Alkaloid profile and antimicrobial activity of <Emphasis Type="Italic">Lupinus angustifolius</Emphasis> L. alkaloid extract

Purpose of the present study was to evaluate alkaloid profile of the aerial parts of Lupinus angustifolius growing in Turkey by capillary gas chromatography-mass spectrometry (GC-MS). Fifteen alkaloids were identified by capillary GC-MS. 13α-Hydroxylupanine (50.78%) and lupanine (23.55%) were determined as the main alkaloids in the aerial parts of L. angustifolius. Ammodendrine, isoangustifoline, tetrahydrorhombifoline, angustifoline, α-isolupanine, 5,6-dehydrolupanine, 11,12-dehydrolupanine, 13α-acetoxylupanine, 13α-isovaleroyloxylupanine, 13α-valeroyloxylupanine, 13α-tigloyloxylupanine, 13α-cis-cinnamoyloxylupanine and 13α-cis-cinnamoyloxy-17-oxolupanine were identified as the minor alkaloids of the plant. Furthermore, antibacterial and antifungal activities of L. angustifolius alkaloid extract were tested against standard strains of the following bacteria; Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus as well as the fungi; Candida albicans and C. krusei. The alkaloid extract showed significant activity on B. subtilis, S. aureus and P. aeruginosa while it was weakly active on E. coli. On the other hand, the extract possessed moderate activity against C. albicans and C. krusei.     

Antibacterial,antifungal, insecticidal,cytotoxicity and phytotoxicity studies on Indigofera gerardiana

The antibacterial, antifungal, acute cytotoxicity, phytotoxicity and insecticidal profile of the crude extract and various fractions of Indigofera gerardiana have been studied. Six bacterial and fungal strains were used, of which Samonella typhi and Microsporum canis were the most susceptible strains with MICs 0.37 mg/mL and 0.09 mg/mL, respectively. The crude extract and the fractions showed low insecticidal activity against Sitophilus oryzae, Rhyzopertha dominica and Callosbruchus analis but no activity against Tribolium castaneum. The brine shrimp lethality assay showed absence of any measurable cytotoxicity of the crude extract and fractions, showing a good safety profile at a preliminary level. All the fractions except crude extract revealed profound and highly significant herbicidal activity against Lemna minor at the concentration of 1000 μg/mL. Indigofera gerardiana was shown by in-vitro assays to be a potential source for natural antifungal, antibacterial and herbicidal agents.     

Antibacterial and antifungal activities of andrachne cordifolia muell

The crude methanolic extract of Andrachne cordifolia Muell. (Euphorbiaceae) and its various fractions in different solvent systems (chloroform, ethyl acetate and n- butanol) were screened for antibacterial and antifungal activities. Crude extract and subsequent fractions demonstrated moderate to excellent antibacterial activities against the tested pathogens. Highest antibacterial activity was displayed by both chloroform and ethyl acetate fractions (100%) followed by the crude extract (68%) against Salmonella typhi. Similarly, crude extract and its subsequent fractions showed mild to excellent activities in antifungal bioassay with maximum (76%) antifungal activity against Microsporum canis by the chloroform fraction followed by the crude extract (65%).     

Antibacterial and antifungal activities of teucrium royleanum (Labiatea)

The crude methanolic extract and subsequent fractions of Teucrium royleanum (Labiatea) were screened for antibacterial and antifungal activities. Against tested pathogens, crude extract and subsequent fractions demonstrated moderate to excellent antibacterial activities. Highest antibacterial activity was displayed by the ethyl acetate fraction against S. typhi (100%), against E.coli (76.7%) and against P. aerugenosa (70.8%) followed by the chloroform fraction against S. typhi (85.7%). Similarly, the crude extract and its subsequent fractions showed mild to excellent activities in the antifungal bioassay with maximum antifungal activity against M. canis (87%) by the chloroform fraction followed by the ethyl acetate (71%) and n-butanol (70%) fractions.     

Antimicrobial activities of Gloriosa superba Linn (Colchicaceae) extracts

The methanol extract of the rhizomes of Gloriosa superba Linn (Colchicaceae) and its subsequent fractions in different solvent systems were screened for antibacterial and antifungal activities. Excellent antifungal sensitivity was expressed by the n-butanol fraction against Candida albicans and Candida glaberata (up to 90%) and against Trichophyton longifusus (78%) followed by the chloroform fraction against Microsporum canis (80%). In the antibacterial bioassay, the crude extract and subsequent fractions showed mild to moderate antibacterial activities. Chloroform fraction displayed highest antibacterial sensitivity against Staphylococcus aureous (88%) followed by the crude extract (59%). The total phenol content of the crude extract and fractions of the plant expressed no significant correlation with the antimicrobial activities.     

Antimicrobial activity in sub-Arctic marine invertebrates

In the marine environment, any living or non-living surface is exposed to bacterial colonization. Many invertebrate species in temperate, tropical and Antarctic regions have demonstrated chemical defences against the formation of microbial films. In the present study, the antimicrobial activity of sub-Arctic invertebrates was investigated for the first time. Crude extracts of abundant invertebrates belonging to several taxonomic groups were tested for their inhibitory effects on the growth of five sympatric phylogenetically diverse bacterial strains. Six out of 18 (33%) crude extracts inhibited bacterial growth at natural extract concentrations. The crude extract of the sponge Haliclona viscosa inhibited growth of all five bacterial strains, suggesting the presence of metabolites with broad-spectrum activity. Three active compounds were isolated from H. viscosa having antibacterial properties similar to those of the crude extract. Our data indicate that antibacterial secondary metabolites are present in sub-Arctic marine invertebrates but are less abundant than in temperate, tropical or Antarctic species.     

川蔓藻内生及根际细菌多样性与抑菌活性研究

该研究采用稀释涂布法结合形态观察、16S rRNA基因序列分析,对广西北海川蔓藻(Ruppia maritima)内生及根际细菌的物种多样性进行了研究,并采用琼脂扩散法和光度计法分析了其粗提物抑制马尔尼菲青霉菌活性。结果表明:从川蔓藻中分离到可培养内生细菌26株,根际可培养细菌31株。分别将内生细菌归属为10科12属13种,根际分离出细菌归属为9科14属19种,其中5株根际细菌可能为潜在新种。获得8株细菌对马尔尼菲青霉菌有抑制活性,总阳性率为25.0%。其中,菌株BGMRC 2015、BGMRC 2059、BGMRC 2043的粗提物表现出较强的抑制马尔尼菲青霉菌效果,其MIC分别为(1.800±0.045)、(1.881±0.061)、(1.604±0.021)mg·m L~(-1)。川蔓藻中可培养细菌具有较高的物种多样性,蕴藏着丰富的新物种资源,且富含抑菌活性良好的菌株。     

Dieter Seidel‐65 Jahre

Fruit extract of Solanum xanthocarpum was evaluated for its toxicity against Alternaria brassicae, the causal agent of Alternaria blight of Indian mustard [Brassica juncea (L.) Czern. &; Coss]. The mass obtained after vacuum drying of the crude methanolic extract was utilised for further sequential fractionation using n-hexane, ethyl acetate, n-butanol and methanol. Among the crude and different fractions tested, methanolic fraction was most effective with a minimum inhibitory concentration (MIC) of 62.5 μg/ml. The methanolic fraction was further fractionated using open column liquid chromatography into five subfractions (I–V) to identify the antifungal bioactive compounds. Among the five subfractions (SFs) tested SF IV was most effective at inhibiting A. brassicae conidial germination and thereby inhibited lesion development of Alternaria blight at a concentration of 15.625 μg/ml or higher. Furthermore, bioautography of SF IV with Alternaria alternata and diagnosis with Dragendorff reagent indicated that SF IV contains a mixture of bioactive alkaloids, namely a₁ (R_f = 0.12) and a₂ (R_f = 0.22). The potential of using S. xanthocarpum as a resource for the development of biofungicides is discussed.     

Antifungal activity of crude extract produced by Bacillus sp. associated with entomopathogenic nematode from media formulated by six nitrogen sources with fructose against Penicillium expansum

A specific symbiotic Bacillus species isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp. was found to produce a number of bioactive compounds. The present study was conducted to determine the effect of six different nitrogen sources in combination with fructose on the production of antifungal crude extract by Bacillus sp. against Penicillium expansum. The yield of crude extract and antifungal activity against the test fungi differed significantly when the nitrogen sources in the fermentation media were changed. The highest yield was recorded for beef extract plus fructose (921?mg/L). The antifungal activity was higher in yeast extract plus fructose [P. expansum (46.5?±?2.12?mm)], followed by beef extract. High performance liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention times indicating that they produced different compounds. When a carbon source was not included in the fermentation medium, the antimicrobial production was substantially reduced almost eight times. Carbon source in the fermentation medium plays a vital role in the production of antimicrobial substances. Yeast extract and fructose as nitrogen and carbon sources in the fermentation medium produced maximum antimicrobial activity.     

Interaction of benzo[c]phenanthridine and protoberberine alkaloids with animal and yeast cells

We compared the effects of four quaternary benzo[c]phenanthridine alkaloids – chelerythrine, chelilutine, sanguinarine, and sanguilutine – and two quaternary protoberberine alkaloids – berberine and coptisine – on the human cell line HeLa (cervix carcinoma cells) and the yeastsSaccharomyces cerevisiae andSchizosaccharomyces japonicus var. versatilis. The ability of alkaloids to display primary fluorescence, allowed us to record their dynamics and localization in cells. Cytotoxic, anti-microtubular, and anti-actin effects in living cells were studied. In the yeasts, neither microtubules nor cell growth was seriously affected even at the alkaloid concentration of 100 μg/ml. The HeLa cells, however, responded to the toxic effect of alkaloids at concentrations ranging from 1 to 50 μg/ml. IC₅₀ values for individual alkaloids were: sanguinarine IC₅₀ = 0.8 μg/ml, sanguilutine IC₅₀ = 8.3 μg/ml, chelerythrine IC₅₀ = 6.2 μg/ml, chelilutine IC₅₀ = 5.2 μg/ml, coptisine IC₅₀ = 2.6 μg/ml and berberine IC₅₀ >10.0 μg/ml. In living cells, sanguinarine produced a decrease in microtubule numbers, particularly at the cell periphery, at a concentration of 0.1 μg/ml. The other alkaloids showed a similar effect but at higher concentrations (5–50 μg/ml). The strongest effects of sanguinarine were explained as a consequence of its easy penetration through the cell membrane owing to nonpolar pseudobase formation and to a high degree of molecular planarity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Antioxidant and antimicrobial actions of the clubmoss <Emphasis Type="Italic">Lycopodium clavatum</Emphasis> L.

Purpose of the present study was to evaluate antioxidant, antibacterial, antifungal, and antiviral activities of the petroleum ether, chloroform, ethyl acetate and methanol extracts as well as the alkaloid fraction of Lycopodium clavatum L. (LC) from Lycopodiaceae growing in Turkey. Antioxidant activity of the LC extracts was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging method at 0.2 mg/ml using microplate-reader assay. Antiviral assessment of LC extracts was evaluated towards the DNA virus Herpes simplex (HSV) and the RNA virus Parainfluenza (PI-3) using Madin-Darby Bovine Kidney (MDBK) and Vero cell lines. Antibacterial and antifungal activities of the extracts were tested against standard and isolated strains of the following bacteria; Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Acinobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis as well as the fungi; Candida albicans and C. parapsilosis. All of the extracts possessed noteworthy activity against ATCC strain of S. aureus (4 μg/ml), while the LC extracts showed reasonable antifungal effect. On the other hand, we found that only the chloroform extract was active against HSV (16–8 μg/ml), while petroleum ether and alkaloid extracts inhibited potently PI-3 (16–4 μg/ml and 32–4 μg/ml, respectively). However, all of the extracts had insignificant antiradical effect on DPPH. In addition, we also analyzed the content of the alkaloid fraction of the plant by capillary gas chromatography-mass spectrometry (GC-MS) and identified lycopodine as the major alkaloid.     

Antimicrobial activity of berberine--a constituent of Mahonia aquifolium

The antimicrobial activity of the protoberberine, alkaloid, berberine, isolated fromMahonia aquifolium, was evaluated against 17 microorganisms including two Gram-negative bacteria—Pseudomonas aeruginosa andEscherichia coli (both resistant and sensitive), two Gram-positive bacteria—Bacillus subtilis andStaphylococcus aureus, Zoogloea ramigera, six filamentous fungi—Penicilium chrysogenum, Aspergillus niger, Aureobasidium pullulans (black and white strain),Trichoderma viride (original green strain and brown mutant),Fusarium nivale, Mycrosporum gypseum, and two yeasts—Candida, albicans andSaccharomyces cerevisiae. The IC₅₀, minimum inhibitory concentration (MIC), minimum microbicidal concentration (MMC) and minimum microbistatic concentration (MMS) varied considerably depending on the microorganism tested, the sensitivity decreasing as follows:S. aureus >P. aeruginosa S (sensitive) >E. coli S>P. aeruginosa R (resistant) >E. coli R>B. subtilis>Z. ramigera>C. albicans>S. cerevisiae>A. pullulans B (black)>A. pullulans W (white)>T. viride Br (brown)>M. gypseum>A niger>F. nivale>P. chrysogenum>T. viride G (green).     

Crude Extracts and Secondary Metabolites of Epichloë bromicola against Phytophthora infestans

Potato late blight caused by Phytophthora infestans is still one of the main factors limiting potato production. Epichloë spp. can provide host plants with various resistances, which makes them show great potential in the biological control of diseases. In this study, we explored the potential biological activity of crude extracts of 20 strains of Epichloë bromicola to control P. infestans. The crude extracts of strains 1 and 8 showed significant antifungal activity with an inhibition rate of 88 % and 81 %, respectively, and showed different effects on the mycelium morphology of P. infestans observed by scanning electron microscopy. Moreover, the two crude extracts demonstrated an interesting therapeutic and protective effect on potato late blight, and none of the extracts had an adverse effect against zebrafish embryos. A total of 13 metabolites were isolated from the crude extract of strain 8, and these tested compounds showed a weak antifungal effect and the inhibition rate was less than 80 %. These findings suggested that strains 1 and 8 have potential for biocontrol of late potato blight.     

Impact of beef extract and six carbon source on antifungal metabolites production by bacterium associated with entomopathogenic nematode against Fusarium oxysporum

A specific symbiotic Bacillus species isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp., was found to produce a number of bioactive compounds. The present study was conducted to determine the effect of six different carbon sources in combination with beef extract on the production of antifungal substances by Bacillus sp. The yield of crude antimicrobial substances and antimicrobial activity against the test microorganism also differed significantly when the carbon sources in the fermentation media were changed. The highest yield was recorded for fructose plus beef extract (956?mg/l). The antifungal activity was significantly high in beef extract plus maltose (21?±?1.5?mm) followed by beef extract plus glucose and beef extract plus fructose. Antifungal activity was significantly reduced in beef extract plus lactose and sucrose. High pressure liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention times indicating that they produced different compounds. When a carbon source was not included in the fermentation media, the antifungal production was substantially reduced. Carbon source in the fermentation medium plays a vital role in the production of antimicrobial substances. Beef extract and maltose as nitrogen and carbon sources in the fermentation medium produced maximum antifungal activity. It is concluded that Beef extract and maltose as nitrogen and carbon sources produced maximum activity which can effectively control the Fusarium oxysporum which causes vascular fusarium wilt in tomato, tobacco, legumes, cucurbits, sweet potatoes, banana, etc.     

Insecticidal and antifeedant effects of two alkaloids from Cynanchum komarovii against larvae of Plutella xylostella L

A bioassay‐guided fractionation of Cynanchum komarovii crude alkaloid extract led to the isolation of two alkaloids. The isolated alkaloids were identified as 7‐demethoxytylophorine (1) and 6‐hydroxyl‐2,3‐dimethoxy phenanthroindolizidine (2) based on the comparison of their spectroscopic characteristics with the literature data. Insecticidal, antifeedant and growth inhibitory effects of these two alkaloids against the 3rd instar larvae of Plutella xylostella L. (Lepidoptera: Plutellidae) were examined. The results showed that alkaloid 1 was more toxic than alkaloid 2 against the 3rd instar larvae of Plutella xylostella L., but both alkaloids were less toxic than the total alkaloid fraction. For antifeedant activity, alkaloid 1 showed AFC₅₀ of 1.82 mg/ml at 24 h after treatment, alkaloid 2 showed 3.89 mg/ml, while total alkaloids showed 1.56 mg/ml. In dipping toxicity test, alkaloids 1 and 2 produced 93.3% and 63.3% mortality at 72 h after treatment, respectively, while total alkaloids produced 96.7% mortality. The LC₅₀ values for alkaloids 1, 2 and the total alkaloids were 3.54, 9.21 and 2.63 mg/ml, respectively. The development of larvae was also inhibited, and the growth inhibition rates at the concentration of 15.00 mg/ml were 92.8%, 78.2% and 98.6% for alkaloids 1, 2 and total alkaloids, respectively, at 72 h after treatment. Compared with antifeedant and dipping effect, the alkaloids 1, 2 and total alkaloid fraction revealed weak feeding toxicity, and their corrected mortality rates at the concentration of 15.00 mg/ml were 60.0%, 40.0% and 63.3% at 7 days after treatment. The LC₅₀ values for alkaloids 1, 2 and total alkaloids were 12.58, 32.37 and 8.88 mg/ml, respectively, at 7 days after treatment.     

Effect of polyacetylenic acids from Prunella vulgaris on various plant pathogens

Aims: This study is aiming at characterizing antifungal substances from the methanol extract of Prunella vulgaris and at investigating those substances’ antifungal and antioomycete activities against various plant pathogens. Methods and Results: Two polyacetylenic acids were isolated from P. vulgaris as active principles and identified as octadeca‐9,11,13‐triynoic acid and trans‐octadec‐13‐ene‐9,11‐diynoic acid. These two compounds inhibited the growth of Magnaporthe oryzae, Rhizoctonia solani, Phytophthora infestans, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. raphani, and Phytophthora capsici. In addition, these two compounds and the wettable powder‐type formulation of an n‐hexane fraction of P. vulgaris significantly suppressed the development of rice blast, tomato late blight, wheat leaf rust, and red pepper anthracnose. Conclusions: These data show that the extract of P. vulgaris and two polyacetylenic acids possess antifungal and antioomycete activities against a broad spectrum of tested plant pathogens. Significance and Impact of the Study: This is the first report on the occurrence of octadeca‐9,11,13‐triynoic acid and trans‐octadec‐13‐ene‐9,11‐diynoic acid in P. vulgaris and their efficacy against plant diseases. The crude extract containing the two polyacetylenic acids can be used as a natural fungicide for the control of various plant diseases.     

Antifungal activity in vitro of Baccharis glutinosa and Ambrosia confertiflora extracts on Aspergillus flavus, Aspergillus parasiticus and Fusarium verticillioides

The aims of this study were to evaluate the antifungal properties of Baccharis glutinosa and Ambrosia confertiflora extracts against Aspergillus flavus, A. parasiticus and Fusarium verticillioides, and to isolate the group of compounds that are responsible for the antifungal activity. Samples of aerial parts from each plant were extracted with 70% methanol and sequentially partitioned with hexane, ethyl acetate, and n-butanol. The partitioned fractions were evaluated in their capacity to inhibit the radial growth of the three species of fungi. The active fraction was used for an assay-guided chromatography of antifungal extracts. The results showed that the extract from B. glutinosa partitioned in ethyl acetate (Bea) showed the highest antifungal activity against the three fungi. Bea completely inhibited the growth of F. verticillioides at 0.8 mg/ml, whereas the radial growth of A. flavus and A. parasiticus was inhibited 70% at 1.5 mg/ml. The purified antifungal fraction from Bea showed 72, 54, and 52% of antifungal activity, respectively.     

Antifungal Activity in Ethanolic Extracts of <Emphasis Type="Italic">Carica papaya</Emphasis> L. cv. Maradol Leaves and Seeds

Bioactive compounds from vegetal sources are a potential source of natural antifungic. An ethanol extraction was used to obtain bioactive compounds from Carica papaya L. cv. Maradol leaves and seeds of discarded ripe and unripe fruit. Both, extraction time and the papaya tissue flour:organic solvent ratio significantly affected yield, with the longest time and highest flour:solvent ratio producing the highest yield. The effect of time on extraction efficiency was confirmed by qualitative identification of the compounds present in the lowest and highest yield extracts. Analysis of the leaf extract with phytochemical tests showed the presence of alkaloids, flavonoids and terpenes. Antifungal effectiveness was determined by challenging the extracts (LE, SRE, SUE) from the best extraction treatment against three phytopathogenic fungi: Rhizopus stolonifer, Fusarium spp. and Colletotrichum gloeosporioides. The leaf extract exhibited the broadest action spectrum. The MIC₅₀ for the leaf extract was 0.625 mg ml⁻¹ for Fusarium spp. and >10 mg ml⁻¹ for C. gloeosporioides, both equal to approximately 20% mycelial growth inhibition. Ethanolic extracts from Carica papaya L. cv. Maradol leaves are a potential source of secondary metabolites with antifungal properties.     

Production and characterization of the milk-clotting protease of<Emphasis Type="Italic"> Myxococcus xanthus</Emphasis> strain 422

The cheese industry is seeking novel sources of enzymes for cheese production. Microbial rennets have several advantages over animal rennets. (1) They are easy to generate and purify and do not rely on the availability of animal material. (2) The production of microbial clotting enzymes may be improved by biotechnological techniques. In this work, the biochemical characterization of a novel milk-clotting extracellular enzyme from Myxococcus xanthus strain 422 and a preliminary evaluation of its cheese-producing ability are reported. Strain 422 was selected from four M. xanthus strains as the best producer of extracellular milk-clotting activity, based on both its enzyme yield and specific milk-clotting activity, which also afforded lower titration values than enzymes from the three other M. xanthus strains. The active milk-clotting enzyme from M. xanthus strain 422 is a true milk-clotting enzyme with a molecular mass of 40 kDa and a pI of 5.0. Highest milk-clotting activity was at pH 6 and 37 °C. The enzyme was completely inactivated by heating for 12 min at 65 °C. The crude enzyme preparation was resolved by anion-exchange chromatography into two active fractions that were tested in cheese production assays of compositional (dry matter, fat content, fat content/dry-matter ratio, and moisture-non-fat content) and physicochemical properties (firmness, tensile strength, pH and Aw) of the milk curds obtained. Purified protein fraction II exhibited a significantly higher milk-clotting ability than either protein fraction I or a total protein extract, underlining the potential usefulness of M. xanthus strain 422 as a source of rennet for cheese production.     

Evaluation of the Antioxidant and Antifungal Actions of the Optimized Crude Extract and Fractions from the Aerial Parts of Acanthospermum hispidum

This study investigated the phytochemical characteristics of the aerial parts of Acanthospermum hispidum, by chromatographic and spectrophotometric methods, and evaluated the antioxidant and antifungal activities of the crude extract and polyphenol-enriched fractions of the species. The phytochemical prospection showed the presence of polyphenols from the groups of hydroxycinnamic derivatives and flavonoids in the crude extract (CE) and fractions of the aerial parts of A. hispidum. In the chromatographic analysis, it was possible to observe that the fractionation process of the CE with hexane and ethyl acetate was efficient in enriching the fractions in phenolic compounds. This enrichment provided an increase in antioxidant activity by the DPPH and ABTS methods, in which it was observed a higher antioxidant activity for EAF in the DPPH test and higher activity against the ABTS radical by the fractions AqF and RAqF. The extract and fractions were effective against Candida non-Candida albicans strains, mainly against C. glabrata, C. parapsilosis and C. krusei, acting predominantly fungicidal. The results indicate that the aerial parts of A. hispidum can serve as a basis for the development of new antioxidant and antifungal products. Moreover, the fractionation process can contribute to increasing the biological potential of the species.