Screening for the Antimicrobial Activity of Lippia nodiflora Leaf Extract against Selective Bacterial Population

 

K.R Jeya1 and M. VeeraPagu2

1 Department of Biotechnology, Bharathidasan University College (w), Orathanadu, Thanjavur(Dt ).

2 Department of Biotechnology, Muthu Mase Arts and Science College, Harur, Dharmapuri(DT)

*Corresponding Author E-mail: krjaya5@gmail.com

ABSTRACT:

Lippia nodiflora plant is used as traditional medicinal plant as diuretic, maturant, useful in fevers and cold, astringent to bowels, stomachic, used in lack of bowel movements, pain in knee joints and in lithiasis. It was found that the plant possesses steroids, alkaloids, carbohydrates, flavonoids, essential oil, tannins and salts of potassium. A study was undertaken to investigate the Indian medicinal plant Lippia nodiflora against various bacterial pathogens Klebsiella pneumonia, Proteus vulgaris, Pseudomonas aeruginosa, Shigella flexneri, Streptococcus pyogenes and Staphylococcus aureus. The acetone, chloroform, ethanol, methanol and petroleum ether extracts of the plant extracts showed different diameters of inhibition zone ranging between 8mm to 16 mm. diameter. Among themselves methanol leaf extract shows highest zone of inhibition against Shigella flexneri. Simultaneously acetone and petroleum ether leaf extract shown to effective against Klebsiella pneumoniae and Staphylococcus aureus.

 

KEYWORDS: Antimicrobial activity, medicinal plants, Lippia nodiflora. Shigella flexneri, Aqueous extract, Klebsiella pneumoniae.

 


 

INTRODUCTION:

Lippia nodiflora Linn. (Verbenaceae ) is commonly called as Poduthuvalai, Poduthalai, Talaipodam in Tamil, Bokkena in Telugu, Bakkan in Hindi. It is found throughout warmer parts of India ascending up to 900m in the hills. It is common in wet places, along irrigation channels, canal edges and river banks. It is a creeping, prostate, much branched perennial herb with branches spreading profusely and rooting at the nodes. In therapeutic treatments, the use of medicinal plants extracts and their phytochemicals with known antimicrobial properties can be of great significance. In different countries, a number of studies have been done to prove such efficiency (Prusti et al.,2008).

 

In India medicinal plants are used directly in folk remedies or indirectly in modern medicine (Srinivasan et al.,2001). Plants are known to contain biologically active components having antimicrobial activities( Branter and Gries, 1994; Beuchat et al., 1990 ). Extracts of different parts of plants inhibit microorganisms (Asha et al., 2001 Chan and Muko ,2001 ; Govindachari et al., 1999 ). Antimicrobial agents derived from plants include phytoalexins, allicin, isothiocynate and phenolic compounds.

 

It was found that the aerial parts are used as anodyne, antibacterial, diuretic, emmenogogue, parasiticide, refrigerant, febrifuge and cooling. According to traditional uses and Unani system of medicine the plant is acrid, hot and dry; diuretic, maturant, useful in fevers and cold, astringent to bowels,stomachic, used in lack of bowel movements, pain in knee joints and in lithiasis. Lippia nodiflora contains flavonoids, sugars, sterol, an essential oil, resin, nonglucosidal bitter substance, tannin, large amount of potassium nitrate and other constituents.

 

In last three decades numbers of new antibiotics potentials. have produced, but clinical efficacy of these existing antibiotics is being threatened by the emergence of multidrug-resistant pathogens (Bandow et al.,2003). In general, bacteria have the genetic ability to transmit and acquire resistance The different parts of plants according to drugs (Cohen, M.L., 1992). The present study was designed to extract leaves of Lippia nodiflora with organic solvents. The leaf extract were used for assay of antibacterial activity against selective microorganisms.

 

MATERIAL AND METHODS :

Plant Material :

The Lippia nodiflora plant which are free of disease were collected from Dharmapuri district. Fresh leaves are collected and are washed three times in running tap water and finally in sterile distilled water. The leaves dried in shade and are powdered.

Extraction :

Ten grams of dried leaf powder was extracted with acetone, chloroform and ethanol, methanol and petroleum ether using a soxhlet extraction unit up to a minimum of six hours. After extraction the solvent was distilled off using rotary vaccum evaporator and concentrated

 

Microorganisms:

In this study common human pathogenic bacteria Klebsiella pneumonia, Shigella fllexneri Proteus vulgaris, Pseudomonas aeruginosa, Streptococcus pyogenes and Staphylococcus aureus were used. These cultures were isolated from various clinical specimen and identified by morphological and biochemical characteristics. The organisms were maintained on agar slope at 40c and subcultured for 24 hours before use.

 

Antibacterial Activity :

Standardized inoculum of 1 2 x107 CFU/ml with 0.5 Mc Farlaand was prepared for Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa, Shigella flexneri, Strptococcus pyogenes and Staphylococcus aureus and introduced onto the sterile Muller Hinton Agar plates separately. Each inoculum was uniformly spread over the medium with the help of sterile glass spreader. Plant leaf extract with different solvent were incorporated into sterile free disc of 6mm with known concentration of 1mg/ml. Each disc was placed aseptically on the Muller Hinton agar plates seeded with six bacteria separately. The plates were examined for the zone of inhibition after 24 hours. Each zone of inhibition was measured with a ruler.

 

RESULT :

The present study reveals that methanol leaf extract of Lippia nodiflora shows highest antimicrobial activity against Shigella flexneri. Acetone extract, Chloroform extract .ethanol extract, methanol extracts and petroleum ether extracts of lippia nodiflora was screened for antibacterial activity against some clinically important gram negative and gram positive bacteria namely Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa, Shigella flexneri, Streptococcus pyogenes and Staphylococcus aureus.


 

Table 1. Antimicrobial Activity of Lippia nodiflora Leaf extract

Solvent

 

Klebsiella

pneumoniae

Proteus

vulgaris

Pseudomonas

aeruginosa

Shigella

flexneri

Streptococcus

pyogenes

Staphylococcus

aureus

Acetone

14

9

8

10

8

9

Chloroform

12

13

9

9

8

8

Ethanol

13

10

12

13

10

8

Methanol

11

11

10

16

9

10

Petroleum ether

11

13

12

11

9

14

Values are mean Zone of inhibition(mm ), Chloramphenicol Standard 30mcg/disc.

 

 

Fig. 1. Antimicrobial Activity of Lippia nodiflora Leaf extract

 


Leaf extracts of five solvents were screened against the six clinically important bacteria. Acetone leaf extract shows maximum zone of inhibition 14mm against Klebsiella pneumoniae.(Table 1 and Fig.1) and shows minimum zone of inhibition 8mm against , Pseudomonas aeruginosa and Streptococcus pyogenes. Chloroform leaf extract shows maximum zone of inhibition 13mm against Proteus vulgaris and shows minimum zone of inhibition 8mm against Streptococcus pyogenes and Staphylococcus aureus. Ethanolic leaf extract shows maximum zone of inhibition 13mm against Klebsiella ppneumoniae and Shigella flexneri, and shows minimum zone of inhibition 8mm against Staphylococcus aureus. Methanolic leaf extract shows maximum zone of inhibition 16mm against Shigella flexneri and shows minimum zone of inhibition 9mm against Streptococcus pyogenes. Petroleum ether leaf extract shows maximum zone of inhibition 16mm against Shigella flexneri and shows minimum zone of inhibition 9mm against Streptococcus pyogenes.

 

DISCUSSION:

Several workers have reported many pharmacological properties including antispasmodic (Bhakuni et. al., 1969), hair afflictions, anti-inflammatory, analgesic and antipyretic (Forestieri et al., 1996), antibacterial, antiHelicobacter pylori activity (Yuan Chuen and Tung Liany 2005 ), hypotensive activity, antinociceptive (Ahmed et al., 2004 ) and antifungal (Pirzada et al., 2005). G. A. Mako and A. A. Noor have reported that crude ethanolic and aqueous extract of lippia nodiflora leaf and stem shows antibacterial activity.

 

Drug resistant human pathogenic microorganism has developed due to indiscriminate use of commercial antimicrobial drugs commonly used in the treatment of infectious diseases. This condition has forced scientists to search for new antimicrobial substances from various sources ([Subramanian et al., 2006). In India, large section of people especially, in villages using the herbal medicine to combat the infectious diseases and disorders. Gradually people move towards the traditional medicine. The reason for that is, trusts on herbal medicine, which improve the diseases conditions, after the herbal medicine treatment. No side effect or fewer side effects is observed due to herbal medicine. Another reason is the cost of the drugs and cost of the treatment is low. People in developing countries now prefer the herbal medicine. (Kandhasamy, M. and K.D. Arunachalam, 2008).

 

Higher plants are much more important in the production of economically important organic compounds, pharmaceuticals and pesticides (Hostettmann, K. and J.L. Wolfender, 1997). Extracts of many higher plants have been reported to exhibit antibacterial, antifungal and insecticidal properties under laboratory trails. Plant metabolites and plant based pesticides appear to be one of the better alternatives as they are known to have minimal environmental impact and danger to consumers in contrast to synthetic pesticides(Varma, J. and N.K. Dubey, 1999). In our present investigation the leaf extract of L.nodiflora possese highest inhibition zone against K.pneumoniae and S.flexneri.

 

The potential for developing antimicrobials from higher plants appears rewarding, as it will lead to the development of a phytomedicine to act against microbes. Many plants have been used because of their antimicrobial traits, which are due to compounds synthesized in the secondary metabolism of the plant such as phenols, terpenoids, alkaloids and flavonoids (Omulokoli et al., 1997). Based on our results, it is concluded that the plant extracts have great potential as antimicrobial compounds against microorganisms and they can be used in the treatment of infectious diseases caused by resistant microorganisms. From the present study it can be inferred that the aerial parts of lippia nodiflora possess antibacterial activity against Klebsiella and Shigella. Further exploration of these plants for isolation of active compounds may be considered.

 

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Received on 21.08.2011 Modified on 30.08.2011

Accepted on 11.09.2011 RJPT All right reserved

Research J. Pharm. and Tech. 4(11): Nov. 2011; Page 1669-1672