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Available online at www.notulaebiologicae.ro

Not Sci Biol, 2011, 3(1)65-69

Print ISSN 2067-3205; Electronic 2067-3264 Notulae Scientia Biologicae

Antibacterial Potential of the Extracts of the Leaves of

Azadirachta indica Linn.Saradhajyothi KOONA, Subbarao BUDIDA

Andhra Uniersity, Department of Botany, Visakhapatnam-530 003(A.P), India;[email protected]

Abstract

Azadirachta indica A. Juss (syn. Meliaazadirachta) is well known in India and popularly known as Indian neem. o evaluateantibacterial potential, the agar well difusion assay was used against Gram-negative and Gram-positive bacteria. Penicillin and Dimethylsuloxide were used as positive and negative controls, respectively. Methanol extract showed the highest and chloroorm extract showedmoderate to good antibacterial activity.Proteus ulgaris and Micrococcus luteus were the most susceptible bacteria whileBacillus subtilis

was more resistant bacterium to the hexane, chloroorm and methanol extracts o neem. Te study recommended or the isolation andseparation o bioactive compounds responsible or the antibacterial activity.

Keywords: agar-well difusion, bacteria, medicinal plants, neem, solvent extracts

Introduction

Plants have been a source o herbal remedies through-out the history o mankind. Various medicinal plants havebeen used or years in daily lie to treat diseases all overthe world (Nimri et al., 1999; Saxena, 1997). Te widespread use o herbal remedies and healthcare preparations,

such as those described in ancient texts like the Vedas andBible, has been traced to the occurrence o natural prod-ucts with medicinal properties. Existence and survival omankind is impossible without plant kingdom, as plantsare the primary procedures and play important role in sus-taining the lie orms on earth. Search o newer drugs rom

plants has been rise; since many o the microorganisms areposing series health related disorders. According to recentestimates by the WHO more than 3.5 billion people inthe developing world rely on plants as source o medicineor various ailments. Over 20,000 plants have medicinal

values and many plants are yet to be explored or their po-tentials. In addition, many o the existing synthetic drugs

cause various side eects. Hence, drug development romplant based compounds could be useul in meeting thisdemand or newer drugs with minimal side eects (Srivas-tava et al., 2000).

Azadirachta indica A. Juss (syn. Meliaazadirachta) iswell known in India and its neighboring countries. It ispopularly known as Indian neem (margosa tree) or Indianlilac. It is an evergreen tree, cultivated in various parts othe Indian subcontinent. Every part o the tree has beenused as traditional medicine or household remedy against

various human ailments. Neem has been extensively used inAyurveda, Unani and homoeopathic medicine and has be-

come a cynosure o modern medicine. Te Sanskrit nameo the neem tree is Arishtha meaning reliever o sickness

and hence is considered as sarbarigabubarini. Te tree isstill regarded as village dispensary in India. Chemical in-

vestigation on the products o the neem tree extensivelyundertaken in the middle o the isolation o nimbin, therst bitter compound isolated rom neem oil, more than135 compounds have been isolated rom dierent parts oneem (Ganguli, 2002). Neem elaborates a vast array o bio-

logically active compounds that are chemically diverse andstructurally complex (Subapriya and Nagini, 2005). Seedscontain a complex secondary metabolite azadirachtin. Temedicinal properties o the plantAzadirachta indicawerestudied by several workers. Tey were Antipyretic (Khat-tak et al., 1985; Okpanyi and Ezeukwk, 1981), anti-ma-larial eect (Rochankij et al., 1985; ella, 1977), anti tu-mour eect (Fujiwara et al., 1982), anti ulcer eect (Pillaiand Santhakumari, 1984), anti diabetic eect (Shukla et

al., 1973), antiertility eect (Sinha et al., 1984), eect oncentral nervous system (Singh et al., 1987) cardiovasculareect (Tompson and Anderson, 1978) and Antioxidantactivity (Bandyopadhyayet al., 2002; Sultana et al., 2007).

Boiled neem lea water makes an excellent antiseptic toclean wounds, soothes, swellings and eases skin problems(Shahidi Bonjar et al., 2004). Oil rom the leaves, seedsand bark possesses a wide spectrum o antibacterial activ-ity action against Gram-negative and Gram-positive mi-croorganisms, includingM. tuberculosis and Streptomycinresistant strains (Chopra et al., 1956).In itro it inhibitsVibrio cholerae, Klebsiella pneumoniae, M. tuberculosis and

M. pyrogenes (Satyavati et al., 1976). Rao et al. (1986) re-ported the antimicrobial activity o the seed oil against avariety o pathogens. Te antiungal eect o lea extractagainst Alternaria alternata (Bhomick and Choudhary,

1982). Antimicrobial eects o neem extract have beendemonstrated against Streptococcus species (Almas, 1999).

Received 11 December 2010; accepted 13 January 2011

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Koona, S. and S. Budida / Not Sci Biol, 2011, 3(1):65-69

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broth were used or inoculation o the nutrient agar plates.An aliquot (0.02 ml) o inoculums was introduced tomolten and cooled to 45C nutrient agar and placed on

petriplate by pour plate technique. Te appropriate wellswere made on respective agar plate by using cork borer. Inagar well diusion method 0.05 ml o hexane, chloroormand methanol extracts were introduced to their respective

wells ollowing an incubation period o 24 to 48 hours at37C. Antibacterial activity was evaluated by qualiyinginhibition zones (IZ) o bacterial growth surrounding the

plant extracts. Te entire antibacterial assay was carried outunder strict aseptic conditions. Penicillin (5 g/disc) wasused as positive control and DMSO as a negative control.riplicates were carried out or each extract against eacho the test bacterium. Te results o screening are shownin the ab. 1.

Results and discussion

Te preliminary screening results o hexane, chloro-orm and methanol extracts are presented in ab. 1. Allthe three extracts o leaves oAzadirachta indica wereexpressed antibacterial activity at least one bacterium.Methanol extract was the most eective against all thetested bacteria. Te chloroorm extract showed good tomoderate where as hexane extract showed low antibacte-rial activity. Te inhibition zone values were interpreted assensitive (18 mm), intermediate (14-17 mm) and resistant(

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nol extract oAzadirachta indica showed airly high degreeo sensitivity (IZ=20-33 mm) (Fig. 1) to all tested bacte-ria, except Bacillus subtilis, which was least susceptible tochloroorm and methanol extracts, while did not showedany activity to hexane extract. Penicillin (5 g) was used asstandard antibiotic reerence (positive control) exhibitedzone o inhibition values range rom 11-20 mm against alltested bacteria (Fig. 2). Tese values were lower than val-ues observed by methanol extract oAzadirachta indica.Hence methanol extract was exhibited a broad spectrumo antibacterial activity. Negative control, DMSO had noeect on microbial growth.

Proteus ulgaris and Micrococcus luteus belongs toGram-negative bacteria and the most susceptible bacteriato hexane, chloroorm and methanol extracts oAzadirach-

ta indica. Tis selective toxicity could be linked to the di-erences in the composition o the lipid bilayer or the twostrains o bacteria. A greater degree o depolarization andhence increased permeability, was expressed in the lipidbilayer o the Gram-negative bacteria or this cembranoid,because they contain more lipids in their cell walls (Staineret al., 1986). Tis depolarization eect is suggested to beassociated with hydrogen bonding on the hydroxyl groupin the carboxylic unctionally situated at the C-19 posi-tion in the diterpene ( Jente et al., 1990).

Te antibacterial activity oAzadirachta indica mightbe due to presence o triterpenoids, phenolic compounds,carotenoids, steroids, valavinoids, ketones and tetra-trit-erpenoids azadirachtin. Earlier studies on Azadirachataclaim that a spermicidal raction o neem oil (NIM-76) is

Fig. 1. Inhibition zones produced by methanol extracts

Fig. 2. Inhibition zones produced by standard antibiotic

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more eective as an antimicrobial agent as compared to theneem oil itsel especially its eect is less onEscherichia coliandKlebsiella pneumonia( Sairam et al., 2000). Antibacte-rial activity o the extracts oAzadirachta indica was eec-tive onEscherichia coli, Klebsiella pneumoniaeandBacillus

subtilis etc. and needs urther conrmation (Jagannadhand Radhika, 2006). Almas (1999) reported antimicro-bial activity o neem oil on Escherichia coli andKlebsiella

pneumonia with methanol but not with chloroorm andhexane extracts as is inuenced by pH o the nal extract.Te present study report also shows very high efcacy ochloroorm and methanol extracts onProteus ulgaris and

Micrococcus luteus and to a lesser extent againstEscherichiacoli, Klebsiella pneumonia, Enterococcus faecalis and Strep-tococcus faecalis. Tis study is also needs urther study toisolate and purication o bioactive compounds respon-sible or the antimicrobial activity.

Conclusions

Although this study investigating the in itro antibac-terial activity, the results showed that the extracts rom

Azadirachta indica leaves possessed good antibacterialactivity, conrming the great potential o bioactive com-

pounds and are useul or rationalizing the use o thisplant in primary health care.In io data may be helpul indetermining the real potential useulness o this plant orthe treatment o inectious diseases.

AcknowledgementTe authors acknowledge UGC-SAP, Department oBotany, Andhra University or nancial support or this

work.

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