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

subtilis

.aureus

.coli

pneumoniae

.luteus

100

200

100

HAE

100

200

Ciprofloxacin

100

200

Dimethyl

formamide

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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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.

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Received on 04.07.2008 Modified on 28.07.2008

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