Open access peer-reviewed chapter

Antimicrobial Potential of Pomegranate Extracts

Written By

Vildan Celiksoy and Charles M. Heard

Submitted: 27 November 2020 Reviewed: 04 January 2021 Published: 11 February 2021

DOI: 10.5772/intechopen.95796

From the Edited Volume

Pomegranate

Edited by Vasiliki Lagouri

Chapter metrics overview

555 Chapter Downloads

View Full Metrics

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 Punica granatum L. tree, is one of oldest recorded edible fruits and it has been used as a folklore medicine since ancient times. There are records of it being used to treat inflammatory diseases and disorders of the digestive tract in the Ayurvedic and Unani systems [3, 4]. In terms of infections, the ancient Egyptians used it in the treatment of tapeworms and other parasites [5], whereas other cultures have used pomegranates to treat diarrhea and dysentery [6, 7, 8], although at the time they would not have known that pathogenic microbes were responsible. In more recent times, the pomegranate has been extensively and scientifically studied for its antimicrobial potential in a diversity of areas such skin infections, dentistry, food preservation etc. [9].

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.

Advertisement

2. Activity against bacteria

Staphyllococcus aureus (S. aureus) and methicillin resistant Staphyllococcus aureus (MRSA) have received the greatest attention as targets for pomegranate extract activity. In 2010, the antibacterial activity of crude and purified extracts of pomegranate peel were assessed by Panichayupakaranant et al. 8 mg crude peel loaded discs showed 20 mm and 30 mm zone of inhibition against clinical isolates of S. aureus and E. coli, respectively. The purified peel extract discs, loaded up to 8 mg, exerted a range of zones of inhibition between 15-20 mm for S. aureus and 20-30 mm for E. coli. Using standardized peel extract, minimum inhibitory concentrations (MIC) values of 0.016, 0.008, and 0.008–0.016 mg/mL were obtained for S. aureus, S. epidermidis and Propionibacterium acnes respectively. Tetracycline was used as a positive control in this study and standardized pomegranate rind extract showed lower activity in zone of inhibition assays, with tetracycline also showing a lower minimum inhibitory concentration (MIC) [14]. A methanolic extract of pomegranate peel inhibited biofilm formation and eradicated pre-formed biofilm of S. aureus, MRSA, E. coli in the concentration range 25 to 150 μg/mL [15]. In the same study, ellagic acid showed biofilm inhibition and eradication activity at somewhat lower concentrations (5–40 μg/mL) than pomegranate peel extract. Furthermore, while pomegranate extract was able to inhibit the growth of S. aureus, it also suppressed enterotoxin production [5].

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: F. nucleatum, P. gingivalis, P. intermedia, S. mutans and A. actinomycetomomitans [16]. In another in vitro study, pomegranate alcoholic extracts have been tested on bacteria which are collected from patients who have tooth decay or periodontitis and inhibited a range of bacteria in both planktonic and biofilm conditions [17]. Synergistic bactericidal activity against S. mutans and R. dentocariosa was reported for pomegranate extract in combination with other plant polyphenolic extracts, honey and myrtle [18].

Moreover, ‘standardized’ pomegranate peel extract showed higher antimicrobial activity than other parts of pomegranate (flower, leaf, stem) and ciprofloxacin (2 mg/mL) against S. mutans, Salmonella mitis and L. acidophilusin in a zone of inhibition assay [19]. Again, pomegranate gel showed an inhibitory activity against S. mutans, Salmonella sanguis, and S. mitis [20]. This gel also showed antiadhesive activity against S. mutans and S. mitis at lower than minimum inhibitory concentrations to a glass surface. In addition to inhibition activity on bacterial growth and biofilm, pomegranate extracts showed antiadhesive activity for S. mutans adherence on tooth surface in orthodontic treated patients [21]. In other clinical studies, the antiplaque effect and prophylactic benefits of pomegranate have been highlighted [22]. Recently, a systematic review and meta-analysis has been carried out by Martins et al. [23], where natural antimicrobial phenolic compounds were compared with synthetic antimicrobials by using 16 clinical studies for qualitative analysis, and 12 studies for meta-analysis. For the meta-analysis, six clinical trials were evaluated for the comparison of natural antimicrobial phenolic compounds, including pomegranate extract mouthwash, and synthetic antimicrobials. It was found that natural antimicrobial phenolic compounds are less effective than chlorhexidine for biofilm control, although it showed similar reduction of the oral microbes count which was sub-grouped as total microorganisms, Streptococcus mutans, and Streptococcus spp. according to type of microorganisms.

Due to its antimicrobial and antioxidant properties, pomegranate extract has been studied for its preservation potential use in the food industry. Kannat et al. [24] did a study to evaluate the antimicrobial activity of pomegranate peel against common food spoilers and potential pathogens. It was shown that pomegranate peel extract increased the shelf-life of chicken and meat products and showed antimicrobial activity against to S. aureus, B. cereus with a minimum inhibitory concentration at 0.01%. However, it did not show antimicrobial activity for E. coli and S. typhimurium even at higher concentrations. Other researchers showed that pomegranate extracts were less effective against Gram-negative compared to Gram-positive bacteria, probably due to the differences in cell wall structure [25, 26]. Moreover, pomegranate has been studied in a novel and smart multi-functional hydrogel (MFH) system as a food packaging material since it is easy to monitor of the color change due to changes in conditions such as pH and temperature. The MFH with pomegranate extract showed promising antimicrobial activity on pasteurized milk and cheese over a 7-day period [27]. Pomegranate peel extract was also added in a film formulation to produce a material for food packaging materials with antimicrobial and antioxidant effects. This film formulation was found to restrict the growth of L. monocytogenes in pork samples inoculated with this bacterium [28].

The activity of pomegranate extract against bacteria is summarized in Table 1.

Part of pomegranate usedFormTest organismsMICsReference
PomElla® (30% punicalagin)Streptococcus mutans,
Fusobacterium nucleatum,
Aggregatibacter actinomycetomcomitans,
Prevotella intermedia
0.8 μg/mL
0.2 μg/mL
0.2 μg/mL
0.8 μg/mL
[16]
PeelAcetonic, Methanolic, Ethanolic PPE and Hydro-alcoholic
PPE
Streptococcus mutans,
Gemella morbillorum,
Enterococcus faecalis,
Staphylococcus epidermis,
Klebsiella oxytoca,
Enterobacter bugandensis,
R. dentocariosa,
Streptococcus mutans
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, stemAqueous and methanolicS. mutans, S. mitis, L. acidhopillus[19]
PeelBasic gel formulation including 540 mg pomegranate peel powderS. mutans,
S. mitis,
S. sanguis,
C. albicans
1:16
1:128
1:16
1:64
[20]
Pomegranate mouthwash (Pomegranate extract were obtained from Verdure science, 30% punicalagin)Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis,
Prevotella intermedia
62.5 mg/mL
>31.25 mg/mL
16.125 mg/mL
[21]
PeelMethanolic PPES. aureus,
MRSA,
E. coli,
C. albicans
250 μg/mL
250 μg/mL
250 μg/mL
1000 μg/mL
[15]
Fruit pericarpMethanolicStaphylococcus aureus,
Streptococcus pyogenes,
Escherichia coli,
Klebsiella pneumoniae,
Proteus vulgaris,
Pseudomonas aeruginosa
640–2560 μg/mL
1280–2560 μg/mL
640–2560 μg/mL
[9]
PeelStandardized extract (13% ellagic acid)Propionibacterium acnes,
Shigella sonnei,
S. aureus,
Staphylococcus epidermidis
15.6 μg/mL
7.81 μg/mL
7.81–15.6 μg/mL
7.81 μg/mL
[14]
PeelWaterB. cereus, S. aureus, P. fluorescens, S. tyriphium, E. coli200–450 ppm[24]
PeelAcetonic, methanolic, waterBacillus cereus,
Bacillus coagulens,
B. subtilis,
Staphylococcus aureus,
E. coli,
Pseudomonas aeruginosa
200–400 ppm
150–500 ppm
200–450 ppm
200–700 ppm
200–400 ppm
[25]
FruitWaterBacillus cereus,
E. coli,
S. aureus
100 mg/mL
100 mg/mL
100 mg/mL
[27]
PeelEthanolic, methanolicL. monocytogenes[28]

Table 1.

Antibacterial activity of pomegranate extracts against different bacteria.

Advertisement

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 Candida species [5, 7, 31], which are part of the normal microbiota of human gastrointestinal, oral, and vaginal mucosae. However, they can cause superficial infections and especially in immunosuppressed patients, they can cause severe infectious problems. In one study, punicalagin showed superior antifungal activity than the conventional fluconazole in an in vitro time-kill assay. In addition, punicalagin caused a significant change in Candida morphology, and alteration in budding pattern and pseudo hyphae when yeasts were treated with a sub-inhibitory concentration of punicalagin [32]. In another study, pomegranate extract showed superior inhibitory action against C. tropicalis while fluconazole and voriconazole, which are commonly prescribed azoles for fungal infections, have been ineffective against C. tropicalis [33].

The potential of pomegranate extract has been studied against fungi in in vitro biofilm assays. Microbes in biofilms have substantially different characteristics to those of their free-living planktonic counterparts [34, 35]. In particular, microbes in biofilms are concealed and therefore protected from antifungal agents and plant extracts [36, 37]. Pomegranate extract and its one of the major components ellagic acid were shown to exert a reduction in biofilm formation and eradicated pre-formed biofilm of C. albicans in an in vitro biofilm study [15]. Spray-dried microparticles containing pomegranate extract showed antifungal activity in in vitro assays under both planktonic and biofilm conditions [38]. In addition, the inhibitory effect of pomegranate extract on the growth of Candida albicans was demonstrated in an in vivo study [31]. In another study, similar effects have been obtained against to Candida mycoderma using different parts of pomegranate, fresh fruit and sterile juice [39].

In addition to Candida species, pomegranate extracts showed inhibitory activity against dermatophytes, which are fungi which use keratin as a source of nutrition and may cause infection in keratinized tissue parts such as nails, skin and hair follicles. Pomegranate peel extract and punicalagin exerted potent antifungal activity against to T. rubrum (125 μg/mL), T. mentagrophytes (125 μg/mL), M. canis (250 μg/mL) and M. gypseum (250 μg/mL). Punicalagin at a concentration of 62.5 μg/mL also inhibited T. rubrum spore germination, and it was further found that punicalagin 62.5 μg/mL and nystatin 0.78 μg/mL showed similar inhibition in hyphal growth of T. rubrum [40]. Moreover, pomegranate extracts have been researched for use as natural preservatives due to their antifungal (in addition to antibacterial) activities [41]. Pomegranate peel extract showed inhibition activity against to Aspergillus niger and Aspergillus parasiticus in a zone of inhibition assays [42]. Pomegranate extract showed inhibitory effects against fungal pathogens which are responsible for fruit and vegetable decay. Punicalagin was proposed as the main compound in the extracts providing the observed antifungal activity and it has been found effective in mycelial growth inhibition against phytopathogenic filamentous fungi such as Fusarium vertillicoides, Mucor indicus, Penicillium citrinum, Rhizopus oryzae and Trichoderma recei [43]. Also, the growth rate of pathogens presented a negative correlation with total punicalagin content, and it has thus been suggested that pure punicalagin may be used as a control agent in storage disease to prevent the excessive use of synthetic fungicides [44, 45].

The activity of pomegranate extract against fungal microbes is summarized in Table 2.

Part of pomegranate usedFormTest organismsMICsReference
PeelCrude extract,
Aqueous fraction,
ethyl acetate fraction,
butanol fraction, punicalagin
C. albicans,
C. parapsilosis
3.9–7.8 μg/mL,
1.9–15.6 μg/mL
[32]
PeelMethanolicC. albicans> 1000 μg/mL[15]
PeelCrude extract,
Crude extract in a spray-dried microparticle formulation
C. albicans,
C. parapsilosis,
C. tropicalis
3.9–15.6 μg/mL[38]
PeelHydroalcoholicTrichophyton rubrum,
Trichophyton mentagrophytes,
Microsporum gypseum,
Microsporum canis
125 g/mL,
125 g/mL,
250 g/mL,
250 g/mL
[40]
PeelAqueousAspergillus niger,
Aspergillus parasiticus
[42]
PeelAqueousA. alternata,
S. botryosum, Fusarium spp.
[43]

Table 2.

Antifungal activity of pomegranate extracts against different fungi.

Advertisement

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 et al. [53], complete inactivation of influenza virus was observed with 1600 μg/mL pomegranate polyphenols, and 400 μg/mL of same extract showed 99% or more titer reduction in only 5 minutes treatment. This result was similar to another study which showed complete inactivation of H3N2 influenza virus within 30 minutes of treatment and a significant viral reduction with approximately 1 μg/mL pomegranate polyphenols. An in vitro study, showed that pomegranate polyphenol extract inhibited viral replication in addition to its virucidal effect – they also obtained same activity for punicalagin and suggested punicalagin is the main compound in pomegranate extract for antiviral activity [54]. In an in vivo mouse model study, pomegranate polyphenols applied to the lung were found to reduce influenza infection, without toxic effect to the host [55, 56].

Hepatitis C virus (HCV) is the main factor in end-stage liver disease and approximately 170 million people are chronically infected with HCV. Pomegranate ellagitannins, punicalagin, punicalin and ellagic acid, blocked and inhibited the NS3/4A protease which is a viral polyprotein responsible for processing and replication in HCV. Moreover, punicalagin and punicalin significantly decreased the HCV replication in an in vitro cell culture system [57]. The more prevalent adenovirus (ADV) in Hep-2 host cells has also shown susceptibility to pomegranate crude extract, fractions, and main phenolic compounds. It has been found that a n-butanol fraction of pomegranate peel extract and gallic acid showed the highest antiviral activity against ADV. Furthermore, the crude extract, n-butanol fraction and gallic acid inhibited ADV replication in the post-adsorption phase [58].

Herpes simplex virus (HSV) is from the Herpes viridae family and infects a high proportion of the populous. HSV-1 is generally responsible for cold sores and encephalitis, whereas HSV-2 is the main causative agent of anogenital infections, which can also infect neonates via the mother [59, 60]. Pomegranate rind extract (PRE) and its major ellagitannin compound, punicalagin, showed virucidal activity against HSV-1. While punicalagin has greater virucidal activity than an equivalent mass of pomegranate rind extract, PRE showed better antiviral activity than punicalagin. Moreover, PRE demonstrated comparable activity to acyclovir against HSV-1 and HSV-2, in addition to antiviral activity against acyclovir-resistant HSV-1 [48]. PRE is thus a promising new alternative treatment for HSV-1 since currently acyclovir is the gold standard treatment in HSV infections [61].

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 usedFormTest organismsMechanism of virus targetReference
RindCrude hydraulic extract,
Punicalagin,
Ellagic acid
Herpes simplex virusVirucidal activity[48]
Juice, peel and pomegranate liquid extractInfluenze A viruses, H1N1, H3N2, H5N1 and coronavirus MHV A59Damage to virion integrity and virucidal activity[53]
JuicePomegranate polyphenol extract, punicalagin, pomegranate liquid extract (from POM Wonderful)Human influenza A (H3N2)Inhibition of viral RNA replication[54]
PeelMethanolic crude extract, punicalin, punicalagin, ellagic acidHepatitis C virus[57]
PeelMethanolic crude extract, ellagic acid, punicalagin, gallic acidAdenovirusInhibition of adenovirus replication[58]

Table 3.

Antiviral activity of pomegranate extracts against different viruses.

Advertisement

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 Punica granatum L., root, stem bark, and rind of fruit, have been used commonly as vermifugal and taenicide agents [63]. The antiprotozoal activity of the pomegranate has been determined and in folkloric medicine, it has been used as anthelminthic especially against tapeworms and for diarrhea [64, 65]. A methanolic extract of pomegranate leaves showed nematicide activity and hepatoprotective activity against carbon tetrachloride induced hepatoxicity [66]. Extracts of pomegranate showed anti-schistosomal activity against Shistosoma mansoni in both in vitro and in vivo conditions [67]. In addition, it caused reduction or complete loss of motor activity, lethality and ultra-morphological changes in adult worms [68]. There is thus potential for the treatment of schistosomiasis.

Al-Musayeib et al. reported the antiparasitic activity of pomegranate rind extract against Plasmodium falciparum [69]. Pomegranate juice was found to exert dose-dependent activity against Leishmania major promastigotes and, at >80 μL/mL, gave significantly greater reduction than the positive control, Pentostam. Furthermore, mice that were orally treated with pomegranate juice, showed significantly reduced cutaneous leishmaniasis lesions compared to untreated mice [70]. Calzada et al. demonstrated pomegranate antiprotozoal activity against Entameoba histolytica and Giardia lamblia that cause diarrheic dysentery [71]. Pomegranate peel suspension also affected C. parvum in different stages and finally caused parasite death in an in vivo murine model; furthermore, pomegranate suspension did not cause any negative change in the mice ileal tissue [72]. In another study, pomegranate extract showed activity against T. vaginalis, both in vitro and clinically. Patients with T. vaginalis infection were treated with pomegranate juice and symptoms were found to have cleared after two months [73]. The activity of pomegranate extract against parasites is summarized in Table 4.

Part of pomegranate usedFormMechanism of organism targetOrganismsReference
SeedMethanolicReduction in gastrointestinal motility[65]
LeaveMethanolicLarvicidal and ovicidal activityM. incognita,
R. reniformis,
P. penetrans,
S. rolfsii
[66]
Peels, juice and leavesMethanolicReduction in viability of parasiteSchistosoma mansoni,
schistosomules
[67]
Leaf, stem barkEthanolicworms separation, reduction of motor activity, lethality, and ultrastructural tegumental alterationsSchistosoma mansoni[68]
PeelPowder form directly given to animal in in vivo studyGrowth inhibition and deathCryptos poridium[72]
JuiceCrude extract was applied by patients in a clinical studyTrichomoniasis vaginalis[73]

Table 4.

Antiparasitic activity of pomegranate extracts against different parasites.

Advertisement

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 Punica granatum L. showed a specific antimicrobial action on dental plaque, which is a complex biofilm on tooth, by inhibiting adherence mechanism of oral microbes to dental surface via disturbing polyglucan synthesis [17, 96, 97]. Moreover, Vasconcelos et al. [98] used Punica granatum L. in a gel formulation using increasing and doubled concentrations of the diluted solutions of the gel with ranging concentrations from 1:1 to 1:1024, and similar results obtained. The gel formulation inhibited the adherence of different bacterial strains and a yeast, C. albicans, in the oral cavity and affected preformed biofilm.

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, Chromobacterium violaceum has been used as a biosensor since it produces violacein, purple pigment color, in response to QS regulation [103]. Pomegranate inhibited the QS of two bacterial strains which are Chromobacterium violaceum (by affecting purple pigment production) and P. aeruginosa (by decreasing bacterial swarming motility) [104, 105]. In another study, different compounds from herbs, fruits and plant extracts have been studied for their QS activity, with resveratrol and pomegranate extract demonstrating the highest inhibition activities. The QS activity of pomegranate has been associated with ellagic acid content of pomegranate extract (85% punicalagin, 7% free ellagic acid) since ellagic acid showed 86% inhibition at a low concentration of 4 μg/mL. However, the anti-QS activity of punicalagin is also believed important in pomegranate extracts [106]. Tannin-rich fraction of pomegranate rind extract showed inhibition of biofilm formation and motility of E. coli and repressed the expressions of curli genes (csgB and csgD) and various motility genes (fimA, fimH, flhD, motB, qseB, and qseC) [107]. Similarly, punicalagin significantly decreased the expression of QS-related genes (sdiA and srgE) of Salmonella [108].

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 et al., ellagic acid, gallagic acid, punicalin and punicalagin were purified and tested for their antiplasmodial and antimicrobial activities. Gallagic acid and punicalagin showed the strongest effects on the growth of bacteria and fungi and it has been suggested that the ellagic acid moiety is not important compared to the gallagyl and hexahydroxydiphenol (HHDP) moieties for the inhibition of microbes [116]. The degradation of punicalagin to ellagic acid, via punicalin and hexahydroxydiphenic acid is shown in Figure 1.

Figure 1.

Reduction of punicalagin, punicalin and HHDP to ellagic acid, adopted from Seeram et al. [3, 12].

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].

Advertisement

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 et al. [122] investigated the potentiated antimicrobial activity of pomegranate rind extract (PRE) in combination with metal ions. In their study, the aim was to exert short term exposure of pomegranate rind extract and ferrous sulfate combination on bacteriophage levels for 3 minutes. This combination showed highly significant synergistic activity and reduced the bacteriophage levels in a short-term exposure. This short screening time was necessary for this experiment due to low stability of pomegranate rind extract/ferrous salt solution which, via a Fenton reaction caused Fe2+ to oxidize to Fe3+ with concomitant solution blackening. To overcome this instability problem, potentiated/synergised antimicrobial activity of pomegranate rind extract has since been examined using alternative metal ions [48, 123, 124].

McCarell et al. [123] investigated the antimicrobial activity of PRE with 4.6 mM FeSO4, CuSO4, MnSO4, ZnO and also studied antimicrobial activity of PRE/metal salt combinations plus vitamin C which was added as a stabilizer. They observed significant synergistic antibacterial activity against E. coli, Ps. Aeruginosa, S. aureus and P. mirabilis when they combined PRE with Cu (II) ions. Moreover, with the addition of vitamin C as antioxidant, the antimicrobial activity increased significantly for PRE/Fe (II) and PRE/Cu (II) combinations against S. aureus. In another study, researchers used the vanillin complexes with different metal ions using the agar diffusion method and it was found that the vanillin and metal salts showed an enhanced activity against S. aureus, E. coli, K. pneumanie, P. aeruginosa and C. albicans. The results from both studies indicated that the addition of metal ions, especially copper salts, can significantly enhance antibacterial activity of a natural product [123, 125].

Significantly enhanced virucidal activity of PRE was later observed against HSV-1, HSV-2 and acyclovir-resistant HSV-1 by Houston et al. [48] in combination with different Zn (II) ion salts, including zinc sulphate, zinc citrate, zinc stearate and zinc gluconate, with a maximum of 6 log reduction observed. Unlike PRE and Fe2+, this activity was not time-limited, and was not associated with blackening. Importantly, this activity was also retained when applied to epithelial surfaces prone to Herpes infection, including buccal and vaginal mucosae [126], indicating potential treatment for cold sores and anogenital Herpes.

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].

Advertisement

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.

Advertisement

Acknowledgments

We would like to thank to Turkish Ministry of Education for supporting Vildan Celiksoy’s PhD project.

Advertisement

Conflict of interest

The authors declare no conflict of interest.

References

  1. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 21. Bhadbhade SJ, Acharya AB, Rodrigues SV, Thakur SL. The antiplaque efficacy of pomegranate mouthrinse. Quintessence International. 2011 Jan 1;42(1).
  22. 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. 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. 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. 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. 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 avellanaL.) cultivars. Food and Chemical Toxicology. 2008 May 1;46(5):1801-7.
  27. 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. 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. 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. 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. 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. 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. 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. 34. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science. 1999 May 21;284(5418):1318-22.
  35. 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. 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. 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. 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. 39. Heber D, Schulman RN, Seeram NP, editors. Pomegranates: ancient roots to modern medicine. CRC press; 2006 Jul 7
  40. 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. 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. 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. 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. 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. 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. 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. 47. Jassim SA, Naji MA. Novel antiviral agents: a medicinal plant perspective. Journal of applied microbiology. 2003 Sep;95(3):412-27.
  48. 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. 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. 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. 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. 52. Kamboj A, Saluja AK, Kumar M, Atri P. Antiviral activity of plant polyphenols. J Pharm Res. 2012 May;5(5):2402-12.
  53. 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. 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. 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. 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. 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. 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. 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. 60. Whitley RJ, Roizman B. Herpes simplex virus infections. The lancet. 2001 May 12;357(9267):1513-8.
  61. 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. 62. Tagboto S, Townson S. Antiparasitic properties of medicinal plants and other naturally occurring products.
  63. 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. 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. 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. 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. 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. 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. 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. 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. 71. Calzada F, Yépez-Mulia L, Aguilar A. In vitro susceptibility of Entamoeba histolytica and Giardia 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. 72. Al-Mathal EM, Alsalem AA. Pomegranate (Punica granatum) peel is effective in a murine model of experimental Cryptosporidium parvum ultrastructural studies of the ileum. Experimental parasitology. 2013 Aug 1;134(4):482-94.
  73. 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. 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. 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. 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. 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. 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. 79. Daglia M. Polyphenols as antimicrobial agents. Current opinion in biotechnology. 2012 Apr 1;23(2):174-81.
  80. 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. 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. 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. 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. 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. 85. Cowan MM. Plant products as antimicrobial agents. Clinical microbiology reviews. 1999 Oct 1;12(4):564-82.
  86. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 109. Ekambaram SP, Perumal SS, Balakrishnan A. Scope of hydrolysable tannins as possible antimicrobial agent. Phytotherapy Research. 2016 Jul;30(7):1035-45.
  110. 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. 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. 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. 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. 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. 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. 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. 117. Compean KL, Ynalvez RA. Antimicrobial activity of plant secondary metabolites: A review. Research Journal of Medicinal Plants. 2014;8(5):204-13.
  118. 118. Silver S. Bacterial resistances to toxic metal ions-a review. Gene. 1996 Jan 1;179(1):9-19.
  119. 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. 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. 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. 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. 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. 124. Houston D. Towards a nanomedicine-based broad-spectrum topical virucidal therapeutic system (Doctoral dissertation, Cardiff University).
  125. 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. 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. 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. 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.

Written By

Vildan Celiksoy and Charles M. Heard

Submitted: 27 November 2020 Reviewed: 04 January 2021 Published: 11 February 2021