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Chemical analysis and antioxidant content of various propolis samples collected from different regions and their impact on antimicrobial activities

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Asian Pacific Journal of Tropical Medicine 2018年第7期11卷 436-442页

作者： Soumaya Touzani Noori Al-Waili Nawal El Menyiy bratko filipic Adriana Pereyra Ilham EL Arabi Wail Al-Waili Badiaa Lyoussi Laboratory of Physiology Pharmacology & Environmental Health Faculty of Science University Sidi Mohamed Ben Abdellah New York Medical Care for Nephrology Institute for Microbiology and Immunology Medical Faculty of Ljubljana University of Ljubljana

Objective: To assess the antioxidant content, antimicrobial and antioxidant activities of various propolis samples. Methods: Seven 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. The antimicrobial activity was assessed against bacterial species, including methicillin resistant Staphylococcus aureus and Candida albicans, and expressed as the minimal inhibitory concentration. Results: The propolis samples showed significant variations in the chemical composition and in the antioxidant or antimicrobial activities even when the samples were collected from the same location. Propolis with high resin and low wax content had high level of antioxidant compounds, and strong antioxidant and antimicrobial activities. Gram-positive bacteria, especially, methicillin resistant Staphylococcus aureus were more sensitive to all propolis samples than Gram-negative bacteria and Candida albicans. Conclusions: The chemical composition and the antioxidant and antimicrobial activities of various propolis samples are different and rely on the geographic and plant origin of propolis collection. Propolis samples with low wax and high resin content might be more suitable to be used in future preclinical or clinical investigations.

关键词： Propolis Antimicrobial Antioxidant Phenol Candida albicans Methicillin resistant Staphylococcus aureus

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Propolis Extracts Enhance the Antimicrobial Activity of Slovenian:Chestnut,Forest and Flower Honeys

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Journal of Agricultural Science and Technology(B) 2019年第3期9卷 203-219页

作者： bratko filipic Klemen Rihar Dunja Exel Gregoric Lidija Gradisnik Adriana Pereyra Eva Ruzic-Sabljic Fabio Galeotti Alfredo Fachini Hrvoje Mazija Croatian Institute for Experimental and Translational Oncology(CIETO) Koledinecka 03Zagreb 10040Croatia Retired Chengdujska 4Ljubljana 1000Slovenia Retired Glavarjeva 19Ljubljana 1000Slovenia Institute of Biomedical Sciences Medical Faculty of MariborTaborska 8Maribor 2000Slovenia Medex D.o.o. Linhartova 49aLjubljana 1000Slovenia Institute for Microbiology and Immunology Medical Faculty of LjubljanaZloska 04Ljubljana 1105Slovenia Department of Life Sciences University of Modena&Regio EmiliaModena 41125Italy BNatural Via Gran SassoCorbettaMI 20011Italy

Because of the overuse and misuse of antibiotic, an increase in antibiotic resistance of pathogenic bacteria is evolving. Attention should be focused on natural alternatives to antibiotics, like propolis and honey. They all have strong antibacterial properties because of the active substances they contain. In the present study, the influence of P1 (water-soluble propolis (WSP) Greit 120) or P2 (phosphate buffer saline (PBS) propolis extract Lj. Center pH = 8.0) on antimicrobial activity of different Slovenian: chestnut honey (CH), forest honeys (FoH), and flower honey (FlH) against various Gram-positive or Gram-negative bacteria and yeast Candida albicans are shown. Gram-positive bacteria used in the experiments were: methicillin-resistant Staphylococcus aureus, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus mutans. The yeast was Candia albicans. Gram-negative bacteria used were: Escherichia coli, Pseudomonas aeruginosa and Acinetobacter baumanii. P1, P2, FoH, CH, FlH and their combinations were prepared in saline as 10% solutions. The composition of both propolis extracts and the three kinds of honey was determined by reverse-phase high-performance liquid chromatography (RP-HPLC). It expressed the antimicrobial activity as a minimal inhibitory concentration (MIC) in mg/mL. Each of the two propolis extracts showed antimicrobial activity when applied alone. Also, the application of each of honey yielded an antimicrobial activity in decreasing order: CH, FoH and FlH honey. The combination of any of the propolis extract with any of the honey showed the additive antimicrobial effect on either Gram-positive or Gram-negative bacteria and on yeast Candida albicans. The effectiveness of P1 or P2 depends on their nature and from the sort of honey and is the highest in the case of CH followed by FoH and FlH.

关键词： Honey propolis flavonoids antimicrobial activity enhancement

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Water Soluble Propolis and Royal Jelly Enhance the Antimicrobial Activity of Honeys and Promote the Growth of Human Macrophage Cell Line

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Journal of Agricultural Science and Technology(B) 2016年第1期6卷 35-47页

作者： bratko filipic Lidija Gradisnik Eva Ruzic-Sabljic Brabara Trtnik Adriana Pereyra Domen Jaklic Rok Kopinc Jana Potokar Almin Puzic Hrvoje Mazija Institute for Microbiology and Immunology Medical Faculty Ljubljana University of Ljubljana Zalogka 4 Ljubljana 1105 Slovenia Institute for Biomedical Sciences Medical Faculty Maribor University of Maribor Taborska 8 Maribor 2000 Slovenia Research and Control Department MEDEX d o.o. Linhartova 49a Ljubljana 1000 Slovenia Veterinary Stations of Mostar Bigke Polje 106 Mostar 88000 Bosnia and Herzegovina Croatian Institute for Experimental and Translation Oncology Koledinedka 3 Zagreb 10040 Croatia

Due to the overuse and misuse of antibiotic, an increase in antibiotic resistance of pathogenic bacteria is evolving. Attention should be focused on natural alternatives to antibiotics, like propolis, royal jelly （R J） and honeys. They all have strong antibacterial properties due to the active substances they contain. This study investigated the effect of combination of water soluble propolis （WSP） Greitl20 or fresh royal jelly （F-RJ）（MiZigoj） and Forest honeys as antibacterial against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Acinetobacter baumanii, Staphylococcus aureus, methicillin resistant Staphylococcus aureus （MRSA）, Streptococcus pyogenes, Streptococcus agalactiae and Candida albicans. These substances are also cell growth promoters for human macrophage （TLT） cell line. WSP Greitl20, F-RJ （M） and different Forest honeys were prepared in saline as 10% solutions. The antimicrobial activity was expressed as the minimal inhibitory concentration （MIC） in mg/mL. The growth promotion activity was measured at optical density （OD） 595 nm. The combination ofWSP Greitl20 with different Forest honeys is better than F-RJ （M） in same combination with different Forest honeys. The best antibacterial/antifungal activity was found with the combination of 10% WSP Greit 120 in the Forest honey （1：10） from Italy or Spain. When measuring the growth promoting activity of TLT cell line, the best activity was detected at the combination of 10% WSP Greitl20 in the Forest honey from Italy （GI3 = 0.796 ± 0.014 and GI5 = 1.133± 0.022）. Antimicrobial and growth promoting activities are correlated and WSP-dependent.

关键词： Propolis royal jelly Forest honey antimicrobial activity human macrophage cell line cell growth promoting activity.

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Additive Effects of Water-Soluble Propolis(Greit 120)and Human Interferon Alfa(HuIFN-αN3)against Influenza Viruses in Vitro

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Journal of Agricultural Science and Technology(A) 2018年第1期8卷 36-47页

作者： bratko filipic Lidija Gradisnik Adriana Pereyra Rok Kopino Klemen Rihar Eva Ruzic-Sabljic Snezana Kramar Damir Dermie Eugen Soos Nicola Volpi Alfredo Fachini and Hrvoje Mazija Croatian Institute of Experimental and Translation Oncology Koledinecka 3Zagreb 10040Croatia Institute of Biomedical Sciences Medical Faculty of MariborTaborska 8Maribor 2000Slovenia Medex D.o.o. Linhartova 49bLjubljana 1000Slovenia Chengdujska 4 Ljubljana 1000Slovenia Institute of Microbiology and Immunology Medical FacultyZaloska 4Ljubljana 1105Slovenia Division of Molecular Biology Ruaer Boskovic InstituteBijenicka 54Zagreb 10000Croatia Trg Sv.Ivana 5 Klostar Ivanic 10312Croatia Department of Life Sciences University of Modena&Reggio EmiliaModena 41125Italy BNatural Via Gran Sasso 33Corbetta-MI 20011Italy

Influenza virus affects the respiratory tract in humans causing a range of distinct manifestations including fever, nasal secretions, cough, headaches, muscle pain and pneumonia, which could become violent and severe. It was found that influenza A viruses remain resistant to amantadine and rimantadin with high level of oseltamvir resistance. Therefore, there is a need for constant improvement of drugs active against resistant influenza viruses. Propolis has anti-influenza activity both in vitro and in vivo. Human leukocyte interferon （HuIFN-αN3） is a multi-subtype protein that displays an antiviral activity against influenza virus. In this study we elucidated the anti-influenza activity of the mixes of water-soluble propolis （WSP）（Greit 120） and HuIFN-αN3 at different ratios. Greit 120 polyphenols, total phenol acids and bioflavonoid were characterized by HPLC-UV-ESI-MS504971 and HuIFN-αN3 by reverse-phase high-performance liquid chromatography （RP-HPLC）. Influenza A and B viruses were separately added to the LLC-MK2 cells treated with WSP （Greit 120） and HuIFN-αN3 alone or in proportions 1：1, 1：2 and 2：1. Plates were incubated and cytopathic effect was determined. The best results （ID50） were obtained with the mix of 10% WSP and HuIFN-αN3 in proportion 1：2, showing ID50 at 12 ± 0.2 μg/mL and 19 ± 0.6 μg/mL for influenza A and B viruses, respectively. When comparing anti-influenza activity of WSP （Greit 120）/HuIFN-αN3 with that of ribavirin, it was found that 1：2 was the optimal ratio for WSP （Greit 120）/HuIFN-αN3 （0.5 and 0.6 for influenza A and B, respectively）. This new formulation of WSP （Greit 120） and HuIFN-αN3, showing better anti-Influenza activity, will definitely improve its application in flu infections.

关键词： Water-soluble propolis Greit 120 HuIFN-αN3 anti-influenza activity ribavirin ID_(50)

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