Putative application of Najas marina L. extracts as a source of bioactive compounds and their antioxidant, antimicrobial, antibiofilm, and genotoxic properties.

In this research paper, the total phenols (TP), flavonoids (TF), and tannins (TT) content in the acetone and ethyl acetate extracts of Najas marina L. and the identification and quantification of phenolic acids and flavonoids from the ethyl acetate extract were performed. Antioxidant, antimicrobial, and antibiofilm properties of the mentioned extracts were investigated in vitro. The genotoxic potential was analyzed in cultured human peripheral blood lymphocytes (PBL). The TP and TF content was higher in the ethyl acetate extract, dominated by quercetin (172.4 µg mg-1) and ferulic acid (22.74 µg mg-1), while the TT content was slightly higher in the acetone extract. Both extracts tested showed limited antioxidant effects compared to ascorbic acid. The strongest antibacterial activity was observed with Gram-positive bacteria, particularly Staphylococcus aureus (MIC and MMC at 0.31 mg ml-1) and S. aureus ATCC 25923 (MIC at <0.02 mg ml-1), while antifungal activity was limited. Both extracts tested showed better activity on preformed biofilms. Acetone extract had no genotoxic activity but showed significant genoprotective activity against mitomycin C-induced DNA damage in cultured PBLs. Results of our research demonstrate the potential for the development of plant-based antibacterial and biofilm agents.

The hydrophobicity of enterobacteria and their co-aggregation with Enterococcus faecalis isolated from Serbian cheese.

In this paper, we investigated the hydrophobicity, ability to adhere to solvents and the pig epithelium and co-aggregation of members of family Enterobacteriaceae and Enterococcus faecalis KGPMF 49. The bacteria used in this study were isolated from traditionally made autochthonous cheese from Southeastern Serbia (Sokobanja). The percentage of adhered bacteria was different in three solvents (chloroform, ethyl acetate and xylene). The highest percentage was detected in the presence of chloroform, and the lowest percentage was detected in the presence of xylene (chloroform < ethyl acetate < xylene). A different degree of co-aggregation of enterobacteria with E. faecalis KGPMF 49 was observed. Klebsiella ornithinolytica KGPMF 8 demonstrated the highest percentage of co-aggregation with E. faecalis KGPMF49 (32.29%). Klebsiella pneumoniae KGPMF 13, K. ornithinolytica KGPMF 9 and Serratia marcescens biogp 1 KGPMF 19 were found to have the ability to adhere to the pig epithelium, whereas Escherichia coli KGPMF 22 showed no such ability. The ability to co-aggregate with other species and the ability to adhere to the pig epithelium are very important characteristics of the isolated bacteria.

Assessment of safety aspect and probiotic potential of autochthonous Enterococcus faecium strains isolated from spontaneous fermented sausage.

OBJECTIVE: The objectives of this research project were isolation, identification, and evaluation of the safety aspect and probiotics properties of 21 Enterococcus faecium strains isolated from sausages originated from southeastern Serbia. RESULTS: Analyzed E. faecium isolates showed tolerance to simulated gastrointestinal conditions. All the examined isolates grew well on media with 0.1% and 0.2% of phenol. None of the tested isolates were histamine-producers, while the synthesis of tyramine was observed for E. faecium sk8-1 and sk8-17. Full resistance to antibiotics was not observed for any examined isolate of E. faecium (penicillin, amoxicillin, and ofloxacin showed the effect on all tested isolates). An inhibition zone against examined pathogens was exhibited by all strains, with the largest inhibition zone against Pseudomonas spp., Proteus spp. and E. coli (12-30 mm/MIC values ranged from 0.5 to 12 mg mL-1). CONCLUSION: The results indicated that E. faecium isolates from spontaneously fermented sausage showed a potential for further investigation and possible application as probiotics.

Autochthonous lactic acid bacteria-presentation of potential probiotics application.

OBJECTIVE: The objective of this study was to evaluate the probiotic potential as well as the ability of adhesion and aggregation of natural and autochthonous lactic acid bacteria, isolated from traditionally made cheese. RESULTS: Lactic acid bacteria from natural food sources can be promising probiotic candidates and they can be used in natural food preservation or like starter cultures. Tested autochthonous isolates showed tolerance to the simulated gastrointestinal condition as well as the sensitivity to clinically relevant antibiotics, especially to ampicillin (MIC at 0.195 µg mL-1 for lactobacilli and from 0.195 to 3.125 µg mL-1 for lactococci). Among isolates, the highest percentage of adhesion was detected with chloroform, while the adhesion ability of selected isolates to pig intestinal epithelium was in the correlation with the results of adhesion ability with solvents. The auto-aggregation ability of isolates was demonstrated, while co-aggregation with Escherichia coli was strain specific. CONCLUSION: The results indicated the potential probiotic properties of the isolates and give evidence for further investigation and potential application in the dairy industry.

Anti-quorum sensing activity, toxicity in zebrafish (Danio rerio) embryos and phytochemical characterization of Trapa natans leaf extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Trapa natans L. (water chestnut or water caltrop) is a widespread aquatic plant, which has been cultivated for food and traditional medicine since ancient times. Pharmacological studies showed that water chestnut exhibits the wide range of biological activities, such as antimicrobial, antioxidative, analgesic, anti-inflammatory, as well as antiulcer. AIM OF THE STUDY: Evaluation of anti-virulence potential and toxicity of T. natans methanol (TnM), acetone (TnA) and ethyl acetate (TnEA) leaf extracts. MATERIALS AND METHODS: The anti-quorum sensing activity of Tn extracts was addressed by measuring their effects on biofilm formation, swarming motility and pyocyanin and elastase production in Pseudomonas aeruginosa. Specific P. aeruginosa biosensors were used to identify which of the signaling pathways were affected. The lethal and developmental toxicity of extracts were addressed in vivo using the zebrafish (Danio rerio) model system. The phenolic composition of T. natans leafs extracts was analyzed by a linear ion trap-OrbiTrap hybrid mass spectrometer (LTQ OrbiTrapMS) and UHPLC system configured with a diode array detector (DAD) hyphenated with the triple quadrupole mass spectrometer. RESULTS: Subinhibitory concentrations of Tn leaf extracts (0.2 MIC) inhibited pyocyanin and elastase production up to 50% and 60%, respectively, and reduced swarming zones, comparing to non-treated P. aeruginosa. TnA inhibited biofilm formation by 15%, TnM showed a stimulatory effect on biofilm formation up to 20%, while TnEA showed no effect. The bioactive concentrations of TnM and TnA were not toxic in the zebrafish model system. Twenty-two phenolic compounds were tentatively identified in TnM, where thirteen of them were identified in T. natans for the first time. Tn extracts, as well as their major components, ellagic and ferulic acids, demonstrated the ability to interfere with P. aeruginosa Las and PQS signaling pathways. CONCLUSIONS: This study demonstrates anti-virulence potential of Tn leaf extracts against medically important pathogen P. aeruginosa and confirms the ethnopharmacological application of this plant against microbial infections.

Low-dimensional compounds containing bioactive ligands. Part VIII: DNA interaction, antimicrobial and antitumor activities of ionic 5,7-dihalo-8-quinolinolato palladium(II) complexes with K+ and Cs+ cations.

Starting from well-defined NH2(CH3)2[PdCl2(XQ)] complexes, coordination compounds of general formula Cat[PdCl2(XQ)] have been prepared by cationic exchange of NH2(CH3)2+ and Cat cations, where XQ are biologically active halogen derivatives of quinolin-8-ol (5-chloro-7-iodo-quinolin-8-ol (CQ), 5,7-dibromo-quinolin-8-ol (dBrQ) and 5,7-dichloro-quinolin-8-ol (dClQ)) and Cat is K+ or Cs+. The cation exchange of all prepared complexes, K[PdCl2(CQ)] (1), K[PdCl2(dClQ)] (2), K[PdCl2(dBrQ)] (3), Cs[PdCl2(CQ)] (4), Cs[PdCl2(dClQ)] (5) and Cs[PdCl2(dBrQ)] (6) was approved using IR spectroscopy, their structures in DMSO solution were elucidated by one- and two-dimensional NMR experiments, whereas their stability in solution was verified by UV-VIS spectroscopy. Interaction of complexes to ctDNA was investigated using UV-VIS and fluorescence emission spectroscopy. The minimum inhibitory concentration and the minimum microbicidal concentration values were detected against 15 bacterial strains and 4 yeast strains to examine the antimicrobial activity for the complexes. The in vitro antitumor properties of the complexes were studied by testing the complexes on leukemic cell line L1210, ovarian cancer cell line A2780 and non-cancerous cell line HEK293. The majority of the prepared compounds exhibited moderate antimicrobial and very high cytotoxic activity.

Low-dimensional compounds containing bioactive ligands. Part VI: Synthesis, structures, in vitro DNA binding, antimicrobial and anticancer properties of first row transition metal complexes with 5-chloro-quinolin-8-ol.

A series of new 3d metal complexes with 5-chloro-quinolin-8-ol (ClQ), [Mn(ClQ)2] (1), [Fe(ClQ)3] (2), [Co(ClQ)2(H2O)2] (3), [Ni(ClQ)2(H2O)2] (4), [Cu(ClQ)2] (5), [Zn(ClQ)2(H2O)2] (6), [Mn(ClQ)3]·DMF (7) and [Co(ClQ)3]·DMF·(EtOH)0.35 (8) (DMF=N,N-dimethylformamide), has been synthesized and characterized by elemental analysis, IR spectroscopy and TG-DTA thermal analysis. X-ray structure analysis of 7 and 8 revealed that these molecular complexes contain three chelate ClQ molecules coordinated to the central atoms in a deformed octahedral geometry and free space between the complex units is filled by solvated DMF and ethanol molecules. Antimicrobial activity of 1-6 was tested by determining the minimum inhibitory concentration and minimum microbicidal concentration against 12 strains of bacteria and 5 strains of fungi. The intensity of antimicrobial action varies depending on the group of microorganism and can be sorted: 1>ClQ>6>3/4>2>5. Complexes 1-6 exhibit high cytotoxic activity against MDA-MB, HCT-116 and A549 cancer cell lines. Among them, complex 2 is significantly more cytotoxic against MDA-MB cells than cisplatin at all tested concentrations and is not cytotoxic against control mesenchymal stem cells indicating that this complex seems to be a good candidate for future pharmacological evaluation. Interaction of 1-6 with DNA was investigated using UV-VIS spectroscopy, fluorescence spectroscopy and agarose gel electrophoresis. The binding studies indicate that 1-6 can interact with CT-DNA through intercalation; complex 2 has the highest binding affinity. Moreover, complexes 1-6 inhibit the catalytic activity of topoisomerase I.

ANTIMICROBIAL AND ANTIBIOFILM EFFECTS OF EXTRACTS FROM TRAPA NATANS L., EVALUATION OF TOTAL PHENOLIC AND FLAVONOID CONTENTS AND GC-MS ANALYSIS.

Research conducted in this study shows the applied in vitro antimicrobial and antibiofilm activity of the four extracts isolated from Trapa natans L. leaves. In this study, different methods were used (microdilution, tissue culture plate, different colorimetric methods, GC-FID and GC-MS analysis). While the water extract didn't show antibacterial activity, the acetone extract showed the strongest one. The same activity in the case of Pseudomonas aeruginosa (MIC was 313 µg/mL) was better than the activity of controls and it matched with antibiofilm activity. The effect of extracts was better on G+ bacteria (MICs were <78-625 µg/mL). For ethanol and ethyl acetate extracts all BIC values were better than MICs. Extracts showed a significant effect on Aspergillus restrictus (MICs were < 78/156 µg/mL). The GC and GC-MS analysis of the ethyl acetate extract revealed the identification of 22 compounds with (all E)-squalene (20.2%), n-alkanes and norlignan hinokiresinol among the most abundant ones. This is the first time that T. natans was studied using these methods.

Extracts of Agrimonia eupatoria L. as sources of biologically active compounds and evaluation of their antioxidant, antimicrobial, and antibiofilm activities.

In this study, we determined the concentration of total phenols, flavonoids, tannins, and proanthocyanidins in the water, diethyl ether, acetone, and ethanol extracts of Agrimonia eupatoria L. We also investigated the antioxidant activity of these extracts using two methods [2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power] and their in vitro antimicrobial (antibacterial and antifungal) activity on some selected species of bacteria and fungi. In addition, the effects of the acetone and water extracts on the inhibition of biofilm formation of Proteus mirabilis and Pseudomonas aeruginosa were investigated using the crystal violet method. The concentration of total phenols was measured according to the Folin-Ciocalteu method and the values obtained ranged from 19.61 mgGA/g to 220.31 mgGA/g. The concentration of flavonoids was examined by the aluminum chloride method and the values obtained ranged from 20.58 mgRU/g to 97.06 mgRU/g. The total tannins concentration was measured by the polyvinylpolypyrrolidone method and the values obtained ranged from 3.06 mgGA/g to 207.27 mgGA/g. The concentration of proanthocyanidins was determined by the butanol-HCl method and the values obtained ranged from 4.15 CChE/g to 103.72 CChE/g. Among the various extracts studied, the acetone extract exhibited good antioxidant activity (97.13%, as determined by the DPPH method). The acetone extract was active in the absorbance value range from 2.2665 to 0.2495 (as determined by the reducing power method). The strongest antimicrobial activity was detected on G+ bacteria, especially on probiotic species, and the acetone extract demonstrated the highest activity. Biofilm inhibitory concentration required to reduce biofilm coverage by 50% values for acetone extract was 4315 µg/mL for P. mirabilis and 4469.5 µg/mL for P. aeruginosa. The results provide a basis for further research of this plant species.

Phytomedical investigation of Najas minor All. in the view of the chemical constituents.

Plants are an abundant natural source of effective antibiotic compounds. Phytomedical investigations of certain plants haven't still been conducted. One of them is Najas minor (N. minor), an aquatic plant with confirmed allelopathy. Research conducted in this study showed the influence of water and ethyl acetate extracts of N. minor on microorganisms, in the view of chemical profiling of volatile constituents and the concentrations of total phenols, flavonoids and tannins. Antimicrobial activity was defined by determining minimum inhibitory and minimum microbicidal concentrations using microdilution method. Influence on bacterial biofilm formation was performed by tissue culture plate method. The total phenolics, flavonoids and condensed tannins were determined by Folin-Ciocalteu, aluminum chloride and butanol-HCl colorimetric methods. Chemical profiling of volatile constituents was investigated by GC and GC-MS. Water extract didn't have antimicrobial activity below 5000 µg/mL. Ethyl acetate extract has shown strong antimicrobial activity on G+ bacteria - Staphylococcus aureus PMFKGB12 and Bacillus subtilis (MIC < 78.13 µg/mL). The best antibiofilm activity was obtained on Escherichia coli ATCC25922 (BIC50 at 719 µg/mL). Water extract had higher yield. Ethyl acetate extract had a significantly greater amount of total phenolics, flavonoids and tannins. As major constituent hexahydrofarnesyl acetone was identified. The ethyl acetate extract effected only G+ bacteria, but the biofilm formation of G-bacteria was suppressed. There was a connection between those in vivo and in vitro effects against pathogenic bacterial biofilm formation. All of this points to a so far unexplored potential of N. minor.

Melilotus albus and Dorycnium herbaceum extracts as source of phenolic compounds and their antimicrobial, antibiofilm, and antioxidant potentials.

Melilotus albus Medic. and Dorycnium herbaceum Vill. (Fabaceae) acetone, ethyl acetate, and ethanol extracts were investigated for their in vitro antimicrobial, antibiofilm, and antioxidant activity with quantification of phenolic compound contents. In general, D. herbaceum extracts showed better antibacterial and antioxidant activity than M. albus extracts. Bacteria Bacillus subtilis, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa, and Proteus mirabilis were the most susceptible with the minimum inhibitory concentrations (MICs), determined by microdilution method, between 1.25-10 mg/mL. Antifungal activity was lower with the detectable MICs at 10 mg/mL and 20 mg/mL. The plant extracts, using the crystal violet assay, inhibit P. aeruginosa biofilm formation in concentration range from 5 mg/mL to 20 mg/mL whereas the effect on mature bacterial biofilm was lower. The antioxidant activity was evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging and reducing power model systems. The intensity of DPPH radicals scavenging activity, expressed as half maximal effective concentration (EC50) values, was from 84.33 µg/mL to >1000 µg/mL. The extracts demonstrated reduced power in a concentration-dependent manner, with ethanol extract as the most active. The total phenols, flavonoids, and proanthocyanidins were determined spectrophotometrically while total extractable tannins were obtained by precipitation method. The phenolic compounds showed differences in their total contents depending on solvents polarities and plant species. Although the plants M. albus and D. herbaceum have not yet been fully explored, these results contribute better understanding of their biotic properties and potential application as antimicrobial and antioxidant agents.

In vitro activity of heather [Calluna vulgaris (L.) Hull] extracts on selected urinary tract pathogens.

Calluna vulgaris L. Hull (Ericaceae) has been used for treatment of urinary tract infections in traditional medicine. In this study we analyzed in vitro antibacterial activity of the plant extracts on different strains of Escherichia coli, Enterococcus faecalis and Proteus vulgaris, as well as the concentrations of total phenols and flavonoids in the extracts. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The concentrations of total phenols were examined by using Folin-Ciocalteu reagent and ranged between 67.55 to 142.46 mg GAE/g. The concentrations of flavonoids in extracts were determined using spectrophotometric method with aluminum chloride and the values ranged from 42.11 to 63.68 mg RUE/g. The aqueous extract of C. vulgaris showed a significant antibacterial activity. The values of MIC were in the range from 2.5 mg/ml to 20 mg/ml for this extract. Proteus vulgaris strains were found to be the most sensitive. The results obtained suggest that all tested extracts of C. vulgaris inhibit the growth of human pathogens, especially the aqueous extract.

Phenolic content, antibacterial and antioxidant activities of Erica herbacea L.

Antibacterial and antioxidant activity, total phenolic and flavonoid concentrations of aqueous, ethanol and ethyl acetate extracts from the leaves and flowers of Erica herbacea L. were studied. In vitro antibacterial activity of the extracts was determined by macrodilution method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) have been determined. Testing was performed on 30 clinical isolates, including different strains of Escherichia coli, Enterococcus faecalis and Proteus vulgaris. The values for MIC were in the range from 2.5 mg/mL to 40 mg/mL. The most sensitive bacterial strains were Proteus vulgaris strains. The aqueous extract from E. herbacea was found the most active. The total phenolic content was determined using Folin-Ciocalteu reagent and ranged between 14.98 and 119.88 mg GA/g. The concentration of flavonoids in extracts was determined using spectrophotometric method with aluminium chloride and obtained results varied from 16.19 to 26.90 mg RU/g. Antioxidant activity was monitored spectrophotometrically using DPPH reagent. The highest capacity to neutralize DPPH radicals was found in the aqueous extract from E. herbacea. The results of the total phenolic content determination of the examined extracts indicate that E. herbacea extracts are a rich source of phenolic compounds and also possess a significant antioxidant activity and moderate antibacterial activity.

Biological evaluation of mechlorethamine-Pt(II) complex, part II: antimicrobial screening and lox study of the complex and its ligand.

The reaction of K(2)PtCl(4) with anticancer-alkylating agent mechlorethamine hydrochloride (CH(3)NH(C(2)H(4)Cl)(2) x HCl = HN2×HCl), in the molar ratio 1 : 2, affords the complex [H2N2](2)[PtCl(4)]. In vitro antimicrobial and lipoxygenase inhibitory activities of the complex and its precursor were evaluated. Antimicrobial activity of the HN2×HCl and [H2N2](2)[PtCl(4)] complex was investigated against 29 species of microorganisms. Testing is performed by microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) have been determined. The difference between antimicrobial activity of precursor and corresponding platinum(II) complex is noticed and the activity of the precursor was higher. Tested compounds demonstrated the high and significant antifungal activity and low to moderate antibacterial activity. It was shown that the gram-positive bacteria were more sensitive than the gram-negative. UV absorbance-based enzyme assays were performed with HN2×HCl and [H2N2](2)[PtCl(4)] complex, in order to evaluate their in vitro inhibitory activity of soybean lipoxygenase (LOX), also. Assay with LOX showed significantly greater inhibitory activity of the complex, than the precursor.

Synergistic antibacterial activity of Salvia officinalis and Cichorium intybus extracts and antibiotics.

Synergistic activity of Salvia officinalis and Cichorium intybus extracts and commonly used antibiotics, amoxicillin and chloramphenicol, were evaluated. Interactions between plant extracts and antibiotics were tested by checkerboard method and interpreted as FIC index. Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and clinical isolates Staphylococcus aureus, Bacillus subtilis, Enterobacter cloacae, Klebsiella pneumoniae, Escherichia coli and Proteus mirabilis were used. Salvia officinalis showed better synergistic capacity than Cichorium intybus. Synergistic interactions were observed between amoxicillin and acetone or ethyl acetate extract of Salvia officinalis and between chloramphenicol and ethyl acetate extract of Salvia officinalis. In the presence of sub-inhibitory concentration (1/4 MIC to 1/32 MIC) of sage extracts, the MIC values of antibiotics were decreased by 2- to 10-fold. Synergism was observed against all test bacteria, except Escherichia coli. The combinations of acetone and ethyl acetate extract from Cichorium intybus and antibiotics resulted in additive and indifferent effects against tested bacteria.

Biological effects, total phenolic content and flavonoid concentrations of fragrant yellow onion (Allium flavum L.).

The antioxidant, antibacterial and antiproliferative activities, total phenolic content and concentrations of flavonoids of A. flavum extracts were determined. The total phenolic content was determined with Folin-Ciocalteu reagent and it ranged between 42.29 to 80.92 mg GA/g. The concentration of flavonoids in various extracts of A. flavum was determined using spectrophotometric method with aluminum chloride and obtained results varied from 64.07 to 95.71 mg RU/g. The antioxidant activity was monitored spectrophotometrically and expressed in terms of IC50 (µg/ml), and its values ranged from 64.34 to 243.34 µg/ml. The highest phenolic content and capacity to neutralize DPPH radicals were found in acetone extract. Antibacterial efficacy was defined by determining minimum inhibitory and minimum bactericidal concentrations using microdilution method. Significant antibacterial activity, especially for ethyl acetate extract, was observed. The best activity was showed against G+ bacteria, Staphylococcus aureus ATCC 25923 and Bacillus subtilis, while Escherichia coli was one of the least sensitive bacteria. Antiproliferative activity of the methanolic extract on HCT- 116 cell line was determined by MTT assay. Results showed that A. flavum has good antiproliferative activity with IC50 values of 28.29 for 24 h and 35.09 for 72 h. Based on these results, A. flavum is a potential source of phenols as natural antioxidant, antibacterial and anticancer substance of high value. Phenolic content of extracts depend on the solvents used for extraction.

Great horsetail (Equisetum telmateia Ehrh.): Active substances content and biological effects.

This paper deals with the antioxidant and antimicrobial activity, total phenolic content and concentrations of flavonoids of Equisetum telmateia extracts. Total phenolic content was determined with Folin-Ciocalteu reagent and it ranged between 129.0 to 262.7 mg GA/g. The concentration of flavonoids in various extracts of E. telmateia was determined using spectrophotometric method with aluminum chloride and obtained results varied from 112.6 to 199.8 mg RU/g. Antioxidant activity was monitored spectrophotometrically and expressed in terms of IC50 (µg/ml), and its values ranged from 33.4 to 982.2 µg/ml. The highest phenolic content, concentrations of flavonoids and capacity to neutralize DPPH radicals were found in the acetone extract. In vitro antimicrobial activity was determined using microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) were also determined. Testing was performed on 22 microorganisms, including 15 strains of bacteria (standard and clinical strains) and 7 species of fungi. There were statistically significant differences in activity between the extracts of E. telmateia. Different effects were noticed against the bacteria and the methanol extract appeared to be most efficient. All the extracts showed significant antibacterial activity against G+ bacteria and weak to moderate activity against other microorganisms.

Biological activities of extracts from cultivated Granadilla Passiflora alata.

Research conducted in this study showed the influence of ethanol, acetone and ethyl acetate extracts of the outgrowth of cultivated Passiflora alata on microorganisms, as well as the antioxidant activity and the concentrations of total phenols, flavonoids and tannins. In vitro antimicrobial activities of extracts were studied on 27 species of microorganisms, of which 17 species of bacteria and 10 species of fungi. The strongest antimicrobial activity was detected on G+ bacteria while the activities on other species were moderate. Ethyl acetate extract showed the strongest effect. The concentrations of total phenols were examined by using Folin-Ciocalteu reagent and the obtained values ranged from 14.04 to 34.22 mg GA/g. By using aluminium chloride method, the concentrations of flavonoids were obtained and the values ranged from 33.19 to 62.30 mg RU/g. In determining the amount of tannins we used the method with buthanol-HCl reagent and the obtained value was 5.1 % of dry matter. The efficiency of antioxidation, which we identified through the reduction of DPPH, was in the range from 808.69 to 1107.79 µg/ml for a particular IC50, and AAI values were between 0.07 and 0.10. The best parameters were shown by ethanol extract. All data were statistically analyzed. Overall, extracts showed potential for further investigation and use.

Immortelle (Xeranthemum annuum L.) as a natural source of biologically active substances.

Antioxidant and antimicrobial effects, total phenolic content and flavonoid concentrations of methanolic, acetone and ethyl acetate extracts from Xeranthemum annuum L. were investigated in this study. The total phenolic content was determined using Folin-Ciocalteu reagent and ranged between 101.33 to 159.48 mg GA/g. The concentration of flavonoids in various X.annuum extracts was determined using spectrophotometric method with aluminum chloride and the results varied from 22.25 to 62.42 mg RU/g. Antioxidant activity was monitored spectrophotometrically using DPPH reagent and expressed in terms of IC50 (µg/ml), and it ranged from 59.25 to 956.81 µg/ml. The highest phenolic content and capacity to neutralize DPPH radicals were found in the acetone extract. In vitro antimicrobial activity was determined by microdilution method. Minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) have been determined. Testing was conducted against 24 microorganisms, including 15 strains of bacteria (standard and clinical strains) and 9 species of fungi. Statistically significant difference in activity between the extracts of X. annuum L. was observed and the acetone extract was found most active. The activity of acetone extract was in accordance with total phenol content and flavonoid concentration measured in this extract. The tested extracts showed significant antibacterial activity against G+ bacteria and weak to moderate activity against other microorganisms. Based on the obtained results, X. annuum can be considered as a rich natural source of polyphenolic compounds with very good antioxidant and antimicrobial activity.