Professional Documents
Culture Documents
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A. Méndez-Vilas (Ed.)
The quest for plants with medicinal properties continues to receive attention as they are known for a range of biological
activities which range from antibiotics to antitumor. Medicinal plants represent one of the most important fields of
traditional medicine all over the world. In light of the recent emergence of bacteria which are resistant to multiple
antimicrobial drugs posing a challenge for the treatment of infections, the need to discover new antimicrobial substances
for use in combating such microorganisms become pertinent. Bioactive compounds are normally accumulated as
secondary metabolites in all plant cells but their concentration varies according to the plant parts. Leaf is one of the highest
accumulatory plant part of such compounds and generally it is preferred for therapeutic purpose. Some of the active
compounds that inhibit the disease causing microbial growth act either singly or in combination. They can inhibit the
growth of the microbes by many mechanisms of actions. It is believed that crude extracts from plants are more effective
than isolated compounds due to their synergistic effect. Here we report the antimicrobial activity of five different solvent
extracts of four Indian nutraceutical plants viz. Manilkara zapota (L.) var. Royen., Psidium guajava L., Punica granatum
L. and Syzygium cumini L. leaves against 14 pathogenic microorganisms. The antimicrobial activity was evaluated by agar
well diffusion method. P. guajava leaves showed best and promising antimicrobial activity, indicating the possibilities of
its potential use in the formula of natural remedies for the treatment of infections.
Keywords antimicrobial activity; solvent extracts; Manilkara zapota; Psidium guajava; Punica granatum; Syzygium
cumini
1. Introduction
Infectious diseases account for a high proportion of health problems and burden in India. Large amounts of antibiotics
are consumed in fighting infections, some of them saving lives, but every use adds antibiotic resistance in bacteria.
Antimicrobial resistance has in fact turned into a global health issue, which hampers the control of infectious diseases;
multi-drug resistant bacteria are thus posing a problem in treatment regimens. Multi-drug resistant organisms are also an
epidemiological concern as they may spread locally, regionally or globally through individual contacts, poor sanitation,
travel or the food chain. During the present millennium, overcoming resistance to antibiotics is one of the major issues
facing WHO [1]. Hence, WHO has declared antimicrobial resistance as the theme for the World Health Day, 2011 [2].
Antimicrobials have been used successfully for over six decades to treat infectious diseases; but antimicrobial resistance
threatens the continued effectiveness of antimicrobials and simultaneously the downward trend in the development of
new antibiotics has serious implications [3].
Resistant bacteria dramatically reduce the possibilities of treating infectious diseases effectively and increase the risk
of complications and fatal outcomes for patients with severe infections. The increasing prevalence of multi - drug
resistant bacteria and fungi, the recent appearance of strains with reduced susceptibility to antibiotics, the side effects
associated with antibiotics, the high costs of antimicrobial drugs and the re-emergence of diseases are the key factors
that obstruct effective management of bacterial infections in many developing countries. Unfortunately, the future does
not look bright in the war against infectious diseases, as multi drug resistant strains of microbes continue to increase at
an alarming rate. In fact, multi drug resistant strains are adapting faster than the introduction of new, more potent
antibiotics. Consequently, this has raised the specter of untreatable microbial infections and adds urgency to the search
for new infection-fighting strategies. This necessitates greater efforts to discover new potent antibiotics. A potential
post-antibiotic era is threatening present and future medical advances.
The development of antibiotic resistance can be natural or acquired. Bacteria usually become resistant to antibiotics
by one of the following mechanisms (Fig. 1). Natural resistance is achieved by spontaneous gene mutation and acquired
resistance is through the transfer of DNA fragments like transposons from one bacterium to another. Bacteria gain
antibiotic resistance due to any of the four reasons namely: (I) by alteration of drug binding or target sites; by alteration
of membrane permeability; (II) direct destruction or in ability of the drug to enter in the cell; (III) by developing the
ability to produce multi resistance pumps, due to this pump drug drained out of the cells before drug can damage or kill
the cell or (IV) replacement of a sensitive pathway and acquisition of a new enzyme to replace a sensitive one [4].
In light of the recent emergence of bacteria which are resistant to multiple antimicrobial drugs causing a challenge
for the treatment of infections, the need to discover new antimicrobial substances for use in combating such
microorganisms becomes pertinent. Thus, there is a constant and urgent need to develop new antimicrobial drugs for the
treatment of infectious diseases from medicinal plants. Naturally occurring compounds have been and continue to be, an
important source of new leads for many pharmaceuticals and agrochemicals. In this context, there is a growing
popularity in nutraceutical products or functional foods.
The traditional and herbal medicines are known as essential resources of nutraceuticals which could provide a variety of
beneficial effects on human health. Nutraceuticals or functional foods are natural bioactive, chemical compounds that
have health promoting, disease preventing or medicinal properties. The secondary metabolites present in them give
them a specific medical benefit other than a purely nutritional. Nutraceuticals have thus dual role to play: as food and
therapeutic agent i.e. aids in prevention and/or treatment of disease and/or disorder. The other benefit is, being natural,
they have no side effects as other therapeutic agents. Nutraceuticals may range from single isolated nutrients, dietary
supplements, or secondary metabolites to genetically engineered designer foods [9]. There are many reports of
nutraceutical plant leaves that are effective as antimicrobials (Table 1).
©FORMATEX 2011
N, N- [28]
(Myrtaceae) P. morganii, Providencia sp.-1, Citrobactor sp.-2, C. freundii, Alcaligenes fecalis, Salmonella
dimethylformamide
typhimurium, Candida sp.-5, C. albicans-2, C. glabrata, C. tropicalis, C. apicola, Cryptococcus
neoformans, C. luteolus, Trichosporan beigelii, Aspergillus flavus, A. candidus A. niger
Rosmarinus officinalis
Crude EtOH/H2O Streptococcus mutans, S. salivarius, S. sobrinus, S. mitis, S. sanguinis, Enterococcus faecalis [29]
L. (Lamiaceae)
Rumex dentatus L. Staphylococcus aureus, S. Setubal, Bacillus subtilis, Micrococcus luteus, Eschericia coli, Pseudomonas
Hexane, methanol [30]
(Plygonaceae) picketii, Bordetella bronchiseptica
Rumex ecklonianus Bacillus cereus, Staphylococcus epidermidis, S. aureus, Micrococcus kristinae, Streptococcus pyogenes,
Acetone, methanol,
Meissner Escherichia coli, Salmonella pooni, Serratia marcescens, Pseudomonas aeruginosa, Klebsiella [31]
water
(Polygonaceae) pneumoniae
Escherichia coli, Proteus mirabilis, Pseudomonas aerugenosa, Salmonella typhi, S. paratyphi, Shigella
Tamarindus indica Water, acetone,
flexnerri, Staphylococcus aureus, Bacillus subtilis, Streptococcus pyogenes, Aspergillus flavus, A. [32]
Linn. (Fabaceae) ethanol
fumigatus, A. niger, Candida albicans
Thymus vulgaris L. Water, ethanol,
Pseudomonas aeruginosa [33]
(Lamiaceae) methanol
Ziziphus mauritiana Aqueous, Staphylococcus aureus, Pseudomonas aeruginosa, S. facecalis, Klebsiella pneumoniae, Escherichia coli,
[34]
Lam. (Rhamnaceae) methanol, saponins Enterobacter faecalis, Enterobacter faecium, Proteus mirabilis
Science against microbial pathogens: communicating current research and technological advances
1253
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A. Méndez-Vilas (Ed.)
Science against microbial pathogens: communicating current research and technological advances
______________________________________________________________________________
A. Méndez-Vilas (Ed.)
2. Objective
The purpose of this work was to investigate the potential of four Indian nutraceutical plant leaves as natural
antimicrobial agents. Therefore leaves of four plants viz. Manilkara zapota L. var. Royen., Psidium guajava L., Punica
granatum L. and Syzygium cumini L. were selected and their antimicrobial potential was explored against some
common pathogenic bacteria and fungi.
3. Experimental methodology
3.1 Chemicals
Petroleum ether, toluene, ethyl acetate, acetone, dimethyl sulphoxide (DMSO), Nutrient broth, Sabouraud dextrose
broth, Muller Hinton agar No. 2 and Sabouraud dextrose agar, agar powder were obtained from Hi-media or Merck. All
reagents used were of analytical grade.
Plant powder
Flow Chart. 1 Systematic representation of different solvent extracts of leaves by sequential extraction.
Manilkara zapota (L.) The seeds are aperients, diuretic, tonic and febrifuge. Bark is antibiotic, astringent and febrifuge. Fruits are
1 var. Royen. Chiku Sapotaceae edible, sweet with rich fine flavour. Chicle from bark is used in dental surgery. Bark is used as tonic and
(PSN429) the decoction is given in diarrhoea and peludism.
The roots are astringent, haemostatic, constipating, antiemetic. Leaves are astringent, anodyne, febrifuge,
antispasmodic, tonic. Flowers are cooling, laxative, tonic. Fruits are sweet, astringent, sour, cooling,
Psidium guajava L. aphrodisiac, laxative, tonic. Roots are used in haemorrhages, diarrhoea, dysentery, ulcers, gingivitis,
2 Jamphal Myrtaceae
(SU/BIO/510/Thakrar) proctoptisis, vomiting. Leaves are used in wounds, ulcers, cholera, diarrhea, vomiting, nephritis, cachexia,
vata, epilepsy, odontalgia, gum boils. Flowers are used in bronchitis, opthalmodynia, colic, ulemorrhagia.
Fruits are used in pitta, dipsia, burning sensation, colic, ulemorrhagia, diarrhoea, dysentery, debility.
The root and stem bark are astringent, cooling and anthelmintic. Flowers are stypic. Fruits are sweet, sour,
astringent, cooling, tonic, aphrodisiac, laxative, diuretic. Seeds are astringent, stomachic, diuretic,
©FORMATEX 2011
cardiotonic. Root and stem bark is used in tape worm infection, vomiting. Flowers are used in vomiting,
Punica granatum L.
3 Dadum Punicaceae pitta, ophalmodynia, ulcers, pharyngodynia, hydrocele. Fruits are used in anaemia, hyperdipsia,
(PSN311)
pharygodynia, opthalmodynia, pectoral diseases, splenopathy, bronchitis, otalgia. Fruit rind is used in
dysentery, diarrhoea and gastralgia. Seeds are used in vomiting, opthalmodynia, pitta, scabies,
hepatopathy, splenopathy.
The bark is astringent, sweet, sour, acrid, refrigerant, carminative, diuretic, digestive, anthelmintic,
febrifuge, constipating, stomachic and antibacterial. The fruits and seeds are sweet, acrid, sour, tonic and
Syzygium cumini L. cooling. The bark is useful in diabetes, leucorrhoea, stomachaigia, fever, gastropathy, strangury and
4 Jambu Myrtaceae
(PSN295) dermatopathy. The tender leaves are used for vomiting. The leaves are used for strengthening the teeth and
gums. The fruits and seeds are used in diabetes, diarrhoea, pharyngitis, splenopathy, urethrorrhoea and ring
worm.
Science against microbial pathogens: communicating current research and technological advances
1255
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A. Méndez-Vilas (Ed.)
Science against microbial pathogens: communicating current research and technological advances
______________________________________________________________________________
A. Méndez-Vilas (Ed.)
Fig. 2 Antimicrobial activity of five different solvent extracts of leaves of Manilkara zapota, Psidium guajava, Punica granatum and
Syzygium cumini.
5. Concluding remarks
The broad spectrum activity exhibited by four nutraceutical plant extracts, establishes the scientific basis of their use as
ethnomedicine and use their potential for development of novel antimicrobial agents effective for treatment of microbial
infectious diseases. Its usefulness in the formulation of antiseptics and disinfectants is also recommended if the active
principles can be isolated and purified. On the basis of the present findings, we conclude that amongst four nutraceutical
plants, P. guajava showed best antimicrobial activity. It might turn out to be a good candidate in the search for effective
and efficient antimicrobial agents.
Acknowledgements The authors thank Prof. S.P. Singh, Head, Department of Biosciences, Saurashtra University, Rajkot, Gujarat,
India for providing excellent research facilities. One of the authors, Mr. Mital Kaneria, is thankful to University Grants Commission,
New Delhi, India for providing financial support as Junior Research Fellow.
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