Evaluation of In Vitro Antibacterial Activity of Roots of Glycyrrhiza glabra Against Multidrug Resistant Bacterial Pathogens of E. coli

 

Charmi P. Shah1*, V.J. Joshi2, Deval M. Patel3, Paras D. Dhami3, Dhruvesh K. Bhavsar3, Manisha N. Trivedi3, Urmila D. Vachhani3 and D.D. Santani3

1Jodhpur National University, Narnadi, Jhanwar Road, Jodhpur, Rajasthan. India.      

2Department of Microbiology, Alkem Lab Ltd., Daman. India.           

3ROFEL, Shri G M Bilakhia College of Pharmacy. Vapi. 396191. Gujarat. India.

*Corresponding Author E-mail: charmishah15@yahoo.co.in

 

ABSTRACT:

Glycyrrhiza glabra is very well known medicinal plant used in Ayuveda. The present study was conducted to evaluate the in vitro antibacterial properties of roots of G. glabra against multidrug resistant clinical isolates of E. coli. The antibacterial activity was carried out with agar well diffusion method. The results of the study revealed that ethanol extract has broad spectrum and strong activity than aqueous extract and ether extract. Its effectiveness against drug resistant bacteria pathogens provides hope that it can serve as an alternative therapeutic agent to treat many diseases.

 

KEYWORDS: Antibacterial activity, Glychrriza glabra, Agar well diffusion method, Zone of inhibition.

 


INTRODUCTION:

More concentration is being paid on bioactive constituents isolated from the plant species used in herbal medicine. The possible reason for the same is the side effects and resistant of antibiotics built up in various pathogenic microbes.1,2 Researchers are turning their attention for the use of and search for drugs and dietary supplements derived from plants to get rid of various microbial ailments.

 

Glycyrrhiza glabra (Licorice) is widely used as a medicinal herb in different parts of world. It is a member of the pea family and is used as a flavoring agent and expectorant in cough and cold preparations.3 Medicinally it is used in various diseases like Addison’s disease, asthma, peptic ulcer, arthritis, bronchitis. It also act as anti-inflammatory, detoxifier and hepatoprotective agent.4-6 A number of active constituents (flavonoids, polysaccharides, sugars, amino acids, of G. glabra have been identified. The main bioactive constituents is glycyrrhizin which possess a sweet taste.7 Antibacterial activities of roots of G. glabra have been studied in many of the previous reports but there are very few reports about its inhibitory effect on selected multi drug resistant pathogens. So here an attempt has made to study antibacterial effect of roots of G. glabra on drug resistant bacterial strains.

 

MATEIRALS AND METHODS:

Preparation of plant extracts:

10g of dried powder of Licorice were macerated with 100 ml of ether, water and ethanol for 24 h.8 The solvent from aqueous and ethanolic extracts were evaporated and to the remaining residue 50 ml of distilled water was added. All the macerates were squeezed through muslin cloth, filtered, centrifuged and sterilized by 0.2µ filters.

 

Clinical isolates and Antibiotic sensitivity test:

The human pathogenic bacterial cultures of E. coli were obtained from the Microbiology laboratory of Bharat Eye Hospital, Bilimora (Guajart).  The antibiotic sensitivity of the clinical isolates of E. coli was studied by Bauer- Kirby disc diffusion method.9 The culture was spread evenly on the surface of each Nutrient agar (HiMedia) plate and antibiotic discs were placed on the surface of agar and the plates were incubated at 370C for 24 h. After incubation the plates were examined for zone of inhibition. Zone were measured and recorded as sensitive, resistant or intermediate referring the zone size interpretive chart.  Agar well diffusion technique was employed to check antibacterial activity of roots of G. glabra against multi drug resistant bacterial cultures.10 Inoculum of 108 CFU/ml was inoculated in the Nutrient agar (HiMedia) and cups were made using cork borer. 100 µl of the prepared extracts were placed in the cups. The plates were incubated at 370C for 24 h and zone of inhibition was measured in mm.

 

 

 


Table 1: Effect of G. glabra on multidrug resistant E. coli

Extract    →

Bacterial strain   ↓

Resistant pattern*

Zone of inhibition in mm

Ether

Aqueous

Ethanol

E. coli 1(EC1)

E, P, K, Co, S, Cp, Gf

15

17

20

E. coli 2(EC2)

Am, A, E, P, K, Cp, Of

-

-

20

E. coli 3(EC3)

Am, A, E, P, Of

14

17

19

E. coli 4(EC4)

E, P, Cp, Co, Of

15

18

20

E. coli 5(EC5)

Am, E, P, K, Cp

13

18

21

E. coli 6(EC6)

E, P, K, Nx, S, Gf

-

-

19

E. coli 7(EC7)

Am, E, K, P, Co

13

16

19

E. coli 8(EC8)

Am, A, P, K, Cp, Co,

-

16

20

E. coli 9(EC9)

Am, E, P, K, T, S

-

16

20

E. coli 10(EC10)

Am, A, E, P, T, Gf, Nx, S

15

17

20

E. coli 11(EC11)

Am, C, P, K, S, G

15

18

19

E. coli 12(EC12)

Am, A, E, P, K, Cp, Co

12

17

21

E. coli 13(EC13)

Am, A, P, Co, S

13

-

19

E. coli 14(EC14)

Am, A, P, K, T, Cp, Co

-

18

19

E. coli 15(EC15)

Am, A, P, K, Cp, Co

14

17

19

E. coli 16(EC16)

Am, E, P, Co, S

13

16

21

E. coli 17(EC17)

Am A, P, Co, Nx

15

16

20

E. coli 18(EC18)

Am, A, P, K, Cp, Co

15

17

20

E. coli 19(EC19)

A, C, P, K, T, Cp, Co

13

17

19

E. coli 20(EC20)

Am, A, P, K, T, Cp, Co

14

18

21

E. coli ATCC 8739

-

15

18

20

*A= Ampicillin, Am= Amoxycillin, C= Chloremphenicol, E= Erythromycin, P= Penicillin, K= Kanamycin, T= Tetracycline, Cp= Ciprofloxacin, Co= Co-trimoxazole, Gf= Gatifloxacin, Nx= Norfloxacin, Of= Oflaxacin, Pf= Pefloxacin, Sc= Sparfloxacin, S= Streptomycin

 

 


RESULTS AND DISCUSSION

E. coli was found to be resistant to more than one antibiotic. E. coli is one of the organisms which act as normal flora of the human colon and is an opportunistic pathogen. Septicemia, urinary tract infections, diarrhea, pyogenic infections are commonly caused by E. coli.10,11 The results of the present study showed remarkable antibacterial activity of all the extracts of G. glabra against E. coli (Table 1). The potency of antibacterial activity of all the plant extracts are in following manner: Ethanol extract > Aqueous extract > Ether extract. These findings are in fair correlation with the study carried out by other authors.12,13 These observations may be attributed to two reasons: firstly, the nature of the biological active components (saponins, tannins, anthraquinone and alkaloids) which could be enhanced in presence of ethanol more easily than other solvents.

 

CONCLUSION:

As G. glabra showed remarkable antibacterial activity against all the pathogenic  multidrug resistant E. coli, for which it can be selected for further studies to discover bioactive natural products that may serve as leads for the development of new pharmaceuticals that address to prospect therapeutic needs.

 

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10.     Shah CP, Joshi BH. Antibacterial Activity of Some Medicinal Plants againts the Pathogens Isolated from Dacryocystitis. VNSGU Journal of Science and Technology. 2009; 1(2): 123-127.

11.     Baldwin TJ. The pathogenicity of enteropathogenic E. coli. J Med Microbiol. 1998; 47: 283.

12.     Manoj MN, Kailas CM, et al. Studies of Antibacterial Activities of Glycyrrhiza glabra Root Extract. International Journal of PharmTech Research.  2010; 2: 899-901.

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Received on 22.12.2010       Modified on 27.01.2011

Accepted on 06.02.2011      © RJPT All right reserved

Research J. Pharm. and Tech. 4(4): April 2011; Page 650-651