Antibacterial activity of pomegranate extracts against different bacteria.
Abstract
The search for plant extracts with efficacious antimicrobial activity remains important, partly due to fears of the side effects associated with conventional antibiotics and to counter the emergence of resistant microorganisms. Pomegranate extracts have been used for millennia for their anti-infective properties, with activity more recently being attributed to its rich composition of ellagitannins and other secondary polyphenolic compounds. This chapter highlights the growing number of publications that have probed the activity of pomegranate extracts against microbes. Research generally supports folklore claims and has shown that pomegranate extracts possess unusual and potent broad-spectrum activities against Gram-positive and Gram-negative bacteria (planktonic and biofilm), fungi, viruses and parasites. Possible pathways/mechanisms of antimicrobial activity of pomegranate extracts are discussed and enhancement/potentiation of such activity using metal ions considered.
Keywords
- antimicrobial
- bacteria
- fungi
- viruses
- parasites
- polyphenols
- pomegranate extracts
- biofilm
- tannins
- punicalagin
1. Introduction
Infectious diseases caused by pathogenic microbes are a fundamental problem and remain one of the major factors behind high morbidity and mortality across the world, especially in developing countries. This is exacerbated by the world-wide emergence of antibiotic-resistant pathogens which has in turn given increased urgency to the discovery of new antimicrobial compounds, including those derived from plants [1, 2].
The pomegranate, fruit of the
The phytochemistry of pomegranate extracts is well described in the literature [10, 11, 12] and they are known to be rich in bioactive compounds especially polyphenolics including anthocyanins and ellagitannins, in particular punicalagin, which is in the highest proportion [13]. As will be seen, it has become apparent that the pomegranate possesses unusual broad-spectrum potency against a wide range of species, which generally correlates with its polyphenol concentration.
In this chapter we aim to summarise published research into pomegranate extracts as antimicrobials and discuss some of the purported mechanisms behind such activity. Finally, the enhancement of antimicrobial activity by co-administration with metal ions is considered.
2. Activity against bacteria
Pomegranate extracts have shown antimicrobial activity against to a range of oral microbes. It has been found that pomegranate extract powder at 1 mg/mL was effective against primary and secondary colonizer bacteria of dental plaque:
Moreover, ‘standardized’ pomegranate peel extract showed higher antimicrobial activity than other parts of pomegranate (flower, leaf, stem) and ciprofloxacin (2 mg/mL) against
Due to its antimicrobial and antioxidant properties, pomegranate extract has been studied for its preservation potential use in the food industry. Kannat
The activity of pomegranate extract against bacteria is summarized in Table 1.
Part of pomegranate used | Form | Test organisms | MICs | Reference |
---|---|---|---|---|
PomElla® (30% punicalagin) | 0.8 μg/mL 0.2 μg/mL 0.2 μg/mL 0.8 μg/mL | [16] | ||
Peel | Acetonic, Methanolic, Ethanolic PPE and Hydro-alcoholic PPE | 0.0125–0.025 mg/mL 12.5–25 mg/mL 0.0125–0.025 mg/mL 0.05–0.4 mg/mL 12.5–25 mg/mL 3.15–100 mg/mL 10 mg/mL 10 mg/mL | [17, 18] | |
Peel, flower, leaf, stem | Aqueous and methanolic | [19] | ||
Peel | Basic gel formulation including 540 mg pomegranate peel powder | 1:16 1:128 1:16 1:64 | [20] | |
Pomegranate mouthwash (Pomegranate extract were obtained from Verdure science, 30% punicalagin) | 62.5 mg/mL >31.25 mg/mL 16.125 mg/mL | [21] | ||
Peel | Methanolic PPE | 250 μg/mL 250 μg/mL 250 μg/mL 1000 μg/mL | [15] | |
Fruit pericarp | Methanolic | 640–2560 μg/mL 1280–2560 μg/mL 640–2560 μg/mL | [9] | |
Peel | Standardized extract (13% ellagic acid) | 15.6 μg/mL 7.81 μg/mL 7.81–15.6 μg/mL 7.81 μg/mL | [14] | |
Peel | Water | 200–450 ppm | [24] | |
Peel | Acetonic, methanolic, water | 200–400 ppm 150–500 ppm 200–450 ppm 200–700 ppm 200–400 ppm | [25] | |
Fruit | Water | 100 mg/mL 100 mg/mL 100 mg/mL | [27] | |
Peel | Ethanolic, methanolic | [28] |
3. Activity against fungi
Treatment of fungal infections is a big challenge because of the eukaryotic nature of fungal cells that have similarity with host cells. While there are some drugs in the treatment of fungal infections available in the clinic, they are limited and there is a need for new alternatives [29, 30]. There are reports showing antifungal activity of pomegranate extracts, especially against
The potential of pomegranate extract has been studied against fungi in
In addition to
The activity of pomegranate extract against fungal microbes is summarized in Table 2.
Part of pomegranate used | Form | Test organisms | MICs | Reference |
---|---|---|---|---|
Peel | Crude extract, Aqueous fraction, ethyl acetate fraction, butanol fraction, punicalagin | 3.9–7.8 μg/mL, 1.9–15.6 μg/mL | [32] | |
Peel | Methanolic | > 1000 μg/mL | [15] | |
Peel | Crude extract, Crude extract in a spray-dried microparticle formulation | 3.9–15.6 μg/mL | [38] | |
Peel | Hydroalcoholic | 125 g/mL, 125 g/mL, 250 g/mL, 250 g/mL | [40] | |
Peel | Aqueous | [42] | ||
Peel | Aqueous | [43] |
4. Activity against viruses
Pomegranate extracts have been examined as an alternative treatment for viral infections [46, 47, 48]. A number of studies have shown that polyphenolic compounds have broad-spectrum antiviral activity, by inhibiting viral DNA and RNA, and directly binding the viral particles. It has also been suggested that polyphenols could provide antiviral activity during intracellular replication [49, 50, 51, 52].
Pomegranate peel extract showed antiviral activity against the influenza virus. In a study by Sundararajan
Studies have suggested that the antiviral activity of pomegranate extract originates from its hydrolysable tannins and polyphenols, especially punicalagin and gallagic acid. However, in one study, four flavonoids, ellagic acid, caffeic acid, luteolin and punicalagin, from pomegranate peel extract were studied against influenza virus and only punicalagin showed an inhibitory effect. The antiviral activity of pomegranate rind extract has been patented in Japan based on pomegranate peel extract ability to prevent the growth and kill viruses on the surfaces [46, 47]. The activity of pomegranate extract against viruses is summarized in Table 3.
Part of pomegranate used | Form | Test organisms | Mechanism of virus target | Reference |
---|---|---|---|---|
Rind | Crude hydraulic extract, Punicalagin, Ellagic acid | Herpes simplex virus | Virucidal activity | [48] |
Juice, peel and pomegranate liquid extract | Influenze A viruses, H1N1, H3N2, H5N1 and coronavirus MHV A59 | Damage to virion integrity and virucidal activity | [53] | |
Juice | Pomegranate polyphenol extract, punicalagin, pomegranate liquid extract (from POM Wonderful) | Human influenza A (H3N2) | Inhibition of viral RNA replication | [54] |
Peel | Methanolic crude extract, punicalin, punicalagin, ellagic acid | Hepatitis C virus | [57] | |
Peel | Methanolic crude extract, ellagic acid, punicalagin, gallic acid | Adenovirus | Inhibition of adenovirus replication | [58] |
5. Activity against parasites
Parasitic infections remain a significant global problem, affecting the health of hundreds of millions of people annually, especially in countries with low economic and social conditions. In addition, the increased world-wide resistance to conventional drugs is making most of currently used drugs less effective. As a result of this situation, the development of new drugs from medicinal plants for parasites is as important as for other microbes [62]. Different parts of
Al-Musayeib
Part of pomegranate used | Form | Mechanism of organism target | Organisms | Reference |
---|---|---|---|---|
Seed | Methanolic | Reduction in gastrointestinal motility | [65] | |
Leave | Methanolic | Larvicidal and ovicidal activity | [66] | |
Peels, juice and leaves | Methanolic | Reduction in viability of parasite | [67] | |
Leaf, stem bark | Ethanolic | worms separation, reduction of motor activity, lethality, and ultrastructural tegumental alterations | [68] | |
Peel | Powder form directly given to animal | Growth inhibition and death | [72] | |
Juice | Crude extract was applied by patients in a clinical study | [73] |
6. Potential mechanisms of antimicrobial activity of pomegranate extracts
From the preceding sections it is clear that there is compelling evidence demonstrating the broad-spectrum antimicrobial activity of pomegranate extracts [74, 75, 76]. However, the precise mechanism behind this activity is not fully understood. The mode of antimicrobial action of polyphenols, in general, is also unknown, although some suggested mechanisms include membrane disruption, toxicity against microorganisms, the ability of complex formation with metal ions and enzyme inactivation [77, 78, 79]. The antimicrobial activity of pomegranate has been associated with polyphenolic tannins, especially punicalagin and ellagic acid content in the extract [80, 81, 82]. However, pomegranate extracts are a complex mixture containing a variety of secondary compounds and interplay between these components may be a factor in antimicrobial activity, with multiple mechanisms operating independently [83].
An antimicrobial mechanism suggested for polyphenolic compounds is based on the precipitation ability of these compounds with bacterial cell membrane proteins which leads to bacterial cell lysis [84]. In addition, polyphenols could inhibit microbial enzymes by reacting with sulfhydryl groups or nonspecific interactions with proteins [85]. In that vein, phenolic compounds can bind the protein sulfhydryl groups and make them unavailable for microbial growth [86]. In addition, it has been reported that polyphenols can damage the microbe respiratory chain by decreasing the oxygen consumption and thus limiting the oxidation of NADH [87].
It has been hypothesized that the antibacterial activity of phenolic acids and flavonoids could cause a decrease in membrane fluidity by giving damage to the bacterial cytoplasmic membrane [88]. Phenolic acids can cause hyper acidification when they interphase with the plasma membrane. This situation would cause an alteration in cell membrane by making it more permeable. This mechanism could explain why phenolic acids show different antimicrobial activity levels against different pathogenic microorganisms [89, 90]. One of the possible mechanisms could be related to hydroxyl groups of polyphenols. The position of OH group in the aromatic ring and the length of saturated side chain could be a cause of antimicrobial activity of polyphenols [91]. Hydroxyl groups can bind to bacteria cell membranes and interfere with processes, such as ion pumping. In addition, OH groups can interact with active site of enzymes and disturb the metabolism of microorganisms [91].
Pomegranate extract exerted an inhibition activity against biofilms, in addition to their planktonic counterparts. Since microbes act differently under biofilm conditions compared to their planktonic form, there are some suggested pathways about polyphenols biofilm eradication and formation inhibition activities, although still unconfirmed. The mechanism behind growth and biofilm inhibition by pomegranate extracts cause protein precipitation and enzyme inactivation [81, 92]. Pomegranate extract could precipitate proteins which play major role in biofilm formation, like adhesins. Moreover, major hydrolysable tannins in pomegranate extract such as ellagic acid can change the surface charge and reduce the cell-substratum interactions and biofilm formation and development on different surfaces [93]. It is well known that tannins have astringency properties, and this feature can play a part in biofilm disruption [94, 95]. Different studies have shown the activity of pomegranate on bacterial attachment and therefore biofilm formation. It has been demonstrated that
There are some reports suggesting that the inhibition of quorum sensing (QS) could play role in the biofilm inhibition activity of pomegranate [99, 100]. QS is a communication system between bacteria in a biofilm, and provides a network involving nutrients, defense against other microorganisms, virulence and biofilm formation. More importantly, QS helps microbes to escape from host immune response [101, 102]. Therefore, inhibition of QS is quite important in order to overcome microbial infectious diseases and resistant pathogenic microbes. For the evaluation of QS inhibitors,
The chemical structure of tannins has importance in bacterial growth inhibition. For example, hydrolysable tannins were found to give lower minimum inhibitory concentration than condensed tannins [109]. The degree of galloylation has an effect on antibacterial activity since a higher degree of galloylation have more protein binding capacity and higher affinity for iron. However, the antibacterial activity is not only correlated to galloyl groups and galloylation, also it is correlated to configuration of the digalloyl or trigalloyl groups that attached to glucose core [110, 111, 112]. In addition, free galloyl groups have a major role in antimicrobial activity of ellagitannins which are abundant secondary compounds in pomegranate extracts [12, 113]. Punicalagin showed the broad-spectrum antimicrobial activity and it has a gallagyl moiety [114]. However another ellgitannin, granatin A, which does not have free galloyl groups, did not show antibacterial activity [115]. In a study done by Reddy
The antimicrobial activity of plants has been studied extensively and many active secondary compounds have been identified. However, it should not be ignored that plant extracts with antimicrobial activities contain potentially many secondary compounds. Therefore, it is not easy to attribute the biological activity of plant extracts to only a single compound or a group of secondary compounds. There is a high possibility that plant extracts show antimicrobial activity due to synergistic effect of different compounds [117].
7. Enhanced antimicrobial activity of pomegranate extracts with metal ions
There are many reports showing the antimicrobial activity of heavy metals such as iron, copper, silver, manganese and zinc, while many bacteria have mechanism for the detoxification of heavy metals [118, 119]. However, although metal ions have a strong antimicrobial effect, they can also be cytotoxic to human cells, therefore, the use of these metals may have limitations in healthcare [120, 121].
Stewart
McCarell
Significantly enhanced virucidal activity of PRE was later observed against HSV-1, HSV-2 and acyclovir-resistant HSV-1 by Houston
The mechanism for the synergistic antimicrobial activity of pomegranate extract in combination with metal ions is not clear, although there are some putative suggested mechanisms for this enhanced antimicrobial activity. For instance, it has been suggested that pomegranate tannins can form a ‘complex’ with metallic ions and this complex could show enhanced toxicity to microbes [127]. Furthermore, PRE could show enhanced activity due to redox cycling of the associated metal ion which increases local levels of reactive oxygen species (ROS). For example some antibiotics e.g. bleomycin showed enhanced ROS production via the ability to bind to ferrous ions which resulted in enhanced toxicity against microbes [128].
The enhancement of antimicrobial activity of pomegranate rind extract with metal ions is important in terms of improved efficacy against antibiotic resistant pathogens, since this enhancement could reduce resistance of microbes by inhibiting their microbial adaptability features [8, 32].
8. Conclusions
The pomegranate has a long history of use as a folklore medicine for its ability to address microbial infections. Published research, as outlined in this chapter, clearly supports this and has shown that pomegranate extracts possess an unusual and potent broad-spectrum of activities against bacteria, fungi, viruses and parasites.
There is some variation in the literature in terms of the levels of antimicrobial activity of pomegranate extracts, which could be attributed to different harvesting time and type of pomegranate cultivars, and use of varying microbial strains. However, it is also apparent that different workers have used a range of approaches to obtain ‘pomegranate extract’, with extraction methods sometimes being poorly described. As such, a lack of standardized test extracts presents a challenge in attempting to make quantitative comparisons. As a complex mixture, pomegranates extracts have the innate ability to inhibit resistance, even more so when used alongside a synergizing metal ion.
Acknowledgments
We would like to thank to Turkish Ministry of Education for supporting Vildan Celiksoy’s PhD project.
References
- 1.
Bereket W, Hemalatha K, Getenet B, Wondwossen T, Solomon A, Zeynudin A, Kannan S. Update on bacterial nosocomial infections. Eur Rev Med Pharmacol Sci. 2012 Aug 1;16(8):1039-1044. - 2.
Savard P, Perl TM. A call for action: managing the emergence of multidrug-resistant Enterobacteriaceae in the acute care settings. Current opinion in infectious diseases. 2012 Aug 1;25(4):371-7. - 3.
Seeram NP, Adams LS, Henning SM, Niu Y, Zhang Y, Nair MG, Heber D. In vitro antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegranate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice. The Journal of nutritional biochemistry. 2005 Jun 1;16(6):360-7. - 4.
Lansky EP, Newman RA. Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. Journal of ethnopharmacology. 2007 Jan 19;109(2):177-206. - 5.
Braga LC, Shupp JW, Cummings C, Jett M, Takahashi JA, Carmo LS, Chartone-Souza E, Nascimento AM. Pomegranate extract inhibits Staphylococcus aureus growth and subsequent enterotoxin production. Journal of ethnopharmacology. 2005 Jan 4;96(1-2):335-9. - 6.
Ahmad I, Beg AZ. Antimicrobial and phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. Journal of ethnopharmacology. 2001 Feb 1;74(2):113-23. - 7.
Voravuthikunchai SP, Sririrak T, Limsuwan S, Supawita T, Iida T, Honda T. Inhibitory effects of active compounds from Punica granatum pericarp on verocytotoxin production by enterohemorrhagic Escherichia coli O157: H7. Journal of health science. 2005;51(5):590-6. - 8.
Chidambara Murthy KN, Reddy VK, Veigas JM, Murthy UD. Study on wound healing activity of Punica granatum peel. Journal of Medicinal Food. 2004 Jun 1;7(2):256-9. - 9.
Dey D, Debnath S, Hazra S, Ghosh S, Ray R, Hazra B. Pomegranate pericarp extract enhances the antibacterial activity of ciprofloxacin against extended-spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) producing Gram-negative bacilli. Food and Chemical Toxicology. 2012 Dec 1;50(12):4302-9. - 10.
Garcia-Villalba R, Espín JC, Aaby K, Alasalvar C, Heinonen M, Jacobs G, Voorspoels S, Koivumaki T, Kroon PA, Pelvan E, Saha S. Validated method for the characterization and quantification of extractable and nonextractable ellagitannins after acid hydrolysis in pomegranate fruits, juices, and extracts. Journal of agricultural and food chemistry. 2015 Jul 29;63(29):6555-66. - 11.
Saad H, Charrier-El Bouhtoury F, Pizzi A, Rode K, Charrier B, Ayed N. Characterization of pomegranate peels tannin extractives. Industrial crops and Products. 2012 Nov 1; 40:239-46. - 12.
Liu Y, Seeram NP. Liquid chromatography coupled with time-of-flight tandem mass spectrometry for comprehensive phenolic characterization of pomegranate fruit and flower extracts used as ingredients in botanical dietary supplements. Journal of separation science. 2018 Aug;41(15):3022-33. - 13.
Zaouay F, Mena P, Garcia-Viguera C, Mars M. Antioxidant activity and physico-chemical properties of Tunisian grown pomegranate ( Punica granatum L.) cultivars. Industrial Crops and Products. 2012 Nov 1; 40:81-9. - 14.
Panichayupakaranant P, Tewtrakul S, Yuenyongsawad S. Antibacterial, anti-inflammatory and anti-allergic activities of standardised pomegranate rind extract. Food Chemistry. 2010 Nov 15;123(2):400-3. - 15.
Bakkiyaraj D, Nandhini JR, Malathy B, Pandian SK. The anti-biofilm potential of pomegranate ( Punica granatum L.) extract against human bacterial and fungal pathogens. Biofouling. 2013 Sep 1;29(8):929-37. - 16.
Avadhani M, Kukkamalla MA, Bhatt KG. Screening of Punica granatum extract for antimicrobial activity against oral microorganisms. Journal of Ayurvedic and Herbal Medicine. 2020;6(2):73-7. - 17.
Benslimane S, Rebai O, Djibaoui R, Arabi A. Pomegranate Peel Extract Activities as Antioxidant and Antibiofilm against Bacteria Isolated from Caries and Supragingival Plaque. Jordan Journal of Biological Sciences. 2020 Jul 1;13(3). - 18.
Sateriale D, Facchiano S, Colicchio R, Pagliuca C, Varricchio E, Paolucci M, Volpe MG, Salvatore P, Pagliarulo C. In vitro Synergy of Polyphenolic Extracts from Honey, Myrtle and Pomegranate Against Oral Pathogens, S. mutans and R. dentocariosa. Frontiers in Microbiology. 2020 Jul 24; 11:1465 - 19.
Rummun N, Somanah J, Ramsaha S, Bahorun T, Neergheen-Bhujun VS. Bioactivity of nonedible parts of Punica granatum L.: a potential source of functional ingredients. International journal of food science. 2013 Jul 8;2013. - 20.
Vasconcelos LC, Sampaio FC, Sampaio MC, Pereira MD, Higino JS, Peixoto MH. Minimum inhibitory concentration of adherence of Punica granatum Linn (pomegranate) gel against S. mutans, S. mitis and C. albicans. Brazilian Dental Journal. 2006;17(3):223-7. - 21.
Bhadbhade SJ, Acharya AB, Rodrigues SV, Thakur SL. The antiplaque efficacy of pomegranate mouthrinse. Quintessence International. 2011 Jan 1;42(1). - 22.
DiSilvestro RA, DiSilvestro DJ, DiSilvestro DJ. Pomegranate extract mouth rinsing effects on saliva measures relevant to gingivitis risk. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2009 Aug;23(8):1123-7. - 23.
Martins ML, Ribeiro-Lages MB, Masterson D, Magno MB, Cavalcanti YW, Maia LC, Fonseca-Gonçalves A. Efficacy of natural antimicrobials derived from phenolic compounds in the control of biofilm in children and adolescents compared to synthetic antimicrobials: A systematic review and meta-analysis. Archives of Oral Biology. 2020 Jul 21:104844. - 24.
Kanatt SR, Chander R, Sharma A. Antioxidant and antimicrobial activity of pomegranate peel extract improves the shelf life of chicken products. International journal of food science & technology. 2010 Feb 1;45(2):216-22. - 25.
Negi PS, Jayaprakasha GK. Antioxidant, and antibacterial activities of Punica granatum peel extracts. Journal of food science. 2003 May;68(4):1473-7. - 26.
Oliveira I, Sousa A, Morais JS, Ferreira IC, Bento A, Estevinho L, Pereira JA. Chemical composition, and antioxidant and antimicrobial activities of three hazelnut ( Corylus avellana L.) cultivars. Food and Chemical Toxicology. 2008 May 1;46(5):1801-7. - 27.
Alpaslan D, Dudu TE, Şahiner N, Aktaşa N. Synthesis and preparation of responsive poly (Dimethyl acrylamide/gelatin and pomegranate extract) as a novel food packaging material. Materials Science and Engineering: C. 2020 Mar 1; 108:110339. - 28.
Cui H, Surendhiran D, Li C, Lin L. Biodegradable zein active film containing chitosan nanoparticle encapsulated with pomegranate peel extract for food packaging. Food Packaging and Shelf Life. 2020 Jun 1; 24:100511. - 29.
Loureiro MM, De Moraes BA, Mendonça VL, Quadra MR, Pinheiro GS, Asensi MD. Pseudomonas aeruginosa: study of antibiotic resistance and molecular typing in hospital infection cases in a neonatal intensive care unit from Rio de Janeiro City, Brazil. Memórias do Instituto Oswaldo Cruz. 2002 Apr;97(3):387-94. - 30.
Morschhäuser J. The genetic basis of fluconazole resistance development in Candida albicans . Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease. 2002 Jul 18;1587(2-3):240-8. - 31.
César de Souza Vasconcelos L, Sampaio MC, Sampaio FC, Higino JS. Use of Punica granatum as an antifungal agent against candidosis associated with denture stomatitis. Mycoses. 2003 Jun;46(5-6):192-6. - 32.
Endo EH, Cortez DA, Ueda-Nakamura T, Nakamura CV, Dias Filho BP. Potent antifungal activity of extracts and pure compound isolated from pomegranate peels and synergism with fluconazole against Candida albicans . Research in Microbiology. 2010 Sep 1;161(7):534-40. - 33.
Rizwan M, Mujtaba G, Memon SA, Lee K, Rashid N. Exploring the potential of microalgae for new biotechnology applications and beyond: a review. Renewable and Sustainable Energy Reviews. 2018 Sep 1; 92:394-404. - 34.
Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science. 1999 May 21;284(5418):1318-22. - 35.
Komiyama EY, Mello de Matos B, Eduardo de Oliveira F, de Souza Reis T, Maynart de Faro H, Balducci I, Almeida JD, Koga-Ito CY. Proposal of Using Ozonated Water to Control Biofilm Formation on Mouth-Related Devices. Ozone: science & engineering. 2011 Sep 1;33(5):417-21. - 36.
Hawser SP, Douglas LJ. Resistance of Candida albicans biofilms to antifungal agents in vitro. Antimicrobial agents and chemotherapy. 1995 Sep 1;39(9):2128-31. - 37.
Sandasi M, Leonard CM, Van Vuuren SF, Viljoen AM. Peppermint (Mentha piperita) inhibits microbial biofilms in vitro. South African Journal of Botany. 2011 Jan 1;77(1):80-5. - 38.
Endo EH, Ueda-Nakamura T, Nakamura CV. Activity of spray-dried microparticles containing pomegranate peel extract against Candida albicans . Molecules. 2012 Sep;17(9):10094-107. - 39.
Heber D, Schulman RN, Seeram NP, editors. Pomegranates: ancient roots to modern medicine. CRC press; 2006 Jul 7 - 40.
Foss SR, Nakamura CV, Ueda-Nakamura T, Cortez DA, Endo EH, Dias Filho BP. Antifungal activity of pomegranate peel extract and isolated compound punicalagin against dermatophytes. Annals of clinical microbiology and antimicrobials. 2014 Dec 1;13(1):32. - 41.
Salahvarzi Y, Tehranifar A, Jahanbakhsh V. Relation of antioxidant and antifungal activity of different parts of pomegranate ( Punica granatum L.) extracts with its phenolic content. Iranian Journal of Medicinal and Aromatic Plants. 2011;27(1). - 42.
Ullah N, Ali J, Khan FA, Khurram M, Hussain A, Rahman IU, Rahman ZU, Ullah S. Proximate composition, minerals content, antibacterial and antifungal activity evaluation of pomegranate ( Punica granatum L.) peels powder. Middle East J Sci Res. 2012;11(3):396-401. - 43.
Glazer I, Masaphy S, Marciano P, Bar-Ilan I, Holland D, Kerem Z, Amir R. Partial identification of antifungal compounds from Punica granatum peel extracts. Journal of agricultural and food chemistry. 2012 May 16;60(19):4841-8. - 44.
Negi PS. Plant extracts for the control of bacterial growth: Efficacy, stability and safety issues for food application. International journal of food microbiology. 2012 May 1;156(1):7-17. - 45.
Viuda-Martos M, Ruiz Navajas Y, Sánchez Zapata E, Fernández-López J, Pérez-Álvarez JA. Antioxidant activity of essential oils of five spice plants widely used in a Mediterranean diet. Flavour and Fragrance Journal. 2010 Jan;25(1):13-9. - 46.
Jassim SA, Denyer SP, Stewart GS, inventors; Merck Patent GmbH, assignee. Antiviral or antifungal composition comprising an extract of pomegranate rind or other plants and method of use. United States patent US 5,840,308. 1998 Nov 24. - 47.
Jassim SA, Naji MA. Novel antiviral agents: a medicinal plant perspective. Journal of applied microbiology. 2003 Sep;95(3):412-27. - 48.
Houston DM, Bugert JJ, Denyer SP, Heard CM. Correction: Potentiated virucidal activity of pomegranate rind extract (PRE) and punicalagin against Herpes simplex virus (HSV) when co-administered with zinc (II) ions, and antiviral activity of PRE against HSV and aciclovir-resistant HSV. Plos one. 2017 Nov 20;12(11): e0188609. - 49.
Sawai-Kuroda R, Kikuchi S, Shimizu YK, Sasaki Y, Kuroda K, Tanaka T, Yamamoto T, Sakurai K, Shimizu K. A polyphenol-rich extract from Chaenomeles sinensis (Chinese quince) inhibits influenza A virus infection by preventing primary transcription in vitro. Journal of ethnopharmacology. 2013 Apr 19;146(3):866-72. - 50.
Das S, Tanwar J, Hameed S, Fatima Z, Manesar G. Antimicrobial potential of epigallocatechin-3-gallate (EGCG): a green tea polyphenol. J Biochem Pharmacol Res. 2014 Sep;2(3):167-74. - 51.
Song JM, Lee KH, Seong BL. Antiviral effect of catechins in green tea on influenza virus. Antiviral research. 2005 Nov 1;68(2):66-74. - 52.
Kamboj A, Saluja AK, Kumar M, Atri P. Antiviral activity of plant polyphenols. J Pharm Res. 2012 May;5(5):2402-12. - 53.
Sundararajan A, Ganapathy R, Huan L, Dunlap JR, Webby RJ, Kotwal GJ, Sangster MY. Influenza virus variation in susceptibility to inactivation by pomegranate polyphenols is determined by envelope glycoproteins. Antiviral research. 2010 Oct 1;88(1):1-9. - 54.
Haidari M, Ali M, Casscells III SW, Madjid M. Pomegranate ( Punica granatum ) purified polyphenol extract inhibits influenza virus and has a synergistic effect with oseltamivir. Phytomedicine. 2009 Dec 1;16(12):1127-36. - 55.
Droebner K, Ehrhardt C, Poetter A, Ludwig S, Planz O. CYSTUS052, a polyphenol-rich plant extract, exerts anti-influenza virus activity in mice. Antiviral research. 2007 Oct 1;76(1):1-0. - 56.
Murzakhmetova M, Moldakarimov S, Tancheva L, Abarova S, Serkedjieva J. Antioxidant and prooxidant properties of a polyphenol-rich extract from Geranium sanguineum L. in vitro and in vivo. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2008 Jun;22(6):746-51. - 57.
Reddy BU, Mullick R, Kumar A, Sudha G, Srinivasan N, Das S. Small molecule inhibitors of HCV replication from pomegranate. Scientific reports. 2014 Jun 24; 4:5411. - 58.
Karimi A, Moradi MT, Rabiei M, Alidadi S. In vitro anti-adenoviral activities of ethanol extract, fractions, and main phenolic compounds of pomegranate ( Punica granatum L.) peel. Antiviral Chemistry and Chemotherapy. 2020 Apr; 28:2040206620916571. - 59.
Whitley RJ, Nahmias AJ, Visintine AM, Fleming CL, Alford CA, Yeager A, Arvin A, Haynes R, Hilty M, Luby J. The natural history of herpes simplex virus infection of mother and newborn. Pediatrics. 1980 Oct 1;66(4):489-94. - 60.
Whitley RJ, Roizman B. Herpes simplex virus infections. The lancet. 2001 May 12;357(9267):1513-8. - 61.
Piret J, Boivin G. Resistance of herpes simplex viruses to nucleoside analogues: mechanisms, prevalence, and management. Antimicrobial agents and chemotherapy. 2011 Feb 1;55(2):459-72. - 62.
Tagboto S, Townson S. Antiparasitic properties of medicinal plants and other naturally occurring products. - 63.
Prakash CV, Prakash I. Bioactive chemical constituents from pomegranate ( Punica granatum ) juice, seed and peel-a review. International Journal of Research in Chemistry and Environment. 2011 Jul;1(1):1-8. - 64.
Asres K, Bucar F, Knauder E, Yardley V, Kendrick H, Croft SL. In vitro antiprotozoal activity of extract and compounds from the stem bark of Combretum molle. Phytotherapy Research. 2001 Nov;15(7):613-7. - 65.
Das AK, Mandal SC, Banerjee SK, Sinha S, Das J, Saha BP, Pal M. Studies on antidiarrhoeal activity of Punica granatum seed extract in rats. Journal of ethnopharmacology. 1999 Dec 15;68(1-3):205-8. - 66.
Emam AM, Ahmed MA, Tammam MA, Hala AM, Zawam S. Isolation and structural identification of compounds with antioxidant, nematicidal and fungicidal activities from Punica granatum L. var. nana. International Journal of Scientific & Engineering Research. 2015;6(11):1023-40. - 67.
Fahmy ZH, El-Shennawy AM, El-Komy W, Ali E, Hamid SA. Potential antiparasitic activity of pomegranate extracts against shistosomules and mature worms of Schistosoma Mansoni: in vitro and in vivo study. Australian Journal of Basic and Applied Sciences. 2009;3(4):4634-43. - 68.
Yones DA, Badary DM, Sayed H, Bayoumi SA, Khalifa AA, El-Moghazy AM. Comparative evaluation of anthelmintic activity of edible and ornamental pomegranate ethanolic extracts against Schistosoma mansoni. BioMed research international. 2016 Jan 1;2016. - 69.
Al-Musayeib NM, Mothana RA, Al-Massarani S, Matheeussen A, Cos P, Maes L. Study of the in vitro antiplasmodial, antileishmanial and antitrypanosomal activities of medicinal plants from Saudi Arabia. Molecules. 2012 Oct;17(10):11379-90. - 70.
Alkathiri B, El-Khadragy MF, Metwally DM, Al-Olayan EM, Bakhrebah MA, Abdel Moneim AE. Pomegranate (Punica granatum) juice shows antioxidant activity against cutaneous leishmaniasis-induced oxidative stress in female BALB/c mice. International Journal of Environmental Research and Public Health. 2017 Dec;14(12):1592. - 71.
Calzada F, Yépez-Mulia L, Aguilar A. In vitro susceptibility of Entamoeba histolytica andGiardia lamblia to plants used in Mexican traditional medicine for the treatment of gastrointestinal disorders. Journal of Ethnopharmacology. 2006 Dec 6;108(3):367-70. - 72.
Al-Mathal EM, Alsalem AA. Pomegranate ( Punica granatum ) peel is effective in a murine model of experimentalCryptosporidium parvum ultrastructural studies of the ileum. Experimental parasitology. 2013 Aug 1;134(4):482-94. - 73.
El-Sherbini GM, Ibrahim KM, El-Sherbiny ET, Abdel-Hady NM, Morsy TA. Efficacy of Punica granatum extract on in-vitro and in-vivo control of Trichomonas vaginalis. Journal of the Egyptian Society of Parasitology. 2010;40(1):229-44. - 74.
Thangavelu A, Elavarasu S, Sundaram R, Kumar T, Rajendran D, Prem F. Ancient seed for modern cure–pomegranate review of therapeutic applications in periodontics. Journal of pharmacy & bioallied sciences. 2017 Nov;9(Suppl 1): S11. - 75.
Alexandre EM, Silva S, Santos SA, Silvestre AJ, Duarte MF, Saraiva JA, Pintado M. Antimicrobial activity of pomegranate peel extracts performed by high pressure and enzymatic assisted extraction. Food research international. 2019 Jan 1;115:167-76. - 76.
Viana GS, Menezes SM, Cordeiro LN, Matos FJ. Biological Effects of Pomegranate ( Punica granatum L.), especially its antibacterial actions, against microorganisms present in the dental plaque and other infectious processes. InBioactive Foods in Promoting Health 2010 Jan 1 (pp. 457-478). Academic Press. - 77.
Papuc C, Goran GV, Predescu CN, Nicorescu V, Stefan G. Plant polyphenols as antioxidant and antibacterial agents for shelf-life extension of meat and meat products: Classification, structures, sources, and action mechanisms. Comprehensive Reviews in Food Science and Food Safety. 2017 Nov;16(6):1243-68. - 78.
Bouarab Chibane L, Degraeve P, Ferhout H, Bouajila J, Oulahal N. Plant antimicrobial polyphenols as potential natural food preservatives. Journal of the Science of Food and Agriculture. 2019 Mar 15;99(4):1457-74. - 79.
Daglia M. Polyphenols as antimicrobial agents. Current opinion in biotechnology. 2012 Apr 1;23(2):174-81. - 80.
Al-Zoreky NS. Antimicrobial activity of pomegranate ( Punica granatum L.) fruit peels. International journal of food microbiology. 2009 Sep 15;134(3):244-8. - 81.
Fan W, Chi Y, Zhang S. The use of a tea polyphenol dip to extend the shelf life of silver carp (Hypophthalmicthys molitrix) during storage in ice. Food chemistry. 2008 May 1;108(1):148-53. - 82.
Tehranifar A, Selahvarzi Y, Kharrazi M, Bakhsh VJ. High potential of agro-industrial by-products of pomegranate ( Punica granatum L.) as the powerful antifungal and antioxidant substances. Industrial Crops and Products. 2011 Nov 1;34(3):1523-7. - 83.
Bassole IH, Ouattara AS, Nebie R, Ouattara CA, Kabore ZI, Traore SA. Chemical composition and antibacterial activities of the essential oils of Lippia chevalieri and Lippia multiflora from Burkina Faso. Phytochemistry. 2003 Jan 1;62(2):209-12. - 84.
Akhtar S, Ismail T, Fraternale D, Sestili P. Pomegranate peel and peel extracts: Chemistry and food features. Food chemistry. 2015 May 1; 174:417-25. - 85.
Cowan MM. Plant products as antimicrobial agents. Clinical microbiology reviews. 1999 Oct 1;12(4):564-82. - 86.
Haslam E. Natural polyphenols (vegetable tannins) as drugs: possible modes of action. Journal of natural products. 1996 Feb 22;59(2):205-15. - 87.
Haraguchi H, Tanimoto K, Tamura Y, Mizutani K, Kinoshita T. Mode of antibacterial action of retrochalcones from Glycyrrhiza inflata. Phytochemistry. 1998 May 1;48(1):125-9. - 88.
Hatano T, Shintani Y, Aga Y, Shiota S, Tsuchiya T, Yoshida T. Phenolic constituents of licorice. VIII. Structures of glicophenone and glicoisoflavanone, and effects of licorice phenolics on methicillin-resistant Staphylococcus aureus. Chemical and Pharmaceutical Bulletin. 2000 Sep 1;48(9):1286-92. - 89.
Miguel MG, Neves MA, Antunes MD. Pomegranate ( Punica granatum L.): A medicinal plant with myriad biological properties-A short review. Journal of Medicinal Plants Research. 2010 Dec 29;4(25):2836-47. - 90.
Lou Z, Wang H, Zhu S, Ma C, Wang Z. Antibacterial activity and mechanism of action of chlorogenic acid. Journal of food science. 2011 Aug;76(6):M398-403. - 91.
Silva-Beltrán NP, Ruiz-Cruz S, Cira-Chávez LA, Estrada-Alvarado MI, Ornelas-Paz JD, López-Mata MA, Del-Toro-Sánchez CL, Ayala-Zavala JF, Márquez-Ríos E. Total phenolic, flavonoid, tomatine, and tomatidine contents and antioxidant and antimicrobial activities of extracts of tomato plant. International journal of analytical chemistry. 2015 Jan 1;2015. - 92.
Naz S, Siddiqi R, Ahmad S, Rasool SA, Sayeed SA. Antibacterial activity directed isolation of compounds from Punica granatum . Journal of food science. 2007 Nov;72(9):M341-5. - 93.
Lei Y, Tang Z, Liao R, Guo B. Hydrolysable tannin as environmentally friendly reducer and stabilizer for graphene oxide. Green chemistry. 2011;13(7):1655-8. - 94.
Gregory WC, Gregory MP, Krapovickas A, Smith BW, Yarbrough JA. Structure and genetic resources of peanuts. Peanuts-culture and uses. 1973:47-133. - 95.
Peng S, Jay-Allemand C. Use of antioxidants in extraction of tannins from walnut plants. Journal of chemical ecology. 1991 May 1;17(5):887-96. - 96.
Ci Z, Kikuchi K, Hatsuzawa A, Nakai A, Jiang C, Itadani A, Kojima M. Antioxidant Activity, and α-Glucosidase, α-Amylase and Lipase Inhibitory Activity of Polyphenols in Flesh, Peel, Core and Seed from Mini Apple. American Journal of Food Science and Technology. 2018 Nov 15;6(6):258-62. - 97.
Pereira JV, Pereira MS, Sampaio FC, Sampaio MC, Alves PM, Araújo CR, Higino JS. In vitro antibacterial and antiadherence effect of Punica granatum Linn extract upon dental biofilm microorganisms. Braz J Pharmacogn. 2006; 16:88-93. - 98.
Vasconcelos LC, Sampaio FC, Sampaio MC, Pereira MD, Higino JS, Peixoto MH. Minimum inhibitory concentration of adherence of Punica granatum Linn (pomegranate) gel against S. mutans, S. mitis and C. albicans. Brazilian Dental Journal. 2006;17(3):223-7. - 99.
O'May C, Tufenkji N. The swarming motility of Pseudomonas aeruginosa is blocked by cranberry proanthocyanidins and other tannin-containing materials. Applied and environmental microbiology. 2011 May 1;77(9):3061-7. - 100.
Sarabhai S, Sharma P, Capalash N. Ellagic acid derivatives from Terminalia chebula Retz. downregulate the expression of quorum sensing genes to attenuate Pseudomonas aeruginosa PAO1 virulence. PLoS one. 2013 Jan 8;8(1): e53441. - 101.
Ni N, Li M, Wang J, Wang B. Inhibitors and antagonists of bacterial quorum sensing. Medicinal research reviews. 2009 Jan;29(1):65-124. - 102.
Rudrappa T, Bais HP. Curcumin, a known phenolic from Curcuma longa , attenuates the virulence of Pseudomonas aeruginosa PAO1 in whole plant and animal pathogenicity models. Journal of Agricultural and Food Chemistry. 2008 Mar 26;56(6):1955-62. - 103.
Morohoshi T, Kato M, Fukamachi K, Kato N, Ikeda T. N-acylhomoserine lactone regulates violacein production in Chromobacterium violaceum type strain ATCC 12472. FEMS microbiology letters. 2008 Feb 1;279(1):124-30. - 104.
Koh KH, Tham FY. Screening of traditional Chinese medicinal plants for quorum-sensing inhibitors activity. Journal of Microbiology, Immunology and Infection. 2011 Apr 1;44(2):144-8. - 105.
Zahin M, Hasan S, Aqil F, Khan M, Ahmad S, Husain FM, Ahmad I. Screening of certain medicinal plants from India for their anti-quorum sensing activity. - 106.
Truchado P, Tomás-Barberán FA, Larrosa M, Allende A. Food phytochemicals act as quorum sensing inhibitors reducing production and/or degrading autoinducers of Yersinia enterocolitica and Erwinia carotovora. Food Control. 2012 Mar 1;24(1-2):78-85. - 107.
Yang Q, Wang L, Gao J, Liu X, Feng Y, Wu Q, Baloch AB, Cui L, Xia X. Tannin-rich fraction from pomegranate rind inhibits quorum sensing in Chromobacterium violaceum and biofilm formation in Escherichia coli. Foodborne pathogens and disease. 2016 Jan 1;13(1):28-35. - 108.
Li G, Yan C, Xu Y, Feng Y, Wu Q, Lv X, Yang B, Wang X, Xia X. Punicalagin inhibits Salmonella virulence factors and has anti-quorum-sensing potential. Applied and environmental microbiology. 2014 Oct 1;80(19):6204-11. - 109.
Ekambaram SP, Perumal SS, Balakrishnan A. Scope of hydrolysable tannins as possible antimicrobial agent. Phytotherapy Research. 2016 Jul;30(7):1035-45. - 110.
Chung KT, Jr SS, Lin WF, Wei CI. Growth inhibition of selected food-borne bacteria by tannic acid, propyl gallate and related compounds. Letters in Applied Microbiology. 1993 Jul;17(1):29-32. - 111.
Engels C, Gänzle MG, Schieber A. Fast LC–MS analysis of gallotannins from mango ( Mangifera indica L.) kernels and effects of methanolysis on their antibacterial activity and iron binding capacity. Food research international. 2012 Jan 1;45(1):422-6. - 112.
Tian F, Li B, Ji B, Zhang G, Luo Y. Identification and structure–activity relationship of gallotannins separated from Galla chinensis. LWT-Food Science and Technology. 2009 Sep 1;42(7):1289-95. - 113.
Farha AK, Yang QQ, Kim G, Li HB, Zhu F, Liu HY, Gan RY, Corke H. Tannins as an alternative to antibiotics. Food Bioscience. 2020 Sep 3:100751. - 114.
Machado TD, Leal IC, Amaral AC, Santos K, Silva MG, Kuster RM. Antimicrobial ellagitannin of Punica granatum fruits. Journal of the Brazilian Chemical Society. 2002 Sep;13(5):606-10. - 115.
Shimozu Y, Kimura Y, Esumi A, Aoyama H, Kuroda T, Sakagami H, Hatano T. Ellagitannins of Davidia involucrata. I. structure of davicratinic acid A and effects of davidia tannins on drug-resistant bacteria and human oral squamous cell carcinomas. Molecules. 2017 Mar;22(3):470. - 116.
Reddy MK, Gupta SK, Jacob MR, Khan SI, Ferreira D. Antioxidant, antimalarial and antimicrobial activities of tannin-rich fractions, ellagitannins and phenolic acids from Punica granatum L. Planta medica. 2007 Oct;53(05):461-7. - 117.
Compean KL, Ynalvez RA. Antimicrobial activity of plant secondary metabolites: A review. Research Journal of Medicinal Plants. 2014;8(5):204-13. - 118.
Silver S. Bacterial resistances to toxic metal ions-a review. Gene. 1996 Jan 1;179(1):9-19. - 119.
Ug A, Ceylan Ö. Occurrence of resistance to antibiotics, metals, and plasmids in clinical strains of Staphylococcus spp. Archives of Medical Research. 2003 Mar 1;34(2):130-6. - 120.
O’Neill MA, Vine GJ, Beezer AE, Bishop AH, Hadgraft J, Labetoulle C, Walker M, Bowler PG. Antimicrobial properties of silver-containing wound dressings: a microcalorimetric study. International journal of pharmaceutics. 2003 Sep 16;263(1-2):61-8. - 121.
Strohal R, Schelling M, Takacs M, Jurecka W, Gruber U, Offner F. Nanocrystalline silver dressings as an efficient anti-MRSA barrier: a new solution to an increasing problem. Journal of Hospital Infection. 2005 Jul 1;60(3):226-30. - 122.
Stewart GS, Jassim SA, Denyer SP, Newby P, Linley K, Dhir VK. The specific and sensitive detection of bacterial pathogens within 4 h using bacteriophage amplification. Journal of applied microbiology. 1998 May 1;84(5):777-83. - 123.
McCarrell EM, Gould SW, Fielder MD, Kelly AF, El Sankary W, Naughton DP. Antimicrobial activities of pomegranate rind extracts: enhancement by addition of metal salts and vitamin C. BMC Complementary and Alternative Medicine. 2008 Dec;8(1):1-7. - 124.
Houston D. Towards a nanomedicine-based broad-spectrum topical virucidal therapeutic system (Doctoral dissertation, Cardiff University). - 125.
Nair MS, Joseyphus RS. Synthesis and characterization of Co (II), Ni (II), Cu (II) and Zn (II) complexes of tridentate Schiff base derived from vanillin and DL-α-aminobutyric acid. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2008 Sep 1;70(4):749-53. - 126.
Houston DM, Robins B, Bugert JJ, Denyer SP, Heard CM. In vitro permeation and biological activity of punicalagin and zinc (II) across skin and mucous membranes prone to Herpes simplex virus infection. European Journal of Pharmaceutical Sciences. 2017 Jan 1; 96:99-106. - 127.
Zhang L, Liu R, Gung BW, Tindall S, Gonzalez JM, Halvorson JJ, Hagerman AE. Polyphenol–aluminum complex formation: implications for aluminum tolerance in plants. Journal of agricultural and food chemistry. 2016 Apr 20;64(15):3025-33 - 128.
Gould SW, Fielder MD, Kelly AF, Sankary WE, Naughton DP. Antimicrobial pomegranate rind extracts: enhancement by Cu (II) and vitamin C combinations against clinical isolates of Pseudomonas aeruginosa. British journal of biomedical science. 2009 Jan 1;66(3):129-32.