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Anti-bacterial and Anthelmintic Effects of Ethanolic Leaf Extract of Aristolochia Indica L.

Farhana Yasmin*
Pharmacy Discipline, Khulna University, Khulna-9208, Bangladesh

Mahin Hossain
Pharmacy Discipline, Khulna University, Khulna-9208, Bangladesh

Rafia Karim
Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Jessore-7408, Bangladesh

Md. Mokaddes Sarder
Pharmacy Discipline, Khulna University, Khulna-9208, Bangladesh

Abstract/ Executive Summary:

Objective: To explore the efficacy of ethanolic leaf extract of Aristolochia indica L. in bacterial and worm infections using experimental models. Methods: Preliminary phytochemical screening was evaluated by following standard procedure. Antibacterial activity was carried out by disc diffusion method and anthelmintic effect of the plant extract was determined based on time taken for paralysis and death of Haemonchus contortus. Results: Phytochemical analysis of the plant extract indicated the presence of carbohydrates, reducing sugars, combined reducing sugars, glycosides, tannins, alkaloids, acidic compounds, steroids, terpenoids and flavonoids. The extract showed activity against the bacterial strains namely Staphylococcus aureus, Staphylococcus epidermidis, Bacillas megaterium, Escherichia coli, Salmonella typhae, and Vibrio cholera species at the dose of 250 μg/disc and 500μg/disc in comparison with standard drug Ciprofloxacin (5 μg/disc). In anthelmintic test, the extract showed significant dose dependent decrease in paralysis time and death time of Haemonchus contortus. Conclusion: These results suggest that the extract possesses significant anthelmintic and antibacterial effects that support ethnopharmacological uses of this plant.

Keywords: Anti-bacterial, Anthelmintic, Haemonchus contortus, Phytochemicals, Aristolochia indica.

Location: Khulna district

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

The present study was undertaken to carry out the possible antibacterial and anthelmintic effects of the ethanolic leaf extract of Aristolochia indica L.

Methodology:

Plant material collection and identification: The leaves of A. indica were collected from the Khulna district on 12th October, 2013 at the daytime. During collection, any type of adulteration was strictly prohibited. The plants were officially authenticated by the experts of Pharmacy Discipline, Khulna University, Bangladesh. Preparation of crude extract: The collected leaves were separated from undesirable materials, shed dried and ground into a coarse powder with the help of a suitable grinder. 150 g of grinded leaves powder was soaked in 700 ml of ethanol in a glass container. After 15 days, the extract was separated from the plant debris by filtration using Whatman filter paper. The extract was concentrated by evaporation. The amount of yield in the extract was 14.67%. Experimental animal: Live parasites Haemonchus contortus (Nematode) were collected from freshly slaughtered cattle at local abattoirs and identified by Dr Md. Royhan Gofur, Lecturer, Department of Animal Husbandry and Veterinary Science, Rajshahi University, Rajshahi, Bangladesh. After washing, parasites were stored in 0.9% phosphate-buffered saline (PBS), pH 7.4 (8.01 g NaCl, 0.20 g KCl, 1.78 g Na2HPO4 and 0.27 g KH2PO4 in 1 L dH20) at 37±1 ºC.2.4. Phytochemical screening test: Preliminary phytochemical analysis of ethanolic leaf extract of A. indica was carried out by employing standard procedure (Ghani 2003).2.5. Evaluation of antibacterial activity by the disc diffusion method: Antibacterial activity of the ethanolic leaf extract of A. indica was measured by disc diffusion method followed by Biswas et al. (2014). Nutrient agar and Muellar-Hinton agar were sterilized in a flask and cooled to 45-50 °C and then taken in sterilized petri dishes with a diameter of 120 mm. The filter paper discs (5 mm in diameter) were impregnated with the crude extract at the concentration of 250 and 500 μg/disc and then placed onto the agar plates previously inoculated with the tested microorganisms. The tested microorganisms included Staphylococcus aureus, Staphylococcus epidermidis, Bacillas megaterium, Escherichia coli, Salmonella typhae, and Vibrio cholera. The Petri dishes were kept at 4 °C for 2 hrs. The plates were incubated at 37 °C for 16 hrs to allow the growth of the microorganisms. The diameters of the zones of inhibition were measured in millimeters using a calibrated scale. All the tests were repeated in triplicate. Blank discs were used as negative control to ensure that the residual solvent’s activity and the filter paper were not active by themselves. Ciprofloxacin was used as standard drug at the dose of 5 μg/disc. Evaluation of anthelmintic activity: Anthelmintic activity of the leaves extract of A. indica was investigated on live parasites H. contortus of cattle as per the method of Akter et al. (2014) with minor modifications. The parasites were divided into different groups consisting of six parasites in each group. Plant extracts at the concentrations of 25, 50 and 100 mg/ml and albendazole at the concentrations of 10 and 15 mg/ml were prepared and transferred to petridishes. Control group was treated with 0.1% tween-80 in PBS. Six parasites were placed in each Petri dish and observed. The time of paralysis was recorded when no movement was observed unless shaken vigorously. The death time was recorded after evaluating that the parasites did not move when shaken vigorously, dipped into warm water (50 ºC) or subjected to external stimuli. Anthelmintic activity was expressed as the time required for paralysis and death of parasites as compared to control..2.7. Statistical analysis: The data were presented as mean ± SEM (n=6). Results were analysed by One-way analysis of variance (ANOVA) followed by Turkey’s multiple comparisons test. Student t test were used to compare between two groups. The significant difference was considered at P < 0.05.

Source of Information: Biosci Bioeng Commun 2016; 2(1): 61-66

Article Link (Online Journal): https://www.journalbinet.com/

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Ethanolic leaf extract of A. indica has potent antibacterial and anthelmintic effect that supports the traditional uses of this plant for therapeutical purposes. Further study is needed to be explored for its phytochemical profile to recognize the bioactive constituents accountable for its versatile activities.

Knowledge Management: Journal