Evaluation of antimicrobial and anthelmintic activities of the aerial parts of Elephantopus scaber L.

 

Lakshmi Kanta Kanthal1*, B.V.V.S. Surya Kiran1, K.Satyavathi1, P. Bhoja Raju1, Subhas Sahoo2

1Koringa College of Pharmacy, Korangi, Tallarevu [M], E. G. Dist. - 533 461, A.P., India

2Pulla Reddy Institute of Pharmacy, Annaram, Jinnaram (M), Medak Dist., A.P., India

.

ABSTRACT:

Herbal medicine is having very old history. Plants are the typical manufacturers of complex drug molecules, which serve as a prototype to develop more effective and less toxic medicines. The antimicrobial activities of medicinal plants can be attributed to the secondary metabolites such as alkaloids, flavonoids, tannins, terpenoids that are present in these plants. Helminthes infections are becoming a cause to disorders like eosinophilia, pneumonia, anaemia, and under nourishment. Anti-helmethic drugs will expel these worms by paralyzing or by killing the worms. In traditional system of medicine a large number of plants are claimed to have anthelmintic activity. Chemical study started from 1960’s and showed that constituents of E. scaber L. include flavonoids, triterpenoids, flavonoid esters and sesquiterpene lactones. Elephantopus scaber L. belongs to the family Asteraceae. The present study was carried out to screen the antimicrobial and anthelmintic activities of Elephantopus scaber L. The activities of extracts of the plant material are compared with the piperazine citrate. The n-hexane, chloroform and methanolic extracts shows concentration dependent and significant anthelmintic activity compared to the piperazine citrate and Albendazole and Amoxicillin and Fluconazole for the case of antimicrobial activity. Thus the anthelmintic and antimicrobial activities of Elephantopus scaber L. are demonstrated for the first time.

 

KEYWORDS: Elephantopus scaber L, antimicrobial activities, Anti-helmethic drugs, Pheritima posthuma

 

 

INTRODUCTION:

The use of medicinal plants as a source for relief from illness can be traced back over five millennia to written documents of the early civilization in China, India and the Near east, but it is doubtless an art as old as mankind. Neanderthals living 60,000 years ago in present day Iraq used plants such as hollyback, these plants are still widely used in ethno medicine around the world1, 2.

 

On a global basis, atleast130 drugs, all single chemical entities extracted from higher plants, or modified further synthetically, are currently in use, though some of them are now being made synthetically for economic reasons3.

 

Medicinal plants represent a rich source of antimicrobial agents. Plants are used medicinally in different countries and are a source of much potent and powerful drugs4.A wide range of medicinal plant parts is used for extract as raw drugs and they possess varied medicinal properties. The different parts used include has root, stem, flower, fruit, twigs exudates and modified plant organs.


While some of these raw drugs are collected in smaller quantities by the local communities and folk healers for local used, many other raw drugs are collected in larger quantities and traded in the market as the raw material for many herbal industries 5.

 

Antimicrobial activities of many plants have been reported by the researchers6,7. The antimicrobial activities of medicinal plants can be attributed to the secondary metabolites such as alkaloids, flavonoids, tannins, terpenoids that are present in these plants8.

 

Elephantopus scaber (Asteraceae) has been used to cause diuresis and antipyresis and to eliminate bladder stones. This genus has been reported to contain the hydroxylated germacranolides molephantin and molephantinin, which also possess cytotoxic and antitumor properties12.Chemical study started from 1960’s and showed that constituents of E. scaber L. include flavonoids, triterpenoids, flavonoid esters and sesquiterpene lactones. Sesquiterpene lactones are most important due to their antitumor activity13. The major phytochemical constitutes of the plant are elephantopin, triterpenes, stigmasterol epifriedelinol and lupeol 14.

 

MATERIALS AND METHODS:

Plant material and extraction procedure:

The fresh aerial parts were collected from natural forests of Korangi, near Kakinada, East Godavari Dist.,  Andhra Pradesh, India during February 2012 and authenticated by Department of Pharmacognosy in Koringa College of Pharmacy, Korangi, A.P.  The aerial part of the plant was dried in shade for 7 days. It w as coarsely powdered and then the powdered plant material was subjected to successive hot soxhlet extraction using n- hexane, chloroform and methanol in their increasing order of polarity. The extract so obtained was concentrated to dryness by evaporating the solvent under reduced pressure using rotary evaporator. Extractive yield of n-hexane, chloroform and methanolic extract were found to be 5.8%, 4.3%, 3.5% of dry plant respectively.

 

Worm collection:

Indian earthworm, Pheretima posthuma was used for evaluating the anthelmintic activity due to its anatomical and physiological resemblance with the intestinal roundworm parasites of human beings 9. The worms are collected from the local water logged soil, washed with normal saline solution and stored in tyrode solution.

 

Antimicrobial activity:

The different plant extracts were screened for their antimicrobial properties against some selected gastrointestinal tract infection causing bacteria Shigella dysenteria (ATCC-3) Bacillus subtilis (UC-564), Salmonella typhi (UC-562), and E. coli (ATCC-2593) few human pathogenic fungi, Aspergillus niger (MTCC-1344) and Candida albicans (MTCC-3018), by agar diffusion cup plate method .For antibacterial studies incubation was carried out at 260C. Incubation period for fungus was carried out at 250C for 72 hours. Ampicillin and Fluconazole were used as standard drug for antibacterial and antifungal respectively. The test solutions were prepared at 500, 1000μg/ml using DMSO (Di Methyl Sulfoxide) as diluting solvent. Inhibition zones were measured and the diameter was calculated in millimetre. The result of zone of inhibition was cited in table-1

 

Anthelmintic activity:

Twelve petridishes of equal sizes were taken and numbered. 50ml of formulation of different concentrations (25, 50, 100 mg/ml) of extracts of the plant were placed in nine petridishes. 50ml of 10 mg/ml solution of piperazine citrate (Glaxo Smith Kline pharmaceuticals) and 50ml of 10 mg/ml solution of albendazole are used as references for paralysis and death respectively. 50ml normal saline solution was taken in another petridish as control. Six earth worms of equal length were introduced into each petridish. The petridishes were placed at room temperature and the time taken for paralysis and death was note. Paralysis was said to occur when the worms do not receive any sense even in normal saline. Death was concluded when the worms lose their motility followed with fading away of their body color, when dipped in warm water (50 0C) 10, 11. The results are shown in Table 2 and expressed as a mean ± SEM of six worms in each group.


 

RESULTS AND DISCUSSION:

Table: 1 Results of antimicrobial activity.

Antibacterial Activity (Zone of Inhibition in mm)*

Antifungal activity

(Zone of Inhibition in mm)

Test Compounds

S.dysenteria

(ATCC-3)

B. Subtilis

(UC-564)

S. typhi

(UC-562)

E. Coli

(ATCC-2593

C. albicans

(MTCC-3018)

A. niger

 (MTCC-1344)

500

μg/ml

1000

μg/ml

500

μg/ml

1000

μg/ml

500

μg/ml

1000

μg/ml

500

μg/ml

1000

μg/ml

500

μg/ml

1000

μg/ml

500

μg/ml

1000

μg/ml

n hexane extract

12

13

14

15

16

17

12

13

12

14

13

15

Chloroform extract

13

13

11

12

17

20

12

13

23

24

14

17

Methanolicextract

13

14

15

18

19

20

15

17

23

25

18

22

Amoxicillin at

10 μg/ml

18

22

19

20

--

--

Fluconazole at

10 μg/ml

--

--

--

--

28

25

DMSO

--

--

--

--

--

--

-

--

-

--

--

--

*Average of three reading.      -- Not Treated.

Table: 2 Results of Anthelmintic activity

Name of the plant

Type of extract

Conc. (mg/ml)

Time taken for paralysis (P) and death (D) of worms in min ± SEM

(P)

(D)

Glycosmis mauritiana

methanolic

25

26 ± 0.8

65 ± 0.7

50

13 ± 0.2

52 ± 0.1

100

07 ± 0.7

40 ± 0.9

n-hexane

25

27 ± 0.6

68 ± 0.3

50

15 ± 0.9

53 ± 0.3

100

08 ± 0.5

46 ± 0.5

chloroform

25

32 ± 0.3

70 ± 0.2

50

17 ± 0.4

57 ± 0.1

100

10 ± 0.8

47 ± 0.4

Piperazine citrate

10

24 ± 0.5

64 ± 0.4

Albendazole

10

30 ± 0.3

51 ± 0.4

Control

-----

-----

-----

Values are Mean ± SEM; n=6 worms in each group *p< 0.05 is considered as significant when compared with standard drug.


 

 


 


Graph showing the details of zone of inhibition (mm)


 


 


Graph showing the details of Time taken for paralysis using different extracts of Elephantopus scaber L. for evaluation of Anthelmintic activity:


 


 


Graph showing the details of Time taken for Death using different extracts of Elephantopus scaber L. for evaluation of Anthelmintic activity


 

Almost all the extracts show antimicrobial activity, but methanolic  and chloroform extract show more active than others, specially against Salmonella typhi (UC-562) (zone were 19 and 20mm at 500 and 1000 μg/ml respectively)and fungus Candida albicans (MTCC-3018)where the zone is 23 and 25mm at 500 and 1000μg/ml respectively comparable to standard Amoxicillin and Fluconazole. The microbial growth inhibition of chloroform extract also found more active especially against Candida albicans, Aspergillus niger and bacteria Salmonella typhi. The results are comparable to standard Fluconazole is 28 and 25mm and amoxicillin is 19 mm at 10μg/ml. The results of antimicrobial agents revealed that better antifungal and antibacterial spectrum of methanolic and chloroform extract of Elephantopus scaber L, which can be further used to isolate the phytoconstituents responsible for the same.

 

All the extracts shows significant anthelmintic activity compared to the standards. They exhibited varying degree of activity in a dose dependent manner and causes paralysis of the worms followed by death. Methanolic extract of Elephantopus scaber L. shows significantly more activity compared to the other extracts.

 

REFERENCES:

1.       Thomson, W.A.R., 1978. Medicines from the Earth. Maidenhead, United Kingdom. McGraw-Hill Book Co.

2.       Stockwell, C., 1988. Nature’s pharmacy. London, United Kingdom. Century Hutchinson Ltd.

3.       Gerhartz, W., Y.S. Yamamota, F.T. Campbell, R. Pfefferkorn and J.F. Rounsaville, 1985. Ullmann’s Encyclopedia of Industrial.

4.       Srivastava, J., J. Lambert and N. Vietmeyer, 1996.. Medicinal plants: An expanding role in development. World Bank Technical Paper. No. 320.

5.       Uniyal, S.K., K.N. Singh, P. Jamwal and B. Lal, 2006.  Traditional use of medicinal plants among the tribal communities of Chhota Bhangal, Western Himalayan.  J. Ethnobiol. Ethnomed, 2: 1-14.

 

6.       Reddy PS, Jamil K and Madhusudhan P. Antibacterial activity of isolates from Piper longum and Taxus baccata. Pharma. Biol. 39, 236-238 (2001).

7.        Ateb DA and ErdoUrul OT Antimicrobial activities of various medicinal and commercial plant extracts. Turk. J. Biol. 27, 157-162 (2003).

8.        Sher A; Antimicrobial activity of natural products from medicinal plants. Gomal J Med Sci.; 7(1):72-78 (2009)

9.       Thorn G.M, Harrison‘s Principles of internal Medicine, 1977, Mc Grew Hill, New York

10.     AA Kale et al; JP Salvekara. J. Chem. Pharm. Res., 2011, 3(2), 298-302.

11.     Y Shivhare et al J. Chem. Pharm. Res., 2011, 3(1), 488-491.

12.     Lee K H, Cowherd C M and Wolo M T. Antitumor agents. XV: deoxyelephantopin, an antitumor principle from Elephantopus carolinianus Willd. J Pharm Sci 1975, 64, 1572–3.

13.     Fuchino H, Koide T, Takahashi M, Sekita S and Satake M. New sesquiterpene lactones from Elephantopus mollis and their leishmanicidal activities. Plant Med 2001, 67, 647–53

14.     Rajkapoor B, Jayakar B and Anandan R. Antitumor activity of Elephantos scaber Linn. Against Dalton’s Ascitic Lymphoma. Indian J Pharm Sci., 2002, 64, 71 – 73

 

Received on 14.04.2013

Modified on 23.04.2013

Accepted on 12.05.2013

© A&V Publication all right reserved

Research J. Pharmacology and Pharmacodynamics. 5(3): May–June 2013, 193-196