Antimicrobial activity of crude extracts from plant parts and corresponding calli of Bixa orellana L

Ethanol extracts from the different parts of B. orellana showed differential antimicrobial activity. It was found that the extracts of in vitro leaves showed maximum activity against Bacillus pumilus followed by the extracts from the roots and hypocotyls. The callus derived from different explants too showed antimicrobial activity. The leaf callus showed maximum activity. The zone of inhibition for the diluted extracts of in vitro hypocotyls and roots and their corresponding calli showed minimum zone of inhibition at concentration 24 mg/ml, whereas the diluted extract of in vitro leaves and leaf derived callus showed minimum zone of inhibition at 16 mg/ml.     

Toxicity of crude extracts from several terrestrial plants to barnacle larvae on mangrove seedlings

This study involved the use of terrestrial plant extracts as substitutes for toxic chemicals to control the major fouling organisms of mangroves, the barnacles. In terms of the solvents used to obtain the plant crude extracts, ethyl acetate was best, ethyl alcohol next and pure water worst, except in the case of Allium cepa (AC), where pure water was better than ethyl alcohol. For different plants, different solvents resulted in different poisonous effects, and plants belonging to different classes showed different levels of activities. In particular, using ethyl acetate crude extracts, Allium sativum (AS), AC and Capsicum annuum (CA) caused massive death of barnacle larvae at a low concentration in the shortest time. The less poisonous organic solvents were able to extract more active materials, and this was advantageous in enhancing the effect. For pure water crude extracts, AC and Zingiber officinale were best, but for 95% ethyl alcohol crude extracts, AS and AC were best. For ethyl acetate crude extracts, the effects of AS, AC and CA were better. Different solvent extracts of AC with 12 h of exposure all had LC₅₀ values <100 mg/L. The concentrations of the effective plant crude extracts used were close to, or even lower than, those used for pesticides referred to in the literature.     

Antifungal metabolites from fungal endophytes of Pinus strobus

The extracts of five foliar fungal endophytes isolated from Pinus strobus (eastern white pine) that showed antifungal activity in disc diffusion assays were selected for further study. From these strains, the aliphatic polyketide compound 1 and three related sesquiterpenes 2-4 were isolated and characterized. Compound 2 is reported for the first time as a natural product and the E/Z conformational isomers 3 and 4 were hitherto unknown. Additionally, the three known macrolides; pyrenophorol (5), dihydropyrenophorin (6), and pyrenophorin (7) were isolated and identified. Their structures were elucidated by spectroscopic analyses including 2D NMR, HRMS and by comparison to literature data where available. The isolated compounds 1, 2, and 5 were antifungal against both the rust Microbotryum violaceum and Saccharomyces cerevisae.     

ATP-citrate lyase activity in fungal extracts

ATP-citrate lyase activity has been demonstrated in crude extracts from several species ofMortierella. This filamentous fungus characteristically stores lipid in the mycelium. The enzyme catalyses an ATP-dependent cleavage of citrate in the presence of CoA to oxaloacetic acid and acetylCoA. A partially purified preparation has been obtained and shown to have properties similar to preparations isolated from other sources. It is highly specific for citrate and ATP and has an optimum pH value for activity of 8.4. When the fungus is grown in the presence of increasing amounts of glucose the specific activity of the enzyme also increases.     

Non-systemic fungal endophytes of grasses

Many fungi behave as endophytes in grasses. Unlike the well known Epichloë/Neotyphodium species, most other endophytes are not capable of systemic colonization of plant organs, or seed transmission. The species diversity of the non-systemic endophytic mycobiota of grasses is large, dominated by ascomycetes. The relative abundance of species is very unequal, a few dominant taxa like Acremonium, Alternaria, Cladosporium, Epicoccum and Penicillium spp., occur in many grasses and locations. In contrast, many rare species are isolated only once in endophyte surveys. The possible ecological functions of endophytes are diverse, and often unknown. Latent pathogens represent a small fraction of endophytic mycobiotas, indicating that many non-pathogenic fungal taxa are able to enter plants overriding defence reactions. Some dominant species behave as latent saprotrophs, sporulating when the host tissue dies. Endofungal viruses and bacteria occur among endophytic species, but their effect in their hosts is largely unknown.     

Taxol from fungal endophytes and the issue of biodiversity

Fungi represent one of the most understudied and diverse group of organisms. Commonly, these organisms make associations with higher life forms and may proceed to biochemically mimic the host organism. An excellent example of this is the anticancer drug, taxol, which had been previously supposed to occur only in the plant genusTaxus (yew). However, taxol has been reported in a novel endophytic fungus—Taxomyces andreanae, but also has been demonstrated to occur in a number of unrelated fungal endophytes includingPestalotia, Pestalotiopsis, Fusarium, Alternaria, Pithomyces, Monochaetia and others. Thus, this report presents information on the presence of taxol among disparate fungal genera, and uses these observations as an additional argument to support efforts to study fungal endophytes and preserve their associated host plants.     

Bioactive natural products from marine sponges and fungal endophytes

This review highlights recent findings of our group on bioactive marine natural products isolated from marine sponges and marine derived fungi. The activated chemical defence of the Mediterranean sponge Aplysina aerophoba is introduced as an example of a dynamic response of marine sponges to wounding. Following tissue disrupture preformed brominated isoxazoline alkaloids are enzymatically cleaved and thereby give rise to aeroplysinin-1 which is believed to protect sponges from invasion of pathogenic bacteria. A preliminary characterization of the membrane bound enzyme(s) involved in the cleavage reaction is presented. Bromotyrosine derived, oxime group bearing peptides, the so called bastadins, obtained from the sponge Ianthella basta and some of their synthetic derivatives were shown to exhibit pronounced antifouling activity against larvae of the barnacle Balanus improvisus. The antifouling activity could be traced to the oxime group as an important pharmacophore that was also found to be present in other sponge derived natural products exhibiting antifouling activity. Marine derived fungi that reside within invertebrates such as sponges or inside Mangrove plants are emerging as a new source of bioactive metabolites as demonstrated for Aspergillus ustus and Alternaria sp. that were isolated from the sponge Suberites domuncula or from the Mangrove plant Sonneratia alba, respectively. The former fungus yielded new moderately cytotoxic sesquiterpenoids of the drimane type whereas the latter was found to produce polyketides such as alternariol that exhibited strong and selective inhibitory activity against several protein kinases, for instance Aurora A and B which are targets for anticancer chemotherapy.     

Antimicrobial effect of crude extracts of Nigella sativa on multiple antibiotics-resistant bacteria

Different crude extracts of Nigella sativa were tested for antimicrobial effectiveness against different bacterial isolates. These isolates comprised 16 gramnegative and 6 grampositive representatives. They showed multiple resistance against antibiotics, specially the gramnegative ones. Crude extracts of Nigella saliva showed a promising effect against some of the test organisms. The most effective extracts were the crude alkaloid and water extracts. Gramnegative isolates were affected more than the grampositive ones.     

Antimicrobial activity of wax and hexane extracts from Citrus spp. peels

Antibacterial and antifungal properties of wax and hexane extracts of Citrus spp. peels were tested using bioautographic and microdilution techniques against three plant pathogenic fungi (Penicillium digitatum, Curvularia sp., and Colletotrichum sp.), two human pathogens (Trichophyton mentagrophytes and Microsporum canis), and two opportunistic bacteria (Escherichia coli and Staphylococcus aureus). Two polymethoxylated flavonoids and a coumarin derivative, were isolated and identified from peel extracts, which presented antimicrobial activity especially against M. canis and T. mentagrophytes: 4',5,6,7,8-pentamethoxyflavone (tangeritin) and 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) from C. reticulata; and 6,7-dimethoxycoumarin (also known as escoparone, scoparone or scoparin) from C. limon.     

Characterization of cellulases of fungal endophytes isolated from Espeletia spp.

Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production.     

Molecular phylogeny and paclitaxel screening of fungal endophytes from Taxus globosa

We studied the endophytic mycoflora associated with Taxus globosa, the Mexican yew. The study localities; Las Avispas (LA), San Gaspar (SG), and La Mina (LM) were three segments of cloud forest within the range of Sierra Gorda Biosphere Reserve, México. Overall, 245 endophytes were isolated and 105 representative Ascomycota (morphotaxons) were chosen for phylogenetic and genotypic characterization. Maximum likelihood analyses of large subunit of ribosomal RNA (LSU) rDNA showed well-supported clades of Dothideomycetes, Eurotiomycetes, Leotiomycetes, Pezizomycetes, and Sordariomycetes. Analyses of ITS rDNA groups showed 57 genotypes (95% sequence similarity), in general consistent with the phylogenetically delimitated taxa based on LSU rDNA sequences. The endophyte diversity measured by Fisher's α, Shanonn, and Simpson indices was ca. three-fold and ca. two-fold greater in LM than in LA and SG respectively. A screening for paclitaxel using a competitive inhibition enzyme immunoassay showed 16 positive isolates producing between 65 and 250 ng l(-1). The isolates included Acremonium, Botryosphaeria, Fusarium, Gyromitra, Nigrospora, Penicillium, three novel Pleosporales, and Xylaria.     

Antimicrobial activity of essential oils and other plant extracts

The antimicrobial activity of plant oils and extracts has been recognized for many years. However, few investigations have compared large numbers of oils and extracts using methods that are directly comparable. In the present study, 52 plant oils and extracts were investigated for activity against Acinetobacter baumanii, Aeromonas veronii biogroup sobria, Candida albicans, Enterococcus faecalis, Escherichia col, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serotype typhimurium, Serratia marcescens and Staphylococcus aureus, using an agar dilution method. Lemongrass, oregano and bay inhibited all organisms at concentrations of < or = 2.0% (v/v). Six oils did not inhibit any organisms at the highest concentration, which was 2.0% (v/v) oil for apricot kernel, evening primrose, macadamia, pumpkin, sage and sweet almond. Variable activity was recorded for the remaining oils. Twenty of the plant oils and extracts were investigated, using a broth microdilution method, for activity against C. albicans, Staph. aureus and E. coli. The lowest minimum inhibitory concentrations were 0.03% (v/v) thyme oil against C. albicans and E. coli and 0.008% (v/v) vetiver oil against Staph. aureus. These results support the notion that plant essential oils and extracts may have a role as pharmaceuticals and preservatives.     

Symbiosis between grasses and asexual fungal endophytes

The symbiosis between vertically transmitted asexual endophytic fungi and grasses is common and generally considered to be mutualistic. Recent studies have accumulated evidence of negative effects of endophytes on plant fitness, prompting a debate on the true nature of the symbiosis. Genetic factors in each of the two partners show high variability and have a range of effects (from positive to negative) on plant fitness. In addition, interacting environmental factors might modify the nature of the symbiosis. Finally, competition and multitrophic interactions among grass consumers are influenced by endophytes, and the effects of plant neighbours or consumers could feedback to affect plant fitness. We propose a mutualism-parasitism continuum for the symbiosis between asexual endophytes and grasses, which is similar to the associations between plants and mycorrhizal fungi.     

Current perspectives on the volatile-producing fungal endophytes

Microbial-derived volatiles are ubiquitous in the environment and actively engaged in bio-communication with other organisms. Recently, some volatile-producing endophytes (VPEs), cryptic fungal symbionts persisting in healthy plant tissues, have attracted great attention due to their strong antibiotic activity or production of carbon chains that are identical to many of those found in petroleum, while other fragrant volatiles can be used in the flavoring industries. From an application-oriented and biotechnological point of view, these findings show significant promise for sustainable development of agriculture, forestry, and industry, especially in the control of fruit postharvest diseases, soil-borne pathogen management, and bio-fuel production. In comparison, the ecological importance of VPEs has only rarely been addressed and warrants further exploration. In this review, we summarize the current knowledge and future directions in this fascinating research field, and also highlight the constraints and progresses towards commercialization of VPEs products.     

DNA ligase activity in crude extracts of fibroblasts and lymphocytes

DNA ligase activity was determined in crude cell extracts using a new assay which measures the retention of double stranded circular phage λ DNA on nitrocellulose filters, and allows accurate determinations of the enzyme activity with cell concentration corresponding to 0.1 μg of proteins. Using this assay, we show that the DNA ligase activity varies greatly among mammalian cell lines. The higher activity is found in actively growing fibroblasts where it is stimulated by dimethyl sulfate pretreatment of the cells, whereas the low activity measured in resting lymphocytes is not modified by dimethyl sulfate. The DNA ligase activity correlates with the cells sensitivity towards ionizing radiations.     

Symbiosis with systemic fungal endophytes promotes host escape from vector-borne disease

Plants interact with a myriad of microorganisms that modulate their interactions within the community. A well-described example is the symbiosis between grasses and Epichloë fungal endophytes that protects host plants from herbivores. It is suggested that these symbionts could play a protective role for plants against pathogens through the regulation of their growth and development and/or the induction of host defences. However, other endophyte-mediated ecological mechanisms involved in disease avoidance have been scarcely explored. Here we studied the endophyte impact on plant disease caused by the biotrophic fungus, Claviceps purpurea, under field conditions through (1) changes in the survival of the pathogen´s resistance structure (sclerotia) during overwintering on the soil surface, and (2) effects on insects responsible for the transportation of pathogen spores. This latter mechanism is tested through a visitor exclusion treatment and the measurement of plant volatile cues. We found no significant effects of the endophyte on the survival of sclerotia and thus on disease inocula. However, both pathogen incidence and severity were twofold lower in endophyte-symbiotic plants than in non-symbiotic ones, though when insect visits were prevented this difference disappeared. Endophyte-symbiotic and non-symbiotic plots presented different emission patterns of volatiles suggesting that they can play a role in this protection. We show a novel indirect ecological mechanism by which endophytes can defend host grasses against diseases through negatively interacting with intermediary vectors of the epidemic process.     

Antimicrobial activity of various extracts and carvacrol from Lippia multiflora leaf extract

This study examines the antimicrobial activity of the hexane, dichloromethane and methanol extracts of Lippia multiflora and carvacrol isolated from the hexane extract. The result shows the hexane extract to be the most active, while the methanol extract exhibited no antimicrobial activity. The isolated carvacrol from the hexane fraction showed tremendous antimicrobial activity. These results confirm the traditional uses of Lippia multiflora in the treatment of disease conditions due to microbes.     

Antimicrobial activity of organic extracts isolated from Aplysina fistularis (Demospongiae: Aplysinidae)

Organic extracts of the sponge Aplysina fistularis (Pallas 1766) were tested for antimicrobial activity against Gram positive bacteria (Staphylococcus aureus) and Gram negative bacteria (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa). The minimal inhibitory concentration (MIC) and toxic activity of extract were determined. Susceptibility trials of organic fractions obtained by VLC: Hexane, EtOAc and CHCl3 showed that EtOAc fraction has antibacterial activity against E. coli, while CHCl3 fraction inhibited E. coli and S. aureus growth. The later refractioning of EtOAc fraction and the biodirected assays showed that fractions F12 and F13 of EtOAc/Hex and EtOAc F14 were bioactive against Gram positive and Gram negative bacteria. Only EtOAc/MeOH Sf2 from subfractionig of EtOAc F14 produced inhibition for E. coli and S. aureus. In Sf2 EtOAc/MeOH, MIC was moderate for S. aureus (MIC > 256 g/ml). F4 CHCl3/MeOH produced a high inhibition in S. aureus (MIC = 0.125 g/ml) and for E. coli (MIC > 16 g/ml). F10 CHCl3/MeOH showed a moderate activity against S. aureus (MIC > 128 g/ml) and low activity against E. coli (MIC = 512 g/ml). F10 CHCL3/MeOH did no present toxic activity against Artemia salina. The fractiorts F4 CHCL3/MeOH and Sf2 EtOAc/MeOH were toxic for this organism when the concentration was higher than 100 microg/ml. LC50 in both cases was 548.4 and 243.4 microg/ml respectively. Secondary metabolites of medium polarity obtained from A. fistularis have a wide spectrum of anti bacterial activity. Toxicity analysis suggests that only F10 CHCL3/MeOH has potential as an antimicrobial agent for clinical use.     

Antimicrobial activity of isocytisoside and extracts of Aquilegia vulgaris L

AIMS: The aim of this study was to analyse the antimicrobial properties extracts of Aquilegia vulgaris, and their principial flavonoid component and to compare the obtained results with the activity of gentamicin and nystatin. METHODS AND RESULTS: The ethanol, acetone and isopropanol extracts as well as the subextracts isolated from the methanol extract together with the main flavonoid: 4'-methoxy-5,7-dihydroxyflavone 6-C-glucoside (isocytisoside) were obtained from the leaves with stems of Aquilegia vulgaris L. All the extracts were analysed by TLC to confirm flavonoids and phenolic acids occurrence. The antimicrobial activity was tested by the method of series dilutions against different Gram-positive, Gram-negative bacteria and also fungi. The results have shown that the extracts, subextracts and isocytisoside inhibit growth of all studied micro-organisms, revealing the greatest activity against Gram-positive Staphylococcus aureus, Staph. epidermidis and the mould Aspergillus niger. CONCLUSIONS: The antimicrobial activity of the tested materials it is possibly related to the content of isocytisoside. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has determined new activity of A. vulgaris and suggested the necessity of further studies.