Anti-Bacterial Activity of Various Crude Extracts of Bryonia seabra

A. Elayaraja* and G. Devala Rao

 

Department of Pharmaceutical Chemistry, KVSR Siddhartha College of Pharmaceutical Sciences, Vijayawada-520010.

* Corresponding Author E-mail: elayaraja80@rediffmail.com

 

ABSTRACT

Extracts of petroleum ether, Benzene, Ethanol and hydro alcohol (50:50) were obtained from leaves of Bryonia seabra and they were evaluated by preparing different concentrations for in vitro antibacterial activity against different microbial strains namely B.subtilis, S.aureus, E.coli, K.pneumonia and M.luteus. The studies showed that both the ethanol and hydroalcohol extracts had significant activity with the standard and their activity had been more than petroleum and benzene extracts. Also they showed an increase in concentration of both the extracts increase the zone of inhibition. Both the extracts had significant broad spectrum antibacterial activity.

 

KEY WORDS Bryonia seabra, Plant extracts, Minimum Inhibitory Concentration

 

INTRODUCTION:

Since animals, human and plants co-evolved together, many of their constituents are similar in nature and therefore, certain plant compounds have been used in managing diseases. Many extracts (saline, acidic and ether) and substances separated from those extracts posses anti-bacterial activity against bacteria and fungi1,2. The  plant of  genus  Bryonia belongs to  the family Cucurbitaceae which consists of 110 genera and 640 species, which are mostly herbs climbing by tendrils, with abundant sap and rapid growth. Those species possess good medicinal properties mainly as diuretics (Cucurbita), purgative (Citrullus), as a bath sponge (Luffa) and as an antiseptic and antibacterial agent (Momordica). Bryonia is one of the genus having four species. The Indian bryonia species is often called as Bryonia seabra (Tam: Musumusukkai), herbal species found in India belongs to the same family. It possesses good therapeutic value in siddha and ayurvedha. Its leaves are used as diet (pathiyam) in the form of cakes. The juice of its leaves is used in treatment of asthma and chronic bronchitis. Other species of the same genera are used as an astringent, aperient, in treatment of rheumatic diathesis, fever and bilious attack3-7. The present study is intended to evaluate the anti-bacterial activity of various extracts obtained from the aerial parts of the plant and this is the first time of evaluating anti-bacterial efficacy of one of the part of this plant.

 

MATERIAL AND METHODS:

The plant material was collected from Cauvery delta region (Thanjavur district), Tamil Nadu, India. The identification of the  plants  including  the  experimental  using  parts  (aerial parts) has been confirmed by using all the official monographic specifications. Then the aerial parts were isolated, washed with water and dried in shade. Then they were  pulverized by using mechanical grinder and  passed through a 40-mesh sieve to obtain fine powder. About one kilogram of the powdered rhizomes was subjected to hot soxhlet extraction for 24 hrs by using petroleum ether (40- 60°C)8.  Then  the  prepared  extract  was  filtered  through muslin cloth by using separating funnel. Then the filterate was concentrated in vacuum by using rotary evaporator to get a semisolid mass. The dried marc was successively subjected to soxhlet extraction by using benzene and ethanol and their extracts were obtained. Finally the dried marc was subjected to cold maceration by using an equal mixture of methanol and water (1:1) to get a hydro alcoholic extract. Then the extract was concentrated to get a semisolid mass. The semisolid mass of all the extracts were redissolved in DMF (Dimethyl formamide) to get various concentrations for evaluating the anti-bacterial efficacy.

 

Bacterial strains namely Bacillus subtilis (NCIM 2063), Staphylococcus aureus (NCIM 2019), Escherchia coli (NCIM  2065),  Klebsiella  pneumoniae (NCIM  2036)  and Micrococcus luteus (NCIM 2043) had been procured from the National Chemical Laboratory, Pune-411008, India for this evaluation. The stock culture was maintained on Mueller Hinton (MH) agar medium at 37°C.

 

The anti-bacterial activity of the two extracts was performed by paper disc diffusion assay method9. The discs of uniform size (6mm) were prepared from Whatmann No.1 filter paper and  were  sterilized  in  hot  air  oven  at  160°C  for 1hr.Then the discs were   impregnated with the MIC (minimum Inhibitory concentration) of different concentrations (25mg/ml, 50mg/ml and 100mg/ml) of various   extracts   and   standard   ciprofloxacin.   The solvent DMF is used as a control. A loopful of each bacterial culture was mixed with MH agar media and poured in to sterilized plates. Then they were allowed to solidify under laminar flow. Then the solidified agar plates were labeled with proper dilutions. Finally the impregnated discs were placed in the proper positions of agar plates and the agar plates were inoculated at 37°C  for  24hrs  and  the   zone  of  inhibition  was observed.  Averages  of  three  independent determinations were recorded.

 

Table I: Minimum inhibitory concentration (MIC in mg/ml) of various extracts of Bryonia Seabra

Various crude

extracts     and various concentration

mg/ml)

Zone of inhibition against bacterial strains

(mm) are average of triplicate experiments

B.

subtilis

S

.aureus

E

.coli

K.

pneumoniae

M

.luteus

PE

25

100

200

 

-

2

2

 

4

6

9

 

-

-

-

 

6

9

11

 

2

5

8

BE

25

50

100

 

-

-

-

 

-

3

7

 

3

5

6

 

-

2

3

 

-

-

-

EE

25

50

100

 

11

17

21

 

13

18

24

 

8

10

12

 

6

9

11

 

9

14

21

HAE

25

100

200

 

10

13

18

 

11

16

22

 

8

10

14

 

9

12

16

 

8

10

13

Ciprofloxacin

25

100

200

 

13

18

24

 

15

22

27

 

11

17

25

 

10

15

21

 

16

19

23

Dimethyl

formamide

5

7

6

5

4

PE- Petroleum ether extract; BE- Benzene extract; EE- Ethanolic extract; HAE- Hydro-alcoholic extract; (-) No activity.

 

RESULTS AND DISCUSSIONS:

The antibacterial bioassays showed that the extracts of ethanol and hydro alcohol of aerial parts of Bryonia seabra were effective against all the tested bacteria in the  present  study.  They  showed  a  significant  anti-bacterial   activity   against   gram+ve   and   gram-ve bacteria, when compared with standard ciprofloxacin. Also by increase in concentrations of both the extracts showed  increase  in  their  anti-bacterial activity.  The zone of inhibition of both the extracts also showed an excellent anti-bacterial activity when comparing to pet- ether and benzene extracts (Table 1). In conclusion, our observations confirmed that B. seabra possesses good antibacterial properties and better efficacy. Further phytochemical studies are needed to identify active constituents responsible for the observed activity.

 

REFERENCES:

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2.      Carrison HJ, Biessell HD and Meller MG. Anti-bacterial substances from plants. J. Bact. 1946; 52: 155-168.

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4.      Nadkarni KM. In: Indian Materia Medica, Popular Prakashan Publications, Bombay. 1993; Vol II, pp.715.

5.      Allen HC. In: Allen’s key notes, Jain Publishers, New Delhi. 2000; 8th edn, pp.66-69.

6.      Das D and Agarwal VS. Discussion on the species In: Fruit Drug  Plants  of  India,  Kalyani  Publishers,  Ludhiana,  India. 1991; pp. 73-74.

7.      Bhattarcharjee   SK.   Citrullus   schard.ex   Eckl.   and   Zeyh. Cucurbitaceae.  In:  Handbook  of  Medicinal  Plants,  Pointer Publishers, Jaipur, India. 2000; pp. 101.

8.      Somchit MN, Reezal I, Nur EI and Mutalib AR. In vitro anti- bacterial activity of ethanol and water extract of cassia alata. J. Ethanopharmacol. 2003; 84: 1-4.

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Received on 04.07.2008    Modified on 28.07.2008

Accepted on 30.08.200 © RJPT All right reserved

Research J. Pharm. and Tech. 1(3): July-Sept. 2008; Page 283-284