Azadirachta indica (neem) has been used for a long time in agricultural and alternative medicine. Neem had been proved effective against certain fungi that could infect human body. This pilot study aims to demonstrate the antifungal effect of Malaysian neem leaf extracts on the pathogenic fungi in otomycosis, Aspergillus niger and Candida albicans. This is a laboratory-controlled prospective study conducted at Universiti Sains Malaysia. The powder form of Malaysian neem leaf was prepared. Ethanol and aqueous extracts of the neem leaf was diluted with sterile water to establish five different concentrations of 50 g/ ml, 25 g/ml, 12.5 g/ml, 6.25 g/ml and 3.125g/ml. The extract was tested on Sabouraud Dextrose Agar suspended with Candida albicans and Aspergillus niger respectively. Well diffusion method was used and zone of inhibition was measured. Growth of the fungi was inhibited in both alcohol and aqueous extract concentrations. The minimum inhibitory concentration (MIC) of Malaysian neem aqueous extract against Candida albicans was 11.91 g/ml, neem ethanol extract against Candida albicans was 5.16 g/ ml, neem aqueous extract against Aspergillus niger was 7.73 g/ml and neem ethanol extract against Aspergillus niger was 9.25 g/ml. Statistical analysis showed that the antifungal activity of Candida albicans is better in alcohol neem than aqueous extract (p<0.001) but aqueous neem extract is better than alcohol extract (p<0.001) for Aspergillus niger.Malaysian neem has significant antifungal effect towards Aspergillus niger, best in aqueous extract and towards Candida albicans, was best in alcohol extract.
Azadirachta

Conventional extraction of oil and azadirachtin, a botanical insecticide, from Azadirachta indica involves defatting the seeds and leaves using hexane followed by azadirachtin extraction with a polar solvent. In order to simplify the process while maintaining the yield we explored a binary extraction approach using Soxhlet extraction device and hexane and ethanol as non-polar and polar solvents at various ratios and extraction times. The highest oil and azadirachtin yields were obtained at 6 h extraction time using a 50:50 solvent mixture for both neem leaves (44.7 wt%, 720 mg(Aza)/kg(leaves)) and seeds (53.5 wt%, 1045 mg(Aza)/kg(seeds)), respectively.
Azadirachta ; Ethanol ; Limonins ; Solvents

The most common cause of pyogenic infection of the skin and soft tissues in children is Staphylococcus aureus, a fast-emerging problem due to its accompanying significant cost and morbidity. The popularity of herbal medications has increased due to the search for cheaper and more accessible alternatives. However, data is still lacking to back up these claims. Although a few in vitro studies have tested Neem leaf extract on S. aureus, there are none done on Methicillin-resistant Staphylococcus aureus (MRSA) despite the fact that it is being marketed for such purposes. Objectives: This study aims to determine if Neem leaf extract (Azadirachta indica) has antibacterial properties against Methicillin-sensitive and Methicillin-resistant Staphylococcus aureus and to compare the anti-staphylococcal properties of Neem leaf extract with oxacillin, vancomycin, mupirocin, and povidone iodine. Methods: An in vitro experimental study was performed using Neem leaf, properly identified and verified, was subjected to ethanol extraction of its active ingredients then diluted to produce 25%, 50%, 75%, and 100% concentrations. Standard strains of Staphylococcus aureus and clinical isolates of MRSA where inoculated on blood agar plates and subjected to the standardized disc susceptibility testing method. Zones of inhibition were measured for each test extract and compared to currently used medications, namely oxacillin, vancomycin, mupirocin, and povidone iodine with the pure diluent as negative control. The data was analyzed using difference of means hypothesis testing; it utilized the student's t-test to determine significance. Results: A trend of increasing antibacterial activity was noted with increasing concentration of the extract. Zones of inhibition started to appear at 50% concentration for S. aureus and 75% for MRSA. The antibiotics were able to produce greater zones of inhibition than the Neem extracts. Conclusion: Data from this study strongly suggest that the ethanol extract from Neem leaves exhibits in vitro antibacterial activity against both Staphylococcus aureus and MRSA with greatest zones of inhibition noted at 100% concentration.
STAPHYLOCOCCUS AUREUS ; AZADIRACHTA ; ANTI-BACTERIAL AGENTS ; METHICILLIN ;

Azadirachtin, as a botanical insecticide, is a highly oxidized limonoid triterpenoid existing in the seeds of Azadirachta indica. However, due to the low content in the seeds, the production of azadirachtin by seed extraction has low yield. Chemical synthesis of azadirachtin is characterized by complex process and low yield. Synthetic biology provides an alternative for the supply of azadirach-tin. In this study, two oxidosqualene cyclases AiOSC1 and MaOSC1 respectively derived from A. indica and Melia azedarach were identified in yeast. A yeast strain producing tirucalla-7,24-dien-3β-ol was constructed by integration of AiOSC1, Arabidopsis thaliana-derived squalene synthase gene(AtAQS2), and Saccharomyces cerevisiae-derived truncated 3-hydroxy-3-methyl-glutaryl coenzyme A reductase gene(PgtHMGR) into the delta site of yeast. Then, the function of MaCYP71BQ5 was successfully verified in yeast after this gene was introduced into the constructed yeast strain. This study not only laid a foundation for the biosynthesis of tirucalla-7,24-dien-3β-ol, but also provided a chassis cell for the functional identification of cytochrome oxidases(CYP450 s) in azadirachtin biosynthesis pathway.
Azadirachta ; Limonins ; Saccharomyces cerevisiae/genetics* ; Triterpenes

OBJECTIVETo examine the acaricidal effects of the essential oil of Cymbopogon citratus leaf extract (lemongrass) and ethanolic Azadirachta indica leaf extract (neem) against house dust mites Dermatophagoides farinae (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus).

METHODSTwenty-five adults mites were placed onto treated filter paper that is soaked with plant extract and been tested at different concentrations (50.00%, 25.00%, 12.50%, 6.25% and 3.13%) and exposure times (24hrs, 48hrs, 72hrs and 96 hrs). All treatments were replicated 7 times, and the experiment repeated once. The topical and contact activities of the two herbs were investigated.

RESULTSMortalities from lemongrass extract were higher than neem for both topical and contact activities. At 50 % concentration, both 24 hrs topical and contact exposures to lemongrass resulted in more than 91% mortalities for both species of mites. At the same concentration and exposure time, neem resulted in topical mortalities of 40.3% and 15.7% against D. pteronyssinus and D. farinae respectively; contact mortalities were 8.0% and 8.9% against the 2 mites, respectively. There was no difference in topical mortalities of D. pteronyssinus from exposure to concentrations of lemongrass and neem up to 12.50%; lemongrass was more effective than neem at the higher concentrations.

CONCLUSIONSGenerally, topical mortalities of D. farinae due to lemongrass are higher than that due to neem. Contact mortalities of lemongrass are always higher that neem against both species of mites.

Acaricides ; chemistry ; pharmacology ; Animals ; Azadirachta ; chemistry ; Cymbopogon ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Pyroglyphidae ; drug effects

Five fungicides such as rovral, bavistin, cupravit, dithane M-45 and thiovit were tested against conidial germination of Fusarium oxysporum. Dithane M-45 was the most effective against the fungus. Rests of the fungicides were more or less effective in the inhibition of conidial germination after 5~30 minutes immersion in 500~2500 ppm concentration. Five plant extracts(leaf extracts of Ocimum sanctum, Lantana camera, Calotropis procera, Azadirachta indica and Vinca rosea) found to be more or less effective against the fungus. 'Dhup' somke have good inhibitory effect on conidial germination of F. oxysporum.
Azadirachta ; Calotropis ; Fungi ; Fusarium* ; Germination* ; Immersion ; Lantana ; Ocimum ; Piper betle* ; Piper* ; Plant Extracts* ; Plants* ; Smoke* ; Vinca

Twenty plant extracts were tested against mycelial growth, sclerotium formatiom and dry weight of mycelium with sclerotia of Sclerotium rolfsii Sacc. The highest(90 mm) mycelial growth was measured in Adhatoda vasica, Tegetes erecta, Allium cepa, and Curcuma longa. The lowest(25 mm) was in Azadirachta indica. No mycelial growth was found in any concentration of cow, buffalo, and goat urine. The highest(90 mm) and the lowest(15 mm) mycelial growth were measured in Biomil and Urea, respectively. No mycelial growth was observed in Zinc. The highest(60 mm) and the lowest(2 mm) mycelial growth were recorded in Macuprex(Dodine; 65% WP) and Boron(100% Boric acid and 17% Boron) respectively. Mycelial growth was totally inhibited in Rovral(Iprodione; 50% WP).
Justicia ; Azadirachta ; Buffaloes ; Curcuma ; Fertilizers* ; Goats ; Mycelium ; Onions ; Plant Extracts* ; Plants* ; Urea ; Zinc

Fifteen limonoids were isolated from 95% ethanol extracts of the dry seeds of neem( Azadirachta indica) by various column chromatography techniques including silica gel,Pharmadex LH-20 gel and ODS resin. Based on spectroscopic analysis,their structures were determined as nimbocinol( 1),17β-hydroxynimbocinol( 2),1α,3α,7α-triacetylvilasinin( 3),7α-benzoyltrichilinin( 4),1,3-diacetyl-7-tigloyl-12-hydroxyvilasinin( 5),3-deacetylsalannin( 6),1-O-acetyl-1-detigloylsalannin( 7),2'( R),3'-dihydrosalannin( 8),2'( S),3'-dihydrosalannin( 9),2,3-dihydronimbolide( 10),6-homodesacetylnimbin( 11),gedunin( 12),7-deacetyl-7-epi-dihydrogedunin( 13),7-deacetoxy-7α-hydroxygedunin( 14) and nimbinene( 15). Compound 7 is a new natural product. 4,8,9,13 and 14 are isolated from the genus Azadirachta for the first time. Compound 2 showed inhibitory activity against Escherichia coli and Staphylococcus epidermidis,with MIC values of 32 and 128 mg·L~(-1),respectively. Compound 10 showed moderate inhibitory activity against S. epidermidis with a MIC value of 64 mg·L~(-1). Compound 11 inhibited the growth of E. coli and Pseudomonas aeruginosa,both with MIC values of 128 mg·L~(-1). Compound 15 exhibited inhibitory activity against P. aeruginosa,with a MIC value of128 mg·L~(-1).
Anti-Bacterial Agents/pharmacology* ; Azadirachta ; Escherichia coli ; Limonins ; Plant Extracts/pharmacology* ; Seeds

Eight limonoids were isolated from 95% ethanol extracts of neem(Azadirachta indica) seeds by various chromatographic methods. By comparison of their spectroscopic data with those reported in the literatures, these limonoids were determined as salannin(1), 1-detigloyl-1-isobutylsalannin(2), salannol-3-acetate(3), salannol(4), spirosendan(5), 1-detigloyloxy-3-deacetylsalannin-1-en-3-one(6), nimbin(7) and 6-deacetylnimbin(8). Compounds 2 and 5 were firstly isolated from this genus and 5 represented the only example of its type. And 6 is a new natural product. 6 showed inhibitory activity against HeLa and HL-60 cells, with IC₅₀ of(21.61±4.37) and(27.33±5.74) μmol·L⁻¹, respectively. Both 7 and 8 mildly inhibited the growth of HeLa cells, with IC₅₀ of (33.15±5.24) and (38.56±6.41) μmol·L⁻¹, respectively.
Azadirachta ; chemistry ; HL-60 Cells ; HeLa Cells ; Humans ; Limonins ; isolation & purification ; pharmacology ; Phytochemicals ; isolation & purification ; pharmacology ; Plant Extracts ; Seeds ; chemistry

OBJECTIVETo investigate the toxic implications of ethanolic stem bark extract of Azadirachta indica (A. indica) at 50, 100, 200 and 300 mg/kg body weight in Wistar rats.

METHODSFifty male rats of Wistar strains were randomly grouped into five (A-E) of ten animals each. Animals in Group A (control) were orally administered 1 mL of distilled water on daily basis for 21 days while those in Groups B-E received same volume of the extract corresponding to 50, 100, 200 and 300 mg/kg body weight.

RESULTSThe extract did not significantly (P>0.05) alter the levels of albumin, total protein, red blood cells and factors relating to it whereas the white blood cell, platelets, serum triacylglycerol and high-density lipoprotein cholesterol decreased significantly (P<0.05). In contrast, the final body weights, absolute weights of the liver, kidney, lungs and heart as well as their organ-body weight ratios, serum globulins, total and conjugated bilirubin, serum cholesterol, low-density lipoprotein cholesterol and computed atherogenic index increased significantly. The spleen-body weight ratio, alkaline phosphatase, alanine and aspartate transaminases, sodium, potassium, calcium, feed and water intake were altered at specific doses.

CONCLUSIONSOverall, the alterations in the biochemical parameters of toxicity have consequential effects on the normal functioning of the organs of the animals. Therefore, the ethanolic extract of A. indica stem bark at the doses of 50, 100, 200 and 300 mg/kg body weight may not be completely safe as an oral remedy and should be taken with caution if absolutely necessary.

Animals ; Azadirachta ; chemistry ; Body Weight ; drug effects ; Ethanol ; Male ; Organ Size ; drug effects ; Plant Bark ; chemistry ; Plant Extracts ; pharmacology ; toxicity ; Plant Stems ; chemistry ; Rats ; Toxicity Tests