Evaluation of antibacterial and anthelmintic activities with total phenolic contents of Piper betel leaves

Document Type : Original Research Article


1 Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3814

2 Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3814, Bangladesh

3 Department of Pharmacy, Noakhali Science and technology University, Sonapur, Noakhali-3814

4 Department of Pharmacy, Noakhali Science and Technology university, Sonapur, Noakhali-3814

5 Department of Pharmacy, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh


Objective: The study was conducted to investigate the antibacterial and anthelmintic activities and to determine total phenolic contents of methanolic extract of Piper betel leaves.
Materials and Methods: The extract was subjected to assay for antibacterial activity using both gram positive and gram negative bacterial strains through disc diffusion method; anthelmintic activity with the determination of paralysis and death time using earthworm (Pheritima posthuma) at five different concentrations and the determination of total phenolic contents using the Folin-ciocalteau method.
Results: The extract showed significant (p<0.01) zone of inhibitions against gram positive Staphylococcus aureus [(6.77±0.25) mm] and Gram negative Escherichia coli [(8.53±0.25) mm], Salmonella typhi [(5.20±0.26) mm], Shigella dysenteriae [(11.20±0.26) mm] compared to positive control Azithromycin (ranging from 20.10±0.17 to 25.20±0.35 mm) while no zone inhibitory activity was found for both the extract and the standard drug against Gram positive Bacillus cereus. The extract also showed potent anthelmintic activity requiring less time for paralysis and death compared to the standard drug albendazole (10 mg/ml). At concentrations 10, 20, 40, 60 and 80 mg/ml, leaves extract showed paralysis at mean time of 9.83±0.60, 8.50±0.29, 6.60±0.17, 6.20±0.44 and 4.16±0.60; death at 11.33±0.88, 9.67±0.33, 7.83±0.17, 7.16±0.60 and 5.16±0.72 minutes, respectively. Whereas the standard drug showed paralysis and death at 19.33±0.71 and 51.00±0.23 minutes respectively. The extract confirmed the higher concentration of phenolic contents (124.42±0.14 mg of GAE /g of extract) when screened for total phenolic compounds.
Conclusion: Asresults confirmed potential antibacterial and anthelmintic activities of Piper betel leaves extract, therefore it may be processed for further drug research.


Main Subjects

Abo-State MAM, Helimish FA, Husseiny ShM, Zickry ARA. 2012. Reduction of Health Hazard of Bacillus Species Contaminating Solution Lenses and Baby Powder by Imipenem and Gamma Radiation. World Appl Sci J, 19: 856-866.
Adate PS, Parmesawaran DS, ChauhanY. 2012. In vitro Anthelmintic Activity of Stem Extracts of Piper betle Linn Against Pheritima posthuma. Pharmacog J, 4: 61-65.
Adhikari P, Chowdhury D, Baneerji J, Chatterjee A. 1998. Antifertility effect of crude alcoholic extract of Piper betel stalk. Ind Jour Phy Allied Sci, 52: 22‐27.
Akhtar MS, Iqbalb Z, Khanb MN, Lateef M. 2000. Anthelmintic activity of medicinal plants with particular reference to their use in animals in the Indo-Pakistan subcontinent. Small Rumin Res,38: 99-107.

Al-Adhroey AH, Nor ZM, Al-Mekhlafi HM, Amran AA, Mahmud R. 2011. Antimalarial activity of methanolic leaf extract of Piper betle L. Molecules, 16: 107-118.

Aruoma OI. 1996. Assessment of potential prooxidant and antioxidant actions. J Am Oil Chem Soc, 73: 1617-1625.
Balaji K, Lisa T, Sarnnia, Tan SK, Mirza B. 2011. Antibacterial Activity of Piper Betel Leaves. Int J Pharm Teach Pract, 2: 129-132.
Baravalia Y, Kaneria M, Vaghasiya Y Parekh J, Chanda S. 2009. Antioxidant and antibacterial activity of Diospyros ebenum Roxb leaf extracts. Turk J Biol, 33: 159-164.
Bate-Smith EC. 1962. The phenolic constituent of plants and their taxonomic significance, dicotylendons. J Linn Soc Bot, 58: 95-103.
Bernardi APM, López-Alarcón C, Aspée A, Rech SB, Von Poser GL, Bridi R, Dutrafilho CS, Lissi E. 2008. Antioxidant Activity in Southern Brazil Hypericum species. J Chil ChemSoc,53: 1658-1662.
Chanda S, Rakholiya K, Dholakia K, Baravalia Y. 2013. Antimicrobial, antioxidant, and synergistic properties of two nutraceutical plants: Terminalia catappa L. and Colocasia esculenta L. Turk J Biol, 37: 81-91.
Chatterjee KD. 1967. Parasitology: Protozoology and Helminthology. 6th Ed, In: Guha Ray Sree Saraswaty Press Ltd., Calcutta.
Eguale T, Tadesse D, Giday M. 2011. In vitro anthelmintic activity of crude extracts of five medicinal plants against egg-hatching and larval development of Haemonchus contortus. J Ethnopharmacol, 137: 108–113.
Elmastas M, Isildak O, Turkekul I, Temur N. 2007. Determination of antioxidant activity and antioxidant compounds in wild edible mushrooms. J Food Compos Anal, 20: 337-345.
Girish HV, Satish S. 2008. Antibacterial activity of important medicinal plants on human pathogenic bacteria – a comparative analysis. World Appl Sci J, 5: 267–271.
Hasanuzzaman M, Ali MR, Kuri S, Islam MS. 2013. Evaluation of total phenolic content, free radical scavenging activity and phytochemical screening of different extracts of Averrhoa bilimbi (fruits). Int Curr Pharm J, 2: 92-96.
Horax R, Hettiarachchy N, Islam S. 2005. Total Phenolic contents and phenolic acid constituents in 4 varieties of bitter melons (Momordica charantia) and antioxidant activities of their extracts. J Food Sci, 70: C275-C280.
Hulin V, Mathot AG, Mafart P, Dufosse L. 1998. Les proprietes anti-microbiennes des huiles essentielles et composes daromes. Sci Aliments, 18: 563-582.
Hurrell R. 2003. Influence of vegetable protein sources on trace element and mineral bioavailability. J Nutr, 133: 2973S–2977S.
Iqbal Z, Nadeem QK, Khan MN, Akhtar MS, Waraich FN. 2001. In vitro anthelmintic activity of Allium sativum, Zingiberofficinale, Cucurbitamexicana, Ficusreligiosa. Int J Agri Biol, 3: 454-457.
Jabbar A, Zaman MA, Iqbal Z, Yaseen M, Shamim A. 2007. Anthelmintic activity of Chenopodium album (L.) and Caesalpinia crista (L.) against trichostrongylid nematodes of sheep. J Ethnopharmacol,114: 86-91.
Jackson F, Coop RL. 2000. The development of anthelmintic resistance in sheep nematodes. Parasitol,120: S95-S107.
Jigna P, Sumitra C. 2006. In-vitro antimicrobial activities of extracts of Launaea procumbens Roxb. (Labiateae), Vitis vinifera L. (Vitaceae) and Cyperus rotundus L. (Cyperaceae). Afr J Biomed Res, 9: 89-93.
Joshi B, Sah GP, Basnet BB, Bhatt MR, Sharma D, Subedi K, Pandey J, Malla R. 2011. Phytochemical extraction and antimicrobial properties of different medicinal plants: Ocimum sanctum (Tulsi), Eugenia caryophyllata (Clove), Achyranthes bidentata (Datiwan) and Azadirachta indica (Neem). J Microbiol Antimicrob, 3: 1-7.
Kaufman BP, Carlson FT, Dayananda P. 1989. Plants: Their Biology and Importance. Harper and Row Publishers, New York.
Kumar A, Lakshman K, Jayaveera KN, Nandeesh R, Manoj B, Ranganayakulu D. 2010.  Comparative in vitro anthelmintic activity of three plants from the Amaranthaceae family. Archives Biol Sci, 62: 185-89.
Kumar GS, Jayaveera KN, Kumar CKA, Umachigi PS, Vrushabendra BMS, Kumar DVK. 2007. Antimicrobial effects of Indian medicinal plants against acne-inducing bacteria. Trop J Pharm Res, 6: 717-723.
Lei D. 2003. Antioxidant and antiplatelet effect of aqueous inflorescence Piper betel extract. J Agri Food Chem, 51: 2083‐2088.
Majumdar B, Chaudhuri SR, Roy A. 2002. Potent antiulcerogenic activity of ethanol extract of leaf of Piper betel Linn by antioxidative mechanism. Ind Jour Clin Bio Chem, 17: 49‐57.
Mali RG, Mehta AA. 2008. A review on anthelmintic plants. Nat Prod Radi, 7: 466-475.
Mali RG, Wadekar RR. 2008. In vitro Anthelmintic activity of Baliospermum Montanum Muell. Arg roots. Indian J Pharm Sci, 70:131-33.
Mohammed AA, Khalil AA, El-Beltagi HES. 2010. Antioxidant and antimicrobial properties of Kaff maryam (Anastatica hierochuntica) and doum palm (Hyphaene thebaica). Grases Y Aceites, 61: 67-75.
Mute VM, Keta A, Patel KS, Mirchandani D, Parth C. Babaria, et.al. 2009. Anthelmintic effect of Tamarind indica Linn leaves juice exract on pheretima posthuma. Int Journal of Pharma Res  Dev, 7: 01-06.
Nair R, Sumitra C. 2008. Antimicrobial activity of Terminalia catappa, Manikara zapota and Piper betel leaf extract. Indian J Pharm Sci, 70: 390-393.
National Committee for Clinical Laboratory Standards. 1997. Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard. Document M27-A. NCCLS, Wayne, PA, USA. 
Obeidat M, Shatnawi M, Al-alawi M, Al-Zu`bi E, Al-Dmoor H, Al-Qudah M, et al. 2012. Antimicrobial activity of crude extracts of some plant leaves. Res J Microbiol, 7: 59-67.
Opara FN, Anuforo HU, Okechuk Wu RI, Mgbemena IC, Akujobi CO, Adjero A. 2012. Preliminary phytochemical screening and antibacterial activities of leaf extracts of Terminalia catappa. J Emerg Trends Eng Appl Sci, 3: 424-428.
Prashant KR, Dolly J, Singh KR, Gupta KR, Watal G. 2008. Glycemic properties of Trichosanthes dioica leaves. Pharm Biol, 46: 894-899.
Rafi KP, Karthikeyan M, Kannan M, Rajasekar S. 2011. Anthelmintic activity of Nerium olender flower extract in Indian adult earthworm.J Nat Prod Plant Res, 1: 40-46.
Rahman MA, Islam MS. 2013. Antioxidant, antibacterial and cytotoxic effects of the phytochemicals of whole Leucas aspera extract. Asian Pac J Trop Biomed, 3: 273-279.
Raju GS, Moghal MMR, Dewan SMR, Amin MN, Billah MM. 2013. Characterization of phytoconstituents and evaluation of total phenolic content, anthelmintic, and antimicrobial activities of Solanum violaceum Ortega. Avicenna J Phytomed, 3: 313-320.
Ramzi AAM, Salah AAA, Hasson S, Althawab FMN, Alaghbari SAZ, Lindequist U. 2010. Antimicrobial, antioxidant and cytotoxic activities and phytochemical screening of some yemeni medicinal plants. eCAM, 7: 323-330.
Rosy BA, Joseph H, Rosalie. 2010. Phytochemical, pharmacognostical, antimicrobial activity of Indigofera spalathoids Vahl. (Fabaceae). Int J Biol Technol, 1: 12–15.
Salhan M, Kumar B, Tiwari P, Sharma P, Sandhar HK, Gautam M. 2011. Comparative anthelmintic activity of aqueous and ethanolic leaf extracts Of Clitoria ternatea.Int J Drug Dev Res 3: 62-69.
Sangeetha J, Soundarya K, Santhoshm K, Sindhura C. 2010. Evaluation of In-vitro Anthelmentic Property of Passiflora edulis Linn. Res J Pharm Biol Chem Sci, 1: 715-718.
Santhakumari P, Prakasam A, Puglendi KV. 2003. Modulation of oxidative stress parameters by treatment with Piper betel leaf in streptozotacin induced diabetic rats. Ind J of Pharmacol, 35: 373‐378.
Sawa T, Akaike T, Maeda H. 2000.Tyrosine nitration by peroxynitrite formed from nitric oxide and superoxide generated by xanthine oxidase. J Biol Chem, 275: 32467-32474.
Sule WF, Okonko IO, Omo-Ogun S, Nwanze JC, Ojezele MO, Ojezele OJ, Alli JA, Soyemi ET, Olaonipekun TO. 2011. Phytochemical properties and in vitro antifungal activity of Senna alata Linn. crude stem bark extract. J Med Plants Res, 5: 176–183.
Sundang M, Nasir SNS, SIpaut CS, Othman H. 2012. Antioxidant activity, phenolic, flavonoid and tannin content of Piper betel and Leukosyke capitella. Malaysian J Fund App Sci, 8: 1-6.
Walsh C. 2000. Molecular mechanisms that confer bacteria drug resistance. Nature, 406: 235-289.