MyMedR

A Combined SEM and TEM Studies of Microsporum canis After in Vitro Treatment with Atratoxin B1 from Holothuria Atra

Darah I., Teo M, Ibrahim C.O.

Sains Malaysiana, 1996;25.

Atratoxin B1 which was extracted from a local sea cucumber, Holothuria atra is a potential antifungal agent against dermatophytes. Exposure of Microsporum canis mycelia inoculated into Sabouraud glucose agar medium to the 10 mg/ml atratoxin B1 solution, resulted in complete suppression of the fungal growth. Prolonged exposure to the atratoxin B] (72 hours) resulted with necrosis in a substantial portion of the existing hyphae. The main anomalies noted were the abnormally shaped hyphae and the alterations in its morphology and cytology.
Atratoksin B] yang diekstrak daripada timun laut tempatan, Holothuria atra, adalah suatu agen antikulat yang berpotensi terhadap dermatofit. Pendedahan miselium Microsporum canis yang diinokulat ke dalam medium agar glukosa Sabouraud kepada larutan atratoksin B] berkepekatan 10 mg/ml, mengakibatkan penindasan lengkap dalam pertumbuhannya. Pendedahan yang lebih lama kepada larutan atratoksin B] (72 jam) mengakibatkan nekrosis berlaku pada sebahagian besar hifanya. Keganjilan yang jelas berlaku adalah ketidaknormalan pada bentuk hifanya dan juga pengubahsuaian yang terjadi pada morfologi dan sitologinya.

Fulltext Antimicrobial activity of Emilia sonchifolia DC., Tridax procumbens L. and Vernonia cinerea L. of Asteracea family: potential as food preservatives

Yoga Latha, L., Darah, I., Sasidharan, S., Jain, K.

Malays J Nutr, 2009;15(2):223-231.

MyJurnal

Chemical preservatives have been used in the food industry for many years. However, with increased health concerns, consumers prefer additive-free products or food preservatives based on natural products. This study evaluated antimicrobial activities of extracts from Emilia sonchifolia L. (Common name: lilac tassel flower), Tridax procumbens L. (Common name: tridax daisy) and Vernonia cinerea L. (Common name: Sahadevi), belonging to the Asteracea family, to explore their potential for use against general food spoilage and human pathogens so that new food preservatives may be developed. Three methanol extracts of these plants were tested in vitro against 20 bacterial species, 3 yeast species, and 12 filamentous fungi by the agar diffusion and broth dilution methods. The V. cinerea extract was found to be most effective against all of the tested organisms and the methanol fraction showed the most significant (p < 0.05) antimicrobial
activity among all the soluble fractions tested. The minimum inhibitory concentrations (MICs) of extracts determined by the broth dilution method ranged from 1.56 to 100.00mg/mL. The MIC of methanol fraction was the lowest in comparison to the other four extracts. The study findings indicate that bioactive natural products from these plants may be isolated for further testing as leads in the development of new pharmaceuticals in food preservation as well as natural plant-based medicine.

In vitro and in situ antiyeast activity of Gracilaria changii methanol extract against Candida albicans

Sasidharan S, Darah I, Jain K

Eur Rev Med Pharmacol Sci, 2011 Sep;15(9):1020-6.

PMID: 22013724

The Gracilaria (G.) sp are widely used in the traditional medicine in Malaysia. The methanol extract of Gracilaria changii B.M. Xia & I.A. Abbott (Gracilariaciae) was evaluated for antiyeast activity against Candida albicans (Berkhout).

Laboratory-scale production of lignin-degrading enzymes by free and entrapped cells ofPhanerochoete chrysosporium in a tubular air-lift bioreactor

Darah I, Ibrahim CO

Folia Microbiol (Praha), 1998;43(2):161-8.

PMID: 18470488 DOI: 10.1007/BF02816503

The production of lignin-degrading enzymes by free and entrapped cells ofPhanerochoete chrysosporium in a tubular air-lift bioreactor was studied. Under optimized cultural conditions the production of lignin peroxidase by free cells, calcium-alginate-entrapped cells and scouring-mesh-entrapped cells was in a ratio of 520ratio720ratio950 mU/mL, while the production of manganese peroxidase was in a ratio of 350ratio480ratio620 mU/mL. The stability of the entrapped cells by fed-batch systems was highest after 3 feeding experiments which is similarly demonstrated in the repeated use of the preparations in batch system.

Polygalacturonase production by calcium alginate immobilized Enterobacter aerogenes NBO2 cells

Darah I, Nisha M, Lim SH

Appl Biochem Biotechnol, 2015 Mar;175(5):2629-36.

PMID: 25547814 DOI: 10.1007/s12010-014-1447-4

Bacterial cells of Enterobacter aerogenes NBO2 were entrapped in calcium alginate beads in order to enhance polygalacturonase production compared to free cells. The optimized condition of 5 % (w/v) sodium alginate concentration, agitation speed of 250 rpm, and 15 beads of calcium alginate with inoculum size of 4 % (v/v; 5.4 × 10(7) cells/ml) produced 23.48 U/mL of polygalacturonase compared to free cells of 18.54 U/ml. There was about 26.6 % increment in polygalaturonase production. However, in this study, there was 296.6 % of increment in polygalacturonase production after improvement parameters compared to before improvement parameters of calcium alginate bead immobilization cells (5.92 U/ml). This research has indicated that optimized physical parameters of calcium alginate bead immobilization cells have significantly enhanced the production of polygalacturonase.

Fulltext Antimicrobial Activity of Emilia sonchifolia DC., Tridax procumbens L. and Vernonia cinerea L. of Asteracea Family: Potential as Food Preservatives

Yoga Latha L, Darah I, Sasidharan S, Jain K

Malays J Nutr, 2009 Sep;15(2):223-31.

PMID: 22691820

Chemical preservatives have been used in the food industry for many years. However, with increased health concerns, consumers prefer additive-free products or food preservatives based on natural products. This study evaluated antimicrobial activities of extracts from Emilia sonchifolia L. (Common name: lilac tassel flower), Tridax procumbens L. (Common name: tridax daisy) and Vernonia cinerea L. (Common name: Sahadevi), belonging to the Asteracea family, to explore their potential for use against general food spoilage and human pathogens so that new food preservatives may be developed. Three methanol extracts of these plants were tested in vitro against 20 bacterial species, 3 yeast species, and 12 filamentous fungi by the agar diffusion and broth dilution methods. The V. cinerea extract was found to be most effective against all of the tested organisms and the methanol fraction showed the most significant (p < 0.05) antimicrobial activity among all the soluble fractions tested. The minimum inhibitory concentrations (MICs) of extracts determined by the broth dilution method ranged from 1.56 to 100.00mg/mL. The MIC of methanol fraction was the lowest in comparison to the other four extracts. The study findings indicate that bioactive natural products from these plants may be isolated for further testing as leads in the development of new pharmaceuticals in food preservation as well as natural plant-based medicine.

Effects of Methanol Extract of Wedelia chinensis Osbeck (Asteraceae) Leaves against Pathogenic Bacteria with Emphasise on Bacillus cereus

Darah I, Lim SH, Nithianantham K

Indian J Pharm Sci, 2013 Sep;75(5):533-9.

PMID: 24403653

The antibacterial activity of the methanol extract of Wedelia chinensis leave was studied and tested against three pathogenic Gram positive bacteria (Bacillus cereus, B. subtilis and Stapylococcus aureus) and three pathogenic Gram negative bacteria (Escherichia coli, Proteus rettgeri and Pseudomonas aeruginosa) by the disk diffusion assay and broth dilution methods. The extract exhibited favourable antibacterial activity against the bacterial cells but was more potent against Gram positive bacteria with the minimum inhibition concentration of 3.12 to 6.25 mg/ml compared to the Gram negative bacteria which had minimum inhibition concentration values of 25 mg/ml. The time-kill study suggested that the extract possessed bactericidal properties at higher concentrations and eradicated the growth of bacterial cells. The major abnormalities occurred to the bacterial cells after exposed to the extract were complete alterations in their morphology and collapsed of the cells beyond repair. The methanol extract of W. chinensis may be an effective antibacterial agent to treat bacterial infections.

Influence of agitation speed on tannase production and morphology of Aspergillus niger FETL FT3 in submerged fermentation

Darah I, Sumathi G, Jain K, Lim SH

Appl Biochem Biotechnol, 2011 Dec;165(7-8):1682-90.

PMID: 21947762 DOI: 10.1007/s12010-011-9387-8

Agitation speed was found to influence the tannase production and fungal growth of Aspergillus niger FETL FT3. The optimal agitation speed was at 200 rpm which produced 1.41 U/ml tannase and 3.75 g/l of fungal growth. Lower or higher agitation speeds than 200 rpm produced lower enzyme production and fungal growth. Based on the SEM and TEM micrograph observation, there was a significant correlation between agitation speed and the morphology of the fungal mycelia. The results revealed an increase of the enzyme production with the change of the fungal growth morphology from filamentous to pelleted growth forms. However, the exposure to higher shear stress with an increasing agitation speed of the shaker also resulted in lower biomass yields as well as enzyme production.

Fulltext Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation

Darah I, Sumathi G, Jain K, Hong LS

Biotechnol Res Int, 2011;2011:897931.

PMID: 21826273 DOI: 10.4061/2011/897931

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO(3), 0.1% KH(2)PO(4), 0.05% MgSO(4) ·7H(2)O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 10(6) spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment.

Effects of Vernonia cinerea less methanol extract on growth and morphogenesis of Candida albicans

Latha LY, Darah I, Jain K, Sasidharan S

Eur Rev Med Pharmacol Sci, 2011 May;15(5):543-9.

PMID: 21744750

Vernonia (V.) cinerea Less (Asteraceae) have many therapeutic uses in the practice of traditional medicine. The methanol extract of V cinerea, was screened for antiyeast activity against pathogenic yeast Candida albicans.

Tannase enzyme production by entrapped cells of Aspergillus niger FETL FT3 in submerged culture system

Darah I, Sumathi G, Jain K, Lim SH

Bioprocess Biosyst Eng, 2011 Sep;34(7):795-801.

PMID: 21347668 DOI: 10.1007/s00449-011-0529-8

The ability of immobilized cell cultures of Aspergillus niger FETL FT3 to produce extracellular tannase was investigated. The production of enzyme was increased by entrapping the fungus in scouring mesh cubes compared to free cells. Using optimized parameters of six scouring mesh cubes and inoculum size of 1 × 10(6) spores/mL, the tannase production of 3.98 U/mL was obtained from the immobilized cells compared to free cells (2.81 U/mL). It was about 41.64% increment. The immobilized cultures exhibited significant tannase production stability of two repeated runs.

Fulltext Production and Optimization of Cellulase Enzyme Using Aspergillus niger USM AI 1 and Comparison with Trichoderma reesei via Solid State Fermentation System

Lee CK, Darah I, Ibrahim CO

Biotechnol Res Int, 2011;2011:658493.

PMID: 21350665 DOI: 10.4061/2011/658493

Novel design solid state bioreactor, FERMSOSTAT, had been evaluated in cellulase production studies using local isolate Aspergillus niger USM AI 1 grown on sugarcane bagasse and palm kernel cake at 1 : 1 (w/w) ratio. Under optimised SSF conditions of 0.5 kg substrate; 70% (w/w) moisture content; 30°C; aeration at 4 L/h · g fermented substrate for 5 min and mixing at 0.5 rpm for 5 min, about 3.4 U/g of Filter paper activity (FPase) was obtained. At the same time, comparative studies of the enzymes production under the same SSF conditions indicated that FPase produced by A. niger USM AI 1 was about 35.3% higher compared to Trichoderma reesei. This shows that the performance of this newly designed SSF bioreactor is acceptable and potentially used as prototype for larger-scale bioreactor design.

Toxicity study of Vernonia cinerea

Latha LY, Darah I, Jain K, Sasidharan S

Pharm Biol, 2010 Jan;48(1):101-4.

PMID: 20645763 DOI: 10.3109/13880200903046203

The methanol extract of Vernonia cinerea Less (Asteraceae), which exhibited antimicrobial activity, was tested for toxicity. In an acute toxicity study using mice, the median lethal dose (LD(50)) of the extract was greater than 2000 mg/kg, and we found no pathological changes in macroscopic examination by necropsy of mice treated with extract. As well as the oral acute toxicity study, the brine shrimp lethality test was also done. Brine shrimp test LC(50) values were 3.87 mg/mL (6 h) and 2.72 mg/mL (24 h), exhibiting no significant toxicity result. In conclusion, the methanol extract of V. cinerea did not produce toxic effects in mice and brine shrimp.

In vitro antimicrobial activity against Pseudomonas aeruginosa and acute oral toxicity of marine algae Gracilaria changii

Sasidharan S, Darah I, Noordin MK

N Biotechnol, 2010 Sep 30;27(4):390-6.

PMID: 20170762 DOI: 10.1016/j.nbt.2010.02.002

Methanol extract of the Gracilaria changii has been screened for antimicrobial activity against Pseudomonas aeruginosa. Antimicrobial activities were carried out using disc diffusion assay and broth dilution method against P. aeruginosa. The methanol extract of G. changii showed a good antimicrobial activity against P. aeruginosa with MIC (Minimum Inhibitory Concentration) value of 6.25mg/ml. Exposure of P. aeruginosa cells to 6.25mg/ml of methanol extract of G. changii resulted in complete inhibition of the bacterial cells. The main abnormalities noted via SEM and TEM studies were the alterations in morphology and cytology of the bacterial cells. The main reason for this deterioration was discussed. The effect of the methanol extract on the growth profile for the bacteria was also done and confirmed the bactericidal effect of the G. changii methanol extract on P. aeruginosa by changing the normal growth profile of P. aeruginosa. In an acute toxicity study using mice, the median lethal dose (LD(50)) of the extract was greater than 2000 mg/kg, and we found no pathological changes in macroscopic examination by necropsy of mice treated with extract. We conclude that G. changii might be safely used as an antimicrobial agent.

Enzymatic deinking of laser printed office waste papers: some governing parameters on deinking efficiency

Lee CK, Darah I, Ibrahim CO

Bioresour Technol, 2007 May;98(8):1684-9.

PMID: 17137782

The protocol for the enzymatic deinking of laser printed waste papers on a laboratory scale using cellulase (C) and hemicellulase (H) of Aspergillus niger (Amano) was developed as an effective method for paper recycling. A maximum deinking efficiency of almost 73% by the enzyme combination of C:H was obtained using the deinking conditions of pulping consistency of 1.0% (w/v) with the pulping time of 1.0min, temperature of 50 degrees C, pH=3.5, agitation rate of 60rpm, pulp concentration of 4% (w/v), concentration of each enzyme of 2.5U/g air dried pulp and the enzyme ratio of 1:1. The deinking efficiency was further enhanced to 95% using the optimized flotation system consisting of pH=6.0, Tween 80 of concentration 0.5% (w/w), working air flow rate of 10.0L/min and temperature of 45 degrees C. The deinked papers were found to exhibit properties comparable to the commercial papers suggesting the effectiveness of the enzymatic process developed.

Fulltext Preliminary isolation and in vitro antiyeast activity of active fraction from crude extract of Gracilaria changii

Sasidharan S, Darah I, Noordin MK

Indian J Pharmacol, 2008 Oct;40(5):227-9.

PMID: 20040962 DOI: 10.4103/0253-7613.44155

OBJECTIVE: To isolate the active fraction from crude extract of Gracilaria changii and to determine its in vitro antifungal activity.

MATERIALS AND METHODS: The active fraction was isolated from the crude extract of G. changii by various purification procedures such as column chromatography, thin layer chromatography, bioauthograph etc. The in vitro antifungal activity (Candida albicans) of the active fraction (1.00, 0.50, and 0.25 mg/ml) was studied by disc diffusion method and the effect of the active fraction on the morphology of yeast was done by scanning electron microscope (SEM) studies.

RESULTS: An active fraction with remarkable antifungal activity was separated from the crude extract. The active fraction was effective as a fungicide against C. albicans and showed a dose-dependent antifungal activity. A Scanning Electron Microscope (SEM) study confirmed the fungicidal effect of G. changii active fraction on C. albicans, by changing the normal morphology of C. albicans.

CONCLUSION: From G. changii crude extract, an active fraction with remarkable in vitro antifungal activity has been isolated.

Antimicrobial effects of Caulerpa sertularioides extract on foodborne diarrhea-caused bacteria

Darah I, Tong WY, Nor-Afifah S, Nurul-Aili Z, Lim SH

Eur Rev Med Pharmacol Sci, 2014;18(2):171-8.

PMID: 24488904

Caulerpa (C.) sertularioides has many therapeutic uses in the practice of traditional medicine in Malaysia. Crude methanolic, diethyl ether extract, ethyl acetate extract and butanolic extract from C. sertularioides were subjected to antimicrobial screening including the three Gram-positive and three Gram-negative diarrhea-caused bacteria.

Fulltext Acute and subchronic toxicity study of Euphorbia hirta L. methanol extract in rats

Yuet Ping K, Darah I, Chen Y, Sreeramanan S, Sasidharan S

Biomed Res Int, 2013;2013:182064.

PMID: 24386634 DOI: 10.1155/2013/182064

Despite Euphorbia hirta L. ethnomedicinal benefits, very few studies have described the potential toxicity. The aim of the present study was to evaluate the in vivo toxicity of methanolic extracts of E. hirta. The acute and subchronic oral toxicity of E. hirta was evaluated in Sprague Dawley rats. The extract at a single dose of 5,000 mg/kg did not produce treatment related signs of toxicity or mortality in any of the animals tested during the 14-day observation period. Therefore, the LD 50 of this plant was estimated to be more than 5,000 mg/kg. In the repeated dose 90-day oral toxicity study, the administration of 50 mg/kg, 250 mg/kg, and 1,000 mg/kg/day of E. hirta extract per body weight revealed no significant difference (P > 0.05) in food and water consumptions, body weight change, haematological and biochemical parameters, relative organ weights, and gross findings compared to the control group. Macropathology and histopathology examinations of all organs including the liver did not reveal morphological alteration. Analyses of these results with the information of signs, behaviour, and health monitoring could lead to the conclusion that the long-term oral administration of E. hirta extract for 90 days does not cause sub-chronic toxicity.

Fulltext Cytotoxicity and genotoxicity assessment of Euphorbia hirta in MCF-7 cell line model using comet assay

Ping KY, Darah I, Chen Y, Sasidharan S

Asian Pac J Trop Biomed, 2013 Sep;3(9):692-6.

PMID: 23998008 DOI: 10.1016/S2221-1691(13)60140-9

To evaluate the cytotoxicity and genotoxicity activity of Euphorbia hirta (E. hirta) in MCF-7 cell line model using comet assay.

Microsporum fulvum IBRL SD3: as novel isolate for chicken feathers degradation

Darah I, Nur-Diyana A, Nurul-Husna S, Jain K, Lim SH

Appl Biochem Biotechnol, 2013 Dec;171(7):1900-10.

PMID: 24013862 DOI: 10.1007/s12010-013-0496-4

Keratinous wastes have increasingly become a problem and accumulate in the environment mainly in the form of feathers, generated mainly from a large number of poultry industries. As keratins are very difficult to degrade by general proteases, they pose a major environmental problem. Therefore, microorganisms which would effectively degrade keratins are needed for recycling such wastes. A geophilic dermatophyte, Microsporum fulvum IBRL SD3 which was isolated from a soil sample collected from a chicken feather dumping site using a baiting technique, was capable to produce keratinase significantly. The crude keratinase was able to degrade whole chicken feathers effectively. The end product of the degradation was protein that contained essential amino acids and may have potential application in animal feed production. Thus, M. fulvum could be a novel organism to produce keratinase for chicken feathers degradation.

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