Antifungal activity of bio-active cell-free culture extracts and volatile organic compounds (VOCs) synthesised by endophytic fungal isolates of Garden Nasturtium.

Antimicrobial resistance in fungal pathogens (both human and plant) is increasing alarmingly, leading to massive economic crises. The existing anti-fungal agents are becoming ineffective, and the situation worsens on a logarithmic scale. Novel antifungals from unique natural sources are highly sought to cope sustainably with the situation. Metabolites from endophytic microbes are the best-fitted alternatives in this case. Endophytes are the untapped sources of 'plants' internal microbial population' and are promising sources of effective bio-therapeutic agents. Fungal endophytes were isolated from Tropaeolum majus and checked for antifungal activity against selected plant and human pathogens. Bioactive metabolites were identified through chromatographic techniques. The mode of action of those metabolites was evaluated through various spectroscopic techniques. The production of antifungal metabolite was optimized also. In particular VOCs (volatile organic compounds) of TML9 were tested in vitro for their anti-phytopathogenic activity. Ethyl acetate (EA) extract of cell-free culture components of Colletotrichum aenigma TML3 exhibited broad-spectrum antifungal activity against four species of Candida and the major constituents reported were 6-pentyl-2H-pyran-2-one, 2-Nonanone, 1 propanol 2-amino. The volatile metabolites, trans-ocimene, geraniol, and 4-terpinyl acetate, produced from Curvularia lunata TML9, inhibited the growth of some selected phyto pathogens. EA extract hampered the biofilm formation, minimised the haemolytic effect, and blocked the transformation of Candida albicans (MTCC 4748) from yeast to hyphal form with a Minimum Fungicidal Concentration (MFC) of 200-600 µg mL-1. Central carbohydrate metabolism, ergosterol synthesis, and membrane permeability were adversely affected and caused the lethal leakage of necessary macromolecules of C. albicans. Volatile metabolites inhibited the growth of phytopathogens i.e., Rhizoctonia solani, Alternaria alternata, Botrytis cinerea, Cercospora beticola, Penicillium digitatum, Aspergillus fumigatus, Ceratocystis ulmi, Pythium ultimum up to 89% with an IC50 value of 21.3-69.6 µL 50 mL-1 and caused leakage of soluble proteins and other intracellular molecules. Citrusy sweet odor volatiles of TML9 cultured in wheat-husk minimised the infections of Penicillium digitatum (green mold), in VOC-exposed sweet oranges (Citrus sinensis). Volatile and non-volatile antifungal metabolites of these two T. majus endophytes hold agricultural and pharmaceutical interests. Metabolites of TML3 have strong anti-Candida activity and require further assessment for therapeutic applications. Also, volatile metabolites of TML9 can be further studied as a source of antifungals. The present investigational outcomes bio-prospects the efficacy of fungal endophytes of Garden Nasturtium.

Broad spectrum bioactivity of a novel ß-glucan rich heteropolysaccharide, Pestalopine isolated from endophytic fungi Pestalotiopsis chamaeropsis CEL6.

A biologically potent exopolysaccharide (EPS), Pestalopine was produced by Pestalotiopsis chamaeropsis CEL6, an endophytic fungal isolate of Chloranthus elatior Sw. Pestalopine is composed of glucose, arabinose, fucose, rhamnose, and galactose in a molar ratio of nearly 10:1:2:2:4 having an Mw ∼ 3.29 × 105 Da. Pestalopine exhibited a radical scavenging effect and significantly increased antioxidant parameters (malondialdehyde, superoxide dismutase, glutathione peroxidase, reduced glutathione oxidized glutathione) in peritoneal macrophage cells in a concentration-dependent manner, with a maximum effect at 500 mg mL-1. Pestalopine is hepatoprotective in nature and improves the liver function profiles-total bilirubin, direct bilirubin, hepatic enzymes: alkaline phosphatase, gamma-glutamyl transferase, aspartate aminotransferase, alanine transaminase of liver fibrosis induced (through thioacetamide) male Wistar rats in comparison to control. Pestalopine-fed rats are reported to have higher counts of beneficial Lactobacillus sp. Present findings suggest that Pestalopine, a novel compound may have promise as a non-toxic exogenous antioxidant with hepatoprotective and probiotic efficacies.

Antifungal activity of volatile and non-volatile metabolites of endophytes of Chloranthus elatior Sw.

Agriculture crops that have fungal infections suffer significant economic losses and reduced crop output. Chemical fungicides are used to tackle the problem, although this has additional detrimental side effects. There is an urgent need for safe and novel antifungals. Volatiles from plant-beneficial endophytic fungi are considered promising alternatives for the biological control of fungal pathogens as a sustainable approach in an agroecosystem. In the present investigation, a volatile-emitting sterile endophytic fungus, Diaporthe sp. CEL3 with bio-fumigation activity, was isolated from leaves of the ethnomedicinal plant Chloranthus elatior Sw., collected from the Passighat forest of North-East India. The camphor odor volatiles of CEL3 showed an inhibitory effect against eight fungal pathogens in vitro and minimized the infections of Monilinia fructicola, a causal agent of cherry fruit rot, in VOC-exposed cherry fruits. Rhizoctonia solani, Botrytis cinerea, Pythium ultimum, and M. fructicola were maximally inhibited up to 51.5%, 55.8%, 61.9%, and 78.5%, respectively, in comparison to control by the volatiles. Another isolate, CEL7, identified as Curvularia sp., synthesized non-volatile, soluble antifungal metabolites in its cell-free extracts and exhibited antifungal action. Bioassay-guided fractionation revealed the presence of imidazole compounds- (2-aminoethyl)-1H-imidazole-2-carbaldehyde, Pyrazole 4, 5 imidazole, 1-formyl 3-ethyl, phenol compounds-Phenol, 4-[2-(methylamino) ethyl]-, 6-Nitro-3-chlorophenol, Phenol, 2,4,6-tri-tert-butyl-, etc., in the cell-free extracts, with a MIC value of 250-2,000 µg ml-1. Optimum VOC emission was achieved in a modified PDA medium with instantly smashed potato (150 g L-1), dextrose (20 g L-1), wheat husk (20 g L-1), and yeast extract (20 g L-1), with additional salts. Interestingly, endophytic CEL3 emitted different types of volatiles, and trans-verbenol (32.25%), geraniol (30.32%), trans-ocimenol (12.90%), and mentha-4,8-diene (5.16%) were the prime ones. These VOCs cause lethal leakage of protein and necessary intracellular molecules from the fungal pathogens. Thus, CEL3 could potentially be used as a bio-fumigating agent to control post-harvest infections caused by fungal pathogens. This study opens a new approach to the use of endophytic fungi in biocontrol.

Broad-Spectrum Antimicrobial Action of Cell-Free Culture Extracts and Volatile Organic Compounds Produced by Endophytic Fungi Curvularia Eragrostidis.

Endophytes are the mutualistic microorganisms that reside within the host plant and promote plant growth in adverse conditions. Plants and their endophytes are engaged in a symbiotic relationship that enables endophytes to access bioactive genes of the ethnomedicinal plants, and, as a result, endophytes are constantly addressed in the sector of pharmaceuticals and agriculture for their multidomain bio-utility. The gradual increase of antimicrobial resistance can be effectively countered by the endophytic metabolites. In these circumstances, in the present investigation, endophytic Curvularia eragrostidis HelS1 was isolated from an ethnomedicinally valuable plant Helecteris isora from East India's forests. The secondary volatile and non-volatile metabolites are extracted from HelS1 and are found to be effective broad-spectrum antimicrobials. A total of 26 secondary metabolites (9 volatiles and 17 non-volatiles) are extracted from the isolate, which exhibits effective antibacterial [against six Gram-positive and seven Gram-negative pathogens with a minimum inhibitory concentrations (MIC) value ranging from 12.5 to 400 µg ml-1] and antifungal (against seven fungal plant pathogens) activity. The secondary metabolite production was optimised by one variable at a time technique coupled with the response surface methodology. The results revealed that there was a 34% increase in antibacterial activity in parameters with 6.87 g L-1 of fructose (as a carbon source), 3.79 g L-1 of peptone (as a nitrogen source), pH 6.75, and an inoculation period of 191.5 h for fermentation. The volatile metabolite production was also found to be optimum when the medium was supplemented with yeast extract and urea (0.2 g L-1) along with dextrose (40 g L-1). Amongst extracted volatile metabolites, 1-H-indene 1 methanol acetate, tetroquinone, N, N-diphenyl-2-nitro-thio benzamide, Trans 1, 2-diethyl-trans-2-decalinol, naphthalene, and azulene are found to be the most effective. Our investigation opens up opportunities in the sector of sustainable agriculture as well as the discovery of novel antimicrobials against dreadful phyto and human pathogens.

Bioactivity study and metabolic profiling of Colletotrichum alatae LCS1, an endophyte of club moss Lycopodium clavatum L.

Endophytes are silent microbial partners of green plants that ensure hosts' survival in odd conditions. They are known as the factories of multipotent metabolites with diverse bioactivities beneficial to modern pharmaceuticals industry. Endophytic fungi have been screened from a variety of plants and it is the first-time endophytes of club moss is being studied for production of antibacterial and antioxidative compounds. The present study reveals that Lycopodium clavatum L. harbors a potent niche of bioactive endophytic fungi and Colletotrichum alatae LCS1 was the prime producer of antibacterial and antioxidative compounds among them. The minimum inhibitory and bactericidal concentrations of ethyl- acetate culture extract ranged from 15.62 to 250 µg/mL against four Gram negative and three Gram positive microorganisms including methicillin resistant Staphylococcus aureus (ATCC-33591). Bio-autogram based screening followed by Gas chromatographic analysis confirmed the occurrence of 17 bioactive compounds and α-bisabolol is known to be the prime one. Alfa bisabolol is a unique and versatile bioactive essential oil and facilitates variety of functions. Killing kinetics data along with leakage of macromolecules into extracellular environment supports the cidal activity of the antibacterial principles at MBC values. Isolate C. alatae LCS1 was optimized by one variable at a time system coupled with response surface methodology for broad spectrum antibacterial production. The organism yielded maximum response (22.66±0.894 mm of zone of inhibition against MRSA) in 250 mL Erlenmeyer flask containing 50 mL potato dextrose broth supplemented with (g/L) glucose, 7.53; yeast extract concentration, 0.47; NaCl, 0.10 with medium pH 6.46; after 134 hours of incubation at 26°C. Optimized fermentation parameters enhanced antibacterial activity up-to more than 50% than the pre-optimized one (10.33±0.57 mm). Endophytic LCS1 was also efficient in free radical scavenging tested by DPPH, ABTS, H2O2 and FRAP assay with an IC50 values of 23.38±5.32 to 82.873±6.479 µg/mL.

Production of Bioactive Compounds with Broad Spectrum Bactericidal Action, Bio-Film Inhibition and Antilarval Potential by the Secondary Metabolites of the Endophytic Fungus Cochliobolus sp. APS1 Isolated from the Indian Medicinal Herb Andrographis paniculata.

Endophytes, being the co-evolution partners of green host plants, are factories of pharmaceutically valuable novel natural products. Cochliobolus sp. APS1, an endophyte of Andrographis paniculata (Green Chiretta), produces a plethora of natural bioactive compounds and the multipotent alkaloid Aziridine, 1-(2-aminoethyl)-, is the prime one among them. The isolate exhibited antibacterial, anti-biofilm, and antilarval potency. The MIC and MBC values of the ethyl-acetate culture extract ranged from 15.62 to 250 µg/mL against ten pathogenic microorganisms (including MRSA and VRSA). Killing kinetics data along with the leakage of macromolecules into the extracellular environment supports the cidal activity of the antibacterial principles. The broad spectrum antibacterial activity of Aziridine, 1-(2-aminoethyl)-, was optimized by a one-variable-at-a-time system coupled with response surface methodology, which led to a 45% enhancement of the antibacterial activity. The maximum response (22.81 ± 0.16 mm of zone of inhibition against MRSA) was marked in 250 mL Erlenmeyer flask containing 90 mL potato dextrose broth supplemented with (g%/L) glucose, 9.7; urea concentration, 0.74; with medium pH 6.48; after 8.76 days of incubation at 26 °C. APS1 strongly inhibited biofilm formation in the tested pathogenic microorganisms and acts as a larvicidal agent against the Dengue-vector Aedes aegypti. This is probably the first report of Aziridine, 1-(2-aminoethyl)-, from any endophytic source. Cochliobolus sp. APS1 possesses industrial importance for the production of bioactive alkaloids.

Drought alleviation efficacy of a galactose rich polysaccharide isolated from endophytic Mucor sp. HELF2: A case study on rice plant.

Endophytes play a vital role in plant growth under biotic and abiotic stress conditions. In the present investigation, a Galactose-Rich Heteropolysaccharide (GRH) with a molecular weight of 2.98 × 105 Da was isolated from endophytic Mucor sp. HELF2, a symbiont of the East Indian screw tree Helicteres isora. OVAT (One Variable at A Time) experiment coupled with RSM (Response Surface Methodology) study exhibited 1.5-fold enhanced GRH production (20.10 g L-1) in supplemented potato dextrose broth at a pH of 7.05 after 7.5 days of fermentation in 26°C. GRH has alleviated drought stress (polyethylene glycol induced) in rice seedlings (Oryza sativa ssp. indica MTU 7093 swarna) by improving its physicochemical parameters. It has been revealed that spray with a 50-ppm dosage of GRH exhibited an improvement of 1.58, 2.38, 3, and 4 times in relative water contents and fresh weight of the tissues, root length, and shoot length of the rice seedlings, respectively "in comparison to the control". Moreover, the soluble sugars, prolines, and chlorophyll contents of the treated rice seedlings were increased upto 3.5 (0.7 ± 0.05 mg/g fresh weight), 3.89 (0.57 ± 0.03 mg/g fresh weight), and 2.32 (1,119 ± 70.8 µg/gm of fresh weight) fold respectively, whereas malondialdehyde contents decreased up to 6 times. The enzymatic antioxidant parameters like peroxidase and superoxide dismutase and catalase activity of the 50 ppm GRH treated seedlings were found to be elevated 1.8 (720 ± 53 unit/gm/min fresh weight), 1.34 (75.34 ± 4.8 unit/gm/min fresh weight), and up to 3 (100 ppm treatment for catalase - 54.78 ± 2.91 unit/gm/min fresh weight) fold, respectively. In this context, the present outcomes contribute to the development of novel strategies to ameliorate drought stress and could fortify the agro-economy of India.

Production, Optimization, Characterization and Drought Stress Resistance by ß-Glucan-Rich Heteropolysaccharide From an Endophytic Fungi Colletotrichum alatae LCS1 Isolated From Clubmoss (Lycopodium clavatum).

Endophytic entities are ubiquitous in nature with all-square bioactivity ranging from therapeutic effects toward animals to growth promoting attributes and stress tolerance activities in case of green plants. In the present study, the club moss Lycopodium clavatum for the first time has been subjected for the isolation of endophytic fungi. An exopolysaccharide (EPS) extracted from Colletotrichum alatae LCS1, an endophytic fungi isolated from L. clavatum Linn., was characterized as a ß-glucan heteropolymer (composed of mannose, rhamnose, arabinose, glucose, galactose, and fucose) which plays a pivotal role in obliterating the drought stress in rice seedlings (Oryza sativa) when applied at an amount of 20, 50, and 100 ppm. The fresh weight contents of rice tissue (39%), total chlorophyll (33%), proline (41%), soluble sugar content (26%) along with antioxidant enzymes such as catalase, peroxidase, and super-oxide dismutase increased (in comparison to control of non-EPS treated seedlings) while malondialdehyde content had reduced markedly after 30 days of regular treatment. The drought resistance of rice seedling was observed at peak when applied at 50 ppm dosage. Vital parameters for EPS production like fermentation duration (5 days), medium pH (6), nutrient (carbon (glucose-7 g%/l), nitrogen (yeast extract-0.4 g%/l), and mineral (NaCl-0.10 g%/l) sources, oxygen requirements (O2 vector or liquid alkane-n-hexane, n-heptane, n-hexadecane), and headspace volume (250 ml Erlenmeyer flask- 50 ml medium, 200 ml-headspace volume) were optimized to obtain an enhanced EPS yield of 17.38 g/L-59% higher than the preoptimized one. The present study, for the first time, reported the ß-glucan rich heteropolysaccharide from Colletotrichum origin which is unique in structure and potent in its function of drought stress tolerance and could enhance the sustainable yield of rice cultivation in areas facing severe drought stress.

Occurrence of a novel tannase (tan BLP ) in endophytic Streptomyces sp. AL1L from the leaf of Ailanthus excelsa Roxb.

The tannase production ability by endophytic actinobacteria and the genetic identity of responsible tannase gene were determined. The studied strains were isolated from surface-sterilized leaf discs of Ailanthus excelsa Roxb. Four strains were found to hydrolyze tannic acid on solid media containing 0.4% tannic acid. The strain AL1L was found as tanBLP indicating production of tannase with diverse of substrate affinity. The tannase production from the potential strain AL1L was performed in liquid tannic acid broth (0.4%, w/v). The strain was later identified as Streptomyces sp. AL1L on the basis of 16S rDNA homology. Highest enzyme activity was observed at 48 h of incubation at the exponential growth phase. The enzyme was purified by ammonium sulfate precipitation followed by dialysis (15 kD cut off). This enzyme, with molecular weight 180 kD shows highest catalytic activity at 35 °C, pH 6 with substrate concentration 0.1 g%. The purified enzyme possesses 1.4 × 10-3 Km and 11.15 U/ml as Vmax. The above study indicates high industrial prospective of endophytic actinobacteria as source of tannase of potential biotechnological applications.

Anti-Infective Metabolites of a Newly Isolated Bacillus thuringiensis KL1 Associated with Kalmegh (Andrographis paniculata Nees.), a Traditional Medicinal Herb.

This study was conducted to isolate endophytic bacteria possessing anti-infective property from Kalmegh (Andrographis paniculata Nees.), a well-known medicinal plant. A total of 23 strains were isolated from this plant among which the strain KL1, isolated from surface-sterilized leaf of this medicinal herb, showed broad-spectrum antagonism against an array of Gram-positive and -negative bacterial pathogens. Ethyl acetate extract of KL1-fermented media yielded a greenish amorphous substance retaining anti-infective property. Solvent-extracted crude material was separated by thin-layer chromatography, and the active ingredient was located by autobiogram analysis. The purified anti-infective compound was found as anthracene derivative as analyzed by ultraviolet and Fourier transform infrared spectroscopy. The strain was identified as Bacillus thuringiensis KL1 from cultural, physiochemical, and molecular aspects. The above results indicate the pharmaceutical potential of the candidate isolate.

Multi-Response Optimization of Granaticinic Acid Production by Endophytic Streptomyces thermoviolaceus NT1, Using Response Surface Methodology.

Streptomyces thermoviolaceus NT1, an endophytic isolate, was studied for optimization of granaticinic acid production. It is an antimicrobial metabolite active against even drug resistant bacteria. Different media, optimum glucose concentration, initial media pH, incubation temperature, incubation period, and inoculum size were among the selected parameters optimized in the one-variable-at-a-time (OVAT) approach, where glucose concentration, pH, and temperature were found to play a critical role in antibiotic production by this strain. Finally, the Box-Behnken experimental design (BBD) was employed with three key factors (selected after OVAT studies) for response surface methodological (RSM) analysis of this optimization study.RSM analysis revealed a multifactorial combination; glucose 0.38%, pH 7.02, and temperature 36.53 °C as the optimum conditions for maximum antimicrobial yield. Experimental verification of model analysis led to 3.30-fold (61.35 mg/L as compared to 18.64 mg/L produced in un-optimized condition) enhanced granaticinic acid production in ISP2 medium with 5% inoculum and a suitable incubation period of 10 days. So, the conjugated optimization study for maximum antibiotic production from Streptomyces thermoviolaceus NT1 was found to result in significantly higher yield, which might be exploited in industrial applications.

Production and structural elucidation of exopolysaccharide from endophytic Pestalotiopsis sp. BC55.

There is a little information on exopolysaccharide production by endophytic fungi. In this investigation endophytic Pestalotiopsis sp. BC55 was used for optimization of exopolysaccharide production. One variable at a time method and response surface methodology were adopted to find out the best culture conditions and medium compositions for maximum exopolysaccharide production. The organism produced maximum exopolysaccharide (4.320 ± 0.022 g/l EPS) in 250 ml Erlenmeyer flask containing 75 ml potato dextrose broth supplemented with (g%/l) glucose, 7.66; urea, 0.29; CaCl2, 0.05 with medium pH 6.93; after 3.76 days of incubation at 24°C. Exopolysaccharide [EPS (EP-I)] produced by this organism have Mw ∼2×10(5)Da with a melting point range of 122-124°C. Structural elucidation of the EPS (PS-I) was carried out after a series of experiments. Result indicated the presence of only (1→3)-linked ß-d-glucopyranosyl moiety. The structure of the repeating unit was established as - →3)-ß-d-Glcp-(1→.

Pueraria tuberosa: a review on its phytochemical and therapeutic potential.

Pueraria tuberosa (Willd.) DC is a perennial herb commonly known as 'vidarikanda', distributed throughout south east Asia. The plant's tuber is widely used in ethanomedicine as well as in traditional systems of medicine, particularly in ayurveda. It has been used in various ayurvedic formulations as restorative tonic, antiaging, spermatogenic and immune booster and has been recommended for the treatment of cardiovascular diseases, hepatosplenomegaly, fertility disorders, menopausal syndrome, sexual debility and spermatorrhoea. Numerous bioactive phytochemicals, mostly isoflavonoids such as puerarin, genistein, daidzein, tuberosin and so on have been identified in the tuber. In vivo and in vitro studies have provided the support against traditional demands of the tuber as spermatogenic, immune booster, aphrodisiac, anti-inflammatory, cardiotonic and brain tonic. However, further studies are required to define the active phytochemical compositions and to validate its clinical utilisation in the herbal formulations for human uses. This review provides an overview of traditional applications, current knowledge on the phytochemistry, pharmacology and toxicology of P. tuberosa. This review also provides plausible hypotheses about how various isoflavones particularly puerarin, genistein and daidzein, individually or collectively, may be responsible for the therapeutic potential against a wide range of ailments.

Muscodor albus MOW12 an Endophyte of Piper nigrum L. (Piperaceae) Collected from North East India Produces Volatile Antimicrobials.

Muscodor albus MOW12, an endophytic fungus isolated from Piper nigrum in Mawlong, Meghalaya, India, resembles some cultural and hyphal characteristics of previous isolates of Muscodor sp. In addition, it possesses about 99 % similarity in its ITS rDNA with other M. albus isolates and thus is nicely centered within the genetic tree to other Muscodor spp. This xylariaceae fungus effectively inhibits and kills certain plant pathogenic fungi by virtue of a mixture of volatile compounds that it produces. The majority of these compounds were identified by gas chromatography/mass spectrometry as small molecular weight esters, alcohols, and acids. The main ester components of this isolate of M. albus in its volatile mixture are acetic acid, ethyl ester; propanoic acid, 2-methyl-, methyl ester and acetic acid, 2-methylpropyl ester. This appears to be the first report of any M. albus strain from India.

Rhodotorula mucilaginosa Keratitis: a rare fungus from Eastern India.

Rhodotorula mucilaginosa rarely cause keratitis in immunocompromised individuals. A 30 year old male with history of minor trauma presented with cotton wool like stromal infiltration and hypopyon in left eye. Microbiological examination of corneal scraping showed fungal hyphae and yeast cells in direct smear. Molecular identification of the organism was performed which showed 100% homology with Rhodotorula mucilaginosa. Management of these cases is difficult often necessitating surgical procedures. However further reports are necessary to understand the disease and establish a treatment protocol.

Optimization of a bioactive exopolysaccharide production from endophytic Fusarium solani SD5.

Endophytic fungi were less investigated for exopolysaccharide production. In this study endophytic Fusarium solani SD5 was used for optimization of exopolysaccharide production. One variable at a time method and response surface methodology were employed to explore the optimum medium compositions and fermentation conditions. The organism produced maximum exopolysaccharide after 13.68 days of incubation at 28 °C in potato dextrose broth supplemented with (g%/l) glucose, 9.8; yeast extract, 0.69; KCl, 0.05; KH2PO4, 0.05 with medium pH 6.46. Use of 50 ml medium in 250 ml Erlenmeyer flask gives highest exopolysaccharide production. The organism produced more than two times higher exopolysaccharide (2.276 ± 0.032 g/l EPS) at optimized condition compared to pre-optimized condition (0.96 ± 0.021). In vivo toxicity test established nontoxic nature of the EPS (≤400 mg EPS/Kg of body weight). The EPS slightly altered intestinal indigenous bacteria and influenced the growth of beneficial Lactobacillus spp.

Schizophyllum commune: a new organism in eye infection.

PURPOSE: We report a case of mycotic keratitis caused by a rare fungus Schizophyllum commune. METHODS: Clinical examination, slit-lamp examination, and microbiological evaluation of the corneal ulcer were done, and its treatment outcome was studied. The fungal etiology was established by conventional microbiological techniques, polymerase chain reaction and speciation by DNA sequencing. RESULTS: Corneal scraping showed the presence of fungal filaments. The fungus was identified as S. commune based on DNA sequence analysis of the internal transcribed spacer region. The organism was susceptible to amphotericin B and voriconazole and demonstrated resistance to anidulafungin, itraconazole, and fluconazole. Therapeutic keratoplasty was performed but there was recurrence of the infection in the graft, which was controlled with topical voriconazole and intracameral amphotericin B. At the end of 3 months, the affected eye had developed phthisis bulbi. CONCLUSION: The best of our knowledge, this is the first reported case of keratitis caused by the rare fungus S. commune. Management of these cases is difficult, and surgical procedures may be needed.

Candida fermentati: a rare yeast involved in fungal keratitis.

PURPOSE: The aim of this study was to report a rare case of fungal keratitis from Eastern India. METHODS: This is a case report. RESULTS: A 52-year-old man with a history of minor trauma presented with a total corneal ulcer and hypopyon in the left eye. Microbiologic examination of corneal scrapings showed yeast cells in direct smear and typical yeast colonies on multiple solid agar media. Identification of the organism isolated in the culture was performed using the D1/D2 region of the large subunit (LSU 28S rDNA)-based molecular technique. Polymerase chain reaction amplified a band with a sequence that was 100% homologous with that of Candida fermentati. The organism was susceptible to amphotericin B and anidulafungin and demonstrated resistance to voriconazole, itraconazole, and fluconazole. Therapeutic keratoplasty was performed, followed by the recurrence of the infection in the graft, which was controlled with topical and intracameral amphotericin B. At the end of 3 months, the affected eye had developed phthisis bulbi. CONCLUSIONS: This is the first report of isolation of C. fermentati, a species closely related to Candida guilliermondii, from keratitis. Molecular diagnostic techniques are helpful in the accurate identification of this organism, which is clinically important in view of an antifungal susceptibility pattern that differs from that of other yeasts and for selection of appropriate therapy.

Fungal exopolysaccharide: production, composition and applications.

Fungal exopolysaccharides (EPSs) have been recognized as high value biomacromolecules for the last two decades. These products, including pullulan, scleroglucan, and botryosphaeran, have several applications in industries, pharmaceuticals, medicine, foods etc. Although fungal EPSs are highly relevant, to date information concerning fungal biosynthesis is scarce and an extensive search for new fugal species that can produce novel EPSs is still needed. In most cases, the molecular weight variations and sugar compositions of fungal EPSs are dependent to culture medium composition and different physical conditions provided during fermentation. An inclusive and illustrative review on fungal EPS is presented here. The general outline of the present work includes fungal EPS production, their compositions and applications. An emphasis is also given to listing out different fungal strains that can produce EPSs.

Evaluation of in vitro antioxidant potency of exopolysaccharide from endophytic Fusarium solani SD5.

A potent endophytic fungus, Fusarium solani SD5 was used for exopolysaccharide (EPS) production. The isolated EPS were purified and major EPS fraction (PS-I); rhamno galactan was used to evaluate anti oxidant activities in vitro. EPS (PS-I) showed significant free radical scavenging effect on DPPH (1,1-diphenyl-2-picrylhydrazyl) and scavenging potency is indicated by IC(50) value 578.541 ± 33.256 µg/ml. EPS (PS-I) significantly induced antioxidant parameters of peritoneal macrophage cells at a concentration dependent manner and at 500 µg/ml it showed maximum protective effect against free radicals [malondialdehyde (MDA) 0.178 ± 0.015; super oxide dismutase (SOD) 41.287 ± 1.051; glutathione peroxidase (GPx) 30.182 ± 1.237; reduced glutathione (GSH) 56.892 ± 1.272; oxidized glutathione (GSSG) 8.458 ± 0.768]. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide] cytotoxicity assay indicated that EPS (PS-I) had no significant cytotoxic effect (concentration up to 500 µg/ml) on macrophage cells. Present findings suggested that the EPS (PS-I) may become a potential nontoxic exogenous antioxidant.