Yeast Diversity in Honey and Pollen Samples from Stingless Bees in the State of Bahia, Brazil: Use of the MALDI-TOF MS/Genbank Proteomic Technique.

(1) Background: The identification of microorganisms includes traditional biochemical methods, molecular biology methods evaluating the conserved regions of rRNA, and the molecular biology of proteins (proteomics), such as MALDI-TOF MS mass spectrometry. This work aimed to identify the biodiversity of yeasts associated with stingless bee species' honey and pollen, Melipona scutellaris, Nannotrigona testaceicornes, and Tetragonisca angustula, from the region of São Gonçalo dos Campos-Bahia (BA) state, Brazil. (2) Methods: Cellular proteins were extracted from 2837 microbial isolates (pollen and honey) and identified via MALDI-TOF MS. The identified yeast species were also compared to the mass spectra of taxonomically well-characterized reference strains, available from the National Center of Biotechnology Information (NCBI) database. (3) Results: Nine yeast species were identified: Candida maltosa, Candida norvegica, Kazachstania telluris, Schizosaccharomyces pombe, Scheffersomyces insectosus, Meyerozyma guilliermondii, Brettanomyces bruxellensis, Kazachstania exigua, and Starmerella lactis-condensi. Nannotrigona testaceicornes pollen had the highest number of yeast colonies. The yeasts Brettanomyces bruxellensis and Kazachstania telluris showed high populations in the samples of Nannotrigona testaceicornes and Melipona scutellaris, respectively. This work shows that there is some sharing of the same species of yeast between honey and pollen from the same beehive. (4) Conclusions: A total of 71.84% of the identified species present a high level of confidence at the species level. Eight yeast species (Candida maltosa, Candida norvegica, Kazachstania telluris, Schizosaccharomyces pombe, Scheffersomyces insectosus, Meyerozyma guilliermondii, Kazachstania exigua, and Starmerella lactis-condensi) were found for the first time in the samples that the authors inspected. This contributes to the construction of new knowledge about the diversity of yeasts associated with stingless bee products, as well as to the possibility of the biotechnological application of some yeast species.

Extraction of Antioxidant Compounds from Brazilian Green Propolis Using Ultrasound-Assisted Associated with Low- and High-Pressure Extraction Methods.

The demand for bee products has been growing, especially regarding their application in complementary medicine. Apis mellifera bees using Baccharis dracunculifolia D.C. (Asteraceae) as substrate produce green propolis. Among the examples of bioactivity of this matrix are antioxidant, antimicrobial, and antiviral actions. This work aimed to verify the impact of the experimental conditions applied in low- and high-pressure extractions of green propolis, using sonication (60 kHz) as pretreatment to determine the antioxidant profile in the extracts. Total flavonoid content (18.82 ± 1.15-50.47 ± 0.77 mgQE·g-1), total phenolic compounds (194.12 ± 3.40-439.05 ± 0.90 mgGAE·g-1) and antioxidant capacity by DPPH (33.86 ± 1.99-201.29 ± 0.31 µg·mL-1) of the twelve green propolis extracts were determined. By means of HPLC-DAD, it was possible to quantify nine of the fifteen compounds analyzed. The results highlighted formononetin (4.76 ± 0.16-14.80 ± 0.02 mg·g-1) and p-coumaric acid (

Technological and Scientific Prospection on Pigments Produced by Microorganisms.

BACKGROUND: Interest in natural pigments has grown due to the negative aspects caused by synthetic options, which trigger damage to the body and the environment. Research with natural pigments produced by microorganisms becomes viable in relation to other pigments extracted from animals or plants, as microorganisms have advantages in terms of versatility and productivity. Thus, production technologies are protected by patents. METHODS: To evaluate the evolution of research and technological development on producing natural pigments by different microorganisms through scientific and technological prospection. Scientific prospecting was done by searching articles published from 2010 to 2020 in the Scopus, Science Direct, and Web of Science databases. Technological prospecting was carried out with patents obtained from the European Patent Office (ESPACENET) database. RESULTS: The searches were based on the keywords "pigment" and "dye" crossed with microalgae, fungi, bacteria and yeast. A total of 2.811 articles and 451 patents were selected. Scientific prospecting has shown interest in alternative cultivation media, and among the microorganisms that produce pigments, fungi and microalgae are the most studied. CONCLUSION: Technological prospecting showed that 375 (83%) recovered patent documents refer to the protection of pigment extraction and production techniques and that the largest patent holders are private companies, followed by Chinese universities.

In vitro antiviral effect of Mexican and Brazilian propolis and phenolic compounds against human coronavirus 229E.

Propolis is a resinous substance collected by bees (Apis mellifera). It is used for its biological properties. This natural product is available as a safe therapeutic option. Herein, we report the antiviral effects of brown propolis extract from Mexico and green and red propolis extracts from Brazil, as well as their phenolic compounds (quercetin, caffeic acid, and rutin) in preventing infection of MRC-5 cells by HCoV-229E. Normal human fibroblast lung cells (MRC-5) were used to determine the cytotoxicity of the compounds. All samples studied showed antiviral activity. Green and brown propolis extracts, and quercetin exhibited the best EC50 values with values of 19.080, 11.240, and 77.208 µg/mL against HCoV-229E, respectively, and with TC50 of 62.19, 29.192, and 298 µg/mL on MRC-5 cells, respectively. These results are the first in vitro study of the effects of propolis on HCoV-229E and provide the basis for the development of natural formulations against other coronavirus strains.

Utilization of Agro-Industrial Residues in the Rearing and Nutritional Enrichment of Zophobas atratus Larvae: New Food Raw Materials.

Edible insects are a potential alternative food source of high feed conversion efficiency and protein content. Zophobas atratus is an edible insect that adapts to different diets, enabling sustainable rearing by adding value to by-products and agro-industrial residues. This study aimed to evaluate the performance and nutritional characterization of Zophobas atratus larvae fed with different proportions of grape residue. Physicochemical analysis of the diets and larvae (AOAC procedures), fatty acid profile (chromatographic techniques), metals and non-metals (inductively coupled plasma optical emission spectrometry), larval mass gain, feed conversion efficiency, and mortality rate were assessed. The replacement of 25% of the conventional diet with grape residue increased lipid, ash, and fiber contents and reduced protein, carbohydrates, and energy. It promoted greater mass gain, lower mortality rate, and reduced larval growth time by 51%. Among the replacements, 25% resulted in the second-highest content of calcium, sodium, magnesium, and zinc, and the lowest content of potassium and phosphorus in the larvae. The 100% replacement resulted in the highest amounts of C18:2n6 (27.8%), C18:3n3 (2.2%), and PUFA (30.0%). Replacing 25% of the conventional diet with grape residue is equivalent to the conventional diet in many aspects and improves several larvae performance indices and nutritional values.

Syrah Grape Skin Residues Has Potential as Source of Antioxidant and Anti-Microbial Bioactive Compounds.

In this study, we evaluated the effects of ultrasound-assisted extraction (UAE) under different time-temperature conditions on the content of bioactive compounds, antioxidant and antimicrobial activities of Syrah grape skin residue. The application of UAE showed a positive effect on the extraction of total flavonoids, and a negative effect on total polyphenols. The temperature of 40 °C and 60 °C without the UAE caused an increase of 260% and 287% of the total polyphenols, respectively. Nineteen individually bioactive compounds were quantified. The anthocyanin concentration (malvidin-3,5-di-O-glucoside 118.8-324.5 mg/100 g) showed high variation, to a lesser extent for phenolic acids, flavonoids, flavonols, procyanidins and stilbenes due to the UAE process. The Syrah grape skin residue has a high concentration of total phenolic compounds of 196-733.7 mg·GAE/100 g and a total flavonoid content of 9.8-40.0 mg·QE/100 g. The results of free radical scavenging activity (16.0-48.7 mg/100 mL, as EC50) and its inhibition of microbial growth (0.16 mg/mL, as EC50 for S. aureus, and 0.04 mg/mL, as EC50 for E. coli) by grape skin extract (UAE 40:20) indicate high antioxidant and antibacterial activity. It was concluded that the use of ultrasound needs further analysis for its application in this context, as it has shown deleterious effects on some compounds of interest. Syrah grape skin residue has potential as a source of bioactive antioxidants, antimicrobial activity and for use as a functional food ingredient.

Activity of antifungal drugs and Brazilian red and green propolis extracted with different methodologies against oral isolates of Candida spp.

BACKGROUND: Oral candidiasis is an opportunistic disease caused by fungi of the Candida genus. The occurrence of Candida spp. resistance to the commercial antifungal drugs points to the search for alternative treatments. Propolis has been successfully used in the treatment of infectious diseases for centuries. It has been proposed that an ultrasound pretreatment in the propolis extraction protocol can enhance the concentrations of molecules with antimicrobial activities in the final extract. Thus, this study aimed to compare the antifungal activity against oral Candida spp. isolates of green and red propolis extracts submitted or not to an ultrasound pretreatment before the extraction procedure. METHODS: Candida spp. were isolated from denture stomatitis lesions and identified by sequencing. Oral Candida spp. isolates and reference strains were submitted to broth microdilution assays using commercial antifungals and Brazilian green and red propolis extracts submitted or not to an ultrasound pretreatment. Minimal Inhibitory Concentrations (MIC) and Minimal Fungicide Concentrations (MFC) were determined and biofilm formation interference was evaluated for resistant isolates. RESULTS: C. albicans, Candida tropicalis and Candida dubliniensis were isolated from denture stomatitis lesions. Growth inhibition was observed in all Candida isolates incubated with all green and red propolis extracts. At lower doses, red propolis extracts presented significant antifungal activity. The ultrasound pretreatment did not promote an increase in the antifungal activity of green or red propolis. Three isolates, which were highly resistant to fluconazole and itraconazole, were susceptible to low doses of red propolis extracts. These same three specimens had their biofilm formation inhibted by red propolis ethanolic extract. CONCLUSIONS: Thus, red propolis can be faced as a promising natural product to be used in the auxiliary antifungal therapy of denture stomatitis.

Activity of Ethanolic and Supercritical Propolis Extracts in Corynebacterium pseudotuberculosis and Its Associated Biofilm.

Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis in small ruminants, a chronic disease characterized by the development of granulomas in superficial and visceral lymph nodes as well as in several organs. An important characteristic of the infection with this bacterium is the formation of a biofilm and the absence of effective antibiotic therapy against the disease. From this scenario, the objective of this study was to evaluate the susceptibility of C. pseudotuberculosis to conventional antibiotics and to red, green, and brown propolis extracts obtained by the supercritical and ethanolic extraction methods as well as its activity in the bacterial biofilm. The results of the sensitivity test using antibiotics indicated a sensitivity of C. pseudotuberculosis strains to the antimicrobial agents. The ethanolic extract of green propolis and the supercritical red propolis extract showed the best antibacterial activities against planktonic C. pseudotuberculosis. A lower antimicrobial activity of the brown propolis extract was identified. Propolis extracts were effective in interfering with the formation of the C. pseudotuberculosis biofilm but had little activity on the consolidated biofilm. In conclusion, propolis extracts are more effective against C. pseudotuberculosis in the planktonic stage, being able to interfere with the formation of bacterial biofilm. However, the action of propolis extracts in a sessile and structured microbial biofilm is reduced.

Effect of Drying Methods on Bioactive Compounds and Antioxidant Capacity in Grape Skin Residues from the New Hybrid Variety "BRS Magna".

The effects of the drying process using the conventional oven and freeze-drying on the thermogravimetric profile, proximate composition, color parameters, individual bioactive compounds, and antioxidant activity in the grape residue (skin) were evaluated. Twenty individual phenolic compounds were identified, where a variation in concentration was observed for flavonols, stilbenes, phenolic acids, flavonoids, procyanidins, and particularly anthocyanins (malvidin-3,5-di-O-glucoside of 253.2-536.9 mg/kg) due to the drying process. Drying in a conventional oven caused a decrease of 23% of the total polyphenols. The skin of the BRS magna grape has a high concentration of total phenolic content of 489.5-148.3 mg.GAE/100 g, total anthocyanin content of 124.9-260.1 mg.CE/100 g, and total flavonoid content of 12.7-26.0 mg.QE/100 g. The results of free radical scavenging activity (1.26-4.91 µg/mL, as EC50) and ferric reducing antioxidant power (82.93-184.10 µmol/g of skin as equivalent to Fe2SO4) indicate high antioxidant activity, independently of the drying process applied. It was concluded that, if the application is directed to anthocyanin compounds, the use of lyophilization is recommended. On the other hand, if the interest is in bioactive compounds that exert antioxidant activity, conventional oven-drying can be used.

Effect of Experimental Parameters on the Extraction of Grape Seed Oil Obtained by Low Pressure and Supercritical Fluid Extraction.

Grape seeds are an important byproduct from the grape process. The objective of this work was to evaluate the effect of experimental parameters (temperature and time of pretreatment with ultrasound) to obtain grape seed oil using low pressure (Soxhlet-Sox and Bligh Dyer-BD) and high pressure (supercritical carbon dioxide-SFE) methods. The best condition for pretreatment of samples was 30 min of sonication at 30 °C before extraction by Sox or BD. Ultrasound pretreatment was efficient to increase oil extraction yield by 32.10 (Sox), 20.31 (BD) and 12.54% (SFE), depending on the extraction method used as well as, and certainly influenced the total phenolic concentration in 311 (Sox), 234 (BD), and 184 (SFE)%. Ten fatty acids were identified in the oils, the major ones being 18:2ω-6cis (linoleic 52.39%-63.12%), 16:0 (palmitic 20.22%-26.80%) and 18:0 (stearic 8.52%-13.68%). The highest epicatechin concentration was identified in the BD sample: 30-30 (150.49 ± 5.98mg/kg), which presented a concentration of ≥3 times compared to the control (56.68 ± 1.81mg/kg). Ultrasound pretreatment also contributed positively (56% and 99% increase) in the α-tocopherol content of the SFE: 30-30 and BD: 30-30 samples, respectively. The results indicate that the ultrasound pretreatment is a suitable technology to improve the quality of the oil from the grape seed.

The improvement of grape juice quality using Thermomucor Indicae-Seudaticae pectinase.

The effect of pectinolytic enzyme preparation (PEP) produced by the fungus Thermomucor indicae-seudaticae-N31 (PEP-N31) on total phenolic content, concentrations of methanol and color of grape juice was studied. Positive results were found when PEP-N31 was used to extract phenolic compounds after the grapes had been blanched for 3 min and macerated for 1 h. The resulting juice had better yield, color characteristics and higher phenolic content (1637.21 mg.L-1, as gallic acid equivalent, or GAE) than the conventionally prepared juice (1422.59 mg GAE.L-1), and it was very similar to the juice obtained through the treatment with a commercial enzyme (1682.10 mg GAE.L-1). The concentration of methanol in the juice produced with the PEP-N31 was less than 200 mg.L-1. These results encourage the use of PEP produced by Thermomucor indicae-seudaticae-N31 by the grape-processing industry.

Antiviral effects of Brazilian green and red propolis extracts on Enterovirus surrogates.

Propolis is a natural product of bees with biological activities that are mainly associated with bee type and geographic origin. Propolis extract has been proposed with several applications in environmental health. The ethanol extracts have shown good antimicrobial activity. The association of this technique with ultrasound-assisted extraction has been studied to improve the characteristics of the obtained extracts. Thus, the objective of this work is to verify the antiviral action against two strains of bacteriophages of two extracts of Brazilian propolis (green and red) obtained by conventional extraction and ultrasonic extraction. The activities of the propolis red and green extracts were confirmed by the significant ~3 and ~4.5 Log 10 PFU/mL reduction in the concentrations of the MS2 and Av-08 bacteriophages, respectively. It was found that ultrasound-assisted extraction is comparable to the maceration process and demonstrated the best antiviral activities. Brazilian red propolis was more effective than green propolis in viral reduction in all treatments.

Brazilian Green Propolis as a Therapeutic Agent for the Post-surgical Treatment of Caseous Lymphadenitis in Sheep.

As antibiotics are ineffective when used to treat caseous lymphadenitis, the surgical excision of lesions is often required. Iodine solution (10%) is currently the choice for the post-surgical treatment; however, it may cause histotoxicity. Propolis are resinous substances composed by a mixture of different plants parts and molecules secreted by bees. As green propolis has already proven to possess anti-bacterial and wound healing properties, this study aimed to evaluate the use of a green propolis-based ointment as a therapeutic agent for the post-surgical treatment of caseous lymphadenitis. The caseous lesions of 28 sheep were surgically excised before dividing animals into two groups: (1) iodine-treated animals and (2) sheep treated with an ointment made with a previously characterized green propolis extract. Clinical data of animals, size of the scar area, the presence of moisture and secretion in the surgical wound, the humoral immune response against the bacterium and the susceptibility of C. pseudotuberculosis clinical isolates to the green propolis extract were analyzed. The green propolis-treated group presented complete healing of the surgical wound 1 week before the iodine-treated group. Additionally, animals treated with the green propolis ointment had fewer cases of wound secretion, but it was not statistically different from the iodine-treated group. No clinical signs indicating green propolis toxicity or other side effects were found, associated with a faster and more organized hair recovery by propolis use. The green propolis extract was able to inhibit the growth of 23 from the 27 C. pseudotuberculosis clinical isolates, with minimum inhibitory and minimum bactericide concentrations ranging from 01 to 08 mg/mL, and did not interfere with the humoral immune response against the bacterium. In addition, green propolis was able to inhibit biofilm formation by four of the C. pseudotuberculosis clinical isolates. We concluded that green propolis is a promising therapeutic agent to be used in the post-surgical treatment of caseous lymphadenitis in small ruminants due to its effects on surgical wound healing, hair recovery, inhibition of wound contamination and bacterial growth.

Grape peel (Syrah var.) jam as a polyphenol-enriched functional food ingredient.

In this study, we evaluated the effects of the storage time on the physicochemical properties, bioactive compound content, and antioxidant capacity of jam prepared from grape peel extract to explore its potential as a supplementary food and/or functional ingredient. The ethanolic extract from Syrah var. grape peel exhibited high bioactive compound concentrations and antioxidant activity. The jam stability (prepared with 8.9% of extract) at 14°C was evaluated at 0, 15, 30, 45, and 60 days. The jam was found to contain high concentrations of polyphenolic compounds (137.0 ± 3.2 mg of gallic acid equivalent/100 g), total flavonoids (128.5 ± 23.0 mg of equivalent/100 g), and total anthocyanins (92.5 ± 4.0 mg of cyanidin equivalent/100 g). However, a large reduction in the flavonoid (70%-90%), anthocyanin (29%-35%), and phenolic (23%-30%) content was observed during storage. The free radical-scavenging activity (DPPH-), ferric reducing antioxidant power (FRAP), and ß-carotene-linoleic acid assays revealed the great antioxidant potential of the jam prepared from grape peel extract, which exhibited significant levels of radical-neutralizing activity, especially as determined by the DPPH method with EC50 values ranging from 2.3 ± 0.1 to 3.9 ± 0.1 µg/ml. High R 2 values (p > 0.90) were obtained for the correlation between the DPPH results and the concentrations of the compounds of interest. In summary, the high bioactive compound contents and antioxidant capacity of the jam produced from grape peel suggest that it may provide health benefits as a source of natural antioxidants upon incorporation to several food industry products.

Susceptibility of Malassezia pachydermatis Clinical Isolates to Allopathic Antifungals and Brazilian Red, Green, and Brown Propolis Extracts.

Clinical mycoses treatment is associated with issues such as negative side effects, high cost, prolonged treatment, and resistant strain selection. Malassezia pachydermatis is the most frequently isolated yeast in cases of canine otitis and dermatitis. The number of fungal strains exhibiting primary resistance to several drugs in vitro is increasing. Propolis has a diverse chemical composition and well-known therapeutic properties against mycoses. An alternative method for producing propolis extracts using supercritical fluid has higher selectivity, yielding extracts with fewer pollutant residues. This study therefore aimed to evaluate the in vitro susceptibility profile of M. pachydermatis clinical isolates to precharacterized supercritical and ethanolic extracts. Three types of Brazilian propolis extracts (green, red, and brown) and commercial allopathic antifungals were used in this investigation. We used the microdilution broth technique to evaluate the susceptibility profile of the yeasts. The minimum inhibitory concentration (MIC) of the brown propolis ethanolic extract was ≥16 µg/mL for all isolates. The MICs of fluconazole, ketoconazole, itraconazole, and amphotericin B ranged from 8 to >64 µg/mL, 0.032-4 µg/mL, 0.0313-16 µg/mL, and 1-2 µg/mL, respectively. The MICs of ethanolic red propolis extracts were lower than those of supercritical red propolis extracts. However, the green propolis ethanolic extract had more pronounced fungicidal activity. Isolates with lower susceptibility to commercial fungicides were inhibited by red and green propolis extracts. These results indicate that propolis can potentially be used in in vivo experiments as a promising therapeutic agent against M. pachydermatis infections.

Chemical characterization and biological activity of six different extracts of propolis through conventional methods and supercritical extraction.

Propolis is a natural product with many demonstrated biological activities and propolis extract has been used in the food, pharmaceutical and cosmetics industries. Different works have showed the variations in the chemical composition, and consequently, on the biological activity of the propolis that are associated with its type and geographic origin. Due to this study evaluated propolis extracts obtained through supercritical extraction and ethanolic extraction (conventional) in three samples of different types of propolis (red, green and brown), collected from different regions in Brazil (state of Bahia). Analyses were performed to determine the humidity, water activity, the content of total ash, proteins, lipids and fiber in raw propolis samples. The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH), catechin, ferulic acid and luteolin and antimicrobial activity against two bacteria (Staphylococcus aureus and Escherichia coli) were determined for all extracts. For the green and red ethanolic extracts the anti-leishmanicidal potential was also evaluated. The physicochemical profiles showed agreement in relation to the literature. The results identified significant differences among the extracts (p>0.05), which are in conformity with their extraction method, as well as with type and botanical origin of the samples. The extraction with supercritical fluid was not efficient to obtain extracts with the highest contents of antioxidants compounds, when compared with the ethanolic extracts. The best results were shown for the extracts obtained through the conventional extraction method (ethanolic) indicating a higher selectivity for the extraction of antioxidants compounds. The red variety showed the largest biological potential, which included the content of antioxidants compounds. The results found in this study confirm the influence of the type of the raw material on the composition and characteristics of the extracts. The parameters analysis were important to characterize and evaluate the quality of the different Brazilian propolis extracts based on the increased use of propolis by the natural products industry.

Antioxidant, antimicrobial, antiparasitic, and cytotoxic properties of various Brazilian propolis extracts.

Propolis is known for its biological properties and its preparations have been continuously investigated in an attempt to solve the problem of their standardization, an issue that limits the use of propolis in food and pharmaceutical industries. The aim of this study was to evaluate in vitro antioxidant, antimicrobial, antiparasitic, and cytotoxic effects of extracts of red, green, and brown propolis from different regions of Brazil, obtained by ethanolic and supercritical extraction methods. We found that propolis extracts obtained by both these methods showed concentration-dependent antioxidant activity. The extracts obtained by ethanolic extraction showed higher antioxidant activity than that shown by the extracts obtained by supercritical extraction. Ethanolic extracts of red propolis exhibited up to 98% of the maximum antioxidant activity at the highest extract concentration. Red propolis extracts obtained by ethanolic and supercritical methods showed the highest levels of antimicrobial activity against several bacteria. Most extracts demonstrated antimicrobial activity against Staphylococcus aureus. None of the extracts analyzed showed activity against Escherichia coli or Candida albicans. An inhibitory effect of all tested ethanolic extracts on the growth of Trypanosoma cruzi Y strain epimastigotes was observed in the first 24 h. However, after 96 h, a persistent inhibitory effect was detected only for red propolis samples. Only ethanolic extracts of red propolis samples R01Et.B2 and R02Et.B2 showed a cytotoxic effect against all four cancer cell lines tested (HL-60, HCT-116, OVCAR-8, and SF-295), indicating that red propolis extracts have great cytotoxic potential. The biological effects of ethanolic extracts of red propolis revealed in the present study suggest that red propolis can be a potential alternative therapeutic treatment against Chagas disease and some types of cancer, although high activity of red propolis in vitro needs to be confirmed by future in vivo investigations.

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

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

Determination of Parameters for the Supercritical Extraction of Antioxidant Compounds from Green Propolis Using Carbon Dioxide and Ethanol as Co-Solvent.

The aim of this study was to determine the best processing conditions to extract Brazilian green propolis using a supercritical extraction technology. For this purpose, the influence of different parameters was evaluated such as S/F (solvent mass in relation to solute mass), percentage of co-solvent (1 and 2% ethanol), temperature (40 and 50°C) and pressure (250, 350 and 400 bar) using supercritical carbon dioxide. The Global Yield Isotherms (GYIs) were obtained through the evaluation of the yield, and the chemical composition of the extracts was also obtained in relation to the total phenolic compounds, flavonoids, antioxidant activity and 3,5-diprenyl-4-hydroxicinnamic acid (Artepillin C) and acid 4-hydroxycinnamic (p-coumaric acid). The best results were identified at 50°C, 350 bar, 1% ethanol (co-solvent) and S/F of 110. These conditions, a content of 8.93±0.01 and 0.40±0.05 g/100 g of Artepillin C and p-coumaric acid, respectively, were identified indicating the efficiency of the extraction process. Despite of low yield of the process, the extracts obtained had high contents of relevant compounds, proving the viability of the process to obtain green propolis extracts with important biological applications due to the extracts composition.