Effect of propolis on Th2 and Th17 cells: interplay with EtxB- and LPS-treated dendritic cells

Dendritic cells (DCs) are antigen-presenting cells that drive the differentiation of T CD4+ cells into different profiles according to the nature of the antigen or immunomodulator. Propolis is a resinous product made by bees that has numerous pharmacological properties, including an immunomodulatory action. To assess whether propolis can modulate the activation of CD4+ T cells by stimulating DCs with heat-labile enterotoxin B subunit (EtxB) or lipopolysaccharide (LPS), we aimed to elucidate the mechanisms affected by propolis in the differential activation of T lymphocytes. Cell viability, lymphocyte proliferation, gene expression (GATA-3 and RORc), and cytokine production (interleukin (IL)-4 and IL-17A) were analyzed. Propolis, EtxB, and LPS induced a higher lymphoproliferation compared with the control. Propolis induced GATA-3 expression and, in combination with EtxB, maintained the baseline levels. Propolis alone or in combination with LPS inhibited RORc expression. EtxB alone and in combination with propolis increased IL-4 production. Propolis in combination with LPS prevented LPS-induced IL-17A production. These results opened perspectives for the study of biological events that may be favored by propolis by promoting Th2 activation or helping in the treatment of inflammatory conditions mediated by Th17 cells.

Antimicrobial activity of propolis and essential oils and synergism between these natural products

In the present study, Apis mellifera propolis and essential oils (EOs) obtained from aromatic plants were evaluated as alternative antimicrobials. We aimed to establish the antimicrobial activity of ethanolic extracts of propolis (EEP) from Apis mellifera and of EOs from Caryophyllus aromaticus, Zingiber officinale, Cinnamomum zeylanicum and Mentha piperita against 32 Staphylococcus aureus and Escherichia coli strains from human clinical specimens. The antimicrobials were diluted in agar and the minimal inhibitory concentrations (MIC) were found whereas MIC90 percent values were calculated. Time-kill curve assays using mixtures containing one quarter of MIC90 percent for EEP with all EOs as well cinnamon EO were performed. The cinnamon EO was found to be the most efficient, with MIC90 percent values of 1.22 and 0.96 mg x mL-1 respectively against S. aureus and E. coli, whereas MIC90 percent of EEP were 1.86 and 20.12 mg x mL-1 respectively against S. aureus and E. coli. The combinations of EEP with ginger and mint EOs, and cinnamon with ginger and clove EOs, showed synergistic effects. Consequently, further studies are necessary to confirm these activities in vivo and to evaluate the phytochemical characteristics of natural products.

Detection of enterotoxin genes of Staphylococcus sp isolated from nasal cavities and hands of food handlers

Food handlers, an important factor in food quality, may contain bacteria that are able to cause foodborne disease. The present study aimed to research coagulase-negative (CNS) and -positive staphylococci (CPS) in 82 food handlers, analyzing nasal and hand swabs, with identification of 62 CNS (75.6 percent) and 20 CPS strains (24.4 percent). Staphylococcal enterotoxins genes were investigated by PCR. In 20 CPS strains, 19 were positive for one or more genes. The percentage of CNS presenting genes for enterotoxins was high (46.8 percent). Despite of the staphylococcal species, the most common gene was sea (35.4 percent), followed by seh and sej (29.2 percent). The detection of new staphylococcal enterotoxins (SEs) genes showed a higher pathogenic potential in this genus. The presence of these gene points out the importance of CNS not only as contaminant bacteria but also as a pathogen.

Propolis effect on streptozotocin-induced diabetic rats

Propolis is one of the hive products that has been used extensively in folk medicine, due to its several biological and pharmaeological properties. Besides, propolis-containing products have been intensely marketed by the pharmaceutical , industry and health-food stores. This work was carried out in order to investigate whether propolis treatment could revert the metabolic alterations of streptozotocin-induced diabetic rats. Animais were kept in metabolic cages and diabetes was induced by a single dose of streptozotocin (35 mg/kg, IV). After a week, rats with glicemia higher than 230 mg/dL were divided into two groups and treated with ethanolic extract of propolis (10 and 90 mg/kg, PO) for seven days. Glycemia and free fatty acids were determined, as well as food and water intake, body weight and, urine were registered weekly. Data showed no significant differences in the analyzed variables. Based on these results, one may conclude that propolis had no effects after diabetes establishment, in our conditions assays. Further assays with different concentrations of propolis and periods of administration should be carried out in order to evaluate its therapeutic potential in this disease

Effects of propolis from Brazil and Bulgaria on Salmonella serovars

Propolis shows biological properties such as antibacterial action. This bee product has a complex chemical composition, which depends on the local flora where it is produced. Salmonella serovars are responsible for human diseases that range from localized gastroenteritis to systemic infections. The aim of the present study was to investigate the susceptibility of Salmonella strains, isolated from food and infectious processes, to the antibacterial action of Brazilian and Bulgarian propolis, as well as to determine the behavior of these bacteria, according to the incubation period, in medium plus propolis. Dilution of ethanolic extract of propolis in agar was the used method. Brazilian and Bulgarian propolis showed an antibacterial action against all Salmonella serovars. The minimal inhibitory concentrations (MIC) of propolis were similar, although they were collected in different geographic regions. Salmonella typhimurium, isolated from human infection, was more resistant to propolis than Salmonella enteritidis.

Susceptibility profile of Salmonella against the antibacterial activity of propolis produced in two regions of Brazil

Propolis antibiotic action has been widely investigated. This assay was carried out in order to observe the in vitro antibacterial activity of propolis against Salmonella enteritidis isolated from food and Salmonella typhimurium isolated from human infections. Propolis was collected by Apis mellifera in two regions of Brazil (Mossoró, Rio Grande do Norte State; and Urubici, Santa Catarina State). Both strains survival percentage decreased with time of incubation in Ethanolic Extracts of Propolis (EEP), demonstrating bactericidal effect after 24 hours. It was also observed that EEP from Mossoró was more effective than that from Urubici. The control of the propolis solvent - 70 percent ethanol - was less effective than EEP, showing only a bacteriostatic effect. We can conclude that propolis shows an activity against Gram-negative bacteria that varies according to the geographical region where it was collected by bees.(AU)

Effect of propolis extract on guines pig lung mast cell

The direct effect of ethanolic extract of propolis on guinea pig lung cell suspension containing mast cells, as well as its influence on the histamine release induced by antigen (ovoalbumin 10 mug/ml) and ionophore A 23187 (3 muM) were investigated. Propolis ethanolic extract (300 mug/ml) increased the histamine release in guinea pig lung suspension containing mast cells by a cytotoxic effect. Lower concentrations of propolis had no effect on histamine release. Our results demonstrated that propolis (3, 10, 30, and 100 mug/ml) shows no significant effect on the histamine release induced by ionophore and antigen. Based on these results, we suggest that propolis could directly activate mast cells, promoting inflammatory mediators release by cytotoxic mechanisms, what could be related to allergic processes in propolis sensitive people.(AU)

Seasonal effect of brazilian propolis on seric biochemical variables

Propolis has been the subject of recent scientific investigation due to its bilogical properties, such as antibiotic, antiinflammatory, anesthetic, healing, immunomodulatory, antioxidant, and carcinostatic. The purpose of this study was to analyze the biochemical profile of propolis-treated rats to observe whether propolis might lead to side effects after administration. Evaluation of total protein, glucose, urea, creatinine, triglycerides, cholesterol, and HDL-cholesterol concentrations and determination of aminotransferases (AST and ALT) and lactic dehydrogenase (LDH) in propolis-treated rat serum were performed. The seasonal effect on propolis activity was also analysed, considering the biochemical variables evaluated. The lack of clinically important changes in seric biochemical variables is probably because propolis showed no biological side effects under these conditions. A possible seasonal effect on the biochemical determinations was not observed.

Absence of seasonal effect on the immunomodulatory action of brazilian propolis on natural killer activity

Propolis, a beehive product widely used in folk medicine as an antiinflammatory agent, has been attracting researchers attention to scientifically elucidate its biological properties and therapeutic activities. The aim of this paper was to study the possible effect of propolis on natural killer activity, since propolis immunomodulatory action has been suggested, especially on non-specific immunity. Propolis was produced by africanized honeybees (Apis mellifera L.), collected throughout a whole year, and pooled by season. Hydroalcoholic solutions of propolis were prepared with each pool and administered to rats by gavage over three days. Natural killer activity of non-adherent spleen cells was evaluated by the 51Cr-release cytotoxicity assay against Yac-1 target cells. Our results indicated that the natural killer activity was increased in spleen cells from propolis-treated animals. There were no significant differences related to the seasonal effect on the immunomodulatory action of propolis.

Seasonal effect of brazilian propolis on Candida albicans and Candida tropicalis

Recently, propolis has been attracting the attention of researchers due to various biological activities and therapeutic properties. In Brazil, propolis is produced all year long, so there may be some seasonal variations. This work was carried out in order to compare propolis collected during the four seasons by its in vitro antimicrobial activity on yeast pathogens isolated from human infections. Propolis was produced by africanized honeybees in Botucatu, Säo Paulo State, collected throughout a year and pooled by season. Hydroalcoholic solutions of propolis were prepared with each pool and diluted in agar, using serial concentrations of propolis from each pool. A determination of minimal inhibitory concentration (MIC) was performed. The results show that Candida tropicalis and Candida albicans were susceptible to low concentrations of propolis, the latter showing a higher susceptibility. No differences were seen in relation to seasonal effects in the minimal inhibitory concentration of propolis

The antibacterial activity of propolis produced by Apis mellifera L. and brazilian stingless bees

This study investigated the antibacterial activity of propolis produced by A. mellifera and Brazilian stingless bees, called "meliponíneos". Susceptibility tests to ethanolic extracts of propolis (EEP) were performed using bacterial strains (Staphylococcus aureus, Enterococcus sp, and Escherichia coli) isolated from human infections. Dilution of EEP in agar (per cent v/v) was used for determination of minimal inhibitory concentration (MIC). The stingless bee species (and common names) were: Nannotrigona testaceicornis ("Irai"), Tetragonisca angustula ("Jataí"), Trigona spinipes ("Arapuá"), Scaptotrigona sp ("Tiúba"), Partamona sp ("Cupira"), Melipona scutellaris ("Uruçu"), Melipona sp ("Manduri"), and Melipona mandaçaia ("Mandaçaia"). EEP inhibitory efficiencies according to bacterial strains were: S. aureus - "Cupira" > "Manduri" = A. mellifera > "Uruçu" > "Mandaçaia" > "Iraí" > "Tiúba" > "Jataí" > "Arapuá" = Ethanol; Enterococcus sp - "Cupira" > "Manduri" > A. mellifera > "Mandaçaia" > "Uruçu" > "Tiúba" > "Jataí" > "Arapuá" = Ethanol; E. coli - "Manduri" > "Jataí" > Ethanol > A.mellifera > "Uruçu" > "Cupira" > "Iraí". Propolis produced by "Cupira" and "Manduri" bees showed higher antibacterial activity than A. mellifera.

Venom production by africanized honeybees (Apis mellifera) and africanized-european hybrids

This study used 15 beehives: five with Africanized queens sisters (Apis mellifera), five with Italian queens sisters (Apis mellifera ligustica), and five with Carniolan queens sisters (Apis meliifera carnica). The queens were fertilized naturally. This experiment was performed in the apiary of the Botucatu School of Veterinary Medicine and Animal Husbandry, UNESP, State of Säo Paulo, Brazil. The following data were obtained from the foraging bees: venom quantity in reservoir, 0.117ñ0.015, 0.139ñ0.020, and 0.147ñ0.024 (mg); venom quantity liberated in extraction apparatus, 0.073ñ0.012, 0.057ñ0.011 and 0.059ñ0.013 (mg); and sting electro stimulus threshold (volts), 10.75ñ1.37, 15.11ñ2.00, and 15.01ñ1.63 for Africanized, Italian x Africanized and Carniolan x Africanized, respectively. The Africanized honeybees possess less venom in reservoir than the European hybrids (Carniolan and Italian). However, they liberated a larger quantity of venom in the extraction apparatus and required lower electro stimulus threshold to promote stinging.

Immunomodulatory action of propolis on macrophage activation

Propolis has been the subject of several recent studies, with the aim of elucidating its biological and pharmacological properties. Propolis has a well-known antimicrobial activity as well as antioxidant, antitumoral, antiinflammatory, and regenerative properties, but literature about its effects on the immunes response in scarce. The goal of this work was to evaluate the propolis effect on macrophage activation by oxygen (H2O2) and nitrogen (NO) metabolite determination. Propolis was produced by africanized honeybees and hydroalcoholic solutions were prepared at different concentrations. Peritoneal macrophages were obtained from male BALB/c mice and culture cells were stimulated in vitro with propolis or interferon-gamma (IFN-gamma). In the in vivo assay, the animals were sacrificed after propolis treatment and cells were stimulated with IFN-gamma. We also investigated the co-stimulant action of propolis associated with IFN-gamma on macrophages. The results show that propolis induces a discreet elevation in H2O2 release and a mild inhibition of NO generation, depending on concentration. Propolis had no co-stimulant activity, diminishing IFN-gamma action on H2O2 and NO production. Data suggest that propolis acts on host non-specific immunity by macrophage activation.

Population analysis of susceptibility to propolis reference strains of Staphylococcus aureus and Escherichia coli

Susceptibility to several ethanolic extracts of propolis (EEP) concentrations was tested with the population analysis technique in reference strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 35218). The results of these tests showed that all bacteria were killed by EEP concentrations approximately equal or higher (2.0 per cent v/v - S. aureus, 10.0 per cent v/v - E. coli) than the respective minimal inhibitory concentrations (MIC). Regarding the susceptibility to propolis, there was a homogeneity of data with the respective time kill curves showing a clear bactericidal effect during 6 to 9 h of exposition.

Serum biochemical determinations of propolis-treated rats

Ethanolic extracts of the bee glue, a resinous substance collected by honeybees called propolis, have been widely used in folk medicine since ancient times. Antibacterial, antifungal and thus antiseptic properties may represent the basis for the historical and present use of these extracts in dermatology, against inflammatory conditions and common colds. This work was carried out in order to verify possible biochemical alterations in some seric parameters of propolis-treated rats. It was shown that propolis possesses an antioxidant property and its administration did not affect either amylase and alanine transaminase activities or total protein concentration.

Nickel chloride effects on erythrocyte generation of superoxide radical

The superoxide anion (O2) is an extremely potent free radical which is produced during the metabolism of aerobic linving cells. (O2) may be involved in lipid peroxidation reactions which occur in a variety of systems. Cu-Zn speroxide dimutase, a metalloprotein, catalyzes the dismutation of the superoxide free radical and protects cells aginst superoxide damage. The ability of NiCl2 to prevent lysis of erythrocytes was tested in rats. NiCl2 administered by intratracheal rouyte prevented hemolysis and decreased total lipids, phospholipids and bilirubin in serum. The protective effect of NiCl2 was linked to an increase in the erytrocyte activity of superoxide dismutase