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Evaluation of the Antimicrobial Potential of Punica Granatum Leaves Hydro-methanolic Extract against Selected Pathogens

Hend Youssef, Abubakr M. El-Mahmoudy


For a long time, extracts of Punica granatumhave been used in alternative and complementary medicine. The object of the present study was to evaluate the activity of Punica granatumleaves hydro-methanolic extract against some Gram-positive, Gram-negative bacteria and Candida albicans using agar diffusion assay.The leaves yielded 34.7% of raw semisolid extract after maceration in hydro-methanol (50:50 v/v) andevaporation.After lyophilization, the dry powder was reconstituted in Muller-Hinton broth to get a concentration of 128 mg/mL; from which further 5lower concentrations have been prepared (64, 32, 16, 8, and 4 mg/mL in Muller-Hinton Broth).The extract exhibitedconcentration-dependent activity against some Gram-positive bacteria, namely,Listeria monocytogenes (20.67 ± 0.82), Clostridium perfringens (17.33 ± 0.52), Staphyllococcus aureus (10.5 ± 0.55), Bacillus cereus (8.83 ± 0.41), and Enterococcus fecalis (6.33 ± 0.52);and against three Gram-negative bacteria, namely, Shigellaflexneri (13.33 ± 0.8), Vibrio parahaemolyticus(12.17 ± 0.42) and Proteus vulgaris (8.5 ± 0.55);as well as against Candida albicans (15.8 ± 0.98); (the values are in mm after 128 mg/mL extract). However, the extract was without any visible activity againstEscherichia coli, Salmonella typhimuriumand Klebsiellaaerogenes.The minimum inhibitory concentrations (MICs) against susceptible organisms were1.06 mg/mL (Clostridium perfringens), 1.11 mg/mL (Vibrio parahaemolyticus), 2.08 mg/mL (Staphyllococcus aureus), 2.16 mg/mL (Bacillus cereus),2.76 mg/mL (Candida albicans), 3.07 mg/mL (Listeria monocytogenes), 3.80 (Shigella flexneri), 12.92 mg/mL (Proteus vulgaris) and 15.55 (Enterococcus faecalis).

These data may indicate that Punica granatumleaves extract is active against some pathogenic bacterial strains and thus may be useful in treatment of disease conditionscaused by these bacteria at least as a complementary medicine.


Antimicrobial; Punica granatum; Complementary medicine

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Iwu M, Duncan AR, Okunji CO. New antimicrobials of plant origin. Perspectives on new crops and new uses. ASHS Press, Alexandria, VA. 1999:457-462

Akinpelu D, Onakoya T. Antimicrobial activities of medicinal plants used in folklore remedies in south-western. African journal of Biotechnology. 2006, 5

Okoko T, Oruambo IF. The effect of hibiscus sabdariffa calyx extract on cisplatin-induced tissue damage in rats. Biokemistri. 2008, 20

Cos P, Vlietinck AJ, Berghe DV, Maes L. Anti-infective potential of natural products: How to develop a stronger in vitro ‘proof-of-concept’. Journal of ethnopharmacology. 2006, 106:290-302

Rios J, Recio M. Medicinal plants and antimicrobial activity. Journal of ethnopharmacology. 2005, 100:80-84

Bagri P, Ali M, Aeri V, Sultana S, Bhowmik M. Evalution of anti-inflammatory and analgesic activity of punica granatum linn. International Journal of Drug Development and Research. 2010, 2

Al-Megrin WA. Efficacy of pomegranate (punica granatum) peel extract against hymenolepis nana in infections mice. Biosciences Biotechnology Research Asia. 2016, 13:103-108

Gheith I, El-Mahmoudy A. Potent anti-oxidant and anti-inflammatory potentials of punica granatum leaf and flower hydromethanolic extracts in vitro. Bioscience Journal. 2017, 33:434-446

Chauhan I, Agrawal S, Goel RK. Status of inflammatory markers and growth factor in gastric ulcer protective effects of punica granatum l. Peel extract in rat. National Journal of Physiology, Pharmacy and Pharmacology. 2018, 8:12-17

Bassiri-Jahromi S. Punica granatum (pomegranate) activity in health promotion and cancer prevention. Oncology reviews. 2018, 12

Al-sallami AS, Al-Bideri AW, Alsaaedi SH. Hepatoprotective effect of pomegranate peel (punica granatum l) against thioacetamide-induced cirrhosis. THE EGYPTIAN JOURNAL OF EXPERIMENTAL BIOLOGY (Zoology). 2018, 14:41-47

Wang D, Özen C, Abu-Reidah IM, Chigurupati S, Patra JK, Horbanczuk JO, Jóźwik A, Tzvetkov NT, Uhrin P, Atanasov AG. Vasculoprotective effects of pomegranate (punica granatum l.). Frontiers in pharmacology. 2018, 9

Mestry SN, Gawali NB, Pai SA, Gursahani MS, Dhodi JB, Munshi R, Juvekar AR. Punica granatum improves renal function in gentamicin-induced nephropathy in rats via attenuation of oxidative stress. Journal of Ayurveda and integrative medicine. 2018,

Harborne J. Phytochemical methods, a guide to modern techniques of plant analysis, jb harborne. 1973,

Bauer A, Kirby W, Sherris JC, turck, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American journal of clinical pathology. 1966, 45:493

Bonev B, Hooper J, Parisot J. Principles of assessing bacterial susceptibility to antibiotics using the agar diffusion method. Journal of antimicrobial chemotherapy. 2008, 61:1295-1301

Friedman M. Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas. Molecular nutrition & food research. 2007, 51:116-134

Nostro A, Germano M, D’angelo V, Marino A, Cannatelli M. Extraction methods and bioautography for evaluation of medicinal plant antimicrobial activity. Letters in Applied Microbiology. 2000, 30:379-384

Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: General principles and contemporary practices. Clinical Infectious Diseases. 1998:973-980

Woods GL. Antibacterial susceptibility tests : Dilution and disk diffusion methods. Manual of clinical microbiology. 1995:1327-1341

Cockerill FR, Clinical, Institute LS. Performance standards for antimicrobial disk susceptibility testing: Approved standard. National Committee for Clinical Laboratory Standards; 2012.

Simor AE, Ofner-Agostini M, Bryce E, McGeer A, Paton S. Laboratory characterization of methicillin-resistant staphylococcus aureus in canadian hospitals: Results of 5 years of national surveillance, 1995–1999. Journal of Infectious Diseases. 2002, 186:652-660

Dahham SS, Ali MN, Tabassum H, Khan M. Studies on antibacterial and antifungal activity of pomegranate (punica granatum l.). Am.–Eurasian J. Agric. Environ. Sci. 2010, 9:273-281

Opara LU, Al-Ani MR, Al-Shuaibi YS. Physico-chemical properties, vitamin c content, and antimicrobial properties of pomegranate fruit (punica granatum l.). Food and Bioprocess Technology. 2009, 2:315-321

Al-Zoreky N. Antimicrobial activity of pomegranate (punica granatum l.) fruit peels. International journal of food microbiology. 2009, 134:244-248

Ryan K. Sherris medical microbiology. 4th ed. New York: McGraw Hill. 2004,

Naziri Z, Rajaian H, Firouzi R. Antibacterial effects of iranian native sour and sweet pomegranate (punica granatum) peel extracts against various pathogenic bacteria. Iranian Journal of Veterinary Research. 2012, 13:282-288

Pradeep B, Manojbabu M, Palaniswamy M. Antibacterial activity of punica granatum l. Against gastro intestinal tract infection causing organisms. Ethnobotanical leaflets. 2008, 2008:143

Nimri LF, Meqdam M, Alkofahi A. Antibacterial activity of jordanian medicinal plants. Pharmaceutical biology. 1999, 37:196-201

Choi J-G, Kang O-H, Lee Y-S, Chae H-S, Oh Y-C, Brice O-O, Kim M-S, Sohn D-H, Kim H-S, Park H. In vitro and in vivo antibacterial activity of punica granatum peel ethanol extract against salmonella. Evidence-Based Complementary and Alternative Medicine. 2011, 2011

Pai MB, Prashant G, Murlikrishna K, Shivakumar K, Chandu G. Antifungal efficacy of punica granatum, acacia nilotica, cuminum cyminum and foeniculum vulgare on candida albicans: An in vitro study. Indian Journal of Dental Research. 2010, 21:334

Abdollahzadeh S, Mashouf R, Mortazavi H, Moghaddam M, Roozbahani N, Vahedi M. Antibacterial and antifungal activities of punica granatum peel extracts against oral pathogens. Journal of Dentistry (Tehran, Iran). 2011, 8:1

Naz S, Siddiqi R, Ahmad S, Rasool S, Sayeed S. Antibacterial activity directed isolation of compounds from punica granatum. Journal of food science. 2007, 72


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