575 Xalapa, Ver., Mexico. The main chemical composition of Sonoran propolis (SP), as well as its antiproliferative activity on cancer cells via apoptosis induction has been reported1. Additionally, the chemical constitution of SP remained qualitatively similar throughout the year, whereas the antiproliferative effect exhibited significant differences amongst seasonal samples2. The goal of this study was to authenticate the botanical source of SP bioactivity by using an approach based on a chemical comparative analysis, antiproliferative activity and cell cycle progression analysis on cancer cell lines. The polyphenolic profile of SP throughout the year resulted to be qualitatively similar to that of Populus fremontii resins (PFR). However, the antiproliferative activity of PFR did not consistently match that exhibited by SP. In addition, SP induced evident morphological modifications (elongation) on treated cells, different to those induced by PFR. Most of cancer cells treated with SP were arrested in G2/M checkpoint (M12.C3.F6: 94.8±1.7 %, and HeLa cells: 70.8±5.7 %), similarly as colchicine did (control drug; 2 µM; 94.8±2.3 % and 72.7±6.4 %, respectively). In contrast, PFR treatment increased cell population at G0/G1 on both M12.C3.F6 (56.9±5.0 %) and HeLa cells (70.3±0.3 %), in comparison with dissolvent control. Interestingly, Ambrosia confertiflora resins induced morphological elongation and cell cycle arrest at G2/M on both M12.C3.F6 and HeLa cells, similarly as SP did. These results suggest that P. fremontii is the main plant origin of SP, nevertheless, A. confertiflora resins participate as a complementary source that enhances its bioactivity. Therefore, SP is a poplar-type propolis from subtropical semi-arid zones. References:1 Alday E, Valencia D, Carreсo AL, Picerno P, Piccinelli AL, Rastrelli L, Robles-Zepeda R, Hernandez J, Velazquez C (2015) Apoptotic induction by pinobanksin and some of its ester derivatives from Sonoran propolis in a B-cell lymphoma cell line. Chemico-Biological Interactions 242: 35–44. 2 Valencia D, Alday E, Robles-Zepeda R, Garibay-Escobar A, Galvez-Ruiz JC, Salas-Reyes M, Jimйnez-Estrada M, Velazquez-Contreras E, Hernandez J, Velazquez C (2012). Seasonal effect on chemical composition and biological activities of Sonoran propolis. Food Chemistry 131: 645–651.


SL 7

Chemical Composition of extracts of geopropolis from Melipona quadrifasciata (Mandaçaia) evaluation of its antioxidant activity

M. B. Gabriel1, M. J.Carneiro1, A. C. H. F. Sawaya2

1 Institute of Biology, State University of Campinas, 13083-970 Campinas - SP, Brazil.2 Faculty of Pharmaceutical Science, UNICAMP, SP, Brazil, [email protected].

Currently, a group of bees called stingless bees, native bees or indigenous bees have attracted interest. These social bees belong to the Apidae family and are divided in three different genera: Melipona, Lestrimelitta and Trigona. These bees have only residual stingers, usually small wings and nests of distinct characteristics that differentiate species 1.These bees are of great importance in the pollination of many cultivable species, being responsible for up to 90% of the pollination of Brazilian native flora. Melipona quadrifasciata produces geopropolis, honey and wax. Geopropolis is the result of the bees mixing the resinous material collected from the plants with wax and soil. Studies of the ethanolic extract of Melipona quadrifasciata geopropolis seek to determine the chemical composition via ESI-MS fingerprinting, as well as their antioxidant activity. Samples collected over one year in Jarguariuna, SP showed a similar chemical profile. Their antioxidant activity was evaluated by the DPPH method and presented similar activity (EC 50 between 1 and 4 µg/mL) of ethanolic solution. These results are comparable propolis of bee species Apis mellifera 2. As these samples were collected over one year, it will be possible to determine if seasonality influences their chemical composition and biological activity.

References1 Cardozo, D. V., Mokochinski, J. B., Machado, C. S., Sawaya, A. C. H. F., Caetano, I. K., Felsner, M. L., & Torres, Y. R. (2015). Variabilidade química de geoprópolis produzida pelas abelhas sem ferrão Jataí, Mandaçaia e Mandurí. Revista Virtual de Química, 7(6), 2456-2474.2 Lima, M. V. D. De. (2015). Geoprópolis produzida por diferentes espécies de abelhas: atividades antimicrobiana e antioxidante e determinação do teor de compostos fenólicos. Dissertação (Mestrado) - Universidade Federal do Pará, Instituto de Ciências da Saúde, Belém. Programa de Pós-Graduação em Ciências Farmacêuticas.


SL 8

Characterized Propolis extracts, Obtained with Standardized extraction method, Show Similar Chemical Profile (HPlC-eSi.mSN) and in vitro Antibacterial, Antioxidant and Anti-inflammatory activity through evaluation of expression of mirnaS, mrnaS and Proteins.

Zaccaria Vincenzo1,2, Galeotti Fabio2,3, Fachini Alfredo2, Passarella Paolo2, Daglia Maria1, Volpi Nicola 3

1 Department of Drug Sciences, Medicinal Chemistry and Pharmaceutical Technology, University of Pavia, Pavia, Italy. 2 B Natural R&D Unit, Milano, Italy.3 Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.

Despite the great number of investigations, the common scientific approaches to study biological activities of propolis present some limitations due to the high natural variability of propolis and different extraction methods used. Therefore, the results obtained so far are often not comparable each other and are poorly reproducible. The aim of this work is the development of a new extraction method to obtain standardized propolis extracts to be studied in vitro for the determination of anti-inflammatory, antioxidant and antibacterial activities.The standardized extraction method was set and posited as patent (Multi Dynamic Extraction method M.E.D.®) and the extracts obtained were characterized; they shown similar chemical profiles (HPLC-ESI-MSn) even if formulated differently (dry, glyceric, glycolic, hydroalcoholic and oily extracts)1. The antioxidant activity of formulated extracts was firstly determined in vitro (TROLOX) and then, to better clarify the intracellular mechanisms behind the antioxidant and anti-inflammatory activities, miRNA, mRNA (RT-qPCR) and their protein validated target (ELISA) changes were evaluated2. Propolis M.E.D.® extracts showed similar antioxidant TROLOX values related to the amount of polyphenols; moreover, they are able to reduce the oxidative stress and inflammation level in HaCat cells acting on expression levels of mRNAs coding for NFE2L2, GPX2 and TNF-α and NFE2L2 protein2. These results highlight a possible molecular mechanism of action of Propolis M.E.D.® behind the antioxidant and anti-inflammatory activity. To test antibacterial activity, different propolis M.E.D.® extracts were tested (MIC) against several strains (ATCC, antibiotics resistant and susceptible) and showed comparable MIC values related to the amount of standardized polyphenols complex.


References: 1 Galeotti F, Maccari F, Fachini A, Volpi N. (2018) Chemical Composition and Antioxidant Activity of Propolis Prepared in Different Forms and in Different Solvents Useful for Finished Products. Foods. 7(3). 2 Zaccaria V, Curti V, Di Lorenzo A, Baldi A, Maccario C, Sommatis S, Mocchi R, Daglia M. (2017) Effect of Green and Brown Propolis Extracts on the Expression Levels of microRNAs, mRNAs and Proteins, Related to Oxidative Stress and Inflammation. Nutrients 9(10).


SL 9

new antiproliferative acyl glycerols from new zealand Propolis and Source Poplar resin

Stephen Bloor1, Owen Catchpole,1,2, Kevin Mitchell1, Rosemary Webby1.

1Callaghan Innovation, 69 Gracefield Road, PO Box 31310, Lower Hutt 5040, New Zealand2Manuka Health NZ Ltd, PO Box 87429, Meadowbank, Auckland 1742, New Zealand.

Previous work has shown a number of phenolic components of NZ propolis demonstrate antiproliferative activity against certain human gastrointestinal cancer cell lines1. Here we report on a series of acyl glycerols isolated from the non-polar fraction of propolis resin, which represent further bioactive constituents unrelated to the more usual phenolic compounds generally found in propolis. NZ propolis is sourced from poplar trees and the acyl glycerols have been shown to be present in the leaves and buds of some common poplars. The compounds are a series of monoglycerides containing 3,8-dihydroxy fatty acids, many of which are further acylated with acetic acid residues. The dihydroxy fatty acids are C18 to C24, with the most abundant being C20 and C22. These acyl glycerols were found to have strong antiproliferative activity against three human gastro-intestinal cell lines, particularly gastric cancer cell line NCI-N87.

O

O

OH

OH

(CH2)nCH33R

8ROHOR

Acyl glycerols 1 (R = H, n = 10), 2 (R = Ac, n = 10), 3 (R = Ac, n = 11) and 4 (R = Ac, n = 12).

References:1Catchpole, O., Mitchell, K., Bloor, S., Davis, P., Suddes, A. (2015). Antiproliferative activity of New Zealand propolis and phenolic compounds vs human colorectal adenocarcinoma cells. Fitoterapia. 106, 3257: 167-174


SL 10

the efficacy of Propolis fluoride in inhibiting dental Caries activity on Primary teeth Risqa Rina Darwita1, Iwany Amalliah1, Sri Angky Soekanto2, Muhamad Sahlan3

1Department of Dental Public Health and Prevention, Faculty of Dentistry, Universitas Indonesia, Jakarta 10430 Indonesia, [email protected] of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta 10430 Indonesia3Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Depok Indonesia

Early childhood caries which is a public health problem is quite large in Indonesia. Therefore, it is necessary to take care of early childhood caries. The Propolis Fluoride is an option to inhibit email or dentinal caries activity. Objective. To determine the efficacy of propolis fluoride in inhibiting email or dentinal caries activity. Method. The respondents were 246 children aged 36-71 month were applied Propolis Fluoride on dental caries surface that has active email or dentinal caries. The respondents were divided into two groups, consists of 149 children were intervention group, they have been treated by propolis fluoride, toothbrushing program and they got a routine Dental Health Education. While, 97 children were as a control group have been treated by propolis fluoride, they had been given Dental Health Education only on the baseline. The evaluation and examination were conducted after 3 months to measure the percentage of arrested email and dentinal caries, and the correlation with plaque index. Result. After 3 months evaluation, the percentage of arrested dental caries for intervention and control groups were 62,44% and 46,18% respectively. There was a significant difference in mean plaque index and the number of active dental caries between two groups. There was a significant correlation between the plaque index and the percentage of arrested dental caries. There was a significant correlation between the behavior score and the percentage of arrested caries. Both correlation has negative and positive value respectively. Conclusion. The efficacy of Propolis fluoride in inhibiting dental caries activity.

Acknowledgements: This study was granted from Ministry of Research & Education Republic of IndonesiaReferences:1 Darwita, R. R., Maharani, D. A., Rahardjo, A., Puspa, D. P., Amalia, R., & Sandy, D. P. (2011). Is joint tooth brushing an effective program for improving dental health among elementary students?: a study from Jakarta, Indonesia. Int J Clin Prev Dent, 7, 149-53.Availablefrom:https://www.researchgate.net/publication/291334634_2 Elham B, Abolghasem H, Tayebeh M. Oral Health Behavior of Parents as a Predictor of Oral Health Status of Their Children. ISRN Dent. 2013;2013. 3 Monse B, Benzian H, Holmgren C, Van Palenstein, Helderman W H-WR. A Silent Public Health Crisis: Untreated Caries and Dental Infections Among 6 and 12 years old Children in Philippine National Oral Health Survey. Asian Pasific J Public Heal. 2016; 4 Sahlan M, Ramadhan. Permen Hard Candy Propolis untuk Kesehatan Gigi Manusia. Depok Univ Indones. 2011;(Teknik Kimia).


SL 11

An Overview of Chemical Studies and Biological activities of Mediterranean Propolis

Konstantia Graikou1, Ioanna Chinou1

1Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece

Knowledge of propolis composition, which depends on the geographical and climatic origin, is extremely valuable with respect to the problem of its standardization. Chemical composition, antioxidant and antimicrobial activity of forty five (45) selected Mediterranean propolis extracts from mainland Greece (22), Greek islands (15), Cyprus (5), Croatia (2), and Algeria (1) were determined. All previously isolated metabolites from propolis by our team, together with their full structural determination through modern spectral means, were used as internal standards in our further GC-MS analyses. This method is proposed as an essential tool in the pursuit of a fast and cheap analytical method of propolis characterization (1, 2).More than 300 constituents have been identified through GC-MS concluded that the Greek propolis samples share characteristics that differentiate them from typical European ones, mainly the presence of diterpenes in significant amounts and the relatively low quantity of phenolic acid esters. Mediterranean propolis could be confirmed as a new type of propolis consisting mainly from diterpenes and produced based to Conifer trees among Cupressaceae and Pinaceae, which are widespread in Mediterranean’s area Flora (3). All studied samples showed a very interesting antimicrobial activity against a panel og eleven human pathogenic microorganisms, probably due to their high terpenic content. Moreover they appeared as rich sources of natural antioxidants which can possibly be exploited as protective agents against various free radical related degenerative diseases. In conclusion, Mediterranean propolis to our view is an adequately defined new propolis type, which can be added to the well accepted existing classes.

Acknowledgments: The author would like to thank the Special Account for Research Grants and the National and Kapodistrian University of Athens for funding their participation in this meeting

References:1 Papachroni D., Graikou K., Kosalec I., Damianakos H., Ingram V., Chinou I. (2015). Phytochemical analysis and biological evaluation of selected African propolis samples from Cameroon and Congo. Natural Product Communication, 10: 67-70.2 Popova M.P., Graikou K., Chinou I., Bankova V.S. (2010). GC-MS profiling of diterpene compounds in mediterranean propolis from Greece. Journal of Agricultural & Food Chemistry, 58: 3167-3176.3 Graikou K., Popova M., Gortzi O., Bankova V., Chinou I. (2016). Characterization and biological evaluation of selected Mediterranean propolis samples. Is it a new type? LWT-Food Science and Technology, 65: 261-267.

http://www.ncbi.nlm.nih.gov/pubmed/?term=Papachroni D%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/?term=Graikou K%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/?term=Kosalec I%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/?term=Damianakos H%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/?term=Ingram V%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/?term=Chinou I%5BAuthor%5D&cauthor=true&cauthor_uid=25920222

http://www.ncbi.nlm.nih.gov/pubmed/25920222

http://www.sciencedirect.com/science?_ob=RedirectURL&_method=outwardLink&_partnerName=27983&_origin=article&_zone=art_page&_linkType=scopusAuthorDocuments&_targetURL=http%3A%2F%2Fwww.scopus.com%2Fscopus%2Finward%2Fauthor.url%3FpartnerID%3D10%26rel%3D3.0.0%26sortField%3Dcited%26sortOrder%3Dasc%26author%3DChinou,%2520Ioanna%2520B.%26authorID%3D7004443591%26md5%3D9a148bbbc8623de0f2331419b267c91d&_acct=C000059642&_version=1&_userid=275166&md5=f3d5b74896e59a6c66b63400553b56b8

http://www.sciencedirect.com/science?_ob=RedirectURL&_method=outwardLink&_partnerName=27983&_origin=article&_zone=art_page&_linkType=scopusAuthorDocuments&_targetURL=http%3A%2F%2Fwww.scopus.com%2Fscopus%2Finward%2Fauthor.url%3FpartnerID%3D10%26rel%3D3.0.0%26sortField%3Dcited%26sortOrder%3Dasc%26author%3DBankova,%2520Vassya%2520S.%26authorID%3D35479237300%26md5%3D7cacbbfec32fa8e4ac62ab4d56bddbe2&_acct=C000059642&_version=1&_userid=275166&md5=a74c8ff5ea10c7239fb6164bc70c419e


SL 12

a Collaborative Study for Performance evaluation of analytical Methods for Propolis. an iHC trial

M. Lopes1, A. Pereyra2, C. Kunert3, G. Beckh4, H. Schreiter5, O. G. Çelemli6, K. Sorkun6, S. Georgé7, L. Paulo8, S. Gardini9, M.T. Sancho10, V. Bankova11, T. Dastan12, C. Tananaki13, L.F. Nunes1, M. Vilas-Boas1

1 Mountain Research Centre (CIMO), Polytechnic Institute of Bragança, Bragança, Portugal2 Medex, Ljubljana, Slovenia3 Intertek Food Services GmbH, Bremen, Germany4 Quality Services International GmbH, Bremen, Germany5 Allwex Food Trading GmbH, Bremen, Germany6 Hacettepe University, Ankara, Turkey7 Centre technique agroalimentaire - CTCPA, Avignon, France8 Associação Centro de Apoio Tecnológico Agro-Alimentar de Castelo Branco (CATAA), Castelo Branco, Portugal9 Council for Agricultural Research and Economics, (CREA-AA), Bologna, Italy10 Universidad de Burgos, Burgos, Spain11 Institute of Organic Chemistry with Centre of Phytochemistry, Sofia, Bulgaria12 Altiparmak, Istanbul, Turkey13 Apiculture-Sericulture Lab, University of Thessaloniki, Thessaloniki, Greece

Propolis is well known for its potential applications but also for its chemical complexity, making it very hard to define quality standards for its commercialization. Every day it is possible to find new publications on this bee product describing a new singularity, a particular substance with high bioactivity never described or a new application. This systematic increment in scientific information, although relevant, do not always enable a clear comparison between samples due to the different methods and procedures used to assess its quality parameters. To strength the scientific studies on propolis and establish a background for future definition of standards for industry, producers and laboratories, the Propolis Working group of the International Honey Commission, performed an inter-laboratory study to harmonize basic analytical methods and evaluate their accuracy and robustness.

The collaborative study was accomplished by 12 laboratories from 9 countries and a company experienced in propolis trade. The samples under analysis, representing fifteen different origins around the globe, where fractionated and distributed to each laboratory, and so all participants analysed the same material. Overall, six parameters were evaluated: ashes, wax and balsamic extract on raw propolis, and total-phenolics, flavones and flavanones on propolis extract. The results were analysed following the international guidelines ISO 57251. Outliers were removed using Cochran’s test, to check the homogeneity of variances at certain levels, and Grubb test’s, to check the consistency of the laboratories average. The results revealed good reproducibility for ash, wax, balsamic content and flavones, while for flavanones the method is clearly unsuitable.


Acknowledgements: Project PDR2020-1.0.1-FEADER-031734: “Diversification and Innovation on Beekeeping Production”

References:1 ISO 5725, 1994. Accuracy (Trueness and Precision) of Measurement Methods and Results.


SL 13

molecular Characterization of Propolis and royal Jelly Components as tool for Study of their Multifunctional therapeutic effects

Bíliková Katarína1, Yamaguchi Yoshihisa2

1 Institue of Forest Ecology, Slovak Academy of Sciences, Zvolen, Laboratory of Molecular Apidology, Bratislava, Slovakia

2 Japan Royal Jelly Institute of Apimedical Science, Tokyo, Japan

Honeybee products, mainly propolis and royal jelly are becoming a basis for development of multipurpose multifunctional drugs, which means increased efficacy compared to previous high-specific synthetic drugs used in human medicine. High efficacy drugs are effective only for a small part of population; they have many uncontrollable side effects and can only be used in limited time. Honeybee products, mainly propolis, offer a variable composition of physiologically active compounds of honeybee origin and secondary metabolites of plants that can exhibit together interesting healing properties as results of synergic effect of all components. The aim of the work is targeted characterization of chemically defined compounds of propolis both honeybee and floral origin and study of their physiological properties separately and in complex. It was found that the water-soluble components of ethanol propolis extract inhibit the transcription of DNA-dependent RNA polymerase by eliminating its binding to the DNA promoter (1). Testing of antimicrobial activity of pinobanksin-3-O-acetate and pinocembrin purified from poplar propolis showed growth inhibition of Paenibacillus larvae strains, the major causative agent of American foulbrood of honey bee larvae disease (2). We present here pharmacological effects of honeybee royal jelly on the nervous system enhanced by flavonoid Nobiletin (3), applicable for prevention and/or delay of neurodegeneration in brains of patients with Alzheimer’s disease. Results are contribution to the practical usage of propolis and royal jelly in human medicine as well as in prevention of honeybee colonies against microbial diseases.

Acknowledgements: This work was supported by JRJ Co., Ltd., Tokyo, Japan

References: 1 Simúth J, Trnovský J, Jeloková J (1986) Inhibition of bacterial DNA-dependent RNA polymerases and restriction endonuclease by UV-absorbing components from propolis. Pharmazie 41(2): 131-1322 Bilikova K, Popova M, Trusheva B, Bankova V (2013) New anti-Paenibacillus larvae substances purified from propolis. Apidologie 44 (3): 278-2853 Fujiwara H, Kogure A, Sakamoto M, Yamakuni T, Mimaki Y, Murata M, Hitomi N, Yamaguchi K, Ohizumi Y (2011) Honeybee Royal Jelly and Nobiletin Stimulate CRE-Mediated Transcription in ERK-Independent and -Dependent Fashions, Respectively, in PC12D Cells. J. Pharmacol Sci 116: 384 - 387

https://www.ncbi.nlm.nih.gov/pubmed/3014571


SL 14

Can Propolis inhibit infection Dynamics of Honey Bee (Apis mellifera l.) viruses in vitro?

Michael Goblirsch1

1 Honey Bee Research Laboratory, Department of Entomology, University of Minnesota, St. Paul, MN USA

Honey bees collect resins from plants and use this material, called propolis, to line their nest cavity. Propolis is a complex mixture of plant secondary metabolites (e.g., terpenes), beeswax, and other compounds, and has been shown to inhibit the growth of pathogenic microbes in vitro, as well as reduce the severity of disease caused by bacterial and fungal honey bee pathogens in the colony setting. Since honey bees from colonies that are challenged by pathogens such as the fungus, Ascosphaera apis (chalkbrood), or the bacterium, Paenibacillus larvae (American foulbrood), actively forage for resin, this behavior parallels those displayed by other organisms that self-medicate in response to disease. Unknown, however, is whether propolis has activity against the numerous viruses that are both abundant and prevalent in honey bee colonies. Most viruses known to cause disease in honey bees are (+)ssRNA viruses. These viruses can persist at high titers without presenting overt symptoms until the host is perturbed by some form of stress, such as pesticide or parasitic mite exposure. Although some evidence suggests propolis has activity against viruses that cause disease in humans, much remains unknown as to whether it can inhibit dynamics of honey bee viruses. The research presented here is a step towards determining whether propolis has activity against honey bee viruses in vitro. Preliminary results are from studies using an in vitro model established from the honey bee embryonic cell line, AmE-711.


SL 15

Study of relationship between geographical location of Collection, Chemical Composition and Biological activity of Propolis by Multivariate Data analysis

Shankar Katekhaye1, Hugo Fearnley2,3, James Fearnley2,3, Anant Paradkar1

1 Centre for Pharmaceutical Engineering Sciences, University of Bradford, Bradford, BD7 1DP, UK2 Nature’s Laboratory, Unit 3b, Enterprise Way, Whitby, North Yorkshire, YO22 4NH, UK3 Apiceutical Research Centre, Unit 3b, Enterprise Way, Whitby, North Yorkshire, YO22 4NH, UK

Propolis contains resinous material collected by honeybees from plants, and has a plethora of biological activities. The geographical location of propolis collection influences its chemical composition, resulting in variations in biological activity. The present study was designed to analyse the relationship of propolis to geographic location, chemical composition and biological activity. We have applied multiple regression analysis using SPSS to examine selected flavonoids and phenolics for their individual concentration as well as total content and biological activities i.e. anti-microbial activity against Staphylococcus aureus and anti-oxidant activity by DPPH assay. Three climatic zones have been classified- temperate, subtropical and tropical. This meta-data analysis revealed some interesting facts about the distribution of specific chemicals (CAPE, pinocembrin, pinobanksin, galangin, naringenin, chrysin, quercetin, apigenin, cinnamic acid, coumaric acid, caffeic acid and ferulic acid), as well as, total phenolic and flavanoid content and biological activity. We observed that, the phenolics have stronger anti-microbial activity as compared to flavonoids and vica-verse for anti-oxidant activity. We also noticed that, total phenolic content is often around 1.5 times higher than flavonoid content in almost all climatic zones. Higher levels of cinnamates were observed in propolis from tropical regions which had better anti-microbial potential, whereas, temperate propolis with higher levels of flavonoids exhibited better anti-oxidant potential.

Acknowledgement: We acknowledge the financial support from Innovate UK and Nature’s Laboratory Ltd. (KTP010490). We also acknowledge the financial support provided by Winston Churchill Memorial Trust (2016-17).


SL 16

interaction of artepillin C with Model Membranes: Optical Absorption and fluorescence Spectroscopy Studies

Isamara Julia Camuri1, Adriano Batista Costa1, Wallance Moreira Pazin2, Amando Siuiti Ito1

1 Department of Physics, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto – SP, Brazil;2 Department of Physics, School of Sciences and Technology, São Paulo State University, Presidente Prudente – SP, Brazil

Brazilian green propolis, collected by the species Apis mellifera, presents antioxidant, antimicrobial, anti-inflammatory and antitumor activities. The major component of green propolis is Artepillin C, a derivative of cinnamic acid with two prenylated groups, that improves the affinity of the compound for lipophilic environment. This may be relevant to the biological activity of green propolis in the cellular environment, in view of the initial contact of the product with the cell membrane. We used optical absorption and fluorescence techniques to study physico-chemical properties of the interaction between Artepillin C and amphiphilic aggregates commonly used as membrane models, namely, micelles and unilamellar vesicles. We examined the interaction of Artepillin C with anionic, cationic and zwitterionic micelles, and with large unilamellar vesicles of neutral and negatively charged phospholipids and positively charged lipid. Optical absorption spectra showed that the protonation state of Artepillin C is dictated by the local pH in the surface of micelles and vesicles. Fluorescence results showed that the polarity around Artepillin C decreased in the presence of micelles and vesicles, and the molecule should be located on the surface region of the model membranes. The negative charge of the compound in deprotonated state favors the interaction with cationic micelles and neutral vesicles. The effects are more prominent when the lipid micelles are in the fluid phase.

Acknowledgments: CAPES, CNPq (305771/2016-7) and FAPESP (2016/09633-4)

References:1 Pazin, W. M. et al. (2017) Interaction of Artepillin C with model membranes. Eur. Biophys. J. 46: 383–393.2 Pazin, W.M. et al (2017) Antioxidant activities of three honeybee stingless propolis and green propolis. Journal of Apicultural Research, 56: 40-49.3 Camuri, I.J. et al (2018) Optical absorption and fluorescence spectroscopy studies of Artepillin C, the major component of green propolis. Spectroch. Acta A 198: 71-77.


SL 17

a new approach to Propolis extraction

Şaban Keskin1, Merve Keskin2, Sevgi Kolaylı3

1 Bilecik Şeyh Edebali University, Faculty of Arts and Science, Department of Chemistry, Bilecik, Turkey.2 Karadeniz Technical University, Institution of Natural Science, Department of Chemistry, Trabzon, Turkey.3 Karadeniz Technical university, Faculty of Science, Department of Chemistry, Trabzon, Turkey.

Propolis, a resinous mixture collected by honey bees, contains many biologically active compounds like phenolic acids, flavonoids and terpenes. Consumption of raw propolis is limited due to its resinous nature. Propolis should be extracted to convert it into consumable form. Although ethanol is reported to be the best solvent for propolis extraction nevertheless ethanol is another limiting factor for propolis consumption. There is an increasing need for new, compatible and healthier solvents for propolis extraction. Volatile oils obtained from different origins contain different compounds and they could be considered as natural green solvent. To the best of our knowledge, this is the first paper describing the extraction of propolis by using volatile oils. Commercially obtained lemon peel oil, peppermint oil and thyme oil extracted with hydro distillation technique were used in this study. For propolis extraction 1:10 (g/v) ratio was used. Chemical composition of volatile oils and volatile oil propolis extracts was separately analyzed by using GC-MS technique. Total polyphenol content and antioxidant capacity of oils and propolis extracts were also determined. It was found that volatile oil propolis extracts were rich in volatile compounds of propolis, phenolic acid derivatives and free fatty acids. It can be concluded that volatile oils could be used for propolis extraction separately or in combination. They could also be used for preparing health promoting products containing propolis.


SL 18

Portuguese Propolis: a Source of valuable Bioactivities

Cunha A1,2,3,4, Alves H1, Araújo C1, Cruz M1, Freitas AS1,2, Gomes AJ1, Gonçalves R1, Marques R1, Moreira B1, Passão C1, Peixoto M1, Pereira H1, Silva-Carvalho R1,5, Valença I1, Ferreira AM6, Baltazar F5, Pinto-Ribeiro F5, Cardoso S7, Oliveira R1,2,3,4, Almeida-Aguiar C1,2,3,4

1 Biology Department, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal3 CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.4 CBMA - Centre of Molecular and Environmental Biology, University of Minho, 4710-057 Braga, Portugal.5 Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 6 Trás-os-Montes e Alto Douro University - 5001-801 Vila Real, Portugal7 QOPNA, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal. Propolis is a complex resinous mixture produced by honeybees and used in the construction, repair and defence of hives. Humans have used propolis in traditional and alternative medicines since ancient times. Subject of intense research, propolis showed to possess many biological and pharmacological properties, namely antimicrobial and antioxidant1. Although clinical studies are still lacking, the broad bioactivities spectrum of propolis, the continuous discovery of new compounds, its long history of use and safety profile suggest promising applications in medicine and pharmacology. The increasing public interest towards natural pharmacological compounds, along with propolis wide antimicrobial activity and synergy with antibiotics, give additional stimulus to propolis research.Portuguese propolis has deserved little attention by the scientific community2,3 making fundamental to study its chemical fingerprints and biological properties. In this work, we present some of the results obtained for Portuguese propolis showing that it is likely to be an important source of valuable bioactivities: it confers antioxidant protection, has antigenotoxic effects, displays broad antimicrobial activity, is cytotoxic against some human carcinoma cells, has anti-inflammatory potential and exhibits phytotoxicity against in vitro flax plants. Such results not only confirmed some widely known propolis activities but also suggested new ones for Portuguese propolis, envisaging potential for several clinical, industrial and environmental applications as well as for its valorisation.

Acknowledgments: To FEDER/COMPETE/POCI– Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013.


References: 1 Silva-Carvalho R, Baltazar F, Almeida-Aguiar C (2015) Evidence-Based Complementary and Alternative Medicine 20152 Falcão SI, Vale N, Gomes P, Domingues MR, Freire C, Cardoso SM, Vilas‐Boas M (2012) Phytochemical Analysis 24(4), 309-318


SL 19

anticancer Properties of aqueous and nonaqueous Propolis extracts

Zbigniev Balion1, Aistė Jekabsone2,3, Kristina Ramanauskiene4, Daiva Majienė1,5

1 Laboratory of Biochemistry, Institute of Neuroscience, Lithuanian University of Health Sciences, Kaunas, Lithuania.2 Laboratory of Molecular Neurobiology, Institute of Neuroscience, Lithuanian University of Health Sciences, Kaunas, Lithuania.3 Laboratory of Pharmaceutical Sciences, Institute of Pharmaceutical Technologies, Lithuanian University of Health Sciences, Kaunas, Lithuania.4 Department of Clinical Pharmacy, Lithuanian University of Health Sciences, Kaunas, Lithuania.5 Department of Drug technology and Social Pharmacy, Lithuanian University of Health Sciences, Kaunas, Lithuania.

Glioblastomas are characterized by the highest degree of invasion and the worst survival prediction because of the lack of efficient therapy that would not be harmful for sensitive and vitally-important surrounding brain tissue. Propolis is a multicomponent substance known for numerous biological effects including antioxidant-related stimulation of immune system and tissue regeneration as well as anticancer activity. The aim of this study was to investigate the effects on viability of glioblastoma (C6 rat glioma cell line) and healthy brain cells (primary cerebellar granule cells) of aqueous (AqEP), polyethylene glycol-aqueous (Pg-AqEP) and ethanolic (EEP) propolis extracts. After 24h treatment, all the extracts tested decreased C6 cell viability in a concentration-dependent manner. Statistically significant increase in the number of dead cells was achieved at the following concentrations of phenolic compounds: 15-18 µg/ml for AqEP, 10 µg/ml for Pg-AqEP and 5 - 7 µg/ml EEP. We have also tested how same propolis extracts influence viability of primary cerebellar granule cell cultures comprised of 76±3 % neurons, 16±2 % astrocytes and 8±2% microglia and found no decrease in cell viability after 24h treatment.

In conclusion, propolis extracts exert selective cytotoxicity against C6 cells while preserving healthy brain cells and could be recommended as adjuvant of glioblastoma therapy as well as safe food supplements for cancer prevention.


SL 20

antimicrobial effect of Commercial Propolis extract (BeeO©)

Aslı Elif Tanuğur1, Sevgi Kolayli2, Merve Keskin2, Şengül Alpay Karaoğlu3

1SBS Scientific Bio Solutions Inc., Istanbul, Turkey2Karadeniz Technical University, Department of Chemistry, Trabzon, Turkey3Recep Tayyip Erdoğan University, Department of Biology, Rize, Turkey

In this study, antimicrobial activities of 7 different commercial bee products formulations of (BEE’O)© consisting of raw honey, propolis and Royal jelly were investigated in terms of antimicrobial properties. A wide screening antimicrobial tests againts Escherichia coli, Yersinia pseudo tuberculosis, Helicobacter pylori, Klebsiella pneumonias subp. pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutants, Streptococcus agalactiae, Bacillus cereus, Lactobacillus acidophilus, Lactobacillus casei, Mycobacterium smegmatis, Candida albicans, Candida tropicalis and Saccharomyces cerevisiae were tested. Agar well dilution and broth-dilution methods were used to measure the activities. The results confirm that all formulations have substantial antimicrobial potentials, and but the pure ethanolic propolis extracts were showed higher antimicrobial potentials than the mixtures. In conclusion, propolis and their mixtures are good antimicrobial agents and they can be used as an alternative to synthetic drugs as a preservative and complementary medicine.


SL 21

new insights into tropical Propolis: Propolis from Pitcairn island

Boryana Trusheva1, Kristina Georgieva1, Milena Popova1, Veselina Uzunova2, Tihomira Stoyanova2, Violeta Valcheva3, 4, Rumiana Tzoneva2, Vassya Bankova1

1Institute of Organic Chemistry with Centre of Phytochemistry, BAS, 1113 Sofia, Bulgaria2Institute of Biophysics and Biomedical Engineering, BAS, 1113 Sofia, Bulgaria3The Stephan Angeloff Institute of Microbiology, BAS, 1113 Sofia, Bulgaria4Laboratory Complex, Sofia Tech Park, 1784 Sofia, Bulgaria

Propolis (bee glue) has a long history of being used in traditional medicine and nowadays it is extensively used in food diet to improve health and prevent diseases such as inflammation, heart disease, diabetes, and even cancer. Because of its broad spectrum of biological activities there is an undying interest in the composition of propolis, which depends on the vegetation of the area from which propolis was collected. There are numerous reports in the literature on the isolation and structural elucidation of biologically active phytochemicals from propolis collected in Europe, South America, Asia and the Pacific region. However there is no data about chemical composition and biological activity of propolis from Pitcairn Islands1. The chemical profiles of Pitcairn propolis extracts were obtained by GC-MS analysis after silylation. The major constituents were terpenoids (mainly diterpenes and less triterpenes). Phenolic components (cardanols, alk(en)ylresorcinols and anacardic acids) are present in significantly lower amounts, as flavonoids and the usually present phenolic acids are completely absent. After detailed chemical investigation of propolis’ dichloromethane extract, four new cycloartane triterpenes with good antimicrobial activity, along with 18 known compounds, were isolated. The biological activity of this extract was also examined and the results showed that it inhibits human breast cancer MDA-MB-231 cells proliferation and induces changes of malignant cells’ morphology, which hint to apoptotic events. Apart from this the studied propolis extract possesses strong antimicrobial activity against some of the most common pathogens causing infections in humans: Staphylococcus aureus, Escherichia coli, and Candida albicans. Thus, the present research provides additional data for the tropical propolis constituents, revealing the distinct character of Pitcairn propolis. Furthermore these findings once again certify the potential role of propolis as a natural chemopreventive agent and a valuable bioactive mixture for a combination therapy approach.

Acknowledgements: The Pitcairn Island Producers’ Co-operative (PIPCO) for providing the sample.

References:1 Georgieva K, Trusheva B, Uzunova V, Stoyanova T, Valcheva V, Popova M, Tzoneva R, Bankova V (2018) New cycloartane triterpenes from bioactive extract of propolis from Pitcairn Island. Fitoterapia 128: 233–241.


SL 22

toothpaste with Propolis “apident” Shows antimicrobial activity in vitro Klemen Rihar1, Dunja Gregorič Exel 2, Adriana Pereyra3, Rok Kopinč3, Bratko Filipič4

1Chengdujska 4, 1000 Ljubljana, Slovenia; 2Glavarjeva 45, 1000 Ljubljana, Slovenia; 3 MEDEX d.o.o., Linhartova 049a, 1000 Ljubljana, Slovenia; 4CIETO, Koledinećka 3, 10040 Zagreb, Croatia

In this study, we evaluated the antibacterial activity of toothpaste with Propolis, used for the treatment of teeth and as a medicine for endodontic treatment. Toothpaste with Propolis was prepared using the water-soluble Propolis extract. Calcium Hydroxide was a control, as well as two commercial and two medical toothpastes. The bacteria on which Propolis toothpaste works are Streptococcus mutans, Staphylococcus aureus, Micrococcus luteus, Escherichia coli, Pseudomonas aeruginosa and yeast Candida albicans. The inhibition of bacterial/yeast growth was evaluated by colouring with resazurine. The inhibition of bacterial/yeast growth is apparent in changing the color of resazurine from blue to pink. MIC (Minimum Inhibitory Concentration) was determined as the maximum dilution in which no minimal color change occurs or a minimal color change occurs. For example, in the seventh hole, the color from pink to blue has changed. Therefore, the color change in the seventh hole at dilution 1:64 means MIC = 1 × 10/64 = 0.156 mg/mL. MIC values of toothpaste with Propolis were against S. aureus: 0.031 mg/mL (0.326 mg/mL of medical toothpaste, 0.187 mg/mL of commercial toothpaste); Micrococcus luteus: 0.003 mg/mL (0.140 mg/mL of medicinal toothpaste, 0.046 mg/mL of commercial toothpaste); Streptococcus mutans: 0.015 mg/mL (0.140 mg/mL medical toothpaste, 0.019 mg/mL of commercial toothpaste); E. coli: 0.015 mg/mL (0.375 mg/mL medical toothpaste, 0.062 mg/mL of commercial toothpaste); Pseudomonas ae ruginosa: 0.062 mg/mL (0.281 mg/mL medical toothpaste, 0.046 mg/mL of commercial toothpaste) and yeast Candida albicans: 0.015 mg/mL (0.140 mg/mL medical toothpaste, 0.281 mg/mL commercial toothpaste ). Comparison of MIC values between toothpaste with Propolis and medical toothpastes demonstrates greater effectiveness of Propolis toothpaste from 4.53 to 25.0 times. For St. aureus, this was 10.51 times; for Str. mutans 9.33 times, E. coli 25.0 times; for Pseudomonas aeruginosa 4.53 times and yeast Candida albicans 9.33 times. Compared with commercial toothpastes, Propolis toothpaste was better: 0.74 to 18.73 times. For St. aureus, was 6.03 times, for Str. mutans 1.26 times, for E. coli 4.13 times; for Pseudomonas aeruginosa 0.74 times and yeast Candida albicans 18.73 times.


SL 23

arC (apiceutical research Centre): Developing a global BeePharma network local Help for local Health.

Hugo Fearnley1

1 ARC (Apiceutical Research Centre), 3B Enterprise Way, Whitby, NORTH YORKSHIRE YO22 4NH, U.K., [email protected]

The discovery of an antitrypanosomal chemical in a sample of propolis from an area where sleeping sickness was present in the local population further stimulated ARC’s growing understanding of how the honey bee may be collecting the plants response to its local health challenges, making it its own and at the same time producing a biochemical cocktail that could help human beings resist those same local health challenges. The BeePharma Africa project was conceived in 2013 as a vehicle for further exploring the idea of local medicines from the beehive by building a network of beekeepers, academics, health professionals and government agencies initially to look at three major local diseases Leishmania, Malaria and Trypanosomiasis – all protozoal infections. In 2016 Hugo Fearnley (Director of ARC) received a research award from the Winston Churchill Memorial Trust to visit 6 African countries to explore this area, collect samples and investigate the potential of rural beekeeping in the local economies. Two new BeePharma projects are now being set up in Jordan and India. ARC now wishes to encourage the creation of other independent BeePharma projects i.e. partnerships between independent groups of beekeepers/academics/health professionals and ARC to further understand how the honey bee can provide Local Help for Local Health.



SL 24

Fatty acid analysis and Biological activity of Jordanian Propolis

Ashok K. Shakya1, Shankar Katekhaye2,3, Ghaleb A. Oriquat1, Rajashri R. Naik1, Anant Paradkar2,4, Hugo Fearnley3,4, James Fearnley3,4

1 Faculty of Pharmacy and Medical Sciences, Al-Ahliyya Amman University, Amman-19328, Jordan. 2Centre for Pharmaceutical Engineering Science, School of Pharmacy, University of Bradford, Bradford, BD7 1DP, UK. 3Nature’s Laboratory, Unit 3b, Enterprise Way, Whitby, North Yorkshire, YO22 4NH, UK4Apiceutical Research Centre, Unit 3b, Enterprise Way, Whitby, North Yorkshire, YO22 4NH, UK

Propolis is a resinous natural product collected by bees (Apis mellifera) from tree exudates which is widely used in folk medicine1. Reports on Jordanian Propolis reveal the presence of new chemical compound 4(Z)-1-3-dihydroxyeupha-7,24-dien-26-oic acid2 along with other compounds like pinobanksin-3-O-acetate, pinocemberin, chrysin3 and lignoceric acid2. The present study was carried out to investigate the fatty acid composition, antioxidant and xanthine oxidase inhibition activity of Jordanian Propolis, collected from Al-Ghour region. The hexane extract of Jordanian Propolis contains different fatty acids, which are reported first time, using GC-FID. The major fatty acid identified were palmitic acid (44.5%), Oleic acid (18:1∆9cis, 24.6% ), Arachidic acid (7.4%), Stearic acid (5.4%), linoleic acid (18:2∆9-12cis, 3.1%), caprylic acid (2.9%), lignoceric acid (2.6%), cis-11,14-eicosadienoic acid (20:2∆11-14cis, 2.4%), palmitoleic acid (1.5%), cis-11-eicosenoic acid (1.2%), α–linolenic acid (18:3∆9-12-15cis, 1.1%), cis-13,16-docosadienoic acid (22:2∆13-16cis, 1.0%), along with minor constituents like saturated fatty acids. Antioxidant properties of the hexane extract were determined via DPPH radical scavenging, β-carotene bleaching assay and NO scavenging assay. The extract produced significant antioxidant activity in-vitro. The extract also exhibit appreciable xanthine oxidase inhibitory activity in-vitro.

References:1 Toreti, V.C., Sato, H.H., Pastore, G.M. and Park, Y.K., 2013. Recent progress of propolis for its biological and chemical compositions and its botanical origin. Evidence-based complementary and alternative medicine, 2013. http://dx.doi.org/10.1155/2013/6973902 Shaheen, S.A., Zarga, M.H.A., Nazer, I.K., Darwish, R.M. and Al-Jaber, H.I., 2011. Chemical constituents of Jordanian propolis. Natural product research, 25(14), 1312-1318.3 Darwish, R.M., Ra‘ed, J., Zarga, M.H.A. and Nazer, I.K., 2010. Antibacterial effect of Jordanian propolis and isolated flavonoids against human pathogenic bacteria. African Journal of Biotechnology, 9(36). 5966-5974


POSter PreSeNtAtiONS


PP 1

Phenolic Compounds of Spain Propolis

Félix Adanero-Jorge1, Mª Camino García-Fernández2, Rosa Mª Valencia-Barrera1 y J. Javier Sanz-Gómez2

1Department of Biodiversity and Environmental Management, Botany, University of León, León, Spain.2ICTAL, Food Science and Technology Institute, University of León, León, Spain.

The phenolic compounds, flavonoids in particular, are very frequent in the plant kingdom with many beneficial properties. Some of these compounds are present in exudates or buds of plants. They appear in propolis as a consequence of the collection and transformation by specialized honey bees in the hive. The aim of the study is the determination1 of phenolic compounds by UPLC-MS/MS and UPLC-PDA in methanolic extracts2 of 134 samples of propolis from Castilla y León (North and Centre of Spain). Such samples were gathered using high density polyethylene mesh for food use which placed in Langstroth beehive from April 2011 to August 2012. The results obtained reveal the interesting source of bioactive compounds of the North and Centre of Spain propolis, being the most significant caffeic acid and its derivatives (CAPE and isoprenyl caffeate), chrysin, pinobanksin, p-coumaric acid, galangin and kaempferol; other compounds identified only in some samples were benzoic acid, trans-ferulic acid, taxifolin, luteolin, sakuranetin, apigenin+genistein and eriodictyol. Furthermore, the content of CAPE and isoprenyl caffeate may restrict its use, for the food and pharmaceutical industry, since they are considered allergens3.

References:1 Kasote, D., Suleman, T., Chen, W., Sandasi, M., Viljoen, A., van Vuuren, S. (2014) “Chemical profiling and chemometric analysis of South African propolis”, Biochemical Systematics and Ecology, 55, pp. 156-163.2 Dias, L. G., Pereira, A. P., Estevinho, L. M. (2012) “Comparative study of different Portuguese samples of propolis: pollinic, sensorial, physicochemical, microbiological characterization and antibacterial activity”, Food and Chemical Toxicology, 50, pp. 4246-4253.3 Walgrave, S. E., Warshaw, E. M., Glesne, L. A. (2005) “Allergic contact dermatitis from propolis”, Dermatitis, 16(4), pp. 209-215.


PP 2

Chemical Profiling of Papua New Guinea Propolis and Assay its Antiprotozoal Activity

Samya Alenezi1, Naif Alenezi Manal1, J Natto1, Harry P. De Koning2, John Igoli2, David G. Watson1

1 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 1RE.2 Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G12 8TA, UK

This study is aimed to investigate the chemical profile and examine the in vitro efficacy of Papua new guinea (PNG) propolis and compounds purified from it purified compounds against the protozoal parasites: Crithidia fasciculate, Trypansoma congolense, Trypanosoma b. brucei and drug resistant Trypanosoma brucei (B48), which cause diseases in humans and other animals as well as insects. In addition. The samples were assayed for their toxicity against human U937 cells in vitro. Chemical profiling was conducted by using negative ion spray ESI (LC-MS) and indicated that the ethanolic extract of PNG contained a series of triterpenes. Principle components analysis (PCA) of the processed LC-MS data collected was demonstrated the uniqueness in chemical composition of the fractions prepared from the sample that were also active against the parasites. Active principles were isolated by bioassay-led fractionation, using medium pressure chromatographic (MPLC). Afterwards, by using spectroscopic and spectrometric means, nine pure compounds were isolated and the their structures were elucidated indicating that eight of them were cycloartane type triterpenes. While the other one was a pentacyclic triterpene, 20 hydroxy betuline that was found to be the most active compound in these assays. Growth curves of drug-resistant T. brucei at concentrations ≥ EC50 showed that this compound was trypanostatic. Overall, the PNG propolis and its fractions showed low toxicity on U937 cell growth and were efficacious against kinetoplastid parasites.


PP 3

Chemical Profile of Greek Arbutus unedo Honey – Biological Properties

Αrgyrο Andreou1, Ioanna Chinou1, Konstantia Graikou1

1 Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece

In the framework of our studies on Greek bee-keeping products (honey, propolis, royal jelly and pollen) and due to an increasing interest in the use of honey as an antioxidant as well as in the treatment of bacterial infections and in traditional Mediterranean diet, we report in this study the chemical analyses, the antimicrobial evaluation and the pollinic spectrum of one honey sample from Greek strawberry-tree (Arbutus unedo L.), which to our best knowledge has never been studied before. Typification of this honey sample, was based on its pollinic spectrum obtained by Louveaux’s quantitative method exerted an approx. 23% Ericaceae pollen. The studied sample was analysed through GC-MS, while metabolites have been also isolated and structurally determined by spectral means. Among the volatile substances: 3,5,5-trimethyl-3-cyclohexen-1-one (β-isophorone), 3,5,5-trimethyl-2-cyclohexen-1-one (α-isophorone) and 3,5,5-trimethylcyclohex-2-ene-1,4-dione (4-oxoisophorone) were detected through GC-MS, as the most characteristic odor constituents1. Moreover, 5-hydroxymethylfurfural (HMF) and homogentistic acid (HGA) have been also determined. HGA is a phenolic metabolite which is responsible for the antioxidant capacity of the strawberry-tree honey itself2, while HMF is a well known, essential parameter of freshness and of good quality of commercial honeys. It is also noteworthy, the isolation of unedone and 2-cis-4-trans-abscisic acid, which according to the international literature are characterized as chemical markers of such European honeys3. Furthermore, the sample exhibited an interesting antimicrobial activity against all assayed human pathogenic microorganisms.

References:1 Bianchi F., Careri M., Musci M. (2005). Volatile norisoprenoids as markers of botanical origin of Sardinian strawberry tree (Arbutus unedo L.) honey. Characterisation of aroma compounds by dynamic headspace extraction and gas chromatography-mass spectrometry. Food Chemistry 89:527-532.2 Rosa A., Tuberoso C.I.G., Atzeri A., Melis M.P., Bifulco E., Dessi M.A. (2011). Anti-oxidant profile of strawberry tree honey and its marker homogentistic acid in several models of oxidative stress. Food Chemistry 129:1045-1053. 3 Tuberoso I.G.C., Bifulco E., Caboni P., Cottiglia F., Cabras P., Floris I. (2010). Floral markers of strawberry tree (Arbutus unedo L.) honey. Journal of Agricultural and Food Chemistry 58:384-389.


PP 4

Propolis included Chewing gum and investigation of its antibacterial activity against Streptococcus mutansİbrahim Palabıyık1, Didem Sözeri Atik1, Esra Bölük1, Sevgi Kolaylı2

1Namık Kemal University, Faculty Of Agriculture, Food Engineering Department, Tekirdag, Turkey2Karadeniz Technical University, Faculty Of Science, Chemistry Department, Trabzon, Turkey

Propolis, or bee glue, is a natural wax-like resinous substance found in bee hives where it is used by honeybees as protective agent. Since antiquity, mankind has been using propolis in different fields, mainly traditional medicine. In addition, propolis has been widely used for its antibacterial, antifungal, antioxidant, and anti-inflammatory properties. The aim of the research is to add propolis into ingredients of chewing gum and to obtain propolis chewing gum and determine antimicrobial effect of this gum against Streptococcus mutans which generate tooth cavity. Chewing gum is the most suitable food product for using propolis wax because it is acting like gum base and so it is constitute a perfect chewing gum mix. For this purpose, propolis samples were collected from oak trees in Kırklareli, Turkey and an extraction method was used to extract propolis from raw propolis. This method include the following steps, 10 g raw propolis and 90 ml of 96 % ethanol were incubated at 60 °C for 24 h and this mixture was filtered after this process was performed, heat treatment (70 °C) was used for extraction. After the extraction, extract that obtained from raw propolis was added to chewing gum mix. Greater inhibitory effect was noted with 4 % propolis extract containing chewing gum compared to that including 10 % propolis containing gum samples, possibly due to different ratio of propolis extract in agreement with other researchs.

Acknowledgements: This study was supported by 117O869 TÜBİTAK Project (3001).


PP 5

Propolis from romania and turkey: Comparative antioxidant and antibacterial activity

Otilia Bobiş1, Merve Keskin2, Şaban Keskin3, Sevgi Kolaylı2, Mihaela Niculae4, Adriana Urcan5, Claudia Paşca5, Liviu Al. Mărghitaş5, Daniel S. Dezmirean5

1Life Science Institure, APHIS Laboratory, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania2Department of Chemistry, Karadeniz Technical University, Turkey3Department of Chemistry, Bilecik Seyh Edebali University, Turkey4Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania5Apiculture and Sericiculture Department, Faculty of Animal Sciences and Biotechnologies, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania

Propolis is an important bee product, used by the bees to seal the cracks and to hygiene the wax combs after brood rearing (Huang et al., 2014). The bioactive compounds of propolis originate from plant resins that bees collect, mix with own secretions and wax. Propolis chemical composition is different according to the geographical location (Mărghitaş et al., 2013), botanical origin and bee specie (Silici and Kutluca, 2005). The aim of the study was to determine the chemical composition of different propolis samples from Romania and Turkey and to correlate the composition with the antioxidant and antibacterial activity of tinctures against Gram-positive and Gram-negative bacteria. Soxhlet extraction, gravimetric measurements, spectrophotometric and chromatographic determinations were used in the study. The antimicrobial properties were evaluated in vitro towards both reference and clinical strains of Staphylococcus aureus, Salmonella enteritis, Salmonella typhimurium, Escherichia coli and Pseudomonas aeruginosa using the disc diffusion assay; the minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations were established by a broth microdilution method and used to calculate the bactericidal index for each extract. The wax content of the samples ranged within the limits provided by the Argentinian and European standards in effect. Balsam content of the samples was above 30%, providing a tincture with high antioxidant properties. High amounts of total polyphenols and flavonoids were determined in all samples, though different for Romanian and Turkish samples. The inhibitory ability was different for samples coming from different geographical origins, depending fundamentally on plant source resins, but also on bacterial species.

References: 1 Marghitas LA, Dezmirean DS, Bobis O (2013) Important developments in Romanian propolis research. Evidence-Based Complementary and Alternative Medicine 2013: 159392.2 Huang MI, Zhang C-P, Wang K, Li GQ, Hu FL (2014) Recent advances in the chemical composition of propolis. Molecules 19: 19610-19632.3 Silici S, Kutluca S (2005) Chemical composition and antibacterial activity of propolis collected by three different races of honeybees in the same region. Journal of Ethnopharmacology 99: 69-73.


PP 6

the Characterization and Bioactive Composition of turkish Propolis

İlknur Coşkun1, Gizem M. Duymaz1, Tuğçe Daştan1, Özge E. Sönmezer1, Sezer Acar1, Emir Akyıldız1, Gamze Düz1, Özlem Yılmaz1, Sinem Raday1

1Balparmak R&D Center, Altıparmak Gıda San. Ve Tic. A.Ş., İstanbul, Turkey

The main aim of this study is to characterise Turkish propolis and to determine the bioactive composition of it. 86 different propolis samples were collected from 25 different provinces in Turkey. Chemical, microscopic and functional properties of Turkish propolis have been determined. Also Turkish propolis was compared propolis types from other countries. Each sample was analyzed for moisture content, beeswax content, total phenolics, antioxidant activity, flavonoid profile, pollen profile and mineral content. Moisture, ash and beeswax content were analyzed by AOAC934.01, IHC methods, respectively. Total phenolic contents, antioxidant activities and flavonoid profile were analyzed by Folin Ciocalteu, DPPH using Spectrophotometer and by HPLC-PDA, respectively. Possible botanical sources of the samples were investigated using microscopic method and mineral contents were determined using ICP-MS. It was determined that the results vary depending on flora, season and climate conditions. It was specified that the flavonoid profile of Turkish propolis showed differences from other types of propolis. With flavonoid profile analysis, totally 38 different bioactive compounds were analyzed. Caffeic Acid, p-Coumaric Acid, Ferrulic Acid, M-Coumaric Acid, 3,4-Dimethoxy Cinnamic Acid, trans-Cinnamic Acid, Pinobanksin, Quercetin, Naringenin, Luteolin, Genistein, Hesperetin, Kaempferol, Apigenin, Pinocembrin, CAPE, Chrysin and Galangin were mainly determined in Turkish Propolis. Possible botanical sources of propolis were determined as Pinus spp., Salix spp., Castanea spp., Eucalyptus spp., Quercus spp. and Populus spp.. Different regions mainly contains Iron, Chromium, Copper, Calcium, Magnesium, Selenium, Manganese and Zinc. It was specified that flavonoid content of Turkish propolis showed differences from other propolis types.


PP 7

effects of Propolis on the Quorum Sensing of Selected Biofilm Producing Bacterial Species

K. Tennick1, S. Wafa1, H. Fearnley2, M. Gomez Escalada1

1 Centre for Biomedical Science Research, School of Clinical and Applied Sciences, Leeds Beckett University, Leeds, UK2 BeeVital, Whitby, UK

The clinical relevance of biofilms has become more apparent, however despite this increased prominence they remain difficult to treat. Biofilm formation relies on bacterial communication mechanisms known as quorum sensing (QS). QS presents a novel target for antimicrobials. It is postulated that QS inhibition would not so readily lead to the development of resistance, a common barrier in the treatment of infectious disease. This study set out to establish whether propolis could have an impact on biofilm formation.Gram positive and negative bacterial species of clinical relevance were tested, namely Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. In addition, Chromobacterium violaceum was used to determine QS inhibition. The effects of an ethanoic extraction of propolis (0-2.5% v/v) on biofilm formation was assessed using colourimetry. Manuka honey was tested as a comparison due to its known impact on biofilm formation and QS. Propolis extract at low concentrations was found to inhibit biofilm formation in all bacteria tested. Effects were similar in both Gram positive and negative bacteria. Whilst Manuka honey also achieved an inhibitory effect on biofilm formation, this was to a lesser extent. Propolis was also found to be particularly potent at inhibiting QS in C. violaceum. Our data demonstrates that propolis has potent anti-biofilm properties. These properties are at least in part due to QS inhibition, which is shown by the inhibition production of homoserine lactone in C. violaceum. This study demonstrates the therapeutic potential of propolis in inhibiting biofilm formation in a range of clinically relevant bacteria.


PP 8

In Vitro evaluation of green and red Propolis extracts against Candida spp.

M. D. Freitas1, G. A. Lopes1, N. A Oliveira1, B. M. Almeida2, S. R. L. Abreu2, R. C. Basques3, N. S. Binda4, S. M. Figueiredo1,3

1Departamento de Alimentos, Escola de Nutrição, Universidade Federal de Ouro Preto. Campus Morro do Cruzeiro, s/n. Ouro Preto. 35400-000. Minas Gerais. Brazil.2NectarPharmaceutical. Rodovia MG 435, Km 2.5. Caeté. Minas Gerais. 34800-000. Brazil.3Instituto de Ensino e Pesquisa da Santa Casa de Belo Horizonte (IEP/SCBH). RuaDomingos Vieira, 590. Belo Horizonte. 30150-240. Minas Gerais. Brazil.4Deparatamento de Farmácia. Escola de Farmácia. Universidade Federal de Ouro Preto. Campus Morro do Cruzeiro. Ouro Preto. 35400-000. Minas Gerais. Brazil

In different countries, propolis has been used as food supplement, which provides organic balance of immune system and as alternative treatment for some diseases1,2. Studies have shown that due to the presence of flavonoids in its chemical composition2, propolis has anti-inflammatory immunomodulator, antiproliferative, antibacterial, antiviral and antioxidant properties2,3. Candidiasis caused by species of Candida genus has high incidence due to the increasing number of immune suppressed people. It has being observed an enhancement of resistance of these fungi to the actual antifungals3. In this work ethanol extracts of green (GrProp) and red propolis (RdProp) were subjected to in vitro assays against six C. albicans ATCC strains and 10 clinical isolates. Minimum inhibitory concentration of extracts was determined. The extracts stood out in relation to the growth inhibition of all ATCC Candida tested, with concentrations varying from 125.0-1000.0μg/mL (25% RdProp) and 500-2000μg/mL (100% GrProp). In relation to inhibition of growth of all tested Candida spp.,best results were found for extract 25% RdProp (125-1000μg/mL) and 100% GrProp (500 -2000 µg/mL). Due to observed anti-Candida activity was inferred that RdProp have potential to be used against candidiasis.

Acknowledgements. To Conselho Nacional de Pesquisas (CNPq) for financial support (Project: 421544/2016-3)

References1 Babaei, S, Rahimi, S, KarimiTorshizi, MA, Tahmasebi, G, KhaleghiMiran, SN (2016). Effects of propolis, royal jelly, honey and bee pollen on growth performance and immune system of Japanese quails.Veterinary Research Forum, 7(1), 13–20.2 Daugsch A, Moraes CS, Fort P, Park YK (2008).Brazilian Red Propolis - Chemical Composition and Botanical Origin.Evid-BasedComplementAlternMed ECAM. 8;5(4):435–441.3 Freitas, M.C.D.; De Oliveira, D.T.; De Miranda, M.B.; Vieira-Filho, S.A.; Caligiorne, R.B. De Figueiredo, S.M. (2017). Biological activities of red propolis: a rewiew. Recent Pat EndocrMetab Immune Drug Discov, 11(1):3-12.



PP9

thirteen Flavonoids from green Propolis from minas Gerais, Brazil, Analyzed for Six YearsM. D. Freitas1, G. A. Lopes1, N. A Oliveira1, B. M. Almeida2, S. R. L. Abreu2, R. C. Basques3, N. S. Binda4, S. M. Figueiredo1,3

1Departamento de Alimentos, Escola de Nutrição, Universidade Federal de Ouro Preto. Campus Morro do Cruzeiro, s/n. Ouro Preto. 35400-000. Minas Gerais. Brazil.2NectarPharmaceutical. Rodovia MG 435, Km 2.5. Caeté. Minas Gerais. 34800-000. Brazil.3Instituto de Ensino e Pesquisa da Santa Casa de Belo Horizonte (IEP/SCBH). RuaDomingos Vieira, 590. Belo Horizonte. 30150-240. Minas Gerais. Brazil.4Deparatamento de Farmácia. Escola de Farmácia. Universidade Federal de Ouro Preto. Campus Morro do Cruzeiro. OuroPreto. 35400-000. Minas Gerais. Brazil

Propolis, a greenish resinous collected by bees, mainly from plants around their habitat, is a Greek word resulting from the prefix “pro” (preservation or defense) with “polis” (hive or community or cluster). There are 13 types of propolis in Brazil, which vary according to the geographical and botanical origin regions and its different chemical compositions2. The green propolis (GrProp) type, derived from plant Baccharis dracunculifolia, popularly known as rosemary-of-field, gained prominence for its varied chemical constitution, mainly related to flavonoids. In this study was evaluated the seasonal effect on the chemical composition of GrProp from Minas Gerais (Brazil) in a period of six years. The GrProp was collected at February, March, May, July, September and October during six years (2008-2013). The chemical composition of thirteen flavonoids of GrProp was evaluated by RP-HPLC and statistical analysis of the results. The relative amounts of the majority of GrProp flavonoids were similar in most analyzed samples p-Coumaric acid, chrysin, galangin and kaempferol showed statistically differences in the analyzes made during the study months and through the period of analysis. Nevertheless, the results indicated that GrProp maintained similar and stable characteristics through the six years of study.

Acknowledgements. To ConselhoNacional de Pesquisas (CNPq) for financial support (Project: 421544/2016-3)

References1 Park, YK, Alencar, SM, Scamparini, ARP, Aguiar, CL (2002). Própolis produzida no sul do Brasil, Argentina e Uruguai: Evidências fitoquímicas de sua origem vegetal. Ciência Rural. 2: 997-1003, 2 De Figueiredo, SM, Binda, NS, Almeida, BM, Abreu, SRL, Abreu, JAS, Pastore, GM, Sato, HH, Toreti, VC, Tapia, EV, Park YK, Vieira-Filho, SA, Caligiorne, RB (2015). Green Propolis: Thirteen Constituents of Polar Extract and Total Flavonoids Evaluated During Six Years through RP-HPLC.Curr Drug DiscovTechnol.12(4):229-239.




PP 10

Water extract of Propolis Might be Safer to use During Pregnancy than ethanol extract

Al Mukhlas Fikri1, Ahmad Sulaeman1, Sri Anna Marliyati1, Mokhamad Fahrudin2

1Department of Community Nutrition, Faculty of Human Ecology, Bogor Agricultural University, Bogor 16680, Idonesia. 2Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor 16680, Idonesia.

Propolis is a natural product that has many health benefits. This study was the first investigation examining the effect of propolis administration on pregnancy. Pregnant mice were used as experimental animals (n = 25) divided in five groups. They were control group (Tween 80 1%), low (380 mg/ kg b.w) and high (1400 mg/ kg b.w) dose of ethanol extract of propolis (EEP), low (380 mg/ kg b.w) and high (1400 mg/kg b.w) dose of water extract of propolis (WEP). Propolis was administered from 0 to 18 days of gestational age. The results showed that high dose of EEP significantly reduced weight and crown-rump of fetuses and increased the amount of resorption (p<0.05). However, low dose of EEP, low and high dose of WEP did not affect weight and crown-rump length of fetuses compare to control group. Low dose of EEP, low and high dose of WEP significantly reduced number of resorption compare to high dose of EEP. There was no difference in liver and kidney weight, serum ureum, creatinine, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and maternal weight gain. No significant difference also was found in placental weight, litter size, number of dead fetuses, embryo mortality before and after implantation. The results of this study indicate that WEP might be safer to use during pregnancy than EEP.

Acknowledgement: Thanks to Ministry of Research, Technology and Higher Education, Republic of Indonesia for research grant through scheme Master and Doctoral Program of Excellent Graduate

References:1 Fikri AM, Sulaeman A, Marliyati SA, Fahrudin M. Antiemetic Activity of Trigona spp. Propolis from Three Provinces of Indonesia with Two Methods of Extraction. Pharmacog J. 2018;10(1):120-2.


PP 11

Differences in Chemical Composition and antioxidant activity of three Propolis Samples Collected in the Same apiary

Ana Sofia P. Freitas1,2, Ana João P. Gomes1, Ana Cunha1,2,3,4, Rui P. S. Oliveira1,2,3,4, Cristina Almeida-Aguiar1,2,3,4

1 Biology Department, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal3 CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.4 CBMA - Centre of Molecular and Environmental Biology, University of Minho, 4710-057 Braga, Portugal.

Propolis, a complex mixture of resinous and balsamic material collected by honeybees from plant material and mixed with bees’ salivary secretions, is composed by different groups of compounds that have been linked with diverse bioactivities. In particular, flavonoids, phenolic acids and their esters were associated to valuable biological properties such as antioxidant and free radical scavenging1,2. Propolis possess substantial variability of chemical composition and bioactive profiles, even from the same origin but collected in different years3. However, Portuguese propolis samples from an apiary named Gerês (G), normally obtained by mixing propolis from three places - Bugalho (b), Toutelo (t) and Felgueiras (f) - show similar chemical and biological profiles over the years (Freitas, under reviewing). Recently, a new propolis place - Roca (r) - was added to the apiary. In this work we have studied three Gerês propolis samples collected in 2017 (G17), but composed by different combinations of source places: G17-r; G17-rt and G17-tf. Total phenolic, total flavonoid and ortho-diphenol contents were determined for the ethanolic extracts (EE) obtained from the three propolis samples and DPPH• scavenging assay was performed to evaluate the in vitro antioxidant capacity of G17.EEs. All propolis samples showed antioxidant capacity, having one of the extracts, namely G17-r.EE, the highest scavenging ability of DPPH•. This extract was also the one with higher content in phenolic, flavonoid and ortho-diphenol compounds.Note: Ana Freitas and Ana Gomes contributed equally to this work.

Acknowledgments: Financial support provided by FCT (PD/BD/128276/2017), under the Doctoral Programme Agrichains - PD/00122/2012.

References:1 Silva-Carvalho, R., Baltazar, F., Almeida-Aguiar, C. 2015. Evidence-Based Complementary and Alternative Medicine 2015.2 Cruz, M., Antunes, P., Paulo, L., Ferreira, A. M., Cunha, A., Almeida-Aguiar, C., Oliveira, R. 2016. RSC Advances 6(55), 49806-49816.3 Veloz, J. J., Saavedra, N., Lillo, A., Alvear, M., Barrientos, L., Salazar, L. A. 2015. BioMed Research International 2015.


PP 12

Combination treatment of Cuban Propolis and nemorosone with Chemotherapeutic agents induce a Synergisitic Cytotoxic effect in Drug-resistant Human Colon Carcinoma Cells

Y. Frión-Herrera1, D. Gabbia1, M. Carrara1

Dept. of Pharmaceutical and Pharmacological Sciences, University of Padova, Italy. [email protected]

Several studies have confirmed that propolis mono-therapy can inhibit the growth of different carcinomas cells1 due to its chemical composition2. At present, there are few studies about the possible use of this natural product in the treatment of chemoresistant tumors3. Combination experiments were carried out in order to study the ability of Cuban propolis extracts (CP) and nemorosone, its main phytocomponent, to increase of antiproliferative efficacy of the traditional cytotoxic chemotherapic drugs doxorubicin (Dox) and oxaliplatin (Oxa) in human colon carcinoma cells (LoVo), doxorubicin-resistant cells (LoVo Dox) and oxaliplatin-resistant cells (LoVo Oxa). The effects of combination therapy were investigated in vitro evaluating cell viability at 24, 48 and 72 hs. Synergism/additivity/antagonism effect was assessed by isobologram/combination index analysis. Molecular mechanism of combination treatment was examined assessing LDH release, Annexin/PI assay, ROS production, mitochondrial membrane potential (∆Ψm) disturbance and ATP intracellular levels. Meanwhile, the expression of genes related to apoptosis P53, BAX and BCL2 were analyzed by qRT-PCR. Our result revealed that combination treatment resulted in a significantly synergistic antiproliferative and cytotoxic effect at 72 hs with respect to monotherapy, in particular in drug-resistant cell lines. Furthermore, combination treatment induced apoptosis through activation of pro-apoptotic genes (P53 and BAX) and downregulation of BCL2 with marked disruption of ∆Ψm, decrease in ATP production and induction of ROS. These findings indicate that combination of CP or nemorosone with classical chemotherapic drugs may represent an intriguing treatment option for drug-resistant human colon carcinoma.

References:1 Oršolić N. A review of propolis antitumour action in vivo and in vitro. Journal of ApiProduct and ApiMedical Science. 2010; 2 (1):1 - 20.2 Marcucci M. Propolis: chemical composition, biological properties and therapeutic activity. Apidologie. 1995;26:83-99.3 Patel S. Emerging Adjuvant Therapy for Cancer: Propolis and its Constituents. Journal of dietary supplements. 2016;13(3):245-


PP 13

Correlation between Phenolic Composition and Biological Properties of Propolis and Heavy Metal Contents

María Inmaculada González-Martín1*, Eddy Valentín Betances-Salcedo1, Isabel Revilla2, Ana María Vivar-Quintana2

1 Department of Analytical and Food Chemistry, Faculty of Chemistry, University of Salamanca, Plaza de la Merced, 37008 Salamanca, Spain. 2 Food Technology. University of Salamanca, E.P.S. de Zamora, Avda. Requejo 33, 49022 Zamora, Spain. *[email protected]

The presence of heavy metals in propolis is associated with the environmental pollution of anthropic origin around the apiaries via various sources such as air, water, plants and soil. Although the presence of these compounds could affect the biological properties of the propolis, the correlation between them has not been previously studied. Then, the aim of this work was to stablish the correlations between the levels of some heavy metals such as lead, chromium, nickel, copper or zinc and phenolic composition and biological properties such as antioxidant or antibacterial activities. For this work, 44 propolis samples of different geographic locations were analyzed: Chile (Biobio region n=10) and Spain (Galicia n=12 and Castilla y León n=22). The mineral composition analysis was performed using ICP-MS (Inductively Coupled Plasma Mass Spectrometry). Antioxidant activity was measured using ABTS, DPPH and inhibiting activity of the linoleic acid/-caroten radical assays. Phenolic acids and their derivatives were determined by HPLC and biocide properties using the Five-Plate test. Pearson correlations were calculated with SPSS statistic package. The results show a negative correlation between antibacterial activity similar to betalactamic antibiotics with nickel (p<0.01) and chromium (p<0.01) levels. The lead content was negatively correlated with phenolic acids and their derivatives. This correlation were statistically significant for the cynamil capheoil ester (p<0.05). The zinc and copper contents were positively correlated with cumaric acid and cinnamic derivatives (p<0.05) respectively. Heavy metal levels were in general negatively correlated with antioxidant activity and these correlations were significant for copper (p<0.05) and lead (p<0.01).


PP 14

Changes in antioxidant and antibacterial activity and in Phenolic Compounds levels Due to the Pesticide residues

María Inmaculada González-Martín1*, Eddy Valentín Betances-Salcedo1, Isabel Revilla2, Ana María Vivar-Quintana2

1 Department of Analytical and Food Chemistry, Faculty of Chemistry, University of Salamanca, Plaza de la Merced, 37008 Salamanca, Spain. 2 Food Technology. University of Salamanca, E.P.S. de Zamora, Avda. Requejo 33, 49022 Zamora, Spain. *[email protected]

Propolis has many known pharmacological properties. However, acaricides insecticides and herbicides are contaminants that can be found in propolis. This residues derive from contamination generated by agricultural practices and the application of pesticides in the hive. As far as we know, there are not works that study if the presence of these contaminants could modify the biological properties of the propolis. Thus, this was the main objective of this research. To do that, 44 samples of propolis from Chile (Biobio region n=10) and Spain (Galicia n=12 and Castilla y León n=22). were analysed. The pesticides were quantified using Gas Chromatography-Mass Spectrometry (GC-MS). Phenolic acid and derivatives were analysed by HPLC and antibacterial activity by Five-Plate tests. Total polyphenols were quantified by Folin-Ciocalteaus method and total flavones and flavonols using AlCl3 colorimetric reaction. Antixidant capacity were determined by ABTS, DPPH and inhibiting activity of the linoleic acid/-caroten radical methods. Results show that 70% of samples contained triadimefon (0.4 a 42.2 mg/Kg) and 7.5% showed residues of dicofol, dichlofluanid, folpet, propham and metazachlor. The phenolic contents and antioxidant and biocide activities of samples with pesticide residues were compared with those free of contaminants. Results show that samples with pesticide residues have lower antioxidant capacity estimated by any of the assayed methods. The biocide properties similar to antibiotics was also lower in samples contaminated with pesticides. A significant decrease of phenolic acids such as cafeic, isoferulic and DMC acids were observed together with a slight decrease of total polyphenols and flavonoids.


PP 15

north aegean greek islands Propolis antibacterial-antifungal activities against Malassezia

Konstantia Graikou1, Aristea Velegraki2, Mirto Varsani1, Ioanna Chinou1

1Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece 2Division of Microbiology, Faculty of Medicine, National and Kapodistrian University of Athens, Greece

The chemical composition of propolis is highly variable and depends mainly on the local flora. Greece is characterized by high biodiversity flora, assuming different propolis composition. Due to its complex chemical constitution and its pharmaceutical and nutraceutical use worldwide, it is necessary to characterize the quality of propolis and to guarantee a reproducible quality, in order to ensure a safe use1. As a part of a systematic research on different propolis, especially from Mediterranean area, we report in this study the chemical analysis of three samples from Northeast Aegean Greek islands (Samos, Chios and Lesvos) performed by GC/MS after silylation. In these samples diterpenes (abietic acid, imbricataloic acid, isoagatholal, agathadiol, communic acid, pimaric acid, 13-epi-cupressic acid, isocupressic acid totarol) have been identified as major constituents. Especially Samos and Chios propolis samples showed high diterpene percentages (58 and 50%, respectively). These findings confirm that both samples belong to the recently defined “Mediterranean type”2, probably due to the presence of conifer trees as main propolis plant sources. All the ethanolic extracts of propolis samples have been evaluated for thei total phenolic content and showed significant antibacterial activity against nine Gram-negative and -positive human pathogenic bacteria and three fungi, probably due to the large amounts of diterpenes. Furthermore they were evaluated for their inhibitory effects against 18 Malassezia strains (reference and clinical)3, concluding that reach diterpene propolis may have interesting applications to control Malassezia fungal-derived diseases.

References:1 Bankova V., de Castro S.L., Marcucci M.C. (2000). Propolis: recent advances in chemistry and plant origin. Apidologie, 31: 3-152 Graikou K., Milena P., Gortzi O., Bankova V., Chinou I. (2016). Characterization and biological evaluation of selected Mediterranean propolis samples. Is it a new type? LWT-Food Science and Technology, 65: 261-267.3 Velegraki A., Alexopoulos E.C., Kritikou S., Gaitanis G. (2004). Use of fatty acid RPMI 1640 media for testing susceptibilities of eight Malassezia species to the new triazole posaconazole and six established antifungal agents by a modified NCCLS M27-A2 microdilution method and E-test. Journal of Clinical Microbiology, 42: 3589-3593.

https://en.wikipedia.org/wiki/Samos_Island

https://en.wikipedia.org/wiki/Chios

https://en.wikipedia.org/wiki/Lesbos

https://en.wikipedia.org/wiki/Samos_Island

https://en.wikipedia.org/wiki/Chios


PP 16

Preliminary Studies: the Potential anti-angiogenic activities of two Sulavesi island (indonesia) Propolis and their Chemical Characterization

Muhammad Iqbal1, Tai-ping Fan1, David Watson2, Sameah Alenezi2, Muhamad Sahlan3

1Department of Pharmacology, University of Cambridge, Cambridge, United Kingdom.2Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.3Department of Chemical Engineering, Universitas Indonesia, Depok, Indonesia.

Several studies have previously reported propolis, or its constituents, to inhibit tumour angiogenesis1. The anti-angiogenic activity of two Indonesian stingless bee propolis extracts from Sulawesi Island on vascular cells were assessed. Sample D01 was obtained from the outer side of bee hives, while D02 was from the inner side of the same hives. Preliminary results of cell viability and multi-channel wound healing assays on HUVECs and placenta-derived pericytes demonstrated that D01 elicited a strong cytotoxic effect and a considerable anti-migratory activity on the vascular cells. Although D02 demonstrated a much weaker cytotoxic effect on the cell lines compared to D01, it elicited a substantial protective effect on the pericytes against CoCl2-induced dropout in an experiment to mimic a micro-environment commonly associated with angiogenesis and tumour growth2. These results demonstrate modulatory effects of these propolis samples in vascular cells, which requires further investigation. The extracts were profiled by using liquid chromatography coupled to high resolution mass spectrometry. The most abundant compounds in Indonesian sample D01 from LC-MS analysis seem to be unusual since they do not immediately fall into a clear class such as flavonoids or terpenoids. Two of the most abundant compounds have elemental compositions matching actinopyrones which are antibiotic compounds isolated from Streptomyces pactum3. Identification on the basis of elemental composition is not definitive but compounds in D01 are possibly due to unusually modified terpenoids. Sample D02 has abundant compounds which include four related diterpenes with differing degrees of oxygenation and some sesquiterpenes. However, again the profile is unusual.

References:1 Daleprane JB and Abdalla DS (2013) Emerging roles of propolis: antioxidant, cardioprotective, and antiangiogenic actions. Evid Based Complement Alternat Med, 2013:175135.2 Raza A, Franklin MJ, and Dudek AZ (2010) Pericytes and vessel maturation during tumor angiogenesis and metastasis. Am J Hematol 85(8):593-8.3 Hayakawa Y, Saito J, Izawa M, and Shin-ya K (2014) Actinopyrone D, a new downregulator of the molecular chaperone GRP78 from Streptomyces sp. J Antibiotics 67: 831–834.


PP 17

the effects of Propolis on growth Performance of Broiler Chickens

Ivana Klaric1, Matija Domacinovic1, Berislav Prakatur2, Ivan Miskulin3, Bozica Lachner2, Dalida Galovic1, Danijela Samac1 1 Faculty of Agriculture in Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 2 The Valpovka Fodder Factory, Valpovo, Croatia 3 Faculty of Medicine Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia Propolis is a resinous and balsamic substance that is collected from buds, leaves and similar parts of trees and plants by honeybees (Apis mellifera) and mixed with wax1. The two main targets in modern poultry production are high growth rate and feed efficiency2. The aim of this study was to determine the effect of diet supplementation with propolis on the growth performance of broiler chickens. This experimental study was conducted on 120 Ross 308 broiler chickens of equally distributed sex, which were randomly divided into three groups: control group (C) and two experimental groups of chickens (P1 and P2). Throughout the whole study (for 42 days) the control group of chickens was fed feed mixture while feed mixture that was fed to the experimental groups of chickens contained propolis in amount of 0,5 g/kg (P1 group) and 1,0 g/kg of feed mixture (P2 group). The average values of body weight of chickens were significantly higher on 14th (p=0.008), 21st (p<0.001), 28th (p=0.001) and 35th (p=0.019) day of fattening in the experimental groups of chickens compared to the control group. The average values of weight gain of chickens were significantly higher on 1st (p=0.017), 3rd (p=0.003) and 4th (p=0.044) week of fattening in the experimental groups of chickens compared to the control group. This study has shown that propolis has significant positive effect on performance parameters in broiler chickens. The administration level of investigated supplement should be further evaluated in order to maximize its efficacy in commercial chicken production. References: 1 Chegini S, Kiani A, Rokni H (2018) Alleviation of thermal and overcrowding stress in finishing broilers by dietary propolis supplementation. Italian Journal of Animal Science 17: 377-385.2 Sugiharto S (2016) Role of nutraceuticals in gut health and growth performance of poultry. Journal of the Saudi Society of Agricultural Sciences 15: 99-111.


PP 18

in vitro assessments of Cytotoxic and Cytostatic effects of Propolis in Cells from the Human Colon Carcinoma Cell line (HCt 116)

Soumaya Touzani1, Sleman Kadan2, Abdalsalam Kmail2, Bashar Saad2, Badiaa Lyoussi1

1 Laboratory of Physiology, Pharmacology & Environmental Health, Faculty of Science, University Sidi Mohamed Ben Abdellah, Fez, Morocco2 Qasemi Research Center and Faculty of Arts and Sciences, Arab American University Jenin, P.O Box 240, Jenin, Palestine.

Based on knowledge from traditional Arab medicine, this in vitro study aims to evaluate cytotoxic and cytostatic effects of traditionally used as anti-cancer propolis from four different regions in Morocco. Propolis samples were collected from different locations in Morocco, which are characterized by different plant predominant vegetations. The resin, wax and balsam of hydroalcoholic extract of propolis content were identified, and the antioxidant content was analyzed with the use of HPLC and colorimetric methods. The antioxidant activity was assessed by DPPH, ABTS and ferric reducing power assays. Cells from the human colon carcinoma cell line (HCT 116) were treated for 24h (cytotoxic effects) and 72h (cytostatic effects) with increasing concentrations (0-1000 µg/ml) of methanol extracts from the four propolis (N1, N2, NC, PJ). Results obtained using the MTT assay indicate that N1, NC and PJ showed no significant difference between their cytotoxic and cytostatic effects with IC50 of 300, 750 and 700 µg/ml, respectively. A clear and highly significant cytostatic effects were measured with N2-extract with IC50 of 750 µg/ml. Furthermore, propolis with high resin and low wax content had high level of antioxidant compounds, and strong antioxidant and anticancer activities. These results indicate that the traditionally known anti-cancer effects of N2-extract might be mediated in part through cytostatic effects.


PP 19

Preliminary evaluation of the Cytotoxic Potential of north-West romanian Propolis

Constantin I. Mates1, Emoke Pall1, Marina Spinu1, Laura Stan2, Mihaela Niculae 1

1Department of Clinics, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania2Department of Food Science, Faculty of Food Science and Technology, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania

The propolis broad spectrum of therapeutic properties is documented by the literature1. Development of propolis based products requires a comprehensive evaluation of both efficacy and safety2. This research was aimed to evaluate the in vitro potential toxicity of North-West Romanian propolis ethanolic extracts previously studied for antimicrobial properties3. The cytotoxic potential was investigated considering propolis biocompatibility on human fibroblasts cell culture (cell line HFL-1) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and evaluating the cell morphology and attachment level. The MTT results expressed as optic density were further used to calculate the viability percentages by dividing the absorbance reading of cells under different propolis concentrations by the absorbance reading of cells under normal growth. The differences between values were analyzed using ANOVA post hoc, followed by Dunnett test (against the control) or by Bonferroni test (against different dilutions). Propolis samples were also characterized using spectrophotometric assays for the quantitative determination of flavonoids (flavones/flavonols, flavanones/dihydroflavonols) and total phenolics (Folin Ciocalteu method).While the spectrophotometric methods indicated the typical poplar composition profile with flavonoids and phenolic acids as main biological active compounds (total phenolics of 38.02% ±2.34%, high amounts of total flavonoids 9±0.3%, with 1.74-9.22% flavones/flavonols and 1.96-4.01% flavanones/dihydroflavonols), MTT test data suggested concentration dependence of propolis-induced effect. The highest dilutions stimulated cell viability (125-131.58%) and did not significantly impact the cell morphology and attachment levels, while the highest tested concentrations had moderate expressed cytotoxicity. Further in vitro and in vivo studies are intended to complete the cytotoxicity profile of the tested propolis samples. References:1 Dantas Silva RP, Machado BA, Barreto GA, Costa SS, Andrade LN, Amaral RG, Carvalho AA, Padilha FF, Barbosa JD, Umsza-Guez MA (2017) Antioxidant, antimicrobial, antiparasitic, and cytotoxic properties of various Brazilian propolis extracts. PLoS One 12(3): e0172585.2 Campos JF, dos Santos UP, Macorini LF, de Melo AM, Balestieri JB, Paredes-Gamero EJ, Cardoso CA, de Picoli Souza K, dos Santos EL (2014) Antimicrobial, antioxidant and cytotoxic activities of propolis from Melipona orbignyi (Hymenoptera, Apidae).Food Chem Toxicol.65: 374-80.3 Niculae M, Stan L, Pall E, Paștiu AI, Balaci IM, Muste S, Spînu M (2015) In vitro synergistic antimicrobial activity of Romanian propolis and antibiotics against Escherichia coli isolated from bovine mastitis. Notulae Botanicae, Horti Agrobotanici. 43(2): 327-334.


PP 20

the effect of Dietary Supplementation with Propolis on Bacteria Colonization Pattern in Gastrointestinal tract of Broiler Chickens

Ivan Miskulin1, Ivana Klaric2, Matija Domacinovic2, Berislav Prakatur3, Mirela Pavic4, Mario Ronta2, Maja Miskulin1

1 Faculty of Medicine Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 2 Faculty of Agriculture in Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 3 The Valpovka Fodder Factory, Valpovo, Croatia 4 Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia

Recent studies have suggested an increase in the production performance in poultry fed diets with propolis1,2. This advantages could be related to effect of propolis on gastrointestinal microbiota which increase the levels of beneficial bacteria and decrease the pathogenic types3. The aim of this study was to determine the total bacterial count, the count of Lactobacillus genera and the count of Enterobacteriaceae genera in ileum and crop of broiler chickens. This experimental study was conducted on 120 Ross 308 broiler chickens of equally distributed sex, which were randomly divided into three groups: control group (C) and two experimental groups of chickens (P1 and P2). Throughout the whole study (for 42 days) the control group of chickens was fed feed mixture while feed mixture that was fed to the experimental groups of chickens contained propolis in amount of 0,5 g/kg (P1 group) and 1,0 g/kg of feed mixture (P2 group). The study revealed statistically significant difference in number of Lactobacillus genera in ileum (p=0.036) between control and experimental groups of broiler chickens while there were no statistically significant differences in total bacterial count (p=0.351) and in number of Enterobacteriaceae genera (p=0.990). The study further showed statistically significant difference in number of Enterobacteriaceae genera (p=0.043) in crop between control and experimental groups of broiler chickens while there were no statistically significant differences in total bacterial count (p=0.066) and in number of Lactobacillus genera (p=0.558). One can conclude that dietary supplementation with propolis influence the colonization patterns of gastrointestinal tract of broiler chickens.

References:1 Zafarnejad K, Nazar A, Mostafa R (2017) Effect of bee glue on growth performance and immune response of broiler chickens. Journal of Applied Animal Research 459: 280-284.2 Khan SH (2017) Recent advances in role of propolis as natural additive in poultry nutrition. Bulgarian Journal of Apicultural Science 61: 167-183.3 Kačániová M, Rovna K, Arpášova H, Čuboň J, Hleba L, Pochop J, Kunová S, Hašèik P (2012) In vitro and in vivo antimicrobial activity of propolis on the microbiota from gastrointestinal tract of chickens. Journal of Environmental Science and Health, Part A: Toxic/Hazardous Substances and Environmental Engineering 47: 1665-1671.


PP 21

the effect of Propolis on Selected Blood Parameters of Broilers

Ivan Miskulin1, Ivana Klaric2, Matija Domacinovic2, Berislav Prakatur3, Zvonimir Steiner2, Vatroslav Seric1, Maja Miskulin1

1 Faculty of Medicine Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 2 Faculty of Agriculture in Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 3 The Valpovka Fodder Factory, Valpovo, Croatia

Recent studies have suggested that propolis is very important factor for the improvement of the performance as well as health of broilers1,2. The aim of this study was to determine the effect of propolis on selected biochemical blood parameters in broilers. This experimental study was conducted on 120 Ross 308 broiler chickens of equally distributed sex, which were randomly divided into three groups: control group (C) and two experimental groups of chickens (P1 and P2). Throughout the whole study (for 42 days) the control group of chickens was fed feed mixture while feed mixture that was fed to the experimental groups of chickens contained propolis in amount of 0,5 g/kg (P1 group) and 1,0 g/kg of feed mixture (P2 group). The study revealed statistically significant differences in average values of blood glucose, blood cholesterol, sodium, chlorides and calcium (p<0.001) as well as in average values of total proteins (p=0.039), globulins (p=0.011), triglycerides (p=0.034) and phosphorus (p=0.011) in chickens of the experimental groups compared to the chickens of the control group on the 21st day of fattening. The study also showed statistically significant differences in average values of blood cholesterol (p<0.001) and sodium (p=0.049) in chickens of the experimental groups compared to the chickens of the control group on the 42nd day of fattening. It can be concluded that the application of propolis as additive in broilers feeding enables the production of more vital and healthier animals, which furthermore, significantly improves the fattening of broilers.

References:1 Madras-Majewska B, Ochnio L, Ochnio M (2015) Use of bee products in livestock nutrition and therapy. Medycyna Weterynaryjna 71: 94-99.2 Shaddel-Tili A, Eshratkhah B, Kouzehgari H, Ghasemi-Sadabadi M (2017) The effect of different levels of propolis in diets on performance, gastrointestinal morphology and some blood parameters in broiler chickens. Bulgarian Journal of Veterinary Medicine 20: 215-224.


PP 22

intestinal Morphology Broiler Chickens Supplemented with Propolis

Maja Miskulin1, Ivana Klaric2, Matija Domacinovic2, Berislav Prakatur3, Mirela Pavic4, Nika Pavlovic1, Ivan Miskulin1

1 Faculty of Medicine Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 2 Faculty of Agriculture in Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 3 The Valpovka Fodder Factory, Valpovo, Croatia 4 Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia

The small intestine is an important organ responsible for digestion and absorption of nutrients from food1. Studies have demonstrated that certain bee products, such as propolis positively affect the small intestine villi making them longer and thicker, which has beneficial effect on the absorption of nutrients from food and consequently performance parameters of fattening chickens2,3. The aim of this study was to determine the influence of dietary supplementation of propolis on the intestinal morphology in broiler chickens. This experimental study was conducted on 120 Ross 308 broiler chickens of equally distributed sex, which were randomly divided into three groups: control group (C) and two experimental groups (P1 and P2). Throughout the whole study (for 42 days) the control group of chickens was fed feed mixture while feed mixture that was fed to the experimental groups of chickens contained propolis in amount of 0,5 g/kg (P1 group) and 1,0 g/kg of feed mixture (P2 group). The duodenal villi of chickens from the experimental groups were significantly higher in comparison to the duodenal villi of chickens from the control group (p=0.034) while there were no statistically significant differences in width of villi’s base and peak between the groups (p=0.114 and p=0.145, respectively). The duodenal villi crypts of chickens from the experimental groups were significantly wider (p=0.001) and deeper (p<0.001) in comparison to those parameters in chickens from the control group on the 42nd day of fattening. Addition of propolis to feed mixtures has significant protective effect on the gut tissue of chickens.

References:1 Toman R, Hajkova Z, Hluchy S (2015) Changes in intestinal morphology of rats fed with different levels of bee pollen. Pharmacognosy Communications 5: 261-264.2 Wang J, Li S, Wang Q, Xin B, Wang H (2007) Trophic effect of bee pollen on small intestine in broiler chickens. Journal of Medicinal Food 10: 276-280.3 Hajkova Z, Toman R, Galik B, Martiniakova M (2014) The effect of bee pollen consumption on functional morphology of small intestine of rats. Mendel Net 138-142.


PP 23

the influence of Propolis on liver Pathology in Broilers

Maja Miskulin1, Ivana Klaric2, Matija Domacinovic2, Berislav Prakatur3, Albina Dumic1, Daniela Cacic Kenjeric4, Ivan Miskulin1

1 Faculty of Medicine Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 2 Faculty of Agriculture in Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia 3 The Valpovka Fodder Factory, Valpovo, Croatia 4 Faculty of Food Technology Osijek, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia

One of the major problems in intensive fattening of chickens is liver damage1. Recent studies have shown that the addition of propolis into the broiler diet protected liver tissue against the adverse effects of various hepatotoxic factors1,2. The aim of this study was to investigate the influence of dietary supplementation of propolis on liver pathology in broilers. This experimental study was conducted on 120 Ross 308 broilers of equally distributed sex, which were randomly divided into three groups: control group (C) and two experimental groups of chickens (P1 and P2). Throughout the whole study (for 42 days) the control group of broilers was fed feed mixture while feed mixture that was fed to the experimental groups of broilers contained propolis in amount of 0,5 g/kg (P1 group) and 1,0 g/kg of feed mixture (P2 group). The study showed that the clusters of lymphocytes in the hepatocytes, the vacuolar degeneration, necrosis of the liver parenchyma, the bile ductule hyperplasia, and the various forms of pathological changes in the liver veins were more frequent in liver tissue samples of the control group compared to liver tissue samples of the experimental groups (p<0.001). The study further showed that the various forms of pathological changes in the liver arteries and steatosis of the liver parenchyma were more frequent in liver tissue samples of the control group compared to liver tissue samples of the experimental groups (p=0.023 and p=0.003, respectively). The supplementation with propolis has a strong protective effect on liver pathology in broilers.

References:1 Klaric I, Pavic M, Miskulin I, Blazicevic V, Dumic A, Miskulin M (2018) Influence of dietary supplementation of propolis and bee pollen on liver pathology in broiler chickens. Animals 8: 54.2 Bhadauria M, Nirala SK (2009) Reversal of acetaminophen induced subchronic hepatorenal injury by propolis extract in rats. Environmental Toxicology and Pharmacology 27: 17-25.


PP 24

Cytotoxic activity of Four Propolis Colombian Samples against Canine Osteosarcoma Cells

Dolly Patricia Pardo Mora1, Oscar Julián Murillo2, Mauricio Rey Buitrago1,2, Mónica Losada1, Jaime Fabian Cruz Uribe1, Karina Basso Santiago3, Bruno José Conti3, Eliza de Oliveira Cardoso3, Rafael María Gutiérrez1, Orlando Torres García1, José Maurício Sforcin3

1 Universidad Antonio Nariño, Bogotá, Colombia2 Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia 3 São Paulo State University (UNESP), Institute of Biosciences, Campus Botucatu, Brazil

The cytotoxicity of propolis against tumor cells has been widely reported1. Propolis from tropical areas present components such as triterpenes, diterpenes and benzophenones2, however, new types of propolis may be recognized and their biological activities need to be elucidated. Therefore, four Colombian propolis samples were chemically characterized by gas chromatography-mass spectrometry (GC–MS) and their cytotoxic action against canine osteosarcoma (OSA) cells was determined, as well as apoptosis induction and changes in gene expression of CASPASE 8 and CASPASE 9. OSA cells were treated with ethanolic extracts of propolis; the lactate dehydrogenase (LDH) assay and the annexin V/propidium iodide kit were used to evaluate the cytotoxic effect and apoptosis induction, respectively. Changes in gene expression were evaluated by quantitative real time PCR (qPCR). The propolis samples were rich in diterpenes, triterpenes and benzophenones. Propolis samples induced a cytotoxic effect towards OSA cells that was correlated to the LDH leakage into the culture medium after cell membrane rupture. The cytotoxic effect increased in a time-concentration manner after incubation with propolis and the cytotoxic concentration 50% (CC50) was lower than 36.24 µg/ml after 72 h of treatment. Colombian propolis samples induced total apoptosis significantly (untreated cells: 10.77 ± 2.22%; treated cells: from 58.403% ±1.39 to 95.97 ± 2.14% after 48 h). All propolis samples increased the expression of CASPASE 8 and three of them induced the expression of CASPASE 9. Colombian propolis samples induced apoptosis in canine OSA cells, suggesting their promising potential as news strategies for cancer therapy.

Acknowledgements: Universidad Antonio Nariño, Bogotá Colombia

References:1 Freires, I. A., de Alencar, S. M., & Rosalen, P. L. (2016). A pharmacological perspective on the use of Brazilian Red Propolis and its isolated compounds against human diseases. European Journal of Medicinal Chemistry, 110, 267-279.2 Castro V, Negri G, Salatino A, Bandeir M, (2011). A new type of Brazilian propolis: prenylated benzophenones in propolis from Amazon and effects against cariogenic bacteria. Food Chemistry, 125, 966-972.


PP 25

Portuguese Propolis: a Potenial Source of environmentally Friendly Fungicides

Catarina Passão 1, Claudia, Rodrigues 1, Cristina Almeida Aguiar 1,2,3,4, Ana Cunha 1,2,3,4

1 Biology Department, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), Biology Department, Minho University, 4710-057 Braga, Portugal3 CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.4 CBMA - Centre of Molecular and Environmental Biology, University of Minho, 4710-057 Braga, Portugal.

The need to increase food production to feed an increasingly growing world population has led to an intensive use of pesticides. Globally, 4.6 million tons of chemical pesticides are sprayed into the environment every year1. Out of the total consumption of pesticides, 17.5% are in the form of fungicides. However, pesticides have undesirable characteristics, such as the development of pathogen resistance and long degradation periods, and negative impacts on the environment, like soil and water pollution, bioaccumulation in the food chain, toxicity to non-target organisms and public health problems2. Therefore, new environmentally friendly natural products, which are effective, safe, biodegradable and leave no harmful residues are needed. Propolis is a natural resinous mixture made by honeybees (particularly Apis mellifera L.) from various plant sources and processed with salivary secretions. More than 300 different compounds have been identified so far in propolis, including several bioactive compounds such as flavonoids and phenolic acids and derivatives, which are characteristic of plant secondary metabolism and are related with diverse biological properties like antimicrobial and antioxidant activities3. In this work we evaluated the antifungal activity of ethanolic extracts (EE) from Portuguese propolis samples against phytopathogenic fungi species responsible for considerable damage in several economically important crops. Results showed that different propolis samples have different fungitoxic efficiencies but also that different species exhibit different susceptibilities. Globally, these findings suggest that propolis EE can be explored as source of biofungicides for application in a more sustainable agriculture management. In order to evaluate this propolis application, its phytotoxicity will be studied using in vitro flax plants (Linum usitatissimum L.).


Acknowledgments: This work is supported by: European Investment Funds by FEDER/COMPETE/POCI– Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013.

References:1 FAO (2017) Available at http://www.fao.org/faostat/en/#home (accessed July 2018).2 Ali EOM, Shakil NA, Rana VS, Sarkar DJ, Majumder S, Kaushik P, Singh BB, Kumar J (2017) Industrial Crops Products 108: 379-387.3 Fokt H, Pereira A, Ferreira AM, Cunha A, Aguiar C (2010) Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology 1: 481-493.

http://www.fao.org/faostat/en/#home


PP 26

Mechanistic Studies of Cytotoxicity induced by a Portuguese Propolis extract, using Saccharomyces cerevisiae as eukaryotic Cell Model

Hugo Alves1, Catarina Passão1, Maria João Sousa1,4, Cristina Almeida Aguiar1,2,3,4, Ana Cunha1,2,3,4 Rui Pedro Oliveira1,2,3,4

1 Biology Department, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), Biology Department, Minho University, 4710-057 Braga, Portugal3 CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.4 CBMA - Centre of Molecular and Environmental Biology, University of Minho, 4710-057 Braga, Portugal.

Propolis is a natural complex mixture produced by honey bees (particularly Apis mellifera L.) by collecting exudates from various plant sources. Characterized by a plethora of chemicals, propolis is generally rich in flavonoids, phenolic acids and terpene derivatives, bioactive compounds associated to it’s antimicrobial, anti-inflammatory, antimutagenic and antioxidant activities1. Previous work had shown that the ethanol extract of a sample from the Portuguese region of Beira Alta exhibited unique dual genotoxic and antigenotoxic effects using the yeast S. cerevisiae eukaryotic model2. In this work we prepared two ethanol extracts (EE) of propolis samples from Pereiro (P) - Beira Alta - collected in 2010 (P10.EE) and 2017 (P17.EE) to investigate the mechanisms of cytotoxicity and genotoxicity using specific S. cerevisiae mutants. While P17.EE didn’t show any toxic effect, yeast cells exposed to P10.EE showed a considerable decreased viability along time, assessed by colony-forming units. Interestingly, the oxidative stress response-defective mutant yap1 was more resistant than the wild type, suggesting that this cytotoxic effect was not mediated by oxidative stress. P.EE’s genotoxicity was also analysed by the nucleus-cytosolic translocation of NHP6A protein, considered a marker of necrosis. P10.EE induced NHP6A protein translocation to the cytoplasm, observed by fluorescence microscopy, suggesting that cytotoxicity of this extract was indeed mediated by necrosis. Although P17.EE didn’t seem to induce necrotic cell death, both extracts induced plasma membrane integrity loss, assessed by flow cytometry, using propidium iodide as marker. As recently observed for erythroleukemic cells with Brazilian propolis3, here we present the first evidence that also Portuguese propolis have necrotic-mediated cytotoxicity in yeast cells.


Acknowledgments: This work is supported by: European Investment Funds by FEDER/COMPETE/POCI– Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013.

References:1 Zabaiou N, Fouache A, Trousson A, Baron S, Zellagui A, Lahouel M, Lobaccaro JMA (2017) Chemistry and Physics of Lipids 207: 214-222.2 Cruz M, Antunes P, Paulo L, Ferreira AM, Cunha A, Almeida-Aguiar C, Oliveira R (2016) RSC Advances 55: 49806-49816.3 Bonamigo T, Campos JF, Alfredo TM, Balestieri JBP, Cardoso CAL, Paredes-Gamero EJ, Santos EL (2017) Oxidative Medicine and Cellular Longevity.


PP 27

direct Visualization of Artepillin C into fibroblast Cells via CarS Microscopy

Wallance M. Pazin1,2, Vita Solovyeva1, Tibebe Lemma2, Bjarke Jørgensen3, Carlos José L. Constantino2, Jonathan Brewer1

1 Department of Biochemistry and Molecular Biology (BMB), University of Southern Denmark, Odense, Denmark2 Department of Physics, School of Sciences and Technology, São Paulo State University, Presidente Prudente – SP, Brazil3 NewTec Engineering A/S, Odense, Denmark

Artepillin C, the major component of the Brazilian green propolis, has become the subject of great biological interest among all the secondary metabolites found in that propolis, especially due to its anticancer property. Biophysical studies have shown that the presence of two prenylated groups in the molecular structure of Artepillin C enhances its affinity for lipophilic environment1, which is in agreement with biological assays reported in the literature, suggesting that Artepillin C is absorbed through cell membranes2. The purpose of this study was to obtain for the first time a direct visualization of Artepillin C internalization into fibroblast cells by means of Coherence Anti-Stokes Raman Scattering (CARS), a powerful microscopy technique with a combination of two pulsed laser beams which induces a vibrational motion of a specific compound, generating an intrinsic image3. A detailed vibrational characterization of Artepillin C has been carried out using Raman microspectroscopy and surface-enhanced Raman scattering (SERS) in combination with quantum chemical calculation based on LANL2DZ/B3LYP level of theory. The success use of CARS microscopy was confirmed by collecting Artepillin C images from dry and hydrated films of the compound, while the intrinsic images of Artepillin C internalized into the cells was compared to confocal fluorescence microscopy images by using a cell-membrane staining compound. With effect, this study shows for the first time a direct visualization of the major compound of green propolis into the cells by means of CARS microscopy.

Acknowledgements: FAPESP (2017/23426-4) and DaMBIC (University of Southern Denmark).

References:1 Pazin, W. M. et al. (2017) Interaction of Artepillin C with model membranes. Eur. Biophys. J. 46: 383–393.2 Paulino, N. et al. 2008 Anti-inflammatory effects of a bioavailable compound, Artepillin C, in Brazilian propolis. Eur. J. Pharmacol. 587: 296–301.3 Evans, C. L. & Xie, X. S. (2008) Coherent Anti-Stokes Raman Scattering Microscopy: Chemical Imaging for Biology and Medicine. Annu. Rev. Anal. Chem. 1: 883–909.


PP 28

Qualitative analysis and Biological evaluation of Propolis from armenia and georgia

Argyro Petropoulou1, Ioanna Chinou1, Konstantia Graikou1

1Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece

Propolis is a bee-keeping product of increasing interest as it has been used widely in food industry as well as in cosmetics; while its chemical composition is highly variable and depends mainly on the local flora as well as the geographic and climatic differences in the collection sites. Therefore it is necessary to characterize propolis, chemistry in order to confirm quality and safe use. As a part of our systematic research on different propolis from all over the world, we report in this study the chemical analyses of two Euroasian samples from Armenia and Georgia respectively, which to the best of our knowledge have not been studied before. All chemical analyses were performed by GC/MS using silylation procedure1. According to our results, both samples showed the characteristic identity profile of a typical propolis from Temperate zone2, as they contained mainly aromatic acids (p-coumaric, caffeic and ferulic acids), esters of phenolic acids (pentenyl and phenylethyl caffeates), flavonoids and chalcones (galangin, pinocembrin, pinobanksin and its 3-O-acetate, as well as pinocembrin, pinobanksin and pinostrobin chalcones). Furthermore, their total phenolic content was determined by the Folin-Ciocalteu method showing comparable high phenolics in both samples, which is in accordance with the results from GC-MS analysis. The ethanolic extracts were also assayed for their antimicrobial activity against nine human pathogenic bacteria and fungi, showing an interesting profile, while they exerted a more specific activity towards the Gram positive studied strains.

Acknowledgments: The authors would like to thank Prof. Alexander Panossian (Head of Research & Development – Swedish Herbal Institute, Sweden) for providing the Armenian propolis sample.

References:1 Popova Μ., Chinou Ι., Marekov Ι. and Bankova V. (2009). Terpenes with antimicrobial activity from Cretan propolis.Phytochemistry, 70: 1262-1271.2 Bankova V., de Castro S.L. and Marcucci M.C. (2000). Propolis: recent advances in chemistry and plant origin.Apidologie 2000, 31, 3-15.


PP 29

Chemical Composition of Selected Propolis Samples from Kyrgyzstan and Kazakhstan

Argyro Petropoulou1, Konstantia Graikou1, Jaroslaw Widelski2, Krystyna Skalicka-Woźniak2, Zuriyadda Sakipova3, Ioanna Chinou1

1Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece2Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Poland3School of Pharmacy, Asfendiyarov Kazakh National Medical University, Almaty, Kazakhstan

Propolis (bee glue) is a well known natural product with healing properties. The chemical composition of propolis is highly variable and depends mainly on the local flora. As a part of a systematic research on bee-keeping products, two propolis samples from Kyrgyzstan and Kazakhstan have been studied for the first time due to their high and very interesting flora. Chemical analysis of propolis samples was performed by GC/MS which is a very useful and highly appreciated analytical tool, after silylation procedure1. The results showed that both samples contain mainly aromatic acids (cinnamic, caffeic and ferulic acids), esters of phenolic acids (pentenyl and phenylethyl caffeates), flavonoids (galangin, pinocembrin, pinobanksin) as well as chalcones of pinostrobin, pinocembrin and alpinon. These results confirm a typical chemical profile characteristic for Temperate zone propolis2 while in both cases appear also an uncommon aliphatic acid content (oleic, linoleic, palmitic and stearic acids) . Furthermore, both samples showed a high TPC (total phenolic content), determined by the Folin-Ciocalteu method. Additionally, the ethanolic extracts were evaluated for their antimicrobial activity, by the agar dilution technique against a panel of human pathogenic microorganisms. The assays revealed broad spectrum of antimicrobial activity specified against Gram positive bacteria as well as against oral pathogenic ones.

References:1 Popova Μ., Chinou Ι., Marekov Ι. and Bankova V. (2009). Terpenes with antimicrobial activity from Cretan propolis.Phytochemistry, 70: 1262-1271.2 Bankova V., De Castro S.L., Marcucci M.C.: Apidologie 2000, 31, 3-15.


PP 30

Caseinates loaded with red Propolis extract

Isabel Cristina Celerino de Moraes Porto1*, Clinston Paulino de Almeida1, Nataly Miranda do Nascimento1, Amanda Barbosa Wanderley1, João V. Lessa de Oliveira1, Felipe J. L. Barbosa dos Santos1, Valdemir da Costa Silva1, Irinaldo Diniz Basílio-Júnior1, Giselda Macena Lira1, Marta Maria da Conceição2, Ana Flávia Oliveira dos Santos3, Ticiano Gomes do Nascimento1*

1 Post-Graduate Program of Pharmaceutical Sciences, School of Nursing and Pharmacy and Post-Graduate Program of Nutrition, College of Nutrition, Federal University of Alagoas, Maceió, Brazil

2 Department of Food Technology, Federal University of the Paraíba, João Pessoa, Brazil3 Department of Pharmacy, University Centre of Maceió/CESMAC, Maceió, Brazil* E-mail: [email protected]; * E-mail: [email protected]

This work aimed to characterize microcapsules of caseinates loaded with red propolis extract (RPE) using different analytical and biological methods. Five compositions of microcapsules of caseinates loaded with RPE at concentrations between 10% and 50% were prepared using sodium casein from bovine milk as encapsulating agent and colloidal silicon dioxide as a flow agent. The caseinates were produced using colloidal dispersion technique and submitted to the spray-drying technique. The caseinates were characterized using SEM analysis, thermal analysis, FTIR, Fluorescence analysis, electrophoresis, dissolution studies, antioxidant and antibacterial activities. SEM images showed spherical particles in the range from 3.24 μm to 20.5 μm. FTIR, NMR spectra in solid-state and thermal analysis also demonstrated the encapsulation of the RPE. Dissolution study revealed limited to poor water-solubility of flavonoids in the compositions CSRPE18% and CSRPE28%, but good water solubility of naringenin in the compositions CSRPE35% and CSRPE50%. Excellent antioxidant activity and antibacterial activity were demonstrated only from CSRPE50% and CSRPE35% compositions. Thermal analysis, water-solubility dissolution assay and antioxidant activity have indicated a high protein binding between sodium casein and flavonoids from RPE in the CSRPE18% and CSRPE28% compositions resulting in a controlled release of the flavonoid. However immediate release of flavonoids in the compositions CSRPE35% and CSRPE50% was detected. The Fluorescence and thermal analysis allied to the dissolution, antioxidant and antibacterial tests were important analytical techniques to elucidate the interaction between flavonoid-protein and flavonoid release kinetic from caseinates soluble complex.

Acknowledgments. The authors are grateful to CNPq, CAPES, FINEP and FAPEAL for financial support.




PP 31

determination of Propolis Origin using Phenolic Composition and Artificial Neural Networks

Isabel Revilla1, Ana María Vivar-Quintana1, Pedro Hernández-Ramos2

1Food Technology. University of Salamanca, E.P.S. de Zamora, Zamora, Spain.2Graphic Expression in Engineering. University of Salamanca, E.P.S. de Zamora, Zamora, Spain.

Different studies have reported that propolis have some specific compounds depending on the origin such as prenylated derivatives of p-coumaric acid in Brazilian propolis. However, the basic composition is quite similar for all of them, 50 % of phenolic compounds, 30 % wax, 10 % essential oils and 5 % pollen. Taking it into account, the aim of this work was to study if it is possible to classify propolis samples according with their origin on the basis of their phenolic composition and antioxidant properties determined by standard methods. To do this, the total contents of flavones and flavonols, flavanones and dihydroflavonols, and the antioxidant capacity by using the methods of ABTS and linoleic acid/β-carotene were analysed in 94 samples of propolis from Spain (Galicia n=14 and Castilla y León n=31 regions) and Chile (Biobio region n=49). For classification purposes, a multi-layered feed-forward network with back-propagation was used for predicting the geographical origin of propolis. The weight, the seed value and bias matrix were randomly initialized. The input layer had seven neurons (analysed parameters) and seventeen or seven neurons in the hidden layer. Two (Chile or Spain) or three (Chile, Castilla y León or Galicia) nominal output variables were used for classification tasks. The results of the ANN showed that 93.6% of the samples were correctly classified according with country of origin. However if we wanted to classify the samples also according the region (Castilla León and Galicia) the number of samples correctly classified were the 82%.


PP 32

Determination of Phenolic acids in raw Propolis using near infrared Spectroscopy

Ana María Vivar-Quintana 1, Isabel Revilla 1, María Inmaculada González-Martín2, Eddy Valentín Betances-Salcedo 2

1 Food Technology. University of Salamanca, E.P.S. de Zamora, Zamora, Spain. 2 Department of Analytical and Food Chemistry, Faculty of Chemistry, University of Salamanca, Salamanca, Spain.

The objective of this study is the assessment of the viability of using Near-Infrared Spectroscopy (NIR) technology to determine the phenolic composition in propolis samples The chemical composition and the floral origin of propolis are closely related, samples from different geographic and climatic areas varies greatly depending on the local flora and site of collection1. The identification and quantification of the phenolic compounds is therefore of great interest as they may affect its biological activities and consequently its clinical properties. HPLC is the most frequently used technique for the characterisation of phenolic compounds2. (NIR) is known for being a multiparametric, rapid, and non destructive technique. It has been used to determine the phenolic composition in different extracts3. The most determinant aspect of this methodology is that it could be developed and applied to any type of raw propolis of varying origins, grinding up the samples without destroying the propolis. The HPLC method was applied to the analysis of phenolic acids in 50 propolis extracts. It was possible to identify and quantify 6 phenolic acids: caffeic acid, p-coumaric acid, cinnamic acid, ferulic acid, Isoferulic acid and 3,4 dimethyl-caffeic acid (DMCA). The results show that we can use the NIR methodology to determine caffeic acid, cinnamic acid, ferulic acid, isoferulic acid and 3,4 dimethyl-caffeic acid (DMCA) with values comparable to those obtained by HPLC.

References:1Frizon, C.N.T., Oliveira, G. A., Perussello, C. A., Peralta-Zamora, P.G., Camlofski, A.M.O., Rossa, Ü.B., & Hoffmann-Ribani. R. (2015). Determination of total phenolic compounds in yerba mate (Ilex paraguariensis) combining near infrared spectroscopy (NIR) and multivariate analysis. LWT - Food Sci. Technol. 60, 795-801.2Gómez-Caravaca, A.M., Gómez-Romero, M., Arráez-Román, D., Segura-Carretero, A., & Fernández-Gutiérrez, A. (2006). Advances in the analysis of phenolic compounds in products derived from bees. J. Pharm. Biomed. Anal. 41, 1220–1234.3Valencia, D., Alday, E., Robles-Zepeda, R., Garibay-Escobar, A., Galvez-Ruiz, J.C, Salas-Reyes, M., Jiménez-Estrada, M., Velazquez-Contreras, E., & Hernández. J. (2012) Seasonal effect on chemical composition and biological activities of Sonora propolis. Food Chemistry 131, 645-651.


PP 33

the Biological activities of indonesian Propolis and it’s Molecular Marker

Muhamad Sahlan1,2, Diah Kartika Pratami3, Safira Candra Asih1, Andrea Devina1, Alfiani Guntari Mahadewi1, Masafumi Yohda4, Siti Farida5,2, Robiatul Adawiyah5,2, Herbert Situmorang5,2, Heri Hermansyah1,2, Anondho Wijanarko1

1 Laboratory of Industrial Bioprocess, Department of Chemical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, Indonesia, [email protected] 2 Research Centre for Biomedical Engineering, Faculty of Engineering, Universitas Indonesia, Depok, Indonesia3 Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia 4 Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Tokyo, Japan. 5 Faculty of Medicine, Universitas Indonesia, Depok, Indonesia

Propolis known have several biological activities1. Recently, we found that the stingless bee cultivated in east Indonesia produced a lot propolis. The source of resin for the propolis production, still unknown. To confirm that the Indonesian propolis have similar activities with other propolis, we performed several study related to the biological activities of the propolis such as, the anti-Candida albicans, anti-inflammatory, and anti-oxidant and it molecular marker, were studied. The results showed that the Indonesian Propolis have anti-Candida albicans2, anti-inflammatory and anti-oxidant activities3. About 7, 7, and 35 molecules are found as molecular marker for their activities, respectively. The molecules are different with other propolis. Acknowledgements: The author would like to thank the financial support from DRPM UI Grant of Indexed International Publication of Student Final Project (Publikasi International Terindeks Untuk Tugas Akhir Mahasiswa/PITTA) 2018.

References: 1 Sonia MF, Marcia CDF, Daiana TO, Marina BM, Sidney AVF, Rachel BC (2018) Biological Activities of Red Propolis: A Review. Recent Patents on Endocrine, Metabolic & Immune Drug Discovery (in press).2 Mahadewi AG, Christina D, Hermansyah H, Rohmatin E, Sahlan M (2018) Selection of discrimination marker from various propolis for mapping and identify anti Candida albicans activity. AIP Conference Proceedings 1933, 0200053 Pratami DK, Mun’im A, Sundowo A, Sahlan M (2018) Phytochemical profile and antioxidant activity of propolis ethanolic extract from tetragonula bee. Pharmacognosy Journal 10 (1) :128-135


PP 34

antioxidant activities of Propolis from aragón (Spain)

M. Teresa Sancho1, Sandra M. Osés1, Andrea Rodríguez-Fernández1, Miguel A. Fernández-Muiño1, Consuelo Pérez-Arquillué2, Regina Lázaro2, Susana Bayarri2

1 Nutrition and Bromatology Unit. Department of Biotechnology and Food Science. Faculty of Sciences. Universidad de Burgos. Plaza Misael Bañuelos s/n. 09001 Burgos, Spain2 Nutrition and Bromatology Unit. Department of Animal Production and Food Science. Faculty of Veterinary Medicine. Instituto Agroalimentario de Aragón -IA2- (Universidad de Zaragoza-CITA). Miguel Servet, 177, 50013 Zaragoza, Spain

Propolis has demonstrated a strong antioxidant activity1. This property, together with other compositional parameters2 could contribute to the difficult standardization of propolis. Antioxidant activity should be determined by methods based on different mechanisms, and against several free radicals3. The purpose of this research was to help characterize propolis from Aragón (Spain) by determining the oxygen radical absorbance capacity (ORAC)4, trolox equivalent antioxidant capacity (TEAC)5, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay6, as well as the radical-scavenging effect on hydroxyl radicals (AOA)5 on hydroalcoholic extracts of eight Aragonese propolis samples. Propolis extracts were obtained with 70% (V/V) ethanol. All extracts showed high antioxidant activities. The following averages were obtained: For ORAC 11070 µmol trolox/g, for TEAC 1040.7 µmol trolox/g, for IC50 DPPH 0.0698 mg/ml and for AOA 0.8783 mmol uric acid/g. At the 95.0% confidence level, significant linear correlations were found between ORAC and TEAC (r= 0.9436), ORAC and AOA (r= 0.9142) and TEAC and AOA (r= 0.9919) and a double reciprocal correlation between TEAC and DPPH (r= 0.7403). The lack of correlation between ORAC and AOA with DPPH results is likely due to the fact that the antioxidant compounds of Aragonese propolis could react very fast with both peroxyl (ORAC assay) and hydroxyl (AOA assay) radicals, and at a slow pace to DPPH due to steric inaccessibility3. Nevertheless, further studies are necessary to corroborate this conclusion.


Acknowledgements: To all the beekeepers who have provided with a free sample of propolis for this study.

References:1 Daleprane JB, Abdalla DS (2013) Emerging Roles of Propolis: Antioxidant, Cardioprotective, and Antiantiogenic Actions. Evidence-Based Complementary and Alternative Medicine 2013: 1-8.2 Bankova V. (2005) Chemical diversity of propolis and the problem of standardization. Journal of Ethnopharmacology 100: 114-117.3 Prior RL, Wu X, Schaich K, (2005) Standardized Methods for the Determination of Antioxidant Capacity and Phenolics in Foods and Dietary Supplements. Journal of Agricultural and Food chemistry 53: 4290-4302.4 Huang D, Ou B, Hampsch-Woodill M, Flanagan JA, Prior RL (2002) High-Throughput Assay of Oxygen Radical Absorbance Capacity (ORAC) Using a Multichannel Liquid Handling System Coupled with a Microplate Fluorescence Reader in 96-Well Format. Journal of Agricultural and Food chemistry 50: 4437–4444.5 Osés SM, Pascual-Maté A, Fernández-Muiño MA, López-Díaz TM, Sancho MT (2016) Bioactive properties of honey with propolis. Food chemistry 196: 1215-1223.6 Miguel MG, Nunes S, Dandlen SA, Cavaco AM, Antunes MD (2010) Phenols and antioxidant activity of hydro-alcoholic extracts of propolis from Algarve, South of Portugal. Food and Chemical Toxicology 48: 3418-3423.


PP 35

Formulations Containing Propolis of Apis mellifera with antibacterial and antitumoral activity

Dos Santos, G.A.1, Lagareiro Netto, A.A.1, Mendes, P.B., 2, Negrão, V.3, Marcucci, M.C.1, Diniz, S.N.1, Carneiro, M.J. 4, Sawaya, A.C.H.F.5

1 Stricto sensu Postgraduate Program, Anhanguera University of São Paulo, SP, Brazil, [email protected],2 Graduate School of the School of Medicine of Santa Casa, SP, Brazil,3 Green Propolis Eucalypt Ltda., SP, Brazil, [email protected],4 Institute of Biology, State University of Campinas, 13083-970 Campinas - SP, Brazil.5 Faculty of Pharmaceutical Science, UNICAMP, SP, Brazil, [email protected].

Propolis, a resin produced by A. mellifera bees, presents several compounds of pharmaceutical interest and, further studies to demonstrate their pharmacological actions is sought. Capsules containing green propolis (BRPX) extracted in flax oil (1 and 2) polyethylene glycol (PEG) (3 and 4) were analyzed and analyzed according to IN 31. The following parameters were evaluated: total phenols and flavonoids, quantification of Artepillin-C (HPLC) confirmed by UHPLC-MS, antitumor activity in MCF-7 (breast code 0162) and B16F10 (melanoma code 0046) and antimicrobial activity (E.coli, C. albicans and S .aureus). The flavonoid content (%) of each capsule was 1: 1.135 ± 0.009, 2: 0.521 ± 0.039, 3: 0.758 ± 0.020, 4: 1.232 ± 0.09. The phenol content (%) of 1: 2,509 ± 0,024, 2: 2,280 ± 0,075, 3: 1,039 ± 0,029, 4: 3,426 ± 0,074. The content of Artepillin-C (mg / capsule) of 1: 1.39 ± 0.12, 2: 1.07 ± 0.05, 3: 0.66 ± 0.01, 4: 0.39 ± 0, 01. The IC50 (μg / mL) for MCF-7 was: 1: 100.49 ± 6.41, 2: 463.65 ± 0.71, 3: 209.03 ± 35.99, 4: 238.84 ± 4 , 16 and for B16F10, 1: 277.53 ± 7.37, 2: 357.48 ± 10.15, 3: 376.15 ± 5.92, 4: 286.87 ± 10.79. Inhibition halos ranged from 10 to 14 mm (E.coli), 8 to 10 mm (C. albicans) and 8 to 9 mm (S.aureus). This study demonstrates the importance of quality control for reliable formulations based on propolis.

References:1 BRASIL (2001). MINISTRY OF AGRICULTURE. Normative Instruction nº 3 - ANNEX VII - Technical regulation for determination of identity and quality of propolis. Diário Oficial da República Federativa do Brasil.


PP 36

Flavonoids Constituents of algeria Propolis

Segueni Narimane1, Jesus. G. Diaz2, Akkal Salah3, Rhouati Salah1

1Laboratoire des produits naturels d’origine végétale et de synthèse organique. Département de chimie, Université Constantine 1. Constantine. Algérie2 Departmento de Quımica Organica y Instituto Universitario de Bio-organica “Antonio Gonzalez”, Universidad de La Laguna, 38206 La Laguna, Spain3Laboratoire de Phytochimie et Analyses Physico-chimiques et Biologiques, Département de chimie, Université Constantine 1. Constantine. Algérie

Propolis is acomplex material collected by honeybees from plant and tree buds, resins and exudates. In the last recent years several research indicated that Algerian propolis possess a large spectra of biological activities such as antibacterial, antifungal, antioxidant and anticancer activities. Few studies on chemical composition of Algerian propolis indicate the presence of two propolis type mainly poplar1 and Mediterranean type2,3 with caffeate esters and flavonoids as main constituents of poplar type while Mediterranean type is rich on labdane and clerodane diterpes1. The objective of the present study is the determination of chemical composition of propolis sample collected from jijel. Chemical investigation allowed the isolation of 12 Compounds. Their structures were identified on the basis of spectral data (1H-NMR and 13C-NMR) and MS data and comparison with literature. All isolated compounds are flavonoids. Among the identified flavonoids pinocembrin, pinostrobin and galangin-3-methyl ether indicate that the studied propolis is a polpar type propolis.

References1 Piccinelli AL, Mencherini T, Celano R, Mouhoubi Z, Tamendjari A, Aquino RP, Rastrelli L (2013) Chemical Composition and Antioxidant Activity of Algerian Propolis. Journal of Agricultural and Food Chemistry. 61: 5080−5088.2 Graikou K, Popova M, Gortzi. O, Bankova V, Chinou I (2016) Characterization and biological evaluation of selected Mediterranean propolis samples. Is it a new type?. LWT - Food Science and Technology 65: 261-267. 3 Velikova M, Vassya Bankova V, Sorkunb K, Houcinec S, Tsvetkovad I, Kujumgiev A (2000) Propolis from the Mediterranean Region: Chemical Composition andAntimicrobial Activity. Z. Naturforsch : 55c, 790-793


PP 37

Antimicrobial efficacy of Some Products with Propolis Hydroalcoholic extract 30% and antiviral Synthesis

Carmen Violeta Popescu*1, 2), Hazem Abbas3), Ştefan Manea1), Lili Ivopol 1)

, Alina

Dune 1), Candice Popiniuc4), Dumitru Lupuleasa3)

1)S.C. Hofigal Export Import S.A., Bucharest, Romania 2)Faculty of Pharmacy, “Vasile Goldiş” Western University, Arad, Romania 3)University of Medicine and Pharmacy „Carol Davila” - Faculty of Pharmacy, Bucharest, Romania 4)Innergy (SC Inner Chi Nature srl), Constanta, Romania - working point, Voluntari, Romania

The aim: The purpose of this study is to verify and prove antimicrobial efficacy of two topical pharmaceutical finished products, a gel and a cream, with propolis hydro alcoholic extract 30% 1 and an antiviral synthesis used as active pharmaceuticals ingredients. These products are intended especially in the treatment of herpes or any of a group of viral diseases caused by herpes viruses, affecting the skin (often with blisters). Methods and Materials: The study of antimicrobial efficacy was performed in accordance with the provisions of the European Pharmacopoeia, 9th edition, version in force, method described in Chapter 5.1.3. Efficacy of antimicrobial preservation and acceptance criteria for preparations for cutaneous use2 and the specialty literature.The sample under study was forcedly contaminated with suspensions (105/mL) of Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus brasiliensis and the times studied were 0, 2, 7, 14, 21, 28 days. Results: Acceptance criteria for antimicrobial efficacy are presented as a logarithmic reduction in the number of viable microorganisms, calculate the ratio between the number of UFC/ mL from baseline (T0) and the number of UFC/ mL determined for each test time and have been fully met. Conclusions: The pharmacopoeia requirements were entirely met for the gel product, and the cream did not prove efficacy on the Candida albicans microorganism.

References1Abbas H., Popescu V.C., Crisan I., Popa M., Chifiriuc M. C., Gradisteanu Pircalabioru G., Lupuliasa D., Farmacia, 2017, Antibacterial Efficiency of five propolis extract on planktonic and adherent microbial strains, Vol. 65, 5: 813-818, ISSN: 0014-8237 (for the Printed Edition)ISSN: 2065-0019 (for the On-Line Edition).2 European Pharmacopoeia 9th ed. (2017), Strasbourg, FR: European Directorate for the Quality of Medicines.


PP 38

“green” Synthesis of gold and Silver nanoparticles with Propolis extract and Rosa damascena Waste

Anton M. Slavov1, Daniela Karashanova2, Biliana Georgieva2, Ivelina Vasileva1, Vassya Bankova3, Rada Dinkova4, Nikoleta Yantcheva1

1 Department of Organic Chemistry and Inorganic Chemistry, Technological Faculty, University of Food Technologies, Plovdiv, Bulgaria2 Institute of Optical Materials and Technologies, Bulgarian Academy of Sciences, Sofia, Bulgaria3 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Plovdiv, Bulgaria4 Department of Canning Technologies, University of Food Technologies, Plovdiv, Bulgaria

Propolis has been used in the traditional medicine for more than 2000 years. It is a treasured medicine due to its antifungal, antiviral, antitumor, immunomodulatory and anti-inflammatory properties. Additionally, due to its antibacterial and antioxidant activity, propolis is a potential candidate for “green” synthesis of nanoparticles (NPs) or/and composite materials with potential medical applications1. Hence, the aim of the present study was focused on biogenic synthesis of gold and silver nanoparticles with propolis extract. A commercial Bulgarian poplar propolis extract was used. The composition of the extract was investigated by GC-MS. The synthesis was followed by visual observations and UV-Vis studies. The morphology and characteristics of the NPs were determined by transmission electron microscopy (TEM). Both, gold and silver nanoparticles were successfully obtained in situ in a simple, one stage process, with the propolis extract. Compared to plant extract from Rosa damascena waste (able to reduce the Ag+ and Au3+ in 2-3 minutes), the visual process of synthesis took 120 and 3 minutes, for silver and gold nanoparticles, respectively. The TEM studies confirmed the АgNPs and AuNPs synthesis with Bulgarian propolis extract. The GC-MS analysis revealed that the propolis was rich in flavonoids (pinobaksin, pinocembrin, chrysin, quercetin, kaempferol and their derivatives). The present study clearly demonstrated the potential of propolis extract for “green” synthesis of gold and silver NPs.

Acknowledgements: We acknowledge the financial support from the National Science Fund of Bulgaria; project DN 17/22 “Valorization and application of essential oil industry wastes for “green” synthesis of metal nanoparticles”.

References:1 Matei PM, Martín-Ramos P, Sánchez-Báscones M, Hernández-Navarro S, Correa-Guimaraes A, Navas-Gracia LM, Rufino CA, Ramos-Sánchez MC, Martín-Gil J (2015) Synthesis of chitosan oligomers/propolis/silver nanoparticles composite systems and study of their activity against Diplodia seriata. International Journal of Polymer Science Article ID 864729, 11 pages, http://dx.doi.org/10.1155/2015/864729


PP 39

Quantitative and Qualitative variations in Propolis Collection

C. Tananaki1, A. Thrasyvoulou1, D. Kanelis1, V. Liolios1

1Laboratory of Apiculture-Sericulture, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, Greece, e-mail: [email protected]

Propolis is a natural material that honeybees produce from resinous secretions of the plants and wax. It is used from bees to protect their hive from adverse external conditions and pathogens. Because of the bioactive compounds included in propolis, it is used as therapeutic agent. The medical applications of this bee hive product led to an increased interest in its production and chemical composition. In this research, propolis samples collected during the whole active beekeeping period for three years and the variations caused to season and type of collection investigated. Parameters such as the yield per hive, the wax content, the dry matter, the total phenols and antioxidant activity of the extracts were evaluated. The results showed a strong seasonal variation, with the maximum production have been recorded between May and July. Bees collect/produce more propolis during the warm periods of the year because of the viscosity of the resins. When they need more product, bees cover the scarcity of the resinous material using more wax. Quantitative and qualitative variations were found between the two different propolis traps and among bee hives in the same apiary. Moreover, propolis samples were collected from different regions of Greece and the antioxidant activity and total phenolic content of their alcoholic extracts were compared. High variance in the studied parameters was observed among the samples. The collection and quality of propolis is related to the local flora as well as to the beekeeping practices.


PP 40

isolated triterpenes from Stingless Bee Lisotrigona furva Propolis in vietnam

Le Nguyen Thanh1, Vu Thi Kim Oanh1, Ha Thi Thoa1, Diep Thi Lan Phuong 2, Nguyen Thi Phuong Lien3, Tran Huu Giap1, Nguyen Thi Minh Hang1, Nguyen Van Hung1, Vassya Bankova4

1 Institute of Marine Biochemistry, Vietnam Academy of Science and Technology, Hanoi,Vietnam 2Department of Chemistry, Quy Nhon University, Binh Dinh, Vietnam3 Institute of Ecological and Biological Resources, Vietnam Academy of Science and Technology, Hanoi,Vietnam4 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria

Stingless bees of the genus Lisotrigona are uncommon species found in India and South East Asian countries. Several Lisotrigona species including L. cacciae, L.carpenteri and L. furva from Vietnam have been described. There has been no information on the chemical constituents and bioactivity of Lisotrigona stingless bee propolis in Vienam. Our goal of this work is to investigate the chemical constituents of the stingless bee Lisotrigona furva propolis collected in Binhdinh province, Vietnam. Using combined chromatographic methods, several triterpenes including mangiferolic acid (1), occotilones I (2) and II (3), dipterocarpol (4), hydroxyhopanone (5) and (13E,17E)-polypoda-7,13,17,21-tetraen-3β-ol (6) were isolated from the ethyl acetate extract of Lisotrigona furva propolis. The chemical structures of isolated compounds were identified by MS, NMR spectral analysis. Compound 5 and 6 were isolated for the first time from stingless bee propolis.

Hydroxyhopanone (13E,17E)-polypoda-7,13,17,21-tetraen-3β-ol

OH

H

H H

OH

H

HHO


Acknowledgements: This research is funded by Vietnam Academy of Science and Technology under grant number VAST.HTQT.BULGARIA.02/17-18.

References: 1 Nguyen, H. X., Nguyen, M. T., Nguyen, N. T., & Awale, S. (2017). Chemical Constituents of Propolis from Vietnamese Trigona minor and Their Antiausterity Activity against the PANC-1 Human Pancreatic Cancer Cell Line. Journal of natural products, 80(8), 2345-2352.2 Sanpa S., Popova M., Bankova V., Tunkasiri T., Eitssayeam S., Chantawannakul P. (2015). Antibacterial compounds from propolis of Tetragonula laeviceps and Tetrigona melanoleuca (Hymenoptera: Apidae) from Thailand. PLoS One, 10(5), e0126886.3 Yoon, N. Y., Min, B. S., Lee, H. K., Park, J. C., & Choi, J. S. (2005). A Potent Anti-complementary acylated sterol glucoside fromOrostachys japonicus. Archives of pharmacal research, 28(8), 892.


PP 41

isolated Xanthones from Lisotrigona furva Propolis in vietnam

Le Nguyen Thanh1, Ha Thi Thoa1, Vu Thi Kim Oanh1, Diep Thi Lan Phuong2, Hoang Thi Van3,4, Nguyen Quynh Chi4, Tran Huu Giap1, Nguyen Thi Tu Oanh1, Nguyen Thi Minh Hang1, Nguyen Van Hung1, Chau Van Minh1, Vassya Bankova5

1 Institute of Marine Biochemistry, Vietnam Academy of Science and Technology, Hanoi,Vietnam 2Department of Chemistry, Quy Nhon University, Binh Dinh, Vietnam3 Department of Pharmacy, Duytan University, Danang, Vietnam4 Hanoi University of Pharmacy, Hanoi, Vietnam5 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria

Propolis from stingless bees in Vietnam has been used in traditional medicine to improve health and prevent diseases. However, there have been few investigations on the chemical constituents and bioactivity of Vietnamese stingless bee propolis. This work studied the chemical composition of the stingless bee propolis collected from the nests of Lisotrigona furva in Binhdinh province, Vietnam using combined chromatographic methods. The chemical structures of isolated compounds were determined by MS, NMR spectral analysis. Several xanthones including cochichinone A, cochichinone I, cochichinone J, cratoxylumxanthone B, α-mangostanin and pruniflorone S were isolated from the ethyl acetate extract of Lisotrigona furva propolis. This is the first report on the chemical constituents of propolis from Lisotrigona species. Cratoxylum cochinchinense (Hypericaceae) is the plant source for the stingless bee Lisotrigona furva propolis.

OOH

O O

OH

Conchinchinone I

OOH

O O

OH

Conchinchinone J


Acknowledgements: This research is funded by Vietnam Academy of Science and Technology under grant number VAST.HTQT.BULGARIA.02/17-18.

References: 1 Sanpa S., Popova M., Bankova V., Tunkasiri T., Eitssayeam S., Chantawannakul P. (2015). Antibacterial compounds from propolis of Tetragonula laeviceps and Tetrigona melanoleuca (Hymenoptera: Apidae) from Thailand. PLoS One, 10(5), e0126886.2 Boonnak N., Karalai C., Chantrapromma S., Ponglimanont C., Fun H. K., Kanjana-Opas A., Chantrapromma K., Kato S. (2009). Anti-Pseudomonas aeruginosa xanthones from the resin and green fruits of Cratoxylum cochinchinense. Tetrahedron, 65(15), 3003-3013.3 Li Z., Song Y. H., Uddin Z., Wang Y., Park K. H. (2018). Inhibition of protein tyrosine phosphatase 1B (PTP1B) and α-glucosidase by xanthones from Cratoxylum cochinchinense, and their kinetic characterization. Bioorganic & medicinal chemistry, 26, 737-746.


PP 42

application of natural Deep eutectic Solvents for green extraction of Bioactive Compounds from Poplar Propolis: a Preliminary Study

Hristo Petkov1, Boryana Trusheva1, Milena Popova1, Vassya Bankova1

1 Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., bl. 9, 1113 Sofia, Bulgaria

At present one of the key issues in the chemistry field is the development of so called “green” technologies that aims to preserve the environment and to reduce the negative influence of human involvement. Among the diverse ways of “green” technology, developing and applying “green” solvents are one of the most important subjects. Recently, a new type of “green” solvents, named deep eutectic solvents (DESs), was developed. DES is a mixture of proton acceptor and a proton donor in solid state, which are joined by intermolecular hydrogen bonds. The resulting mixture is eutectic, i.e. it has a lower melting point than that of any of its individual components and it is liquid even at very low temperatures. In a particular combination and ratio natural compounds in solid state (primary metabolites) may also be converted into liquids to form so called natural deep eutectic solvents (NADESs), which are present in nature. They are biocompatible and play a role in all kinds of cellular processes of living organisms. NADESs have several advantages as solvents: easy and inexpensive preparation, biodegradability and in addition, the precursors used for their production are natural, non-toxic, renewable compounds, which are abundant in our daily foods. It was found that NADESs have a number of interesting applications in electrochemistry, functional materials, organic synthesis, catalytic transformations and pretreatment processes. Recently, the interest in NADESs increases due to their successful application in the extraction of bioactive plant metabolites, including non-water soluble. Besides, the extracted compounds can be easily recovered from NADES. This predicts a great potential for NADESs as solvents in the extraction processes of valuable secondary metabolites, further implemented in the food or pharmaceutical industry. Still the most effective and widely used way for exhaustively extraction of all biologically active components from poplar type propolis, is extraction with 70% ethanol in water. The use of alcohol, however, limits the application of propolis as it is contraindicated in a number of diseases, in young children and pregnant women. In order to protect both human health and the environment it is necessary to seek alternative ways of extraction of this valuable biologically active complex. Therefore the present study aims to apply for the first time a new approach for extraction of biologically active compounds from poplar type propolis based on the use of NADESs and to evaluate their extraction efficiency.

Acknowledgment: The financial support of the National Science Fund of Bulgaria, project DN19/4, is gratefully acknowledged


PP 43

the impact of Honeybee Origin on the Quality of Propolis

Gomes da Silva Cahango1, Soraia I. Falcão1, Miguel Vilas-Boas1

1 Mountain Research Centre (CIMO), Polytechnic Institute of Bragança, Campus de Sta. Apolónia, 5300-253 Bragança, Portugal Propolis is a complex mixture of resinous material collected by bees from tree branches, bushes and leaves and transported to the beehive, where secretions, wax and pollen are added. Honeybees use this mixture with mechanical and biological purposes, to repair the hive, moisture management and to prevent the entrance of intruders1. The chemical composition of propolis depends on the botanical source around the apiary but also on the geographical and climatic conditions, with bees preferring specific resin sources. In temperate regions, such as Portugal, the main botanical sources of the resin are poplar, gum rockrose, oak, willow and acacia2.This research outlines the study of how the origin of honeybees can influence the quality of propolis, comparing its physicochemical parameters. Honeybees used on this assay came from two distinct regions of Portugal, from the south (Vila-do-Bispo) and north (Bragança). Propolis production was carried out in two apiaries geographically distinct, Bragança and Vila-do-Bispo and honeybees from the two regions were established in both sites.The results showed variability between the propolis collected in the apiaries, observing a greater content on wax and ashes in the apiary samples from the south. As regards to the phenolic composition, the content of total phenol and flavonoids was higher on the samples from the north side. In respect to the impact of the geographical origin of honeybees, it is found that the physical parameters and the phenolic characteristics are similar within the same apiary and no significant differences were observed considering the origin of honeybees.

References:1 Bankova VS, De Castro SL, Marcucci MC (2000) Propolis: recent advances in chemistry and plant origin. Apidologie 31: 3-15.2 Falcão SI, Tomás A, Vale N, Gomes P, Freire C, Vilas-Boas M (2013) Phenolic quantification and botanical origin of Portuguese propolis. Industrial Crops and Products 396: 887-897.


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antituberculosis activity of Propolis.

Jarosław Widelski1, Joanna Golus2, Piotr Okińczyc3, Rafał Sawicki2, Grażyna Ginalska2, ,Konstantia Graikou4, Zuriyadda Sakipova5, Ioanna Chinou4, Krystyna Skalicka-Woźniak1, Tomasz Mroczek1

1Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Poland2Department of Biochemistry and Biotechnology, Faculty of Pharmacy, Medical University of Lublin3Department of Pharmacognosy, Wrocław Medical University, Wrocław, Poland4Division of Pharmacognosy & Chemistry of Natural Products, Faculty of Pharmacy, National and Kapodistrian University of Athens, Greece5School of Pharmacy, Asfendiyarov Kazakh National Medical University, Almaty, Kazakhstan

Tuberculosis an infectious disease caused by Mycobacterium tuberculosis is one of the leading causes of human morbidity and mortality. In 2013 the WHO reported one and half million of deaths and nine millions new cases of active tuberculosis caused by TB1.The increase in the incidence of clinical tuberculosis is associated with increasing reports of new cases of multi drug resistant (MDR-TB) and extensively multidrug resistant (XDR-TB) strains1.Propolis (bee glue) is well known for its antibacterial and antifungal properties.More than 30 ethanol-water extracts obtained from different propolis samples were tested for antituberculosis activity.Minimal Inhibitory Concentrations (MIC) values for the EO were established with 96-well micro plate method with AlamarBlue (Invitrogen). Reference strain of Mycobacterium tuberculosis H37Ra inoculum in Middlebrook 7H9 broth (Difco) was 5x105cfu/ml per well, accordingly to CLSI standards. Serial twofold dilutions of propolis extracts ranged from 64 to 8 µg/ml. As the internal control of the method serial twofold dilutions of four first line antibiotics dedicated to the tuberculosis treatment: isoniazid (INH), rifampicin (RMP), ethambutol (EMB) and streptomicin (SM) were used2,3.Acknowledgments. The work was financed from grant No 4/POLTUR-1/2016.

References1 WHO (2014). Global tuberculosis report (www.who.int/tdr/news/2014/global-TB-report/en/).World Health Organization (2010). Multidrug and extensively drug resistant TB (M/ XDR-TB) Global report on surveillance and response. Geneva: World Health2 Palomino, J.C., Martin, A., Camacho, A., Guerra, H., Swings, J. ,Portaels, F.(2002). Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis. Antimicrobial Agents and Chemotheraphy, 46, 2720-27223Wayene, P.A. (2011). Susceptibility Testing of Mycobacteria, Nocarediae, and Other Aerobic Acinomycetes; Approved Standard-Second Edition. CLSI document M24-A2. USA. Clinical and Laboratory Standards Institute.


PP 45

GC-mS and uPlCA-PdA-tOf Profile of Polish and eurasian Propolis

Jarosław Widelski1, Piotr Okińczyc2 , Antoni Szumny3, Jakub Szperlik4, Anna Kulma4, Zbigniew Sroka2, Krystyna Skalicka-Woźniak1, Tomasz Mroczek1

1Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, Poland2 Department of Pharmacognosy, Wrocław Medical University, Wrocław, Poland3Department of Chemistry, Wrocław University of Environmental and Life Sciences4Faculty of Biotechnology, Wrocław University, Poland

Propolis, is natural resinous substance used in beehive as a protective barrier against bee’s enemies among them bacteria and fungi. For this properties were used in folk medicine all over the world since centuries for treatment of different types of wounds, burns and many others applications.The chemical composition of propolis is strongly connected with type of plant material available to the bees in different regions and climate. Chemical profile of propolis sample determines all biological activity of this complicated mixture and it is god indicator of its origin. We analyzed chemical composition of propolis samples from Poland and Eurasia (Ukraine and Russia) by two complementary techniques. Dichloromethane extracts, rich in apolar components, were investigated by GC-MS. More polar substances, which was presented in 70 ethanol in water extracts, were analyzed by UPLCA-PDA-TOF. In a results, main components of GC-MS analysed was wax alcohols and acids, some flavonoid aglicones (chrysin, pinocembrin and tectochrysin) and trace of essential oils substances. UPLC-PDA-TOF allowed to tracking free phenolic acid (mainly p-comuric, caffeic and ferrulic acids), their monoesters and glycerides and flavonoids. According to samples composition, most of them exhibited black poplar, aspen or mixed origin.


PP 46

effect of Brazilian Propolis-containing Ointment on genital itching in Menopausal Women

Hiroshi Miura1, Yasuko Miura1, Yuki Shimoda1, Satoko Kagabu2, Hiromitsu Tsubaki3, Yukihiro Terada1

1 Division of Obstetrics and Gynecology, Department of Reproductive and Developmental Medicine, Akita University School of Medicine, Akita, Japan2 Department of Obstetrics and Gynecology, Nakadori General Hospital, Akita, Japan3 Department of Obstetrics and Gynecology, Ogachi Central Hospital, Akita, Japan

Introduction: Menopausal women can develop genital pruritus due to skin contraction and drying associated with estrogen deficiency. The causes of pruritus include contact dermatitis, fungal dermatitis, dry dermatitis, and others. A definite cause often cannot be identified. Propolis has reported antibacterial, antifungal, and antipruritic effects, and can inhibit histamine release. Accordingly, application of a propolis-containing agent to the genital region in menopausal women may treat many causes of pruritus. Methods: Postmenopausal women with genital pruritus were randomly divided into 3 treatment groups: Group A (n=9) was treated with a 1% Brazilian propolis-containing ointment, Group B (n=6) with an antihistamine-containing ointment, and Group C (control, n=5) with a Vaseline ointment. Each group applied ointment to the vulva twice a day. After 14 days, improvement in pruritus was quantified with the visual analog scale (VAS). The study protocol was approved by the ethics committee of Akita University. Results: The average VAS score was 4.7 for Group A, 3.0 for Group B, and 9.5 for Group C. There was a significant difference between Group C and the other 2 groups. Conclusion: The 1% Brazilian propolis-containing ointment may be not as effective as the antihistamine ointment, but did have some antipruritic effect.

Acknowledgements:This research was supported by Yamada Research Grant.


PP 47

yoga Bee, a new approach for Health

Catherine Flurin1

1 Ballot-Flurin Apiculteurs, Maubourguet

Catherine Flurin founded a new holistic and unique in the world method to cure with bees: Gentle beekeeping. It is also named Yoga des Abeilles, Yoga Bee. This is a new practice. Benefits are for children, adults of all ages and conditions, healthy or with all kind of diseases. Progress with a deep change in the beekeeping business. Based on the responsibility of beekeepers and on full respect of the ecosystem: bees, their environment, consumers, and the economic system. Bees have been living on Earth for so many years. They have a deep knowledge, intelligence of life, and a special energy. Bees collapse is the sign of human errors about the way they manage their life and the world. Gentle beekeeping teaches us how to learn from the bees. Gentle Beekeeping is the fruit of 30 years research at the innovative Ballot Flurin Beefactory located in a small village of South West of France. Bees, Nature and Humans are connected to live healthy and rich together. To promote this new era in beekeeping and in communication with bees, join our movement.


PP 48

Correlation Between Chemical Composition and antibacterial activity of Propolis from Different locations in transilvaniaErzsébet-Timea Domokos1, Adriana Urcan1, Liviu Alexandru Margitas1, Daniel Severus Dezmirean1 and Otilia Bobis2*

1Department of Apiculture and Sericulture, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania.2Life Science Institute “King Michael I of Romania”, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania.

Propolis is a resinous substance collected by honeybees from buds and leaves of trees and plants, mixing with pollen as well as enzymes secreted by bees. It is used by bees as a glue, general-purpose sealer, and draught extruder for beehives.Propolis contains a wide variability of active principles (flavonoids and phenolic acids). Variations in the chemical composition of propolis are mainly assigned to the difference in the preferred regional plants visited by honeybees. Flavonoid and esters of phenolic acids are generally regarded to be responsible for the antimicrobial activity of propolis.The aim of the present study was to investigate the antibacterial activity of propolis samples collected from 5 counties in Transylvania (Romania) against different bacterial strains: three Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes), two Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) and, also different bacteria responsible for different bee diseases. The methodology used for the determination was agar diffusion technique. Enroxil was used as positive control and alcohol as negative control. The tested ethanolic propolis extracts are rich sources of polyphenolic substances (high amounts of phenolic acids and flavonoids were quantified in the samples) and show a good antibacterial activity against Gram-positive bacteria. As for the Gram-negative bacteria the results obtained are very different. It can be concluded that a positive correlation exists between antibacterial activity and the content of polyphenols and, also between antibacterial activity of extracts and some specific compounds from propolis.


PP 49

Comparative Study of Commercial Propolis Based Products – antibacterial activity and Bioactive Compounds

Mihaela Niculae1, Laura Stan*2, Adriana Urcan3, Timea Domokos3, Daniel S. Dezmirean3, Otilia Bobiş4

1Infectious Diseases Department, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania2Sensory Analysis Department, Faculty of Food Sciences and Technology, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania3Apiculture and Sericulture Department, Faculty of Animal Sciences and Biotechnologies, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania4Life Science Institute, APHIS Laboratory, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania

Propolis is one of the best natural antibiotic, with high amounts of polyphenols, derived from plant’s resins from where the bees collect, mix with own substances and wax, and preserve it in the hive. High variability among propolis samples was noticed, depending on the geographical origin, plant source, harvesting procedures and methods of evaluation1. One of the major problems in labeling the propolis that is commercially available is the real bioactive compounds concentration. The study was aimed to evaluate the in vitro antibacterial activity and the chemical composition of different commercially available propolis based products (tinctures, ethanolic, glycolic and hydroglycolic extracts, products for veterinary and human use). Spectrophotometric and chromatographic determinations were used to characterize the propolis products. The in vitro antibacterial activity was evaluated against reference strains of Staphylococcus aureus, Staphylococcus pseudintermedius, Bacillus cereus, Enterococcus faecalis, Salmonella enteritis, Salmonella typhimurium and Escherichia coli using the disc diffusion assay. The results indicated intense antibacterial properties in case of both tinctures and ethanolic extracts, especially against Bacillus cereus, Staphylococcus aureus and Staphylococcus pseudintermedius. While the inhibition zone diameters were significantly smaller for the Gram negative bacteria, no efficacy was recorded towards Enterococcus faecalis. Total polyphenols varied from 18-26% and flavonoids (6-10%) were determined in all samples. The total amount of polyphenols and flavonoids depended both on the concentration of propolis in the extracts and the solvent type used for extract preparations and its concentration. These data underline the complex chemical composition and antimicrobial potential of certain propolis based products.

References: 1. Bankova V (2005) Chemical diversity of propolis and the problem of standardization. Journal of Ethnopharmacology 100: 114-117


P1 Phenolic compounds of Spain popolisFélix Adanero-Jorge, Spain

P2 Chemical profiling of Papua New Guinea propolis and assay of its antiprotozoal activitySamya Alenezi, UK

P3 Chemical profile of Greek Arbutus unedo Honey– biological propertiesΑrgyrο Andreou, Greece

P4 Propolis included chewing gum and investigation of its antibacterial activity against Streptococcus mutansDidem Sözeri Atik, Turkey

P5 Propolis from Romania and Turkey: Comparative antioxidant and antibacterial activityOtilia Bobiş, Romania

P6 The characterization and bioactive composition of Turkish propolisİlknur Coşkun, Turkey

P7 Effects of propolis on the quorum sensing of selected biofilm producing bacterial speciesMargarita Gomez Escalada, UK

P8 In Vitro evaluation of green and red propolis extracts against Candida spp.Sonia Figueiredo, Brazil

P9 Thirteen flavonoids from green propolis from Minas Gerais, Brazil, analyzed for six yearsSonia Figueiredo, Brazil

P10 Water extract of propolis might be safer to use during pregnancy than ethanol extractAl Mukhlas Fikri, Indonesia

P11 Differences in chemical composition and antioxidant activity of three propolis samples collected in the same apiary Ana Sofia P. Freitas, Portugal

P12 Combination treatment of Cuban propolis and nemorosone with chemothgerapeutic agents induce a synergisitic cytotoxic effect in drug-resistant human colon carcinoma cellsYahima Frión-Herrera, Italy

P13 Correlation between phenolic composition and biological properties of propolis and heavy metal contentsMaría Inmaculada González-Martín, Spain

P14 Changes in antioxidant and antibacterial activity and in phenolic compounds levels due to the pesticide residuesMaría Inmaculada González-Martín, Spain

list of poster presentations


P15 North Aegean Greek Islands propolis: Antibacterial-antifungal activities against MalasseziaKonstantia Graikou, Greece

P16 Preliminary studies: the potential anti-angiogenic activities of two Sulavesi Island (Indonesia) propolis and their chemical characterizationMuhammad Iqbal, UK

P17 The effects of propolis on growth performance of broiler chickensIvana Klaric, Croatia

P18 In vitro Assessments of Cytotoxic and Cytostatic Effects of Propolis in Cells from the Human Colon Carcinoma Cell Line (HCT 116)Badiaa Lyoussi, Morocco

P19 Preliminary evaluation of the cytotoxic potential of North-West Romanian propolisConstantin I. Mates, Romania

P20 The effect of dietary supplementation with propolis on bacteria colonization pattern in gastrointestinal tract of broiler chickensIvan Miskulin, Croatia

P21 The effect of propolis on selected blood parameters of broilersIvan Miskulin, Croatia

P22 Intestinal morphology broiler chickens supplemented with propolisMaja Miskulin, Croatia

P23 The influence of propolis on liver pathology in broilers Maja Miskulin, Croatia

P24 Cytotoxic activity of four propolis Colombian samples against canine osteosarcoma cellsDolly Patricia Pardo Mora, Colombia

P25 Portuguese propolis: A potential source of environmentally friendly fungicidesCatarina Passão, Portugal

P26 Mechanistic studies of cytotoxicity induced by a Portuguese propolis extract, using Saccharomyces cerevisiae as eukaryotic cell modeCatarina Passão, Portugal

P27 Direct visualization of Artepillin C into fibroblast cells via CARS microscopyWallance M. Pazin, Denmark

P28 Qualitative analysis and biological evaluation of propolis from Armenia and GeorgiaArgyro Petropoulou, Greece

P29 Chemical composition of selected prtopolis samples from Kyrgyzstan and KazakhstanArgyro Petropoulou, Greece


P30 Caseinates loaded with red propolis extractIsabel Cristina Celerino de Moraes Porto, Brazil

P31 Determination of propolis origin using phenolic composition and artificial neural networksIsabel Revilla, Spain

P32 Determination of phenolic acids in raw propolis using near infrared spectroscopyIsabel Revilla, Spain

P33 The biological activities of Indonesian propolis and it’s molecular MarkerMuhamad Sahlan, Indonesia

P34 Antioxidant activities of propolis from Aragón (Spain)M. Teresa Sancho, Spain

P35 Formulations containing propolis of Apis mellifera with antibacterial and antitumoral activityAlexandra Christine Helena Frankland Sawaya, Brazil

P36 Flavonoids constituents of Algeria propolisSegueni Narimane, Algeria

P37 Antimicrobial efficacy of Some Products with Propolis Hydroalcoholic Extract 30% and Antiviral Synthesis Carmen Violeta Popescu, Romania

P38 “Green” synthesis of gold and silver nanoparticles with propolis extract and Rosa damascena wasteAnton M. Slavov, Bulgaria

P39 Quantative and qualitative variations in propolis collectionC.Tananaki, Greece

P40 Isolated triterpenes from stingless bee Lisotrigona furva propolis in VietnamLe Nguyen Thanh, Vietnam

P41 Isolated xanthones from Lisotrigona furva propolis in VietnamLe Nguyen Thanh, Vietnam

P42 Application of natural deep eutectic solvents for green extraction of bioactive compounds from poplar propolis: A preliminary studyBoryana Trusheva, Bulgaria

P43 The impact of honeybee origin on the quality of propolisMiguel Vilas-Boas, Portugal

P44 Antituberculosis activity of propolisJarosław Widelski, Poland

P45 GC-MS and UPLCA-PDA-TOF profile of Polish and Eurasian propolisJarosław Widelski, Poland


P46 Effect of Brazilian Propolis-containing Ointment on Genital Itching in Menopausal WomenHiroshi Miura, Japan

P47 Yoga Bee, a New Approach for HealthCatherine Flurin, France

P48 Correlation Between Chemical Composition and Antibacterial Activity of Propolis from Different Locations in TransilvaniaErzsébet-Timea Domokos, Romania

P49 Comparative Study of Commercial Propolis Based Products – Antibacterial Activity and Bioactive CompoundsMihaela Niculae, Romania

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