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1.  Antibacterial and antifungal activities from leaf extracts of Cassia fistula l.: An ethnomedicinal plant 
This study was carried out with an objective to investigate the antibacterial and antifungal potentials of leaves of Cassia fistula Linn. The aim of the study is to assess the antimicrobial activity and to determine the zone of inhibition of extracts on some bacterial and fungal strains. In the present study, the microbial activity of hydroalcohol extracts of leaves of Cassia fistula Linn. (an ethnomedicinal plant) was evaluated for potential antimicrobial activity against medically important bacterial and fungal strains. The antimicrobial activity was determined in the extracts using agar disc diffusion method. The antibacterial and antifungal activities of extracts (5, 25, 50, 100, 250 μg/ml) of Cassia fistula were tested against two Gram-positive—Staphylococcus aureus, Streptococcus pyogenes; two Gram-negative—Escherichia coli, Pseudomonas aeruginosa human pathogenic bacteria; and three fungal strains—Aspergillus niger, Aspergillus clavatus, Candida albicans. Zone of inhibition of extracts were compared with that of different standards like ampicillin, ciprofloxacin, norfloxacin, and chloramphenicol for antibacterial activity and nystatin and griseofulvin for antifungal activity. The results showed that the remarkable inhibition of the bacterial growth was shown against the tested organisms. The phytochemical analyses of the plants were carried out. The microbial activity of the Cassia fistula was due to the presence of various secondary metabolites. Hence, these plants can be used to discover bioactive natural products that may serve as leads in the development of new pharmaceuticals research activities.
PMCID: PMC3217694  PMID: 22171301
Cassia fistula; in vitro antibacterial activity; antifungal activity; secondary metabolites
2.  Microbial evaluation of Limnophila rugosa Roth. (Merr) leaf 
Ayu  2014;35(2):207-210.
Limphonia rugosa Roth. (Merr.), family-Scrophulariaceae is considered as a botanical source of classical Ayurvedic drug Bhringaraja by the traditional practitioners of Odisha and is being used for the management of various disorders.
To study the antimicrobial activity of leaf of L. rugosa.
Materials and Methods:
Methanol extract of L. rugosa leaf (LRLM) has been studied, at various (5, 25, 50, 100, 250 μg/ml) dilutions, against medically important human pathogenic bacteria (two Gram-positive Staphylococcus aureus, Streptococcus pyogenes and two Gram-negative-Escherichia coli, Pseudomonas aeruginosa) and two fungal strains (Aspergillus niger, A. clavatus, Candida albicans) by using the agar disc diffusion method. A zone of inhibition of extract was compared with that of different standards such as ampicillin, ciprofloxacin, norfloxacin and chloramphenicol for antibacterial activity and nystatin and griseofulvin for antifungal activity.
The antibacterial and antifungal activities of the LRLM increased linear with the increase in concentration of extracts. When compared with standard drugs, the results revealed that, for bacterial activity S. pyogenes and S. aureus were more sensitive and in fungal activity C. albicans was more inhibited. The range of growth inhibition zone for all the sensitive bacteria was 11-20 mm and 13-19 mm for fungal strains.
Methanolic extract of L. rugosa leaf is having antibacterial and antifungal activities.
PMCID: PMC4279330  PMID: 25558169
Antifungal activity; Bhringaraja; Gandhamardan hills; in vitro antibacterial activity; Limnophila rugosa leaf; microbial load
3.  Antimicrobial and antifungal activities of Cordia dichotoma (Forster F.) bark extracts 
Ayu  2011;32(4):585-589.
Cordia dichotoma Forst.f. bark, identified as botanical source of Shlesmataka in Ayurvedic pharmacopoeias. Present study was carried out with an objective to investigate the antibacterial and antifungal potentials of Cordia dichotoma bark. Antibacterial activity of methanol and butanol extracts of the bark was carried out against two gram negative bacteria (Escherichia coli, and Pseudomonas aeruginosa) and two Gram positive bacteria (St. pyogenes and Staphylococcus aureus). The antifungal activity of the extracts was carried out against three common pathogenic fungi (Aspergillus niger, A.clavatus, and Candida albicans). Zone of inhibition of extracts was compared with that of different standards like Amplicilline, Ciprofloxacin, Norfloxacin and Chloramphenicol for antibacterial activity and Nystain and Greseofulvin for antifungal activity. The extracts showed remarkable inhibition of zone of bacterial growth and fungal growth and the results obtained were comparable with that of standards drugs against the organisms tested. The activity of extracts increased linearly with increase in concentration of extract (mg/ml). The results showed the antibacterial and antifungal activity against the organisms tested.
PMCID: PMC3361940  PMID: 22661859
Antibacterial; antifungal; Cordia dichotoma; gram positive; gram negative; in vitro
4.  In vitro antioxidant activity of hydro alcoholic extract from the fruit pulp of Cassia fistula Linn 
Ayu  2013;34(2):209-214.
The present study is aimed to investigate antioxidant activity of the extracts of Cassia fistula Linn. (Leguminosae) fruit pulp. Cassia fistula Linn., a Indian Laburnum, is widely cultivated in various countries and different continents including Asia, Mauritius, South Africa, Mexico, China, West Indies, East Africa and Brazil as an ornamental tree for its beautiful bunches of yellow flowers and also used in traditional medicine for several indications. The primary phytochemical study and in vitro antioxidant study was performed on hydro alcoholic extract of fruit pulp. Phytochemical screening of the plant has shown the presence of phenolic compounds, fatty acids, flavonoids, tannins and glycosides. Phenolic content was measured using Folin-Ciocalteu reagent and was calculated as gallic acid equivalents. Antiradical activity of hydro alcoholic extract was measured by DPPH (2,2-diphenyl-1- picrylhydrazyl) assay and was compared to ascorbic acid. Ferric reducing power of the extract was also evaluated by Oyaizu method. In the present study, three methods were used for evaluation of antioxidant activity. First two methods were for direct measurement of radical scavenging activity and third method to evaluate the reducing power. Results indicate that hydro alcoholic fruit pulp extracts have marked amount of total phenols which could be responsible for the antioxidant activity. These in vitro assays indicate that this plant extract is a significant source of natural antioxidant, Cassia fistula fruit pulp extract shows lower activity in DPPH and total phenol content as compared with standard which might be helpful in preventing the progress of various oxidative stresses.
PMCID: PMC3821253  PMID: 24250133
Antioxidants; Aragwadha; Cassia fistula; free radical; phenols
5.  The Antibacterial Activity of Cassia fistula Organic Extracts 
Cassia fistula, is a flowering plant and a member of Fabaceae family. Its leaves are compound of 4 - 8 pairs of opposite leaflets. There are many Cassia species around the world which are used in herbal medicine.
This study was designed to examine in vitro anti-bacterial activity of methanolic and ethanolic extracts of C. fistula native to Khuzestan, Iran.
Materials and Methods:
The microbial inhibitory effect of methanolic and ethanolic extracts of C. fistula was tested on 3 Gram positive: Bacillus cereus, Staphylococcus aureus and S. epidermidis and 5 Gram negative: Salmonella Typhi, Kelebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis bacterial species using disc diffusion method at various concentrations. The minimum inhibitory and bactericidal concentrations (MIC and MBC) were measured by the tube dilution assay.
The extract of C. fistula was effective against B. cereus, S. aureus, S. epidermidis, E. coli and K. pneumoniae. The most susceptible microorganisms to ethanolic and methanolic extracts were E. coli and K. pneumoniae, respectively. Also B. cereus and S. aureus showed the least sensitivity to ethanolic and methanolic extracts, respectively. The MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration) of ethanolic extracts against S. aureus, E. coli, S. epidermidis and K. pneumoniae were also determined.
With respect to the obtained results and regarding to the daily increase of the resistant microbial strains to the commercial antibiotics, it can be concluded that these extracts can be proper candidates of antibacterial substance against pathogenic bacterial species especially S. aureus, E. coli, K. pneumoniae and S. epidermidis.
PMCID: PMC4138669  PMID: 25147664
Cassia fistula; Anti-bacterial Activity; Disc Diffusion Antibacterial Test; Minimum Inhibitory Concentration; Minimum Bactericidal Concentration
6.  Investigation on Antibacterial and Antioxidant Activities, Phenolic and Flavonoid Contents of Some Thai Edible Plants as an Alternative for Antibiotics 
This study was aimed to examine the antibacterial and antioxidative properties of seven edible plants from Thailand to develop alternative antibiotics as feed additives. The plants include Citrus aurantifolia Swingle (Lime) fruits and its leaves, Sesbania grandiflora L. (Agati sesbania) leaves, Piper sarmentosum Roxb (Wild betal) leaves, Curcuma domestica Valeton (Turmeric) roots, Morinda citrifolia L. (Beach mulberry) leaves, Cassia siamea britt (Siamea cassia) leaves, and Cocos nucifera L. (Coconut) peels. The plants were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. Antibacterial activities with minimum inhibitory concentration (MIC) were determined by agar diffusion assay against Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, and Clostridium perfringens that were considered pathogenic strains in livestock infection. Methanol extracts of C. aurantifolia Swingle fruits and leaves showed the broadest spectrum of antibacterial activities except for C. perfringens. Butanol extract of S. grandiflora L. leaves showed the strongest activity against Burkholderia sp. with MIC, 135 μg/mL. P. sarmentosum Roxb leaves showed antibacterial activities against E. coli, Burkholderia sp. and H. parasuis. Ethyl acetate and water extracts from C. domesitca Valeton roots showed MIC of 306 μg/mL and 183 μg/mL, respectively against only C. perfringens. Antioxidative activity was determined by 2-diphenyl-2-picryl hydrazyl photometric assay. The methanol extracts of C. aurantifolia Swingle fruits and P. sarmentosum Roxb leaves showed the highest antioxidant activity among all the extracts with 3.46 mg/mL and 2.70 mg/mL effective concentration 50% (EC50) values, respectively. Total contents of phenolics and flavonoids were measured from the plant extracts. Methanol extracts of S. grandiflora L. and chloroform extracts of C. domestica Valeton were found to have the highest amount of total phenolics, 41.7 and 47.8 μg/mL, respectively. Flavonoid content of methanol extracts in S. grandiflora L. T was 22.5 μg/mL and the highest among plant extracts tested. These results indicated that C. aurantifolia Swingle, S. grandiflora L., P. sarmentosum Roxb, and C. domestica Valeton have antibacterial and antioxidant activities and can be used as alternative antibiotics or potential feed additives for the control of animal pathogenic bacteria.
PMCID: PMC4150179  PMID: 25178298
Plant Extract; Antibacterial Activity; Antioxidant Activity; Polyphenol; Flavonoid
7.  Antibacterial activities and phytochemical analysis of Cassia fistula (Linn.) leaf 
Cassia fistula Linn. which belongs to family Leguminosae is a medium-sized tree and its different parts are used in ayurvedic medicine as well as home remedies for common ailments. Sequential extraction was carried out using solvents viz. petroleum ether, chloroform, ethanol, methanol and water from leaf of the plant were investigated for preliminary phytochemical and antibacterial property. Results of the study showed that all the extracts had good inhibitory activity against Gram-positive test organism. Although all five extracts showed promising antibacterial activity against test bacterial species, yet maximum activity was observed in ethanol extract. The minimum inhibitory concentration ranged in between 94 to 1 500 μg/ml. Evaluation of phytochemicals such as alkaloids, flavonoids, carbohydrates, glycosides, protein and amino acids, saponins, and triterpenoids revealed the presence of most of constituents in polar extracts (ethanol, methanol, and aqueous) compared with nonpolar extracts (petroleum ether and chloroform). Furthermore, the ethanol extract was subjected to TLC bioautography and time-kill study against Staphylococcus epidermidis. All the findings exhibit that the leaf extracts have broad-spectrum activity and suggest its possible use in treatment of infectious diseases.
PMCID: PMC3217677  PMID: 22171295
Cassia fistula; human pathogenic bacteria; minimum inhibitory concentration; Similipal Biosphere Reserve; TLC bioautography
8.  Study of the synergistic anti-inflammatory activity of Solanum xanthocarpum Schrad and Wendl and Cassia fistula Linn 
Nonsteroidal and steroidal drugs are generally used as a part of drug therapy in inflammation. However, these drugs have severe side-effects like nausea and vomiting. Therefore, there is a need to identify anti-inflammatory compounds that will be effective with a better safety profile. Solanum xanthocarpum Schrad and Wendl and Cassia fistula Linn has many therapeutic uses mentioned in Ayurveda and therefore we aimed to study its anti-inflammatory activity both alone and in combination.
Materials and Methods:
The water extract of dried fruits of Solanum xanthocarpum Schrad and Wendl and dried pulp of Cassia fistula Linn was prepared. The anti-inflammatory activity of these extracts was investigated using the carragenan-induced paw edema model in rats individually and in two different combinations. ED50 of both the extracts singly and in combination were calculated by dose-response curves, and this information was then plotted on the isobologram. The interaction index of the extracts was also investigated to determine whether both the extracts in combination show synergistic or antagonistic or additive effects.
It was observed that extracts of dried fruits of Solanum xanthocarpum showed more anti-inflammatory activity than dried fruits of Cassia fistula Linn. Both the extracts showed maximum anti-inflammatory activity at 500 mg/kg dose. Among the different dose combinations of both the extracts, the 1:1 combination at the 500 mg/kg dose showed maximum percentage inhibition of 75%, which was comparable with the positive control, diclofenac sodium, which showed 81% inhibition.
As revealed by the isobolograms, both the combinations fell below the additivity line, which indicates synergistic interactions between Solanum xanthocarpum and Cassia fistula extracts. Interaction indices of both combinations were observed to be <1, which re-demonstrated the synergistic effects of the combination.
PMCID: PMC2996575  PMID: 21170209
Anti-inflammatory activity; Cassia fistula linn; isobolographic analysis; Solanum xanthocarpum Schrad and Wendl
9.  HPLC Quantitative Analysis of Rhein and Antidermatophytic Activity of Cassia fistula Pod Pulp Extracts of Various Storage Conditions 
BioMed Research International  2013;2013:821295.
Cassia fistula is well known for its laxative and antifungal properties due to anthraquinone compounds in the pods. This study quantitatively analyzed rhein in the C. fistula pod pulp decoction extracts kept under various storage conditions using HPLC. The antifungal activity of the extracts and their hydrolyzed mixture was also evaluated against dermatophytes. The contents of rhein in all stored decoction extracts remained more than 95% (95.69–100.66%) of the initial amount (0.0823 ± 0.001% w/w). There was no significant change of the extracts kept in glass vials and in aluminum foil bags. The decoction extract of C. fistula pod pulp and its hydrolyzed mixture containing anthraquinone aglycones were tested against clinical strains of dermatophytes by broth microdilution technique. The results revealed good chemical and antifungal stabilities against dermatophytes of C. fistula pod pulp decoction extracts stored under various accelerated and real time storage conditions.
PMCID: PMC3792547  PMID: 24171175
10.  Minimum inhibitory concentrations of medicinal plants used in Northern Peru as antibacterial remedies 
Journal of ethnopharmacology  2010;132(1):101-108.
The plant species reported here are traditionally used in Northern Peru to treat bacterial infections, often addressed by the local healers as “inflammation”. The aim of this study was to evaluate the Minimum Inhibitory Concentration (MIC) of their antibacterial properties against Gram-positive and Gram-negative bacteria.
Materials and methods
The antimicrobial activity of ethanolic and water extracts of 141 plant species was determined using a deep-well broth microdilution method on commercially available bacterial strains.
The ethanolic extracts of 51 species inhibited Escherichia coli, and 114 ethanolic extracts inhibited Staphylococcus aureus. In contrast, only 30 aqueous extracts showed activity against E. coli and 38 extracts against S. aureus. The MIC concentrations were mostly very high and ranged from 0.008 to 256mg/ml, with only 36 species showing inhibitory concentrations of <4mg/ml. The ethanolic extracts exhibited stronger activity and a much broader spectrum of action than the aqueous extracts. Hypericum laricifolium, Hura crepitans, Caesalpinia paipai, Cassia fistula, Hyptis sidifolia, Salvia sp., Banisteriopsis caapi, Miconia salicifolia and Polygonum hydropiperoides showed the lowest MIC values and would be interesting candidates for future research.
The presence of antibacterial activity could be confirmed in most species used in traditional medicine in Peru which were assayed in this study. However, the MIC for the species employed showed a very large range, and were mostly very high. Nevertheless, traditional knowledge might provide some leads to elucidate potential candidates for future development of new antibiotic agents.
PMCID: PMC2956840  PMID: 20678568
South America; Medicinal Plants; ethnobotany; Minimum Inhibitory Concentration (MIC); antibacterial; Escherichia coli; Staphylococcus aureus
11.  Gas chromatography-mass spectroscopy analysis of bioactive petalostigma extracts: Toxicity, antibacterial and antiviral activities 
Pharmacognosy Magazine  2014;10(Suppl 1):S37-S49.
Petalostigma pubescens and Petalostigma triloculare were common components of pharmacopeia's of multiple Australian Aboriginal tribal groupings which traditionally inhabited the areas in which they grow. Among these groups, they had a myriad of medicinal uses in treating a wide variety of bacterial, fungal and viral infections. This study was undertaken to test P. pubescens and P. triloculare leaf and fruit extracts for the ability to inhibit bacterial and viral growth and thus validate Australian Aboriginal usage of these plants in treating bacterial and fungal diseases.
Materials and Methods:
P. pubescens, and P. triloculare leaves and fruit were extracted and tested for antimicrobial, antiviral activity and toxicity. The bioactive extracts were further examined by RP-HPLC and GC-MS to identify the component compounds.
The methanol, water and ethyl acetate leaf and fruit extracts of displayed potent antibacterial activity. The methanol and ethyl acetate extracts displayed the broadest specificity, inhibiting the growth of 10 of the 14 bacteria tested (71%) for the leaf extract and 9 of the 14 bacteria tested (64%) for the fruit extracts. The water extracts also had broad spectrum antibacterial activity, inhibiting the growth of 8 (57%) and 7 (50%) of the 14 bacteria tested, respectively. All antibacterial extracts were approximately equally effective against Gram-positive and Gram-negative bacteria, inhibiting the growth of 50-75% of the bacteria tested. The methanol, water and ethyl acetate extracts also displayed antiviral activity in the MS2 plaque reduction assay. The methanol and water extracts inhibited 26.6-49.0% and 85.4-97.2% of MS2 plaque formation, respectively, with the fruit extracts being more potent inhibitors. All ethyl acetate extracts inhibited 100% of MS2 plaque formation. All extracts were also non-toxic or of low toxicity. Analysis of these extracts by RP-HPLC showed that the P. triloculare ethyl acetate fruit extract was the least complex of the bioactive extracts. Subsequent analysis of this extract by GC-MS revealed that it contained 9 main compounds: acetic acid; 2,2-dimethoxybutane; 4-methyl-1,3-dioxane; decane; unadecane; 2-furanmethanol; 1,2-benzenediol; 1,2,3-benzenetriol; and benzoic acid.
These studies validate Australian Aboriginal therapeutic usage of Petalostigma species and indicate their medicinal potential.
PMCID: PMC4047571  PMID: 24914307
Antibacterial; antiviral; Australian medicinal plants; euphorbiaceae; Petalostigma pubescens; Petalostigma triloculare; quinine bush
12.  Antimicrobial potential of Ricinus communis leaf extracts in different solvents against pathogenic bacterial and fungal strains 
To investigate the in vitro antimicrobial activities of the leaf extract in different solvents viz., methanol, ethanol and water extracts of the selected plant Ricinus communis.
Agar well diffusion method and agar tube dilution method were carried out to perform the antibacterial and antifungal activity of methanol, ethanol and aqueous extracts.
Methanol leaf extracts were found to be more active against Gram positive bacteria (Bacillus subtilis: ATCC 6059 and Staphylococcus aureus: ATCC 6538) as well as Gram negative bacteria (Pseudomonas aeruginosa: ATCC 7221 and Klebsiella pneumoniae) than ethanol and aqueous leaf extracts. Antifungal activity of methanol and aqueous leaf extracts were also carried out against selected fungal strains as Aspergillus fumigatus and Aspergillus flavus. Methanolic as well as aqueous leaf extracts of Ricinus communis were effective in inhibiting the fungal growth.
The efficient antibacterial and antifungal activity of Ricinus communis from the present investigation revealed that the methanol leaf extracts of the selected plant have significant potential to inhibit the growth of pathogenic bacterial and fungal strains than ethanol and aqueous leaf extracts.
PMCID: PMC3621469  PMID: 23593573
Antibacterial; Antifungal; Relative percentage inhibition; Ricinus communis; Methanol; Ethanol
13.  Phytochemical screening, antioxidants and antimicrobial potential of Lantana camara in different solvents 
To evaluate the antioxidant activity, hydrogen peroxide radicals scavenging activity, reducing power, the total phenolic and flavonoids contents, and antimicrobial and antifungal activities of methanol, ethanol and water extracts of leaves of Lantana camara (L. camara).
Methanol, ethanol and water extracts were evaluated against four Gram positive and Gram negative bacterial isolates (Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus subtilis) and two fungal strains (Aspergillus fumigatus and Aspergillus flavus). Methanol extract at different concentrations was tested for antioxidant potential and phytochemicals were determined by using spectrophotometric method.
The total phenolic content was (40.859±0.017) mg gallic acid/g in the leaves of L. camara, while the total flavonoids was (53.112±0.199) mg/g dry weight. Methanol leaf extract of L. camara showed maximum antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa and was also effective against other bacterial strains as compared to ethanol and aqueous extracts of leaves. The methanol leaf extract of L. camara exhibited significant inhibition (71%) and (66%) against Aspergillus fumigatus and Aspergillus flavus respectively.
The methanol extract of the L. camara leaves is effective against selected bacterial and fungal strains. Its phytochemical contents have broad antimicrobial properties and the plant might be a novel source of antimicrobial drug.
PMCID: PMC4027341
MIC; Antioxidants; Phytochemicals; Antimicrobial; Lantana camara
14.  Bactericidal Activity of Methanol Extracts of Crabapple Mangrove Tree (Sonneratia caseolaris Linn.) Against Multi-Drug Resistant Pathogens 
The crabapple mangrove tree, Sonneratia caseolaris Linn. (Family: Sonneratiaceae), is one of the foreshore plants found in estuarine and tidal creek areas and mangrove forests. Bark and fruit extracts from this plant have previously been shown to have an anti-oxidative or cytotoxic effect, whereas flower extracts of this plant exhibited an antimicrobial activity against some bacteria. According to the traditional folklore, it is medicinally used as an astringent and antiseptic. Hence, this investigation was carried out on the extract of the leaves, pneumatophore and different parts of the flower or fruit (stamen, calyx, meat of fruit, persistent calyx of fruit and seeds) for antibacterial activity using the broth microdilution method. The antibacterial activity was evaluated against five antibiotic-sensitive species (three Gram-positive and two Gram-negative bacteria) and six drug-resistant species (Gram-positive i.e. Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium and Gram-negative i.e. Extended-spectrum beta-lactamase-Escherichia coli, multidrug-resistant–Pseudomonas aeruginosa and Acenetobacter baumannii). The methanol extracts from all tested parts of the crabapple mangrove tree exhibited antibacterial activity against both Gram-positive and Gram-negative bacteria, but was mainly a bactericidal against the Gram-negative bacteria, including the multidrug-resistant strains, when compared with only bacteriostatic on the Gram-positive bacteria. Using Soxhlet apparatus, the extracts obtained by sequential extraction with hexane, dichloromethane and ethyl acetate revealed no discernable antibacterial activity and only slightly, if at all, reduced the antibacterial activity of the subsequently obtained methanol extract. Therefore, the active antibacterial compounds of the crabapple mangrove tree should have a rather polar structure.
PMCID: PMC3574533  PMID: 23441048
Antimicrobial activity; bactericidal; crabapple mangrove tree; drug resistant bacteria; Sonneratia caseolaris Linn
15.  Antimicrobial Activity and Phytochemical Constituents of Leaf Extracts of Cassia auriculata 
Plants produce a wide variety of phytochemical constituents, which are secondary metabolites and are used either directly or indirectly in the pharmaceutical industry. ‘For centuries, man has effectively used various components of plants or their extracts for the treatment of many diseases, including bacterial infections. In the present study methanol, chloroform and aqueous extracts of Cassia auriculata leaf were subjected for antimicrobial activity by well-diffusion method against six bacterial strains namely Bacillus cereus, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus mirabilis. The results revealed that the methanol and chloroform extracts exhibited strong inhibitory activity against all the tested organisms (zone of inhibition of 12-20 mm), except Pseudomonas aeruginosa (zone of inhibition 10 mm or nil). The aqueous extracts showed moderate activity by ‘Zone of inhibition ≤12 or nil). The extracts were screened for their phytochemical constituents by standard protocols’ and were shown to contain carbohydrates, proteins, alkaloids, flavonoids, steroids, saponins and tannins. The antibacterial activity of these extracts is possibly linked to the presence of flavonoids, steroid, saponins and/or tannins. Further studies are needed to determine the precise active principles from Cassia auriculata.
PMCID: PMC3719143  PMID: 23901174
Antibacterial activity; Cassia auriculata; phytochemical constituents
16.  A study on trypsin, Aspergillus flavus and Bacillus sp. protease inhibitory activity in Cassia tora (L.) syn Senna tora (L.) Roxb. seed extract 
Proteases play an important role in virulence of many human, plant and insect pathogens. The proteinaceous protease inhibitors of plant origin have been reported widely from many plant species. The inhibitors may potentially be used for multiple therapeutic applications in viral, bacterial, fungal diseases and physiological disorders. In traditional Indian medicine system, Cassia tora (Senna tora) is reportedly effective in treatment of skin and gastrointestinal disorders. The present study explores the protease inhibitory activity of the above plant seeds against trypsin, Aspergillus flavus and Bacillus sp. proteases.
The crushed seeds of Cassia tora were washed thoroughly with acetone and hexane for depigmentation and defatting. The proteins were fractionated by ammonium sulphate (0-30, 30-60, 60-90%) followed by dialysis and size exclusion chromatography (SEC). The inhibitory potential of crude seed extract and most active dialyzed fraction against trypsin and proteases was established by spot test using unprocessed x-ray film and casein digestion methods, respectively. Electrophoretic analysis of most active fraction (30-60%) and SEC elutes were carried employing Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Gelatin SDS-PAGE. Inhibition of fungal spore germination was studied in the presence of dialyzed active inhibitor fraction. Standard deviation (SD) and ANOVA were employed as statistical tools.
The crude seeds' extract displayed strong antitryptic, bacterial and fungal protease inhibitory activity on x-ray film. The seed protein fraction 30-60% was found most active for trypsin inhibition in caseinolytic assay (P < 0.001). The inhibition of caseinolytic activity of the proteases increased with increasing ratio of seed extract. The residual activity of trypsin, Aspergillus flavus and Bacillus sp. proteases remained only 4, 7 and 3.1%, respectively when proteases were incubated with 3 mg ml-1 seed protein extract for 60 min. The inhibitory activity was evident in gelatin SDS-PAGE where a major band (~17-19 kD) of protease inhibitor (PI) was detected in dialyzed and SEC elute. The conidial germination of Aspergillus flavus was moderately inhibited (30%) by the dialyzed seed extract.
Cassia tora seed extract has strong protease inhibitory activity against trypsin, Aspergillus flavus and Bacillus sp. proteases. The inhibitor in Cassia tora may attenuate microbial proteases and also might be used as phytoprotecting agent.
PMCID: PMC3144011  PMID: 21749682
17.  Phytochemical screening, antimicrobial and antioxidant efficacy of different extracts of Rumex dentatus L. - A locally used medicinal herb of Kashmir Himalaya 
To elucidate the antimicrobial and antioxidant activities of Rumex dentatus L. (R. dentatus) along with its phytochemical analysis.
Agar disk diffusion method for antimicrobial activity and DPPH, riboflavin photo-oxidation, deoxyribose and lipid peroxidation assay for antioxidant activity.
The antimicrobial and antioxidant activities of different concentrations of five R. dentatus extracts were tested against different clinical bacterial strains (Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhimurium) and fungal strains (Aspergillus versicolor, Aspergillus flavus, Accremonium spp., Penicillium dimorphosporum, Candida albicans, Candida parapsilosis, and Candida kruesie). Among all extracts, the butanol extract showed strong antibacterial activity against Klebsiella pneumoniae (inhibition zone diameter of 20 mm) and aqueous extract showed no activity against any of the bacterial strains. While as in case of the fungal strains, the maximum antifungal activity was observed against Aspergillus flavus by aqueous extract. The antioxidant activity revealed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals. The phytochemical tests carried out with the crude extracts of R. dentatus showed the presence of flavonoids, terpenoids, alkaloids, saponins, tannins, anthraquinones and cardiac glycosides in it. The total phenolic content of these extracts was estimated quantitatively from standard calibration curve of gallic acid and it varied from 145 µg/mg in butanol extract to 45 µg/mg in petroleum ether extract.
It can be concluded that the plant has got a broad spectrum antimicrobial and antioxidant activity and could be used as a potential alternative for treating various diseases.
PMCID: PMC4027333
Antimicrobial activity; Antioxidant activity; Rumex dentatus; Medicinal herb
18.  Assessment of phytochemicals, antimicrobial and cytotoxic activities of extract and fractions from Fagonia olivieri (Zygophyllaceae) 
In Pakistan Fagonia olivieri (Zygophyllaceae) is commonly used in the indigenous system of medicine for treatment of conditions like diabetes, cancer, fever, asthma, toothache, stomach troubles and kidney disorders. This study evaluated the crude methanol extract of F. olivieri (FOM) and its derived fractions for their antimicrobial and cytotoxic activities as well as the classes of phytochemical.
Dried powder of whole plant of F. olivieri was extracted with methanol (FOM) and the resultant was fractionated to give n-hexane fraction (FOH), chloroform fraction (FOC), ethyl acetate fraction (FOE), n-butanol fraction (FOB) and residual aqueous fraction (FOA). Methanol extract and its derived fractions were subjected to phytochemical screening using standard procedures. Also the extract and fractions were assayed for antibacterial, antifungal and cytotoxic activities using agar well diffusion technique, agar tube dilution method and brine shrimps lethality test, respectively.
The results obtained for phytochemical analysis indicate the presence of saponins and alkaloids in all the tested extract and fractions while anthraquinones were not detected. The results showed that all the bacterial strains tested in this study were susceptible to at least one of the fractions tested. However, FOE and FOB were the best antibacterial fractions and showed antibacterial activity against maximum number of bacterial strains. The results showed that Escherichia coli was the most sensitive bacterium while Bordetella bronchiseptica and Enterobacter aerogenes were less susceptible against various fractions. Maximum percent inhibition for growth was recorded for the fungus Aspergillus flavus with FOE whereas growth of Aspergillus fumigatus and Fusarium solani was inhibited by FOM and its all derived fractions. Minimum LC50 (24.07 mg/L) for brine shrimp assay was recorded for FOE followed by LC50 of FOC (26.1 mg/L) and FOB (30.05 mg/L) whereas maximum LC50 was exhibited by FOH (1533 mg/L).
These results indicated the use of F. olivieri to treat infections with emphasis to isolate and characterize the active principle responsible for antibacterial, antifungal and cytotoxic activities and its exploitation as therapeutic agent.
PMCID: PMC3717109  PMID: 23842440
Antibacterial activity; Brine shrimps lethality assay; Cytotoxicity; Fagonia olivieri; Phytochemicals
19.  Antibacterial and Antifungal Activity of Holothuria leucospilota Isolated From Persian Gulf and Oman Sea 
Emergence of antimicrobial resistance toward a number of conventional antibiotics has triggered the search for antimicrobial agents from a variety of sources including the marine environment.
The aim of this study was to evaluate the antimicrobial potential of Holothuria leucospilota from Qeshm and Kharg Islands against some selected bacteria and fungi.
Materials and Methods:
In this investigation, sea cucumbers from two coastal cities of Persian Gulf were collected in March and May 2011 and identified by the scale method according to the food and agriculture organization of the United Nations. Antibacterial activity of hydroalcoholic extracts of the body wall, cuvierian organs and coelomic fluid, methanol, chloroform, and n-hexane extracts of the body wall were evaluated by the spot test. In addition, their antifungal activity was assessed by the broth dilution method.
The displayed effect was microbiostatic at concentrations of 1000 and 2000 µg/mL rather than microbicidal. The highest activity of hydroalcoholic extracts was exhibited by body wall, cuvierian organs and coelomic fluid against Escherichia coli, Salmonella typhi, Staphylococcus aureus and Pseudomonas aeruginosa; Aspergillus niger, A. fumigatus, A. flavus and A. brasilensis. However, none of the methanol, chloroform and n-haxane extracts showed appreciable effects against Shigella dysenteriae, Proteus vulgaris, Bacillus cereus, S. epidermidis and Candida albicans. Moreover, cuvierian organs did not possess any antifungal potential.
Our data indicated that water-methanol extracts from the body wall of H. leucospilota possess antibacterial and antifungal activity. However, additional and in-depth studies are required to isolate and identify the active component(s).
PMCID: PMC4138674  PMID: 25147657
Holothuria leucospilota; Anti-Infective Agents; Persian Gulf
20.  Antimicrobial activity of some ethnomedicinal plants used by Paliyar tribe from Tamil Nadu, India 
Antimicrobial activity of 18 ethnomedicinal plant extracts were evaluated against nine bacterial strains (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Ervinia sp, Proteus vulgaris) and one fungal strain (Candida albicans). The collected ethnomedicinal plants were used in folk medicine in the treatment of skin diseases, venereal diseases, respiratory problems and nervous disorders.
Plants were collected from Palni hills of Southern Western Ghats and the ethnobotanical data were gathered from traditional healers who inhabit the study area. The hexane and methanol extracts were obtained by cold percolation method and the antimicrobial activity was found using paper disc diffusion method. All microorganisms were obtained from Christian Medical College, Vellore, Tamil Nadu, India.
The results indicated that out of 18 plants, 10 plants exhibited antimicrobial activity against one or more of the tested microorganisms at three different concentrations of 1.25, 2.5 and 5 mg/disc. Among the plants tested, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Punica granatum and Syzygium lineare were most active. The highest antifungal activity was exhibited by methanol extract of Peltophorum pterocarpum and Punica granatum against Candida albicans.
This study evaluated the antimicrobial activity of the some ethnomedicinal plants used in folkloric medicine. Compared to hexane extract, methanol extract showed significant activity against tested organisms. This study also showed that Toddalia asiatica, Syzygium lineare, Acalypha fruticosa and Peltophorum pterocarpum could be potential sources of new antimicrobial agents.
PMCID: PMC1621080  PMID: 17042964
21.  Estimation of flavoniods, antimicrobial, antitumor and anticancer activity of Carissa opaca fruits 
Carissa opaca Stapf ex Hanes fruits is traditionally used in the treatment of asthma, hepatitis and microbial infections. The present study was arranged to investigate the antimicrobial, cytotoxic and antitumor activity of various fractions of C. opaca extract and its bioactive metabolites responsible for that activity.
To characterize various fractions of C.opaca antibacterial, antifungal, cytotoxic and antitumor assays are used. Eight strains of bacteria including Bacillus subtilis, Enterobactor aerogenes, Escherichia coli, Klebsiella pneumoniae, Micrococcus luteus, Pseudomonas aeroginosa, Salmonella typhy, and Staphylococcus aureus and four strains of fungal viz: Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger and Fusarium solani are used. Brine shrimps and potato dics are used for anticancer and antitumor potency of extract. High performance liquid chromatography (HPLC) is utilized for determination of bioactive metabolites responsible for the activity.
HPLC chromatogram revealed the presence of orientin, isoquercetin, myricetin and apigenin. Various fractions of C. oapca showed significant antibacterial, antitumor and anticancer activity. In case of C. opaca fruit inhibition growth of Aspergillus niger was ranged between 23.2 ± 1.36% to 43.3 ± 2.39%, Aspergillus flavus ranged between 27.6 ± 1.39% to 65.6 ± 3.44%, Aspergillus fumigatus ranged between 13.2 ± 1.00% to 52.4 ± 1.54% and Fusarium solani ranged between 10.5 ± 1.02% to 14.6 ± 1.74%.
It can be concluded that, various fractions of C.opaca are accessible source of ethno pharmacy as they are consumed in different areas of Pakistan with ultimate health compensations.
PMCID: PMC3893608  PMID: 24373124
Carissa opaca; High performance liquid chromatography; Escherichia coli; Anticancer
22.  Antioxidant, Antimicrobial and Phytochemical Analysis of Cichoriumintybus Seeds Extract and Various Organic Fractions 
This study was carried out to evaluate the antimicrobial and antioxidant effectiveness of methanolic extract and different fractions (n-butanol, ethyl acetate, chloroform and n-hexane) of C.intybus seeds. The antimicrobial activity was determined by the disc diffusion method and minimum inhibitory concentration (MIC) against a panel of microorganisms (four bacterial strains, i.e. P. multocida, E. coli, B. subtilis and S. aureus and three fungal strains, i.e A. flavus, A. niger and R. solani). The results indicated that seeds extract and fractions of C. intybus showed moderate activity as antibacterial agent. While Antifungal activity of C. intybus seeds extract/fractions was very low against A. flavus and A. niger while mild against R. solani. The C.intybus seeds extract/fractions contained appreciable levels of total phenolic contents (50.8-285 GAE mg/100g of Dry plant matter) and total flavonoid contents (43.3-150 CE mg/100g of Dry plant matter). The C. intybus seed extract/fractions also exhibited good DPPH radical scavenging activity, with IC50 ranging from 21.28-72.14 μg/mL. Of the C .intybus seeds solvent extract/fractions tested, 100% methanolic extract and ethylacetate fraction exhibited the maximum antioxidant activity. The results of the present investigation demonstrated significant (p < 0.01) variations in the antioxidant and antimicrobial activities of C. intybus seeds solvent extract/fractions.
PMCID: PMC3813158  PMID: 24250548
C.intybus; Antimicrobial activity; Resazurin; A. flavus; Minimum inhibitory concentration
23.  Assessment of phytochemical content, polyphenolic composition, antioxidant and antibacterial activities of Leguminosae medicinal plants in Peninsular Malaysia 
Many medicinal plants from Leguminosae family can be found easily in Malaysia. These plants have been used as traditional medicines by local ethnic groups, where they are prepared as decoction, pastes for wound infections, and some have been eaten as salad. This paper focused on the assessment of antioxidant potential, antibacterial activity and classes of phytochemicals of nine plants from the Leguminosae family.
Acacia auriculiformis, Bauhinia kockiana, Bauhinia purpurea, Caesalpinia pulcherrima, Calliandra tergemina, Cassia surattensis, Leucaena leucocephala, Peltophorum pterocarpum, and Samanea saman were extracted with aqueous methanol and dichloromethane:methanol mixture to test for antioxidant and antibacterial activities. The Folin-Ciocalteu assay was conducted to quantify the total phenolic content and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay was used to determine the free radical quenching capacity. Antibacterial activity was assessed using disc diffusion (Kirby-Bauer) assay. Screening for major classes of phytochemical was done using standard chemical tests.
B. kockiana flowers and C. pulcherrima leaves contained high total phenolic content (TPC) and strong DPPH radical scavenging ability with TPC of 8280 ± 498 mg GAE/100 g, IC50 of 27.0 ± 5.0 μg/mL and TPC of 5030 ± 602 mg GAE/100 g, IC50 of 50.0 ± 5.0 μg/mL respectively. Positive correlation was observed between TPC and free radical scavenging ability. Most extracts showed antibacterial activity against Gram positive bacteria at 1 mg, while none showed activity against Gram negative bacteria at the same dose. All extracts (except Samanea saman flower) showed antibacterial activity against two strains of methicillin resistant Staphylococcus aureus (MRSA) with MID values ranging between 100 μg/disc and 500 μg/disc.
The potential source of antioxidant and antibacterial agents, especially for MRSA infection treatments were found in B. kockiana, C. pulcherrima, C. tergemina and P. pterocarpum. These preliminary results would be a guide in the selection of potential candidates for further pharmacological study and in search of new drug candidate in treating MRSA infections.
PMCID: PMC3045388  PMID: 21306653
24.  Antimicrobial activities of the rhizome extract of Zingiber zerumbet Linn 
To investigate antimicrobial effects of ethanolic extract of Zingiber zerumbet (Z. zerumbet) (L.) Smith and its chloroform and petroleum ether soluble fractions against pathogenic bacteria and fungi.
The fresh rhizomes of Zingiber zerumbet were extracted in cold with ethanol (4.0 L) after concentration. The crude ethanol extract was fractionated by petroleum ether and chloroform to form a suspension of ethanol extract (15.0 g), petroleum ether fraction (6.6 g) and chloroform soluble fraction (5.0 g). The crude ethanol extract and its petroleum ether and chloroform fractions were evaluated for antibacterial and antifungal activity against thirteen pathogenic bacteria and three fungi by the disc diffusion method. Commercially available kanamycin (30 µg/disc) was used as standard disc and blank discs impregnated with the respective solvents were used as negative control.
At a concentration of 400 µg/disc, all the samples showed mild to moderate antibacterial and antifungal activity and produced the zone of inhibition ranging from 6 mm to 10 mm. Among the tested samples, the crude ethanol extract showed the highest activity against Vibrio parahemolyticus (V. parahemolyticus). The minimum inhibitory concentration (MIC) of the crude ethanol extract and its fractions were within the value of 128-256 µg/mL against two Gram positive and four Gram negative bacteria and all the samples showed the lowest MIC value against V. parahemolyticus (128 µg/mL).
It can be concluded that, potent antibacterial and antifungal phytochemicals are present in ethanol extract of Z. zerumbet (L).
PMCID: PMC3614197  PMID: 23569803
Zingiber zerumbet (L.) Smith; Antibacterial; Antifungal; Minimum inhibitory concentration (MIC); Antimicrobial activity; Rhizome; Antifungal activity; Phytochemicals; Disc diffusion method; Soluble fraction; Pathogenic bacteria
25.  In Vitro Antimicrobial Potential of the Lichen Parmotrema sp. Extracts against Various Pathogens 
Objective(s): The ongoing increasing antibiotic resistance is one of the biggest challenges faced by global public health. The perennial need for new antimicrobials against a background of increasing antibiotic resistance in pathogenic and opportunistic microorganisms obliges the scientific community to constantly develop new drugs and antimicrobial agents. Lichens are known prolific sources of natural antimicrobial drugs and biologically active natural products. This study was aimed to explore in vitro antimicrobial activity of lichen Parmotrema sp.
Material and Methods: The methanol and aqueous extracts of lichen Parmotrema sp. was extracted using Soxhlet extractor. Antibiotic assessment of methanol and aqueous extracts was done against eight bacterial (Escherichia coli, Staphylococcus aureus, Proteus mirabilis, Salmonella sp., Shigella sp., Enterococci faecalis, Pseudomonas aeruginosa, Klebsiella pneumoniae,) clinical pathogens and five plant pathogenic fungal strains (Aspergillus terreus strain JAS1, Scedosporium sp. JAS1, Ganoderma sp. JAS4, Candida tropicalis and Fusarium sp.) by Kirby-Bauer method.
Results: The methanol lichen Parmotrema sp. extract inhibited all the test organisms. The highest antibacterial activity was found against Pseudomonas aeruginosa and Staphylococcus aureus. The weakest activity was manifested in Salmonella sp. and Scedosporium sp. JAS1. Strong antifungal effect was found against Ganoderma sp. JAS4 and Fusarium sp. The aqueous lichen Parmotrema sp. extract revealed neither antibacterial nor antifungal activity.
Conclusion: The present study shows that tested lichen Parmotrema sp. extracts demonstrated a strong antimicrobial effect. That suggests the active components from methanol extracts of the investigated lichen Parmotrema sp. can be used as natural antimicrobial agent against pathogens.
PMCID: PMC3758061  PMID: 23997920
Antimicrobial agents; Kirby-Bauer Method; Lichens; Multi-drug resistance

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