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Antibacterial and Antioxidant Potential of Arisaema Jacquemontii Blume from Manali, Himachal Pradesh

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Antibacterial and Antioxidant Potential of Arisaema Jacquemontii Blume from Manali, Himachal Pradesh Bulletin of Pure and Applied Sciences. Print version ISSN 0970 4612 Vol.38 B (Botany), No.1. Jan-Jun 2019: P.23-33 Online version ISSN 2320 3196 DOI: 10.5958/2320-3196.2019.00004.1 Original Article Available online at www.bpasjournals.com Antibacterial and Antioxidant Potential of Arisaema jacquemontii Blume from Manali, Himachal Pradesh Kiran Bala1*, Jagriti Rana2, Anand Sagar3 Author’s Affiliation *Corresponding Author: 1,2,3Department of Biosciences, Himachal Kiran Bala Pradesh University, Shimla-171005, India Department of Biosciences, Himachal Pradesh University, Shimla- 171005, India E-mail: [email protected] Received on 21.01.2019, Accepted on 04.03.2019 Keywords: Abstract Arisaema jacquemonti Blume, Manali is not only a famous tourist place but also a treasure of many Antibacterial activity, traditional medicines. The antibacterial and antioxidant potential of leaves, Agar-well diffusion tubers and fruit of Arisaema jacquemonti Blume were evaluated in different method, extracts viz. acetone, chloroform, distilled water and methanol. The samples Percent inhibition of were collected from the area of Manali. Using Agar-well diffusion method bacterial growth, and different concentrations of extracts (25%, 50%, 75% and 100%), the Antioxidant potential, antibacterial activity was tested against three Gram-negative bacteria DPPH free radical (Salmonella typhi, Pseudomonas aeruginosa and Shigella dysenteriae) and two scavenging assay. Gram-positive bacteria (Lysteria monocytogenes and Staphylococcus aureus). Streptomycin (5 µg/ml) was taken as a standard antibiotic. Percent inhibition of bacterial growth was calculated for each tested extract. The tuber acetone, fruit methanol and leaf methanol extract were found to be more effective against some the tested bacteria than the standard antibiotic used. The chloroform and distilled water extract showed negligible results. IC50 value was calculated after 2, 2 Diphenyl-1-picrylhydrazyal (DPPH) free radical scavenging assay to investigate the antioxidant potential at different concentrations (20, 40, 60, 80 and 100 µg/ml). Ascorbic acid at similar concentrations was taken as standard (IC50 = 31.33µg/ml). The methanol- leaf extract was found to exhibit highest antioxidant potential with IC50 value of 47.89µg/ml followed by methanol-tuber, water- fruit, acetone-fruit and methanol-fruit, acetone-tuber, acetone-leaf and water-leaf extracts. The results can be taken as a base for further analysis and isolation of active compounds of this plant. INTRODUCTION Infectious diseases affect about 50,000 people every day1. Staphylococcus aureus is typical bacterium causing number of skin disorders. Food borne illnesses are one of the major problems in the world which are mostly contributed by bacteria like Salmonella spp. and Listeria monocytogenes. Pseudomonas aerugenosa is one of the top three contributors of opportunistic human infections2. Shigella dysenteriae is Kiran Bala et al. / Antibacterial and antioxidant potential of Arisaema jacquemonti Blume from Manali, Himachal Pradesh associated with the poor sanitation and overcrowded areas of the world. Beside these issues, the imprudent use of commercial and synthetic antimicrobial drugs for the treatment of infections has developed multiple drug resistance in pathogenic organisms3,4. In 2003, Finch5 reported S. typhi attaining MDR to the first line of antibiotics. Synthetic antibiotics are also associated with allergy, hypersensitivity and immune suppression6. With changing life style and environmental conditions, several chemical molecules such as free radicals are exposed which are not appropriate for human health. Being extremely unstable, these radicals draw electrons out from other molecules by deteriorating them to attain stability. Oxygen is vitally important but its toxicity to living tissues can’t be denied. Number of Reactive Oxygen Species (ROS) such as hydroxyl radicals, superoxide anion radicals and hydrogen peroxide radicals are the crucial part of metabolism. In adequate concentration they are essential for energy supply, detoxification, chemical signalling and immune responses7. But increased concentration may lead to the consequences like oxidative stress relating to the initial symptoms of variety of suffering including cancer, inflammations, age related diseases and Neuro-degradation8-12. Plants have tremendous potential in the field of medicines because they are effective against infections and also, they mollify the side effects commonly associated with the synthetic drugs13. Plants produce large number of secondary metabolites such as phenols, flavonoids, anthocyanins, carotenoids, dietary glutathionine and vitamins which are the main components of defence system14. These molecules present in the herbal remedies have the ability to combine with many other inactive substances and thus provides a plant much efficiency and superiority to its isolated and pure active compounds15. These metabolites also act as antioxidants and function as singlet or triplet oxygen quenchers, peroxide decomposers, enzyme inhibitors and synergists16. Between 1981 and 2002, about 61% of the developed drugs in the field of infectious disease and cancer were based on natural products17. Even today, 80% of the world population is dependent on traditional remedies for the treatment of various diseases. Traditionally, specific plants are being used as medicine for specific diseases18. Arisaema jacquemontii Blume of Araceae family is native to sub-alpine Himalayan region, distributed at the heights between 2,500-4,000 m in Afganistan, Pakistan, India (Jammu and Kashmir, Himachal Pradesh, Uttrakhand, Sikkim and Arunachal Pradesh), Nepal, Tibet and Bhutan19. In Himachal Pradesh, this plant is reported from Kinnaur, Kullu, Mandi and Shimla districts, preferring the cervices between stones under the shade of oak and deodar trees on loamy, peaty and well drained but moist soil full of humus. In Kulluvi dialect, it is known as Basair. In traditional medicine of local area, tubers of this plant are being used for dermatological infections and ringworms20. This plant is also used for boils, blisters, psychic and nervous disorders in other areas where this plant grows21-23. Whole plant is also being used against snake bites, microbial infections, swelling, throat infection, obstructions, infertility and uterus diseases24-25. In Tibetan therapy, the flower, tuber and tuber is used against scabies, toothache, swellings, chest infection, menstrual disorders and throat problems26. Keeping this ethno medical knowledge and above mentioned challenges in mind, the present investigation was carried out to find out the antibacterial and antioxidant potential of the plant in different solvents using different concentrations. MATERIALS AND METHODS Collection of plant material The study material was collected from the Solang Valley at an altitude of 2353–2543m in Manali, Kullu district, Himachal Pradesh. The leaves were collected at the end of July 2018 and tuber and fruit were collected at the first week of September. The plant was identified at Department of Biosciences, HPU. Collected leaves, fruit and tuber were washed properly under tap water to remove dirt and soil particles and were surface sterilized using 0.1% of mercuric chloride solution. This material was dried and ground coarsely in mortar- pestle for further use. The experiments were conducted in the month of October and November. Bulletin of Pure and Applied Sciences/ Vol.38-B –Botany (No.1)/ January-June 2019 24 Kiran Bala et al. / Antibacterial and antioxidant potential of Arisaema jacquemonti Blume from Manali, Himachal Pradesh Preparation of plant extracts Acetone, chloroform, methanol and water extracts of dried fruit, tubers and leaves were prepared to screen antimicrobial activity. 3 g of material was taken in Erlenmeyer flask to which 30 mL of required solvent was added. The flask was covered with aluminium foil and placed at safe place for 3-5 days for extraction. Material was filtered using Whatman filter paper no.1 and the extract was evaporated at 40°C using rotary evaporator. The extract was collected and weighed. At last, a stock solution of 50 mg/mL concentration was prepared. Procurement of bacteria Different human Pathogenic bacteria (Listeria monocytogenes, Pseudomonas aureginosa, Salmonella typhi, Shigella dysenteriae and Staphylococcus aureus,) have been procured from IGMC, Shimla and Department of Biotechnology, HPU, Shimla for screening antibacterial properties of different plant extracts. The collected pathogens were revived fortnightly in nutrient broth and were stored at 4°C in refrigerator. Pure cultures of all the bacteria were maintained on nutrient agar medium slants and preserved in refrigerator at 4°C. Sub-culturing was done at regular intervals in order to maintain the cultures. Screening of prepared extracts for antibacterial activity Different extracts (methanol, acetone, chloroform and water) of plant were screened using Agar-well diffusion method. Nutrient agar medium (Beef extract 1 g, Yeast extract 2 g, Sodium Chloride 1 g, Peptone 5 g, Agar 20 g, Distilled Water 1,000 mL) was prepared and autoclaved at 121.6oC for 30 minutes. 20 mL of hot, liquid nutrient agar medium was poured into each of the Petri plates and let it to set for 20 minutes. Bacteria were grown in nutrient broth for 24 hours prior to screening experiment. Bacteria were spread on solid nutrient agar plates with the help of Spreader. Five wells of 8 mm diameter were bored with the help of sterilized
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