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Qualitative analysis of an ethanolic extract from Trametes versicolor and biological screening against Leishmania amazonensis
Abstract
This study was carried out with basidiomycete mushroom Trametes versicolor. The fruiting body of the mentioned mushroom was extracted with alcohol and submitted to a qualitative phytochemical screening for secondary metabolites identification. This experiment afforded the presence of secondary metabolites kind of triterpenes, steroids, flavonoids, alkaloids, and in a small amount cardenolides. Quinones were not found within tested metabolites. In addition, extracts of this mushroom fresh fruiting body of different polarity (ranging from n-hexane, dichloromethane, ethyl acetate until ethanol) were evaluated against promastigotes of Leishmania amazonensis, a widespread tropical and subtropical disease produced by around 20 species of protozoa from Leishmania genus. The ethyl acetate and the ethanol extract exhibited anti-leishmanial activity, with IC50 values of 101.8 +/- 4.2 mu g/mL and 97.4 +/- 2.0 mu g/mL, respectively. The cytotoxicity against peritoneal macrophage from BALB/c mice from those extracts was reported.
... Another extract that showed a strong positive reaction was LF007 (Laccaria fraterna); Matsuda et al. (1996) described the alkaloid Laccarin for Laccaria vinaceoavellanea Hongo, and Spiteller and Schrey (2019) described the E-Zproxamidine, two isomeric alkaloids with a highly unusual core structure for Laccaria proxima (Boud.) Pat.; TC009 (Trametes cubensis) showed positive results for alkaloids, previous reports by Bian et al. (2017) for the species Trametes trogii Berk., reported the presence of Trametramide (a pyrimidic alkaloid), qualitative studies revealed the presence of alkaloids in T. versicolor (Leliebre-Lara et al., 2015). ...
... Another positive species was PA008; Lobo et al. (1983Lobo et al. ( , 1988, Baumert et al. (1997), Zamuner et al. (2005) and Márquez-Fernandez (2013) described several types of sterols for Pisolithus tinctorius (Pisolithus arhizus) among them ergosterol, lanosterol and derivatives, agnosterol, piscolactone and the pisosteral. Regarding TC009, the positive results coincide with those of Leliebre-Lara et al. (2015) who obtained positive results for the species T. versicolor; with respect to AG001 the extract indicated the presence of terpenoid compounds and sterols, previous studies of the genus revealed the presence of terpenoid compounds (Juma et al., 2016); with regard to GS002, positive results agree with the work by Cateni et al. (2015), who described two triterpene acids for Gloeophyllum odoratum (Wulfen) Imazeki, and 10 new ergoste-roids, (gloeophyllins A-J) for Gloeophy-llum abietinum (Bull.) P. Karst (Han et al., 2015). ...
The discharge of industrial and urban wastewater without prior treatment is a growing problem in Paraguay. The search of technologies to reduce the environmental impacts generated by this problem, is an area of interest in recent times. Phytoremediation is a remediation alternative that proposes the application of artificial or constructed wetlands where a complex system of plants, microorganisms, and substrate act together to remove contaminants. The objective of this work was to describe the performance and efficiency
of two macrophytes in constructed wetlands of horizontal subsurface flow for the treatment of urban wastewater in the city of San Lorenzo. A septic tank was used as primary treatment, followed by two wetlands, one with Cyperus giganteus and other with Typha dominguensis. The results of water analyzes show that both wetlands built at pilot scale proved to be effective for the treatment used, demonstrating a significant reduction for each parameter studied with respect tothe entrance to the system. The removal (%) of
ammoniacal nitrogen was 42% (CG) and 65% (TD); for total phosphorus it was 45% (CG) and 58% (TD), the biochemical oxygen demand was 64% (CG) and 81% (TD), while for chemical oxygen demand efficiency it was 40% (GC) and 61% (TD), respectively.
... This lead to the identification of each compound (TE1 and TG1) as cerevisterol (ergosta-7,22E-diene-3 5 ,6 -triol). Cerevisterol (TE1 and TG1) isolated from T. elegans and T. gibbosa is not unusual since other studies have found cerevisterol in fungi, and most especially mushrooms [42,43]. Though Leliebre-Lara et al. [43] reported the presence of cerevisterol in the genus Trametes (such as T. versicolor), this is being reported for the first time for T. elegans and T. gibbosa. ...
... Cerevisterol (TE1 and TG1) isolated from T. elegans and T. gibbosa is not unusual since other studies have found cerevisterol in fungi, and most especially mushrooms [42,43]. Though Leliebre-Lara et al. [43] reported the presence of cerevisterol in the genus Trametes (such as T. versicolor), this is being reported for the first time for T. elegans and T. gibbosa. Cerevisterol has been reported to exhibit pharmacological activities such as anti-inflammatory [13] and antimicrobial [13,14]. ...
Abstract:
Background: The development of resistance by pathogenic organisms has renewed the worldwide search for novel antimicrobial agents. Mushrooms have been of recent interest because a wide variety of biologically active compounds are being isolated from them. Objective: This study isolated antimicrobial compound (s) from two wood decaying mushrooms, Trametes gibbosa and Trametes elegans, and determined the resistance modifying activities of the isolated compound (s). Methods: Bioactivity guided isolation of active principles from the methanol extract of T. gibbosa and T. elegans was performed using column and preparative high performance liquid chromatography. The structures of isolated compounds were elucidated using nuclear magnetic resonance spectroscopy. Broth micro-dilution assay was used to determine the antimicrobial and resistance modifying activities of the isolated compounds against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella typhi, Streptococcus pyogenes, Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, Candida albicans, Aspergillus niger, Aspergillus flavus and Aspergillus tamarii. Results: Bioactivity guided isolation lead to the isolation of cerevisterol (ergosta-7,22E-diene-3β5α,6β-triol) from both T. gibbosa and T. elegans. Cerevisterol inhibited the growth of S. typhi, S. aureus and A. niger with MICs of 25 µg/mL each and 50 µg/mL against E. faecalis. The MBCs of cerevisterol against S. typhi S. aureus, E. faecalis and A. niger were 50, 100, 200 and 100 µg/mL, respectively. The sub-inhibitory concentration (3 µg/mL) of cerevisterol modified the activity of erythromycin, ampicillin, ciprofloxacin, tetracycline and amoxicillin either by potentiating or reducing their activities. Conclusion: Cerevisterol possess antimicrobial and resistance modifying activities.
Keywords: Trametes gibbosa, Trametes elegans, cerevisterol, antibiotic resistance modifying activities
... Previously, we studied the in vitro antileishmanial activity of different extracts from T. versicolor, and the n-hexane extract turned out to be moderately active [22]. In the present work, we report the phytochemical investigation of this extract that resulted in the isolation of three triterpenes: two ergostane types and a lanostanoid derivative. ...
... Although the n-hexane crude extract only showed moderate activity in a preliminary in vitro leishmanicidal assay [22], trametenolic acid B 3 as one of its constituents showed a strong inhibitory activity. This could be due to the low concentration of the sterol in the crude n-hexane extract or to an antagonism among constituents. ...
Two ergostanes, 5α,8α-epidioxy-22E-ergosta-6,22-dien-3β-ol (1) and 5α-ergost-7,22-dien-3β-ol (2), and a lanostane, 3β-hydroxylanostan-8,24-diene-21-oic acid (trametenolic acid) (3), were isolated from an n-hexane extract prepared from the fruiting body of Trametes versicolor (Bres. Rivarden). The activity of the isolated sterols was evaluated against promastigotes and amastigotes of Leishmania amazonensis Lainson and Shaw, 1972. The lanostane, compound (3), showed the best inhibitory response (IC50 promastigotes 2.9 ± 0.1 μM and IC50 amastigotes 1.6 ± 0.1 μM). This effect was 25-fold higher compared with its cytotoxic effect on peritoneal macrophages from BALB/c mice. Therefore, trametenolic acid could be regarded as a promising lead for the synthesis of compounds with antileishmanial activity.
... The secondary metabolite content of Trametes versicolor mushroom extract using an ethanol solvent was found to contain alkaloids, flavonoids, terpenoids, and steroids (Leliebre-Lara et al. 2015). After extraction using a water solvent, a phytochemical test on secondary metabolite compounds of T. versicolor mushrooms detected saponins, tannins, flavonoids, steroids, and terpenoids (Chengo et al. 2016). ...
Herawati E, Ramadhan R, Ariyani F, Marjenah, Kusuma IW, Suwinarti W, Mardji D, Amirta R, Arung ET. 2021. Phytochemical screening and antioxidant activity of wild mushrooms growing in tropical regions. Biodiversitas 22: 4716-4721. This study aims to determine the presence of secondary metabolites which generally have bioactivity that is antioxidant with phytochemical tests and antioxidant tests in Auricularia auricula, Schyzophyllum commune, Microporus xanthopus, and Trametes versicolor fungi that grow in the wild. Phytochemical analysis was carried out by testing color changes and modifications, while antioxidant tests were carried out by using a UV-Vis spectrophotometer and a DPPH solution (1,1-diphenyl-2-picrylhydrazyl). This research found that edible wood fungus that grows, wild such as A. auricula, contains secondary metabolites, namely flavonoids, triterpenoids, saponins, and tannins, but does not contain steroids, alkaloids, carotenoids, or coumarin, while S. commune contains secondary metabolites, namely flavonoids, steroids, tannins, and coumarin, but does not contain triterpenoids, alkaloids, carotenoids, or saponins and non-edible wood fungus, such as M. xanthopus, contains secondary metabolites, namely, flavonoids, saponins, tannins, coumarin, and steroids, but does not contain triterpenoids, alkaloids, or carotenoids, while T. versicolor contains compounds, secondary metabolites, flavonoids, saponin, tannins, coumarin, and triterpenoids, but does not contain steroids, alkaloids, or carotenoids. The antioxidant test results are based on IC50 calculations; it is known that the IC50 value of A. auricula is 499.25 g/mL, while S. commune is 121.37 g/mL, M. xanthopus is 251.20 g/mL, and T. versicolor is 493.04 g/mL. Based on the research results, edible and inedible wood mushroom extracts contain several phytochemicals and show potential for antioxidant activity.
... 1-phenyl-3-methylpyrazolones oxidoreductase mediators [297] aryl tetral226in lignans bioactive compounds [298] podophyllotoxin bioactive compounds [298] Trametes versicolor funatrol D bioactive compound [299] isodrimenediol sesquiterpenes (anticancer) [299] tramspiroins sesquiterpenes (anticancer) [299] triterpenes bioactive compounds [300] Database on potential toxigenic capacities of microorganisms used for industrial production trichoderiol A, C [304] trichodimerol insecticide activity [302] Trichoderma harzianum 1,8-dihydroxy-3-methyl-anthraquinone antifungal [307][308][309] 1-hydroxy-3-methylanthraquinone antifungal [307][308][309] Database on potential toxigenic capacities of microorganisms used for industrial production ...
Fungi produce several bioactive metabolites, pigments, dyes, antioxidants, polysaccharides, and industrial enzymes. Fungal products are also the primary sources of functional food and nutrition, and their pharmacological products are used for healthy aging. Their molecular properties are validated through the use of recent high-throughput genomic, transcriptomic, and metabolomic tools and techniques. Together, these updated multi-omic tools have been used to study fungal metabolites structure and their mode of action on biological and cellular processes. Diverse groups of fungi produce different proteins and secondary metabolites, which possess tremendous biotechnological and pharmaceutical applications. Furthermore, its use and acceptability can be accelerated by adopting multi-omics, bioinformatics, and machine learning tools that generate a huge amount of molecular data. The integration of artificial intelligence and machine learning tools in the era of omics and big data has opened up a new outlook in both basic and applied researches in the area of nutraceuticals and functional food and nutrition.
Key Points
•Multi-omic tool helps in the identification of novel fungal metabolites
•Intra-omic data from genomics to bioinformatics
•Novel metabolites and application in human health
Introduction: Leishmaniasis is a parasitic disease caused by several species of the genus Leishmania. These protozoan parasites do not circulate autochthonously in Cuba, but imported cases may be found. There are three main clinical forms, and various methods are used to make the diagnosis. Among these are parasitological, serological and more recently molecular methods. The treatment of choice is pentavalent antimonials, though they have drawbacks which have led to the search for new therapeutic alternatives. Objectives: summarize the work results obtained from the study of Leishmania for twenty years and explain the reasons fostering training, professional development and research about this condition at Pedro Kouri Tropical Medicine Institute. Methods: a bibliometric study was conducted and an analysis was performed of all the results obtained by the Leishmania group at Pedro Kouri Tropical Medicine Institute in twenty years. All the materials published by the group were consulted, and determination was made of the fundamental results obtained in the main research lines. Results: a description is provided of the most important contributions in research areas such as DNA immunization, molecular diagnosis and the search for new therapeutic alternatives, and readers are directed to the publications containing the details of each study. Mention is made of the main forms of cooperation with national and foreign institutions. Comments are provided on the future perspectives of the work of the research group. Conclusions: the group has engaged in research lines of interest, and is responsible for the greatest contribution to the country in the field of research about the study parasite, in keeping with its mission. A challenge for the years to come is to maintain the continuity of the research, contribute to the solution of some problems still to be solved concerning this parasitic disease, and the application of the results obtained.
La leishmaniasis es una enfermedad parasitaria causada por varias especies del género Leishmania, protozoo parásito que no circula en Cuba de forma autóctona. Sin embargo, se presentan casos importados. Existen tres formas clínicas principales y diversos métodos para realizar el diagnóstico, que incluyen los parasitológicos, serológicos y más recientemente, los moleculares. El tratamiento de elección son los antimoniales pentavalentes, los cuales presentan inconvenientes que han estimulado la búsqueda de nuevas alternativas terapéuticas. Se realizó un estudio bibliométrico y se analizaron todos los resultados de trabajo del grupo de Leishmania del Instituto de Medicina Tropical Pedro Kourí durante 20 años, se consultaron todas las publicaciones realizadas en el grupo y se determinaron los resultados fundamentales en las principales líneas de investigación. Se describen los aportes más importantes en áreas de investigación como la inmunización con ADN, el diagnóstico molecular y la búsqueda de nuevas alternativas terapéuticas, y se remite al lector a consultar las publicaciones que contienen los detalles de cada estudio. Se mencionan las colaboraciones principales con centros nacionales y extranjeros. Se comentan las perspectivas futuras del trabajo del grupo de investigación. El grupo ha acometido líneas de investigación de interés y es responsable del mayor aporte del país en el campo de investigación de este parásito, en correspondencia con su misión. La continuidad de la investigación, contribuir a la solución de algunos de los problemas aún no resueltos de esta parasitosis y la aplicación de los resultados obtenidos, será un reto para los próximos años.
The in vitro activity of parthenolide against Leishmania amazonensis was investigated. Parthenolide is a sesquiterpene lactone purified from the hydroalcoholic extract of aerial parts of Tanacetum parthenium. This isolated compound was identified through spectral analyses by UV, infrared, H-1 and C-13 nuclear magnetic resonance imaging, DEPT (distortionless enhancement by polarization transfer), COSY (correlated spectroscopy), HMQC (heteronuclear multiple-quantum coherence), and electron spray ionization-mass spectrometry. Parthenolide showed significant activity against the promastigote form of L. amazonensis, with 50% inhibition of cell growth at a concentration of 0.37 mug/ml. For the intracellular amastigote form, parthenolide reduced by 50% the survival index of parasites in macrophages when it was used at 0.81 mug/ml. The purified compound showed no cytotoxic effects against J774G8 macrophages in culture and did not cause lysis in sheep blood when it was used at higher concentrations that inhibited promastigote forms. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis with gelatin as the substrate showed that the enzymatic activity of the enzyme cysteine protease increased following treatment of the promastigotes with the isolated compound. This finding was correlated with marked morphological changes induced by parthenolide, such as the appearance of structures similar to large lysosomes and intense exocytic activity in the region of the flagellar pocket, as seen by electron microscopy. These results provide new perspectives on the development of novel drugs with leishmanicidal activities obtained from natural products.
Prior to 1985, cultures at the Center for Forest Mycology Research were maintained on 1.5% malt extract agar test-tube slants. This system not only made it necessary to transfer the entire collection ev-ery year but also permitted genetic change because continual growth occurred. In 1985, the method of storing fungal cultures in sterile distilled water in cryovials was introduced. This study reports on the use of this method for long-term fungal storage. For varying periods up to 7 years, 151 miscellaneous spe-cies of wood-decaying Basidiomycotina were stored in sterile distilled water. Water storage has numerous advantages: culture viability or growth rate is not sig-nificantly influenced; isolates can be stored longer; genetic stability is greater; the method is quick, easy, and inexpensive, and requires less space. The Center for Forest Mycology Research (CFMR) is home to the largest culture collection of wood-rotting Basidiomycotina in the world. The collection, started in 1932, contains more than 11000 secondary my-celium isolates of more than 1600 species and more than 3200 monobasidiosporous isolates taken from many of the same specimens as the dikaryons. Prior to 1985, the cultures were maintained on 1.5% malt extract agar test-tube slants, making it necessary to transfer the entire collection at least once every year.
During a revision of some polypores from Cuba (Basidiomycetes, Aphyllophorales), the type specimen of Navisporus ortizii was re-examined and found to be conspecific with Perenniporia martius. The latter is compared with Navisporus sulcatus and its presence in Cuba is briefly discussed. An annotated review of Perenniporia species reported from Cuba is added.
The MTT assay for cell viability and cell proliferation has been modified to improve its reproducibility and accuracy. The modified test is performed on MultiScreen filtration plates, which permits the removal of the culture medium prior to formazan solubilisation, without loss of cells or formazan crystals. A 1:1 mix of DMSO and ethanol is used as the solvent, since this has the same optical refraction index as the filters, making it possible to measure the optical densities directly on the MultiScreen plate. Data obtained in the assay method using the MultiScreen plate were compared with data from studies employing the normal flat-bottomed plate, by using cells which grow in suspension. Two cell lines were used in the study. CTLL-2, which are IL-2 dependent cells of murine T cell origin, and Jurkat E.6.1 which are IL-2 producing cells of human lymphoma origin. CTLL-2 cells and the modified MTT assay were also used for evaluating the effects of different IL-2 concentrations on cell proliferation.
The crude extracts of the European wood-rotting fungus Polyporus versicolor contain cytotoxic principles. Two of these have been isolated by monitoring the purification by tests on hepatoma cells grown in vitro. They are polyoxygenated derivatives of ergosterol, characterized by the partial structure Δ7, 9α-OH.
Antibacterial activity of the methanol extract of three polypore macrofungi Phellinus rimosus, Ganoderma lucidum and Navesporus floccosa were studied. The activity was evaluated by hole-plate diffusion and microtitre plate dilution methods using Escherichia coli, Pseudomonas aeuroginosa, Staphylococcus aureus, Salmonella typhimurium, and Bacillus subtilis. The methanol extract of P. rimosus and N. floccosa showed activity against all the strains at a concentration of 800 µg/well and 1 mg/well, respectively. The methanol extract of G. lucidum showed activity against E. coli, S. typhimurium and B. subtilis at a concentration of 1 mg/well. The minimum inhibitory concentration (MIC) of P. rimosus and N. flocossa were found to be 500 µg/well and 1 mg/well, respectively. The MIC of G. lucidum was also found to be 1 mg/well. Chemical analysis of the methanol extracts of these fungi indicated the presence of polyphenols, flavonoids, quinones and terpenes.