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      Antimicrobial activity against oral pathogens and immunomodulatory effects and toxicity of geopropolis produced by the stingless bee Melipona fasciculata Smith

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          Abstract

          Background

          Native bees of the tribe Meliponini produce a distinct kind of propolis called geopropolis. Although many pharmacological activities of propolis have already been demonstrated, little is known about geopropolis, particularly regarding its antimicrobial activity against oral pathogens. The present study aimed at investigating the antimicrobial activity of M. fasciculata geopropolis against oral pathogens, its effects on S. mutans biofilms, and the chemical contents of the extracts. A gel prepared with a geopropolis extract was also analyzed for its activity on S. mutans and its immunotoxicological potential.

          Methods

          Antimicrobial activities of three hydroalcoholic extracts (HAEs) of geopropolis, and hexane and chloroform fractions of one extract, were evaluated using the agar diffusion method and the broth dilution technique. Ethanol (70%, v/v) and chlorhexidine (0.12%, w/w) were used as negative and positive controls, respectively. Total phenol and flavonoid concentrations were assayed by spectrophotometry. Immunotoxicity was evaluated in mice by topical application in the oral cavity followed by quantification of biochemical and immunological parameters, and macro-microscopic analysis of animal organs.

          Results

          Two extracts, HAE-2 and HAE-3, showed inhibition zones ranging from 9 to 13 mm in diameter for S. mutans and C. albicans, but presented no activity against L. acidophilus. The MBCs for HAE-2 and HAE-3 against S. mutans were 6.25 mg/mL and 12.5 mg/mL, respectively. HAE-2 was fractionated, and its chloroform fraction had an MBC of 14.57 mg/mL. HAE-2 also exhibited bactericidal effects on S. mutans biofilms after 3 h of treatment. Significant differences (p < 0.05) in total phenol and flavonoid concentrations were observed among the samples. Signs toxic effects were not observed after application of the geopropolis-based gel, but an increase in the production of IL-4 and IL-10, anti-inflammatory cytokines, was detected.

          Conclusions

          In summary, geopropolis produced by M. fasciculata can exert antimicrobial action against S. mutans and C. albicans, with significant inhibitory activity against S. mutans biofilms. The extract with the highest flavonoid concentration, HAE-2, presented the highest antimicrobial activity. In addition, a geopropolis-based gel is not toxic in an animal model and displays anti-inflammatory effect.

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          Most cited references55

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          Antimicrobial activity of flavonoids

          Flavonoids are ubiquitous in photosynthesising cells and are commonly found in fruit, vegetables, nuts, seeds, stems, flowers, tea, wine, propolis and honey. For centuries, preparations containing these compounds as the principal physiologically active constituents have been used to treat human diseases. Increasingly, this class of natural products is becoming the subject of anti-infective research, and many groups have isolated and identified the structures of flavonoids possessing antifungal, antiviral and antibacterial activity. Moreover, several groups have demonstrated synergy between active flavonoids as well as between flavonoids and existing chemotherapeutics. Reports of activity in the field of antibacterial flavonoid research are widely conflicting, probably owing to inter- and intra-assay variation in susceptibility testing. However, several high-quality investigations have examined the relationship between flavonoid structure and antibacterial activity and these are in close agreement. In addition, numerous research groups have sought to elucidate the antibacterial mechanisms of action of selected flavonoids. The activity of quercetin, for example, has been at least partially attributed to inhibition of DNA gyrase. It has also been proposed that sophoraflavone G and (−)-epigallocatechin gallate inhibit cytoplasmic membrane function, and that licochalcones A and C inhibit energy metabolism. Other flavonoids whose mechanisms of action have been investigated include robinetin, myricetin, apigenin, rutin, galangin, 2,4,2′-trihydroxy-5′-methylchalcone and lonchocarpol A. These compounds represent novel leads, and future studies may allow the development of a pharmacologically acceptable antimicrobial agent or class of agents.
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            Antibacterial, antifungal and antiviral activity of propolis of different geographic origin.

            Propolis samples from different geographic origins were investigated for their antibacterial (against Staphylococcus aureus and Escherichia coli), antifungal (against Candida albicans) and antiviral (against Avian influenza virus) activities. All samples were active against the fungal and Gram-positive bacterial test strains, and most showed antiviral activity. The activities of all samples were similar in spite of the differences in their chemical composition. In samples from the temperate zone, flavonoids and esters of phenolic acids are known to be responsible for the above mentioned activities of bee glue; tropical samples did not contain such substances but showed similar activities. Obviously, in different samples, different substance combinations are essential for the biological activity of the bee glue. It seems that propolis has general pharmacological value as a natural mixture and not as a source of new powerful antimicrobial, antifungal and antiviral compounds.
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              Chemical diversity of propolis and the problem of standardization.

              Chemical variability of propolis is discussed with respect to the problem of standardization. Several chemical types of propolis are formulated, based on their plant source. Reliable criteria for chemical standardization of different propolis types are needed but such generally accepted criteria do not yet exist. The chemical profile of "poplar" propolis, typical for the temperate zone, can be characterized by the following parameters: total flavone and flavonol content, total flavanone and dihydroflavonol content, and total phenolics content. These parameters correlate better with the biological activity and are more informative that the quantification of individual components. There is still a lot of work to be done to achieve standardization of other propolis types. Working with standardized material will allow scientists to connect a particular chemical propolis type to a specific type of biological activity and formulate recommendations for mainstream practitioners.
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                Author and article information

                Journal
                BMC Complement Altern Med
                BMC Complementary and Alternative Medicine
                BioMed Central
                1472-6882
                2011
                4 November 2011
                : 11
                : 108
                Affiliations
                [1 ]Laboratório de Imunofisiologia, Universidade Federal do Maranhão, Av. dos Portugueses, SN, CEP: 65.080-040, São Luís, Maranhão, Brazil
                [2 ]Departamento de Odontologia, Universidade Federal do Maranhão, Av. dos Portugueses, SN, CEP: 65.080-040, São Luís, Maranhão, Brazil
                [3 ]Laboratório de Microbiologia, Universidade Federal do Maranhão, Av. dos Portugueses, SN, CEP: 65.080-040, São Luís, Maranhão, Brazil
                [4 ]Laboratório de Farmacognosia, Universidade Federal do Maranhão, São Luís, Av. dos Portugueses, SN, CEP: 65.080-040, Maranhão, Brazil
                [5 ]Laboratório de Microbiologia, Centro Universitário do Maranhão, Rua Josué Montello No. 1, CEP: 65.075-120, São Luís, Maranhão, Brazil
                Article
                1472-6882-11-108
                10.1186/1472-6882-11-108
                3225302
                22053900
                85ebe430-8e9a-4116-aeb8-59110e238a1f
                Copyright ©2011 Liberio et al; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 23 June 2011
                : 4 November 2011
                Categories
                Research Article

                Complementary & Alternative medicine
                Complementary & Alternative medicine

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