Antioxidant activities of Salvia miltiorrhiza and Panax notoginseng

Traditional Chinese medicinal herb Salvia miltiorrhiza (SM) and Panax notoginseng (PN) have been widely used for the prevention and treatment of vascular diseases in the clinics. To better understand their mechanisms of pharmacological actions, the in vitro antioxidant activities of extract of Salvia miltiorrhiza (ESM) and extract of Panax notoginseng (EPN) were evaluated with different antioxidant testing systems. Their activities of scavenging superoxide anion radicals, DPPH radicals, hydroxyl radicals, and hydrogen peroxide, chelating Ferrous ion, and ferric ion reducing power were assessed. The results showed that the mechanisms of their antioxidant effects were distinct and diverse. ESM possessed strong reducing power and high scavenging activities against free radicals including superoxide anion, hydroxyl and DPPH radicals, but a weaker scavenging activity for hydrogen peroxide. ferrous ion chelating activity of ESM was undetectable at the tested concentrations. In contrary, EPN exhibited strong ferrous ion chelating activity and high scavenging activities against hydrogen peroxide, hydroxyl radicals, and a weak activity against superoxide anion and DPPH free radicals. EPN did not show any ferric ion reducing power. Since their antioxidant mechanisms are complementary, the combined use of ESM and EPN might be even more beneficial. These antioxidant properties of SM and PN are likely part of the reasons that they are effective in the prevention and treatment of vascular diseases.     

Superoxide and hydroxyl radical scavenging actions of botanical extracts of Wagatea spicata

Wagatea spicata, candy corn plant, a woody legume shrub, widespread medicinal plant found in Western Ghat of India has significant abilities to scavenge highly reactive free radicals. Shade dried leaf, bark and flower powder of this plant has been extracted with water and fractionated with different solvents. Extracts and their solvent fractions were found to be good scavengers of superoxide and hydroxyl radicals. Free radical scavenging action of W. spicata is due to its rich phenolic and flavonoid contents. Bark and leaf extracts showed significant scavenging action against superoxide radicals, where as flower extracts efficiently inhibited hydroxyl radicals.     

Insecticidal activities of aromatic plant extracts and essential oils against Sitophilus oryzae and Callosobruchus chinensis

Methanol extracts from 30 aromatic medicinal plant species and five essential oils were tested for their insecticidal activities against adults of Sitophilus oryzae (L.) and Callosobruchus chinensis (L.), using direct contact application and fumigation methods. Responses varied with plant material, insect species, and exposure time. In a test with a filter paper diffusion method at 3.5 mg/cm², potent insecticidal activity against both species was produced by an extract from Cinnamomum cassia bark, cinnamon (C. cassia) oil, horseradish (Cocholeria aroracia) oil, and mustard (Brassica juncea) oil within 1 day after treatment. Over 90% mortality at 3 or 4 days after treatment was achieved using extracts of Acorus calamus var. angustatus rhizome, Acorus gramineus rhizome, Illicium verum fruit, and Foeniculum vulgare fruit. An extract from Cinnamomum sieboldii root bark gave 100% mortality at 2 days after treatment. At 0.7 mg/cm², extracts from C. cassia, C. sieboldii, and F. vulgare as well as cinnamon oil, horseradish oil and mustard oil were highly effective against both species. In a fumigation test with S. oryzae adults, the oils described were much more effective in closed containers than in open ones, indicating that the insecticidal activity of the oils was attributable to fumigant action. The plant extracts and essential oils described could be useful for managing field populations of S. oryzae and C. chinensis.     

In vitro antibacterial and antioxidant activities of Orthosiphon stamineus Benth. extracts against food-borne bacteria

Extracts of the Orthosiphon stamineus plant were tested for antimicrobial and antioxidant activities against selected food-borne bacteria in vitro. Whole O. stamineus plants (powdered) were extracted using various concentrations (0%, 25%, 50%, 75%, and 100%) of methanol. O. stamineus extracted with 50% methanol, 75% methanol and fraction 5 of a 50% methanolic extract demonstrated inhibitory activity against Vibrio parahaemolyticus. The inhibition observed with these O. stamineus extracts was comparable to the inhibition seen with the natural food preservative 5% lactic acid; this is likely due to the high concentration of rosmarinic acid found in the O. stamineus extracts. This study showed that the highest concentration of rosmarinic acid had the best antibacterial and free radical scavenging activities. This suggests that rosmarinic acid content is closely associated with antibacterial and free radical scavenging activities of O. stamineus extracts.     

Antimicrobial activities of some thymol derivatives from the roots of Inula hupehensis

In a search for some naturally occurring antimicrobial compounds found in medicinal plants, herbs and spice extracts, four thymol derivatives were isolated from the roots of Inula hupehensis (a medicinal plant) and identified as the new compound 9-O-β-d-glucopyranosyl-9-hydroxythymol (1), together with 8,9-epoxy-10-isobutyryloxythymyl isobutyrate (2), 8-hydroxy-9, 10-diisobutyloxythymol (3) and 8,9,10-trihydroxythymol (4). These thymol derivatives, with a semi-synthetic compound 9,10-dihydroxy-8-methoxythymol (5), were tested for their antimicrobial activities against three bacteria and six plant pathogenic fungi. Within the series of thymol derivatives tested, compound 3 was the most active, particularly displaying moderate antibacterial activities against Staphylococcusaureus, Methicillin-resistant S. aureus and Escherichia coli with MICs of 62.3, 62.8, and 250 μg/ml. Moreover, this compound exhibited inhibitory activities against three plant pathogenic fungi: Rhizoctonia solani, Phytophthora melonis and Peronophythora litchi, with EC₅₀ values of 157, 180 and 141 μg/ml. To our knowledge, this is the first report that these five thymol derivatives show inhibitory activity against plant pathogenic fungi.     

Integrated management of insect vectors of Aspergillus flavus in stored maize, using synthetic antioxidants and natural phytochemicals

The purpose of this study was to investigate the insecticidal activity of two benzoic acids 2(3)-tert-butyl-4 hydroxyanisole (BHA) and 2,6-di(tert-butyl)-p-cresol (BHT); two phenolic acids 3-phenyl-2-propenoic acid (CA) and trans-4-hydroxy-3-methoxycinnamic acid (FA) and two essential oils of Eugenia caryophyllata (clove tree) and Thymus vulgaris (thyme) against Sitophilus zeamais, Tribolium confusum and Rhyzopertha dominica, vector carriers of aflatoxigenic fungi in stored maize. The susceptibility of insects, the frequency of isolation of Aspergillus section Flavi in insects and maize, and the analysis of aflatoxin B₁ in maize were determined. BHA, BHT, BHA/BHT mixture and the natural phytochemicals AF and AF/AC mixture showed the highest insecticidal activity against S. zeamais, T. confusum and R. dominica after 120 days of incubation. The insecticidal efficacy of the volatile fraction of essential oils of clove and thyme showed less inhibition. There was no contamination of Aspergillus section Flavi in dead and live insects collected from maize treated with BHA. No aflatoxin B₁ accumulation was detected in the control and treatments. The information obtained shows that these substances have the potential to control pest insect vectors of aflatoxigenic fungi in stored maize in microcosms during 120 days.     

Chemical composition and biological activities of Algerian Thymus oils

The compositions of essential oils isolated from nine samples of three Thymus species (Thymus algeriensis, Thymus pallescens and Thymus dréatensis) were analysed by GC and GC–MS, and a total of 114 components were identified. T. pallescens collected from various regions showed a great similarity in their compositions and were characterised by carvacrol (44.4–57.7%), p-cymene (10.3–17.3%) and γ-terpinene (10.8–14.2%) as the major components for four samples; only one sample was thymol-rich (49.3%) with a small amount of carvacrol (9.0%). On the other hand, T. algeriensis showed a chemical polymorphism, even for samples from the same location, and two new chemotypes for this species were proposed. Oxygen-containing monoterpenes were the predominant class (76.3%) in T. dreatensis oil, with linalool (30.4%), thymol (20.2%) and geraniol (19.6%) as the principal constituents. The oils were screened for their possible antioxidant activities by four complementary assays, namely DPPH free radical scavenging, hydroxyl radical scavenging, inhibition of lipid peroxidation and reducing power. The two new chemotypes of T. algeriensis exhibited strong hydroxyl radical scavenging (IC₅₀ = 2.2–3.3 μg/ml), but were not or only slightly active against the other radicals and exhibited a weak reducing power. Despite their chemical similarity, T. pallescens oils sometimes produced significant differences in their antioxidant activities. The essential oils were also screened for their antimicrobial activity against five bacteria (three Gram-positive and two Gram-negative) and one yeast (Candida albicans). The tested essential oils showed antimicrobial activity against the microorganisms used, in particular against two important pathogens, C. albicans and Helicobacter pylori.     

Antimicrobial activity of extract and two alkaloids from traditional Chinese medicinal plant Stephania dielsiana

Two alkaloids, stephanine and crebanine, were isolated from tubers of the traditional Chinese medicinal plant Stephania dielsiana, using an activity-directed isolation method, and inhibitory activity of methanol extract, stephanine and crebanine against ten animal pathogenic bacteria and eight plant pathogenic fungi was evaluated in vitro. The results showed that extract from S. dielsiana exhibited high inhibitory activity against five gram-positive and four gram-negative animal pathogenic bacteria, with MIC values of 0.625–7.5 g/l; stephanine and crebanine had high inhibitory activity against gram-positive animal pathogenic bacteria, with MIC values of 0.078–0.312 g/l, but low inhibitory activity against gram-negative animal pathogenic bacteria. Methanol extract, stephanine and crebanine also inhibited hyphal growth of the plant pathogens Cercospora kaki, Gymnosporangium haraeanum, Pyricularia oryzae, Rhizoctonia solani and Colletotrichum graminicola, and spores germination of Thielaviopsis paradoxa, Fusarium oxysporum f. sp. niveum, Sphaceloma fawcettii and G. haraeanum.     

Polyphenols and triterpenes from Chaenomeles fruits: Chemical analysis and antioxidant activities assessment

Mugua, fruit of the genus Chaenomeles, is a valuable source of health food and Chinese medicine. To elucidate the bioactive compounds of five wild Chaenomeles species, extracts from fresh fruits were investigated by HPLC-DAD/ESI-MS/MS. Among the 24 polyphenol compounds obtained, 20 were flavan-3-ols (including catechin, epicatechin and procyanidin oligomers). The mean polymerisation degree (mDP) of procyanidins was examined by two acid-catalysed cleavage reactions; the mDP value was the highest in Chaenomeles sinensis and the lowest in Chaenomeles japonica. Total polyphenol content (TPC) reached 46.92 and 46.28 mg/g gallic acid equivalents (GAE) in Chaenomeles speciosa and Chaenomeles thibetica, respectively, although their main bioactive compounds were different (being epicatechin and procyanidin B2 in the former, and catechin and procyanidin B1 in the latter). These two species also exhibited equally strong free radical scavenging activities. Our results further showed that the antioxidant ability of Chaenomeles fruits was significantly correlated to their total polyphenol contents. Two triterpenes (oleanolic acid and ursolic acid) were the highest quantity in Chaenomeles cathayensis and C. thibetica, respectively.     

Biological activities of flavonoids from pathogenic-infected Astragalus adsurgens

The present study was aimed at the identification of biological components from pathogenic-infected Astragalus adsurgens with an activity-guided purification process. Fifteen flavonoids were obtained from the most active ethyl acetate fraction and identified as astradsurnin (1), a new chalcone derivative, together with fourteen known flavonoids (2-15). These compounds were tested for antibacterial activities against five bacteria and cytotoxic activities against two selected cancer cells. Within the series of flavonoids tested, compounds 4 and 15 were the most active against Escherichia coli, Bacillus cereus, Staphylococcus aureus, Erwinia carotovora and Bacillus subtilis with MICs ranging from 7.8 to 31.3 μg/ml. Moreover, compounds 4 and 5 exhibited moderate activity against HL-60 and SMMC-7721 cells with IC₅₀ values between 5 and 10 μg/ml. To our knowledge, this is the first time that it has been reported on the biological and chemical study of A. adsurgens infected by Embellisia astragali.     

Anthraquinone profile,antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, R. fallax, R. intermedia and R. pumila

The quantity of phenols, as well as antioxidant and antimicrobial activities, were investigated in bark of Rhamnus alaternus L., R. fallax Boiss., R. intermedia Steud. et Hochst., and R. pumila Turra from natural stands in Croatia. The most abundant anthraquinones in the investigated extracts were chrysophanol in R. alaternus (3.14 mg/g), emodin in R. pumila (0.339 mg/g), and physcion in R. fallax (2.70 mg/g) and R. intermedia (0.285 mg/g). The species exhibiting the highest antioxidant activity were R. fallax and R. pumila. A positive correlation was observed between total phenolic and flavonoid levels of the extracts and antioxidant activity in some of the assays. All species showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, Aspergillus niger and Microsporum gypseum with minimal inhibitory concentrations equal to or below 2.500 mg/mL. The results indicate that the investigated Rhamnus species are a source of anthraquinones and other phenols, which act as multifunctional antioxidants with antimicrobial activity.     

Essential oils of Zingiber officinale var. rubrum Theilade and their antibacterial activities

The essential oils obtained by hydrodistilation of the leaves and rhizomes of Zingiber officinale var. rubrum Theilade were analysed by capillary GC and GC–MS. Forty-six constituents were identified in the leaf oil, while 54 were identified in the oil from the rhizomes. The leaf oil was clearly dominated by β-caryophyllene (31.7%), while the oil from the rhizomes was predominantly monoterpenoid, with camphene (14.5%), geranial (14.3%), and geranyl acetate (13.7%) the three most abundant constituents. The evaluation of antibacterial activities using the micro-dilution technique revealed that both the leaf and rhizome oils were moderately active against the Gram-positive bacteria Bacilluslicheniformis, Bacillus spizizenii and Staphylococcus aureus, and the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae and Pseudomonas stutzeri.     

Evaluation of antioxidant,antibacterial and anti-tyrosinase activities of four Macaranga species

The methanolic fresh leaf extracts of Macaranga gigantea, Macaranga pruinosa, Macaranga tanarius and Macaranga triloba were screened for their antioxidant properties (AOP), tyrosinase inhibition and antibacterial activities. Total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ferric-ion reducing power (FRAP), ferrous-ion chelating (FIC) and lipid peroxidation inhibition (LPI) activities were used to evaluate the AOP. Modified 3,4-dihydroxy-L-phenylalanine (L-DOPA) method was used to determine tyrosinase inhibition activity, whereas antibacterial activity was determined using the disc-diffusion technique. TPC screening of the same species from different collection sites showed no significant difference between sites. M.triloba showed the highest ascorbic acid equivalent antioxidant activity (AEAC), FRAP and LPI values. M. tanarius, which showed the lowest TPC, AEAC, FRAP and LPI activities, exhibited the best FIC activity. M. pruinosa showed the best tyrosinase inhibition activity, whereas M. triloba showed the best antibacterial activity against Gram-positive bacteria species, with minimal inhibition dosage (MID) values as low as 10 μg/disc.     

Legume type and temperature effects on the toxicity of insecticide to the genus Callosobruchus (Coleoptera: Bruchidae)

Three bruchid pest species, Callosobruchus maculatus, Callosobruchus chinensis and Callosobruchus rhodesianus, were studied for their response to insecticide toxicity taking into account the separate and interactive effects of temperature and pre-adult food. The food types used were cowpea (Vigna unguiculata) and mungbean (Vigna radiata). Callosobruchus maculatus was the most tolerant to malathion and the least affected by temperature change while C. rhodesianus was the least tolerant. Over a 4 °C range (23°, 25°, 27 °C), there was generally a significant impact of temperature on the tolerance of the three species to the insecticide. The food type on which the insects developed influenced considerably the degree of insecticide tolerance. Callosobruchus maculatus and C. chinensis populations reared on mungbean had higher tolerance to malathion than their counterparts reared on cowpea, but the opposite was observed in C. rhodesianus populations. The food influence in this study suggested an ancestral cause or fitness cost depending on the species. The interaction of food-by-temperature had no significant effect on malathion toxicity to this genus. Correlation analysis showed C. chinensis to be relatively less sensitive to insecticide concentration over the range studied compared with the other two species.     

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.     

In vitro activity of methanol extracts of plants used as spices against Mycobacterium tuberculosis and other bacteria

This study determined the phenolic composition of two thyme species (Lamiaceae), Origanum minutiflorum O. Schwarz and P.H. Davis and Thymbra spicata L. var. spicata, and assessed their antibacterial and antimycobacterial activities. “Kekik” is a collective term used in Turkey for plants that smell like thyme. O. minutiflorum, (locally “Sutculer kekigi”, endemic) and T. spicata var. spicata (locally “Karakekik”) are widely used in Turkey and are important export commodities.     

Characterisation of esterolytic activity from two wild mushroom species, Amanita vaginata var. vaginata and Tricholoma terreum

Characterisation of esterase activities from the edible mushroom species, Amanita vaginata var. vaginata and Tricholoma terreum, were investigated. Native electrophoresis of the crude extracts prepared from both mushroom samples showed the presence of esterolytic activities. The extracts had the greatest activity in the presence of p-nitrophenyl butyrate (pNPB) as a substrate. pH and temperature optima were found to be 8.0 and 30 °C for both enzymes, respectively. V_max and K_m values were determined as 14.2 U/l and 71 μM for A. vaginata var. vaginata and 34.6 U/l and 9.6 μM for T. terreum, respectively. The pH-stability profile showed a stationary line between 3.0 and 10.0 for both enzymes. The esterolytic activities from the extracts were maintained between 10 and 40 °C for 4 h and started to decrease at 50 °C. The effects of EDTA, NaN₃, DTT and PMSF on the enzyme activity were also investigated.     

Susceptibility of commercial oat cultivars to Cryptolestes pusillus and Oryzaephilus surinamensis

Susceptibility to two storage insect pests [(Cryptolestes pusillus (Schönherr) and Oryzaephilus surinamensis (L.)] of eight commercial oat cultivars from the United States was determined in laboratory studies. Duration of insect development was shorter and number of progeny produced was greater on cracked than on whole oats. Simulations based on data from the study showed that insect populations would reach the threshold level for treatment in 2-3 months of storage at 30°C on cracked oats. Insect population development was slowest on the hulless cultivar Paul when the oat kernels were cracked. Simulations also indicated that all cultivars of whole oats tested could be stored for at least 1 yr at 30°C without reaching the threshold for treatment when infested with these two species of insects, and insect populations would decrease over time on the cultivars Don, Jerry, Milton, NewDak, Otana, and Valley. Analyses of oat grain quality characteristics, including kernel weight, groat hardness, and groat composition, provided little insight into the mechanism of observed differences in insect development among cultivars. Hardness of the kernels (as indicated by % broken groats after dehulling) may be related to near immunity to these two species of insects in whole Otana. Steaming whole oats to inactivate hydrolytic enzymes in the trichomes of the pericarp did not increase susceptibility to these two species of insects, suggesting that enzymes in the trichomes were not responsible for insect population development being slower on whole oats than on cracked oats. Although we were unable to identify the factors that determined relative susceptibility in this study, the results will be useful for selecting commercial oat cultivars for planting that will be less susceptible to insect pests in storage and suggest that the economics of cleaning oats before storage to reduce insect population growth should be investigated.     

In vitro antimicrobial activity of essential oil from endemic Origanum minutiflorum on ciprofloxacin-resistant Campylobacter spp.

This study evaluated the antimicrobial activities of an essential oil of Origanum minutiflorum (O. Schwarz and P.H. Davis) against ciprofloxacin-resistant Campylobacter spp., by broth microdilution and agar well-diffusion methods. Moreover, O. minutiflorum oil was analyzed by gas chromatography/mass spectrometry (GC/MS). Twenty-nine components were identified, representing 98.7 of the oil. The oil yield from the plants was 4.0–4.4% v/w. The major components of O. minutiflorum oil were carvacrol (73.9%) and p-cymene (7.20%). The oil has lower contents of carvacrol methyl ether (0.05%), heptadecanol (0.06%) and carvacryl acetate (0.06%). Twenty-one Campylobacter spp. (12 C. jejuni, 5 C. lari and 4 C. coli) strains using in this study were selected among 300 isolates according to their resistance to ciprofloxacin. The minimum inhibitory concentration (MIC) values for bacterial strains, which were sensitive to the essential oil of O. minutiflorum, were in the range of 7.8–800 μg/ml. The essential oil obtained showed strong antimicrobial activity against all of the tested ciprofloxacin-resistance Campylobacter spp. These results suggest that the essential of O. minutiflorum may be used as a natural preservative in food against food-born disease, such as Campylobacteriosis.     

Composition and biological activities of essential oils from four Heracleum species

The composition of essential oils from aerial parts of Heracleum persicum, a widely used medicinal plant, and three other Heracleum species growing wild in Iran were analysed by GC and GC–MS. Myristicin (53.6%), (E)-anethole (25.0%), hexyl butanoate (29.7%) and elemicin (41.1%) were the major compounds of Heracleum pastinacifolium, H. persicum, Heracleum rechingeri and Heracleum transcaucasicum, respectively. Cytotoxic activity assessed on three human cancer cell lines (HeLa, LS180 and Raji), showed that essential oils from H. transcaucasicum (IC₅₀ values; 0.362–0.594 mg/ml) followed by H. pastinacifolium (0.497–1.398 mg/ml) had moderate antitumoral activities. In the DPPH radical scavenging assay, H. pastinacifolium and H. persicum oils showed the highest activities with IC₅₀ values of 7.3 and 7.4 mg/ml, respectively. Antioxidant activity correlated well with the total phenolic content of the oils. None of the essential oils showed significant antimicrobial activities.