ANTIPROTOZOAL AND ANTIMICROBIAL ACTIVITY OF SELECTED

015Vol 4, Issue 05, 2 www.wjpps.com

1720

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

ANTIPROTOZOAL AND ANTIMICROBIAL ACTIVITY OF

SELECTED MEDICINAL PLANTS GROWING IN UPPER EGYPT,

BENI-SUEF REGION

Shimaa Mohammed Abdel Gawad1, 2

*, Mona Hafez Hetta1, Samir Anis Ross

2 and

Farid Abd El- Reheim Badria3

1Pharmacognosy Department, Faculty of Pharmacy, Beni-Suef University, 62514, Egypt.

2National Center for Natural Products Research, and Department of Bio Molecular Sciences,

School of Pharmacy, University of Mississippi, MS 38677, USA.

3Pharmacognosy Department, Faculty of Pharmacy, Mansoura University, 35516, Egypt.

ABSTRACT

Alcoholic extracts of fifty six plants cultivated in Beni-Suef

Governorate (Egypt) were screened in vitro for their antiprotozoal and

antimicrobial activities. Emblica officinalis, Quercus infectoria and

Punica granatum were the most active as antimalarial with IC50 4.92,

2.51, 10.61µg/mL respectively against the chloroquine sensitive strain

of Plasmodium falciparum and IC50 3.1, 2, 7.4µg/mL respectively

against chloroquine resistant strain of Plasmodium falciparum. The

extracts of Ricinus communi, Corchorus olitorius and Psidium guajava

were the most active as antileishmanial and the percentage of

inhibition were 91.4%, 90.9% and 90.3% respectively. The extract of

Emblica officinalis, Punica granatum, Quercus infectoria, Ricinus

communi, Tamarix nilotica, Camellia sinensis and Curcuma aromatic were active against

Candida glabrata with IC50 <8, <8, <8, 52.25, 17.12, 45.3, 26.91 µg/mL respectively while

the extract of Emblica officinalis, Quercus infectoria galls and Curcuma aromatica were the

most active against Cryptococcus neoformans with IC50 10.8, <8, 50.6 µg/mL respectively. A

good antibacterial activity against MRSA was shown by the ethanol extracts of Spinacia

oleracea, Corchorus olitorius, Cyperus alopecuroids, and Sesamum indicum with IC50 13.5,

45.31, 18.73 and 19.32µg/mL respectively. Tannins and phenolic acids constituents of these

plants proposed to be responsible for the activity through carbonic anhydrase inhibition.

WWOORRLLDD JJOOUURRNNAALL OOFF PPHHAARRMMAACCYY AANNDD PPHHAARRMMAACCEEUUTTIICCAALL SSCCIIEENNCCEESS

SSJJIIFF IImmppaacctt FFaaccttoorr 55..221100

VVoolluummee 44,, IIssssuuee 0055,, 11772200--11774400.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357

Article Received on 15 March 2015,

Revised on 06 April 2015,

Accepted on 27 April 2015

*Correspondence for

Author

Shimaa Mohammed

Abdel Gawad

Pharmacognosy

Department, Faculty of

Pharmacy, Beni-Suef

University, 62514, Egypt.


1721


KEYWORDS: Medicinal plants; Beni-Suef; Egypt; Antiprotozoal; Antimicrobial; Carbonic

anhydrase inhibitors.

INTRODUCTION

Natural products were used by humans in many countries such as Egypt, China and India as

drugs for thousands of years.[1]

It was reported in a recent study conducted by the information

and decision support centre in Egypt that 23% of the Egyptian use medicinal plants as a

remedy.[2]

There are more than 250,000 species of higher plants in the world among them

only a small percentage of them have been investigated for their potential value as drugs.[3]

There are several approaches for selecting plants as candidate for drug discovery such as

random selection followed by chemical or biological screening, follow up of ethnomedical

uses of plants and biological activity reports.[4,5]

There is no previous study could be traced on plants growing in Beni-Suef Governorate as

antiprotozoal and antimicrobial agents, therefore the present study focused on a systemic

evaluation of a selection of commonly growing plant species in this area.

Infectious diseases either protozoal such as malaria and leishmania or microbial infections are

considered as the major killing factors in the third world countries and the most important

causes of premature death.[6]

Difficulty of controlling the sources of infection, the high cost

of treatment/prevention, poor compliance, low efficacy, poor safety and drug resistance are

the major factors that may retard the treatment of these diseases. The drug resistance has

further complicated the treatment of infectious diseases in immune-compromised AIDS and

cancer patients. Therefore, there is always need for the development of new and more

effective drugs. In this respect, natural products offer good sources for new drug discovery.[6]

Various antiparasitic drugs have been developed from natural sources, including Quinine,

Artemisinine and Atovaquone as antimalarials and Amphotericin B as antileishmanial drug.[7]

The Centers for Disease Control and Prevention[8]

received information that 19 locally

transmitted malaria cases have been reported in one village in Aswan Governorate in Egypt.

These are the first malaria cases seen in Egypt since 1998.


1722


MATERIALS AND METHODS

Plant material

The fifty six plants used in this study (Table 1) were collected at the period from September

to December 2010 from different regions: Beni-Suef fields and herbal markets. The plants

were kindly authenticated by Prof. Dr. Lotfy Boules, Prof. Dr. Mohammed El Gibali, (senior

botanists, Faculty of Science, Cairo University), Dr. Mahmoud Omar (lecturer at plant

taxonomy Department, Faculty of Science, Beni-Suef University) and Mrs Thérèz Labib

(botanist specialist at Orman garden, Giza, Egypt). Voucher specimens (BUPD-35: BUPD-

90) were kept at the Herbarium of the Department of Pharmacognosy, Faculty of Pharmacy,

Beni-Suef University.

Preparation of extracts

The air-dried powder of each plant material (100 g) was extracted with 75% ethanol (500 mL

×3). Each extract was filtered off and concentrated using a rotatory evaporator. Each residue

was weighed and stored in a refrigerator at −4 °C until used.

Microbial strains used

Antimalarial activity was tested in vitro against Chloroquine sensitive (D6, Sierraleon)

and Chloroquine resistant (W2, Indo-China) strains of Plasmodium falciparum.

Antileishmanial activity was tested against Leishmania donovani promastigotes.

The used fungal strains were Candida albicans (ATCC 90028), Candida glabrata

(ATCC 90030), Candida krusei (ATCC 6258), and Aspergillus fumigates (ATCC 90906).

The used bacterial strains were methicillin-resistant Staphylococcus aureus (ATCC

33591), Cryptococcus neoformans (ATTC 90113), Staphylococcus aureus (ATTC

29213), Escherichia coli (ATCC 35218), Pseudomonas aeruginosa (ATCC 27853), and

Mycobacterium intracellulare (ATCC 23068).

Standards used

For the antimalarial activity, Chloroquine and Artemisine were used as positive controls

while Pentamidine and Amphotericin B were used as positive controls in the antileishmanial

testing. Amphotericin B was used also as a positive control in the antifungal activity and

Ciprofloxacine in the antibacterial. All reference drugs were obtained from (ICN

Biomedicals, Ohio).


1723


Biological Assays

Antimalarial screening[9]

Crude extracts were initially tested in duplicate against D6 Plasmodium falciparum strain as

a primary screen at 15.867 µg/mL and percent inhibition (% inhibition) was calculated

relative to negative and positive controls. The extracts which showed more than 50%

inhibition proceeded to the secondary assay. In the Secondary antimalarial assay, the samples

were dissolved to 20 mg/mL and tested at 47.6, 15.867, and 5.289 µg/mL and IC50s in µg/mL

were calculated against both D6 and W2 strains. In addition to P. falciparum strains, samples

were tested in the VERO mammalian cell line as an indicator of general cytotoxicity. The

selectivity indices (SI) – ratio of VERO IC50 to D6 or W2 IC50- were calculated. All IC50

were calculated using XLFit fit curve fitting software.

Antileishmanial screening[10, 11]

A culture of Leishmania donovani promastigotes was grown in RPMI 1640 medium

supplemented with 10% GIBCO fetal calf serum at 26°C. Growth of leishmanial

promastigotes was determined by the Alamar Blue assay (BioSource International, Camarillo,

CA). Standard fluorescence was measured by a Fluostar Galaxy plate reader (excitation

wavelength, 544 nm; emission wavelength, 590 nm). Crude extracts were initially tested in

duplicate as a primary screen at 80µg/ mL and % inhibitions were calculated.

Antifungal and antibacterial screening[9]

Crude extracts were initially tested in duplicate as a primary screen at 50µg/ mL and %

inhibitions were calculated relative to negative and positive controls. The extracts which

showed more than 50% inhibition were proceeded to a secondary assay. In the latter, the

samples were dissolved to 20mg/ mL and tested at 200, 40, 8µg/mL and then IC50 were

determined.

RESULTS

Antimalarial activity (Table 1)

Results showed full inhibition with Emblica officinalis and Quercus infectoria (100%

inhibition) followed by Punica granatum (96 % inhibition) with IC50 values of 4.92, 2.51,

10.61µg/mL respectively against chloroquine-sensitive (D6) strain of P. falciparum and IC50

values of 3.1, 2.0, 7.4 µg/mL respectively against chloroquine- resistant (W2) strain of P.

falciparum compared to Artemisine (IC50 value of 0.0269 µg/mL ; D6, 0.0165; W2) and


1724


Chloroquine (IC50 of 0.0098 µg/mL ; D6, 0.187; W2) without showing any cytotoxicity to

the mammalian cells (Table 2).

Antileishmanial activity (Table1)

Ricinus communi, Corchorus olitorius and Psidium guajava showed the best activity as

antileishmanial among the selected plants and their % of inhibition were 91.4%, 90.9% and

90.3% respectively.


1725


Table 1. Results of antimalarial and antileishmanial activity of selected plants collected from Beni-Suef Governorate, Egypt.

Plant Common name Family Part used Antimalarial D6

(%Inhibition)

Antileishmanial

(%Inhibition)

Alhagi graecorum Boiss Camel thorn, manna tree Papilionaceae Leaves 47 0

Amaranthus lividus L. Pigweed Amaranthaceae Herb 46 6.5

Anastatica hierochuntica L.

Dinosaur plant, Jericho rose, Mary’s

flower, Palestinian tumbleweed,

resurrection plant

Cruciferae Herb 44 0

Artemisia Absinthium L. Absinthe wormwood, green ginger Asteraceae Leaves ,

Flowers 52 25.1

Aster squamatous Sprengel Aster, Starwort Compositae Herb 45 52.4

Beta vulgaris var.cicla L. Spinach beet Chenopodiaceae Leaves 32 9.6

Camellia sinensis (L.)

Kuntze Green tea plant Theaceae Leaves 44 48.5

Cartagena ipecacuanha Brot. Cartagena Ipecacuanha, Rio ipecac Rubiaceae Root 70 84.9

Chenopodium murale L. Nettleleaf goosefoot Chenopodiaceae Herb 39 6.3

Cichorium endivia L. cultivated endive Asteraceae Leaves 44 0

Cichorium intybus L. Chicory Asteraceae Roots,

Leaves 42 0

Cinnamomum cassia (Nees

& T.Nees) Farw. Chinese cassia, Chinese cinnamon Lauraceae Bark 44 0

Citrus reticulate Blanco West African Cherry Orange,

Omuboro Rutaceae Leaves 33 75.1

Conyza dioscoridis (L.) Desf Horseweed, butterweed, fleabane Compositae Herb 38 60.2

Corchorus olitorius L. Jew's mallow Tiliaceae Leaves 37 90.9

Curcuma aromatic Salisb. Curcuma Zingebracea Rhizomes 52 44.1

Cymbopogon Proximus

Spreng. Halfa bar, Lemongrass Poaceae Leaves 47 6.1

Cyperus alopecuroides Foxtail flatsedge Cyperaceae Leaves, 28 63

http://en.wikipedia.org/wiki/Lauraceae


1726


Rottb. Flowers

Cyperus Rotundus L. Nut grass, Tiririca Cyperaceae Leaves,

Flowers 44 0

Daucus carota L. Wild carrot, Queen Anne's lace Apiaceae Leaves 41 0

Desmostachia bipinnata (L.)

Stapf

Halfa grass, big cordgrass, salt reed-

grass Poaceae Herb 44 0

Emblica officinalis (L.) Kurz Indian gooseberry Phyllanthaceae Herb 100 64.1

Eruca sativa Mill. Brassicaceae Leaves 33 0

Ficus carica L. Common fig Moraceae Leaves 36 0

Glycyrrhiza glabra L. Liquorice Fabaceae

Roots and

rhizomes 49 0

Hibiscus sabdariffa L. Roselle, karkadeh Malvaceae

Flowers

Calyx and

epi-calyx

34 0

Hyphaene thebaica (L.)

Mart. Doum palm Arecaceae Fruits 35 0

Lawsonia inermis L. Loose strife, Henna Lythraceae Leaves 0 0

Lupinus termis L. Lupine Fabaceae Seeds 0 0

Malva parviflora L. Cheeseweed Malvaceae Leaves 50 68.3

Mentha longifolia (L.) Huds. Horsemint Labiatae Herb 47 28.4

Morus alba L. White mulberry Moraceae Leaves 29 81.3

Opuntia ficus indica (L.)

Mill. Prickly pear Cactaceae Leaves 52 0

Origanum majorana L. Sweet marjoram Lamiaceae Herb 41 31.1

Peganum harmal L. Harmal, Syrian rue Nitrariaceae Seeds 70 83

Phaseolus vulgaris L. French bean Papilionaceae Leaves 41 79.3

Phragmites communis

(Cav.) Trin. ex Steud. Common reed Poaceae

Leaves ,

Flowers 0 0

Pimpinella anisum L. Aniseed Umbelliferae Fruits 44 10.1

Psidium guajava L. Guava Myrtaceae Leaves 50 90.3

Punica granatum L. Pomegranate Lythraceae Fruit 96 13.9

http://en.wikipedia.org/wiki/Fabaceae


1727


pericarp

Quercus infectoria gall Aleppo oak Fagaceae Galls 100 74.6

Ricinus communi L. Castor-oil plant Euphorbiaceae Leaves 40 91.4

Salix subserrata Willd. Salix Salicaceae Leaves 0 49.2

Sesamum indicum L. Sesame Pedaliaceae Leaves 0 75.9

Sesbania sesban (L.) Merr. Riverhemp Leguminosae Leaves ,

Flowers 0 0

Sisymbrium irio L. London rocket Brassicaceae Herb 3 0

Solenostemma argel (Delile)

Hayne Argel Apocyanaceae Leaves 43 1.7

Spinacia oleracea L. Spinach Chenopodiaceae Leaves 2 81

Tamarindus indica L. Tamarind Fabaceae Fruit and

Seeds 0 0

Tamarix nilotica L. Nile tamarisk Tamaricaceae Herb 8 0

Thymus vulgaris L. Common thyme Lamiaceae Herb 0 0

Tilia cordata Mill. Small leaved lime Tiliaceae Leaves ,

Flowers 3 31.6

Trifolium alexandrinum L. Egyptian clover Leguminosae Leaves 0 58.5

Withania somnifera (L.)

Dunal. Ashwagandha Solanaceae Leaves 15 35

Zingiber officinale Roscoe Ginger Zingebracea Rhizome 38 88.2

Zizyphus Spina-christi (L.)

Desf. Christ’s Thorn Jujube Rhamnaccae Leaves 0 24.9

Standard

Chloroquine 97 -

Amphotericin B - 98.7


1728


Table 2. IC50 ( µg/mL) of most active plants as anti-malarial

Plant Plasmodium falciparum (D6) D6 (SI) Plasmodium

falciparum (W2) W2 (SI) VERO

Emblica officinalis (L.)

Kurz 4.92 >9.7 3.1 >15.4 No cytotoxicity

Quercus infectoria gall 2.51 >19 2 >22.9 No cytotoxicity

Punica granatum L. 10.61 >4.5 7.4 >6.4 No cytotoxicity

Standard

Chloroquine 0.0098 >24.4 0.187 >1.3 No cytotoxicity

Artemisine 0.0269 >8.8 0.0165 >14.4 No cytotoxicity

IC50s were calculated based on 2 replicates, SI: selectivity index, D6: Chloroquine sensitive strainW2: Chloroquine resistant strain.


1729


Antimicrobial activity (Table 3)

The ethanol extract of Emblica officinalis, Punica granatum, Quercus infectoria, Ricinus

communi, Tamarix nilotica, Camellia sinensis and Curcuma aromatic were active against

Candida glabrata with IC50 values of <8, <8, <8, 52.25, 17.12, 45.3, 26.91 µg/mL

respectively when compared to Amphotericin B (IC50 value of 0.283 µg/mL) (Table 4).

The ethanol extract of Emblica officinalis, Quercus infectoria (galls) and Curcuma

aromatica were the most active against Cryptococcus neoformans with IC50 values of 10.8,

<8, 50.6µg/mL respectively when compared to Amphotericin B (IC50 Value of 0.269

µg/mL) (Table 4).

The ethanol extract of Spinacia oleracea, Corchorus olitorius, Cyperus alopecuroids and

Sesamum indicum were the most active against MRSA with IC50 values of 13.5, 45.31,

18.73 and 19.32µg/mL respectively when compared to Ciprofloxacin (IC50 value of

0.091µg/mL) (Table 4).

The ethanol extract of Quercus infectoria was active against E. coli with IC50 value of

40µg/mL when compared to Ciprofloxacin (IC50 value of 0.003µg/mL) (Table 4).

The ethanol extract of Quercus infectoria was the most active extract against P. aeruginosa

with IC50 = <8µg/mL when compared to Ciprofloxacin (IC50 = 0.092 µg/mL) (Table 4).


1730


Table 3. Results of antimicrobial activity (% inhibition) of selected plants collected from Beni-Suef Governorate, Egypt

Plant Fungi Bacteria

Ca Cg Ck Af Cn Sa MRSA Ec Pa Mi

Alhagi graecorum Boiss 0 0 0 0 58 1 0 4 0 4

Amaranthus lividus L. 0 1 0 1 59 2 0 4 0 0

Anastatica hierochuntica L. 0 8 0 5 56 0 0 17 3 0

Artemisia Absinthium L. 0 21 0 2 41 3 4 1 0 0

Aster squamatous Sprengel 0 7 0 3 17 14 9 9 0 0

Beta vulgaris var.cicla L. 0 1 0 3 7 0 0 3 0 2

Camellia sinensis (L.) Kuntze 0 95 0 12 15 16 31 30 8 0

Cartagena ipecacuanha Brot. 11 17 17 9 58 7 4 18 0 1

Chenopodium murale L. 0 0 0 14 60 6 2 11 0 0

Cichorium endivia L. 0 4 0 10 40 0 0 9 0 0

Cichorium intybus L. 0 13 1 8 42 0 0 16 0 0

Cinnamomum cassia (Nees & T.Nees)

Farw. 0 7 0 5 61 0 1 6 1 0

Citrus reticulate Blanco 0 9 0 10 29 4 10 4 0 0

Conyza dioscoridis (L.) Desf 0 10 0 8 24 0 7 14 0 0

Corchorus olitorius L. 1 15 0 10 33 34 77 0 0 0

Curcuma aromatic Salisb. 1 80 3 3 77 9 2 16 1 0

Cymbopogon Proximus Spreng. 1 11 3 9 10 7 1 10 3 0

Cyperus alopecuroides Rottb. 0 5 0 6 45 26 74 0 0 0

Cyperus Rotundus L. 11 5 1 11 57 7 2 12 1 0

Daucus carota L. 3 6 0 11 22 0 4 11 1 0

Desmostachia bipinnata (L.) Stapf 5 6 4 10 21 1 5 18 0 0

Emblica officinalis (L.) Kurz 0 100 25 12 99 24 19 42 13 0

Eruca sativa Mill. 2 3 0 10 14 0 0 23 0 0

Ficus carica L. 0 2 0 3 63 1 1 13 0 0

Glycyrrhiza glabra L. 1 7 3 5 34 21 12 21 0 0

Hibiscus sabdariffa L. 0 11 0 3 43 0 14 27 2 0


1731


Hyphaene thebaica (L.) Mart. 7 6 0 8 8 4 3 18 2 0

Lawsonia inermis L. 0 38 0 8 2 7 3 4 6 0

Lupinus termis L. 0 13 6 8 33 5 2 21 0 0

Malva parviflora L. 1 4 6 12 14 14 9 16 0 0

Mentha longifolia (L.) Huds. 1 6 0 10 7 4 6 25 0 0

Morus alba L. 8 12 4 13 48 12 14 17 0 0

Opuntia ficus indica (L.) Mill. 3 1 0 8 22 3 1 8 0 0

Origanum majorana L. 0 7 0 10 1 2 4 22 0 0

Peganum harmal L. 0 0 0 6 0 0 2 12 7 14

Phaseolus vulgaris L. 0 0 0 5 8 18 7 8 4 0

Phragmites communis

(Cav.) Trin. ex Steud. 0 0 0 5 6 0 1 0 2 3

Pimpinella anisum L. 0 6 0 3 51 0 0 12 3 0

Psidium guajava L. 4 18 0 14 24 40 36 5 3 0

Punica granatum L. 0 100 3 4 56 7 29 43 32 0

Quercus infectoria gall 0 100 62 5 99 40 36 53 74 0

Ricinus communi L. 0 100 0 5 64 20 47 0 0 0

Salix subserrata Willd. 0 0 0 5 9 12 22 17 0 3

Sesamum indicum L. 0 0 0 7 2 33 71 27 1 0

Sesbania sesban (L.) Merr. 0 6 0 4 2 0 1 9 2 0

Sisymbrium irio L. 0 0 0 6 2 0 0 8 4 1

Solenostemma argel (Delile) Hayne 0 0 0 5 24 0 0 6 0 0

Spinacia oleracea L. 0 2 0 3 5 10 80 18 7 4

Tamarindus indica L. 0 0 0 2 1 0 1 4 11 0

Tamarix nilotica L. 0 99 0 7 1 0 0 28 3 0

Thymus vulgaris L. 0 16 0 10 0 0 1 22 3 0

Tilia cordata Mill. 0 10 0 9 4 14 37 16 1 1

Trifolium alexandrinum L. 0 6 0 10 3 14 19 18 4 0

Withania somnifera (L.)Dunal. 0 5 0 7 3 0 6 13 1 0

Zingiber officinale Roscoe 6 22 13 11 41 2 8 10 0 0

Zizyphus Spina-christi (L.) Desf. 0 3 0 6 4 0 0 9 2 5


1732


Candida albicans (Ca), Candida glabrata (Cg), Candida krusei (Ck), and Aspergillus fumigates (Af) and the bacteria methicillin-resistant

Staphylococcus aureus (MRSA), Cryptococcus neoformans (Cn), Staphylococcus aureus (Sa), Escherichia coli (Ec), Pseudomonas aeruginosa

(Pa), and Mycobacterium intracellulare (Mi).

IC50s were calculated based on 2 replicates

Candida glabrata (Cg), Cryptococcus neoformans (Cn), methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli (Ec) and

Pseudomonas aeruginosa (Pa).

Standard

Amphotericin B (for fungi) 100 99 100 99 100 - - - - -

Ciprofloxacin (for bacteria) - - - - - 89 96 98 97 85

Table 4. IC50 (µg/mL) of some of the active plants in primary antimicrobial screening

Plant Fungi Bacteria

Cg Cn MRSA Ec Pa

Camellia sinensis 45.3 - - - -

Corchorus olitorius - - 45.31 - -

Curcuma aromatic 26.91 50.6 - - -

Cyperus alopecuroids - 18.73 - -

Emblica officinalis ˂ 8 10.8 - - -

Punica granatum ˂ 8 9.8 - - -

Quercus infectoria ˂ 8 ˂ 8 - 40 < 8

Ricinus communi 52.25 - - - -

Sesamum indicum - - 19.32 - -

Spinacia oleracea L. - - 13.5 - -

Tamarix nilotica 17.12 - - - -

Standards

Amphotericin B (for fungi) 0.283 0.269 - - -

Ciprofloxacin (for bacteria) - - 0.091 0.003 0.092


1733


DISCUSSION

Based on the activity and selectivity, twelve plant extracts among the tested plants could be

considered as promising and interesting to be further elaborated through purification and

biological evaluation on an individual compound basis. Camellia sinensis, Corchorus

olitorius, Curcuma aromatica, Cyperus alopecuroids, Emblica officinalis, Psidium guajava,

Punica granatum, Quercus infectoria galls, Ricinus communi, Sesamum indicum Spinacia

oleracea, and Tamarix nilotica.

Emblica officinalis, Quercus infectoria and Punica granatum were found to be the most active

antimalarial plants. The activities of Punica granatum[12]

and Emblica officinalis[13]

were

previously reported but it is the first report on the activity of Quercus infectoria galls as

antimalarial plant. Ricinus communi, Corchorus olitorius and Psidium guajava were reported

to have significant inhibitory effect as antileishmanial agents. The antileishmanial activity of

Ricinus communi was previously reported[14]

but it is the first report on the activity of

Corchorus olitorius and Psidium guajava as antileishmanial plants.

Reviewing the antifungal and antibacterial screening results; it could be assessed that the

activity of Quercus infectoria,[15]

Ricinus communi,[16]

Punica granatum,[16]

and Camellia

sinensis[17]

against C. glabrata were previously reported but it is the first report on the activity

of Emblica officinalis, Curcuma aromatic and Tamarix nilotica as antifungal agents against

this strain. Quercus infectoria was previously reported against Cryptococcus neoformans,[18]

but it is the first report on the activity of Emblica officinalis and Curcuma aromatic as

antifungal agents against Cryptococcus neoformans. Activity of Corchorus olitorius against

MRSA was previously reported[19]

but it is the first report on the activity of Spinacia oleracea,

Cyperus alopecuroids and Sesamum indicum. The activity of Quercus infectoria against E.

coli was previously reported.[20]

The activity of Quercus infectoria against P. aeruginosa was

previously reported.[21]

Emblica officinalis, Quercus infectoria galls and Punica granatum showed prominent activity

as antimicrobial and antiprotozoal against Plasmodium falciparum, Candida glabrata and

Cryptococcus neoformans.


1734


On studying the reported phytochemical constituents of these plants (Table 5) we noticed that

the major active constituents in most of them are tannins and phenolic acids and their esters

and these could be related to their biological activity.

In a trial to correlate the activity to the phytochemical constituents of these plants, carbonic

anhydrase inhibition activity was studied. It has been noticed that gallic and ellagic acids and

their derivatives are common constituents in the these plants.

Carbonic anhydrase inhibitors (CAIs) had many uses such as topically acting antiglaucoma,

anticonvulsants, antiobesity and antitumor agents. Thirteen catalytically active isoforms of CA

enzyme were present and belong to alpha, beta, gamma-, delta-, and zeta families and found in

many organisms.[22]

CA inhibition was studied ultimately for some pathogenic protozoa

(Plasmodium falsiparum) α-CA,[23]

fungi β-CA (Cryptococcus neoformans,[24]

Candida

albicans,[22]

Candida glabrata[25]

and some Sacharomyces cerevisiae,[22]

and bacteria α-, β-,

and/or γ-CA (Helicopacter pylori, Mycobacterium tuberculosis and Brucella suis), β-class

enzymes from E. coli and M. tuberculosis [26]

.

In addition to sulfonamides and sulfamate, novel chemotypes of CAIs as coumarins, phenols

and fullerenes were also recently reported.[22]

A series of phenolic acids and phenol natural

products, such as p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, ferulic acid, gallic

acid, syringic acid, quercetin, and ellagic acid showed inhibitory effects against the

metalloenzyme carbonic anhydrase against all isozymes.[27]

Ellagitannins as inhibitors against carbonic anhydrase have been previously isolated from the

pericarps of Punica granatum L.[28]


1735


Table 5. The reported active constituents of the active antiprotozoal and antimicrobial plants

Medicinal plant Active constituents

Camellia sinensis Catechin-based flavonoids such as epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG), epigallocatechin

(EGC) and epicatechin (EC).[17]

Corchorus olitorius Chlorogenic acid, 3, 5-dicaffeoylquinic acid, quercetin 3-galactoside, quercetin 3-glucoside, quercetin 3- (6-

malonylglucoside), and quercetin 3- (6 malonyl galactoside) and cardiac glycosides.[29, 30]

Curcuma aromatica Zederonecurdione, neocurdione, curcumol, tetramethyl pyrazine, 1, 2-hexadecanediol, 9-oxo-neoprocurcumenol),

neoprocurcumenol and curcumin.[31]

Cyperus alopecuroids Alopecuquinone (benzoquinone) [32]

.

Vicenin 2, orientin, diosmetin, quercetin 3, 3'-dimethyl ether and its 3, 4'-dimethyl ether (flavonoids).[32]

Emblica officinalis - Emblicanin A and B, punigluconin, pedunculagin (hydrolysable tannins).[33]

- Gallic acid, ellagic acid, chebulinic acid, chebulagic acid, emblicanin A, emblicanin B, punigluconin, pedunculagin,

citric acid, ellagotannin, trigallayl glucose, pectin, 1-O-galloyl-β-D-glucose, 3,6-di-O-galloyl-D-glucose, chebulagic acid,

corilagin, 1,6-di-O-galloyl-β-D-glucose, 3 ethylgallic acid (3 ethoxy 4,5 dihydroxy benzoic acid), and isostrictiniin.[33]

- Phyllantine and phyllantidine (alkaloids).[33]

Psidium guajava - Gallic acid, caffeic acid and ellagic acid (phenolic acids).[34]

- Guavanoic acid, guavacoumaric acid, 2 -hydroxyursolic acid, jacoumaric acid, asiatic acid and isoneriucoumaric acid

(triterpene acids).[35]

Punica granatum - Punicalagin, punicalin, gallic acid, ellagic acid and ellagic acid derivative such as ellagic acid,3, 3'-di-O-methyl,ellagic

acid,3,3',4'-tri-O-methyl,ellagicacid,3'-O-methyl-3,4-methylene (hydrolysable tannins).[36]

- Pedunculagin, punicacortein A–D, granatin A and B, punicafolin, punigluconin and corilagin (Phenolic compounds).[36]

- Quercetin-3-O-rutinoside (flavonoid).[36]

Quercus infectoria Tannic acid (gallotannic acid, the principal constituent 50-70%), gallic acid, syringic acid, ellagic acid,

hexagalloylglucose and polygalloylglucose.[11]

Ricinus communi - Ricinine (0.55%) and N-demethyl ricinine (0.016%) (Alkaloids) [37]

. Quercetin, rutin.[37]

- Six flavones glycosides.[37]

Gallic acid, gentisic acid, epicatechin and ellagic acid (phenolic compounds).[37]

Sesamum indicum Tannins, saponins, flavonoids, terpenes and cardiac glycosides as phytoconstituents.[38]

Spinacia oleracea L. - para-coumaric acid, ferulic acid and ortho- coumaric acid (phenolic compounds).[39]

- Querecetin; myricetin; kampeferol; apigenin; luteolin; patuletin; spinacetin (flavonoids).[39]

Tamarixnilotica Methyl ferulate 3-O-sulphate, coniferyl alcohol 4-O-sulphate , kaempferol 4′-methyl ether , tamarixetin and quercetin 3-

O-β-D-glucupyranuronide.[40]


1736


CONCLUSIONS

Natural products have been the most productive source of leads for new drugs. There is a need

for developing new antiprotozoal and antimicrobial agents from natural sources to overcome

resistance and safety problems of the currently existing drugs. As a part of our research

programme of investigating some medicinal plants cultivated in Beni-Suef Governorate

(Egypt) as antiprotozoal and antimicrobial agents, twelve plant extracts among the tested

plants could be considered as promising and interesting to be further elaborated through

purification and biological evaluation on an individual compound basis; Camellia sinensis,

Corchorus olitorius, Curcuma aromatica, Cyperus alopecuroids, Emblica officinalis, Psidium

guajava, Punica granatum, Quercus infectoria galls, Ricinus communi, Sesamum indicum

Spinacia oleracea, and Tamarix nilotica. The majority of studies still presented only

preliminary screening data and there is a need for further studies on the standardization or

chemical characterization of the extracts used and more description about the mechanisms of

action.

ACKNOWLEDGMENTS

This work was supported by research grants from the ParOwn program 2012 (Project No.

0911), Ministry of Higher Education at Egypt and in part from National Center for Natural

Products Research, School of Pharmacy, University of Mississippi.

We are grateful to the Egyptian Government, AI 27094 program from the NIH NCRR

(antifungal), 58-6408-1-603 program from USDA Agricultural Research Service Specific

Cooperative Agreement (antibacterial) and 58-6408-1-603 program from USDA Agricultural

Research Service Specific Cooperative Agreement (antimalarial and antileishmanial).

Thanks go to the National Center for Natural Products Research, also thanks to Dr. Melissa

Jacob, Dr. Babu Tekwani and Dr. Shabana Khan for carrying out the antimicrobial,

antileishmanial and antimalarial testing.

CONFLICT OF INTERESTS

The authors declare no conflict of interests.

REFERENCES

1. Balandrin N F KAD and Farnsworth N R. Human Medicinal Agents from Plants;. In A. D.

B. Kinghorn, N. F., Eds. (Ed.), ACS Symposium Series 534:1993, pp. 2-12.

2. AbouZid S F and Mohamed A A. Survey on medicinal plants and spices used in Beni-

Sueif, Upper Egypt. Journal of ethnobiology and ethnomedicine, 2011; 7(18): 1-6.‏


1737


3. Grifo F, Newman D J, Fairfield A S, Bhattacharya B, Grupenhoff J T. The Origin of

Prescription Drugs, Grifo F. and Rosenthal J. Eds., Island Press, Washington D.C: 1997, p

131.

4. Khafagi IK and Dewedar A. The efficiency of random versus ethno-directed research in

the evaluation of Sinai medicinal plants for bioactive compounds. Journal of

Ethnopharmacology, 2000; 71(3): 365-376.

5. Fabricant D S and Farnsworth N R. The value of plants used in traditional medicine for

drug discovery. Environmental health perspectives, 2001; 109: 69.

6. Singh M, Khatoon S, Singh S, Kumar V, Rawat A K, Mehrotra S. Antimicrobial screening

of ethnobotanically important stem bark of medicinal plants. Pharmacognosy Res., 2010;

2(4): 254.

7. Al-Musayeib N M, Mothana R A, 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; 17(10): 11379-11390.

8. Centers for Disease Control and Prevention (CDC), June 26, 2014,

http://wwwnc.cdc.gov/travel/notices/watch/malaria-in-egypt#fn1

9. Bharate SB, Khan SI, Yunus NA, Chauthe SK, Jacob MR, Tekwani BL, Khan IA, Singh

IP. Antiprotozoal and antimicrobial activities of O-alkylated and formylated

acylphloroglucinols. Bioorganic and Medicinal Chemistry, 2007; 15: 87-96.

10. Ma G, Khan S I, Jacob M R, Tekwani B L, Li Z, Pasco D S, Walker L A, Khan I A.

Antimicrobial and antileishmanial activities of hypocrellins A and B. Antimicrobial agents

and chemotherapy, 2004; 48(11): 4450-4452.

11. Hamid H, Kaur G, Abdullah S T, Ali M, Athar M, Alam M S. Two New Compounds from

the Galls of Quercus infectoria with nitric oxide and superoxide inhibiting ability.

Pharmaceutical Biology, 2005; 43(4): 317-323.

12. Ferreira D, Khan S, Jacope M, Gupta S, Reddy M. Antioxidant, antimalarial and

antimicrobial activities of tannin-rich fractions, ellagitannins and phenolic acids from

Punica granatum L. Planta Med., 2007; 73(5): 461.

13. Bagavan A, Rahuman A A, Kaushik N K, Sahal D. In vitro antimalarial activity of

medicinal plant extracts against Plasmodium falciparum. Parasitology research, 2011;

108(1): 15-22.

14. Zahir A A, Rahuman A A, Pakrashi S, Ghosh D, Bagavan A, Kamaraj C, et al. Evaluation

of antileishmanial activity of South Indian medicinal plants against Leishmania donovani.

Experimental Parasitology, 2012; 132(2): 180-184.

http://wwwnc.cdc.gov/travel/notices/watch/malaria-in-egypt#fn1


1738


15. Hassan H F. Study the Effect of Quercus infectoria Galls Extracts on Growth of Candida

albicans and Candida glabrata In Vitro Which Isolated from Vaginal Swabs. Iraqi J. Vet.

Med., 2011; 35(2): 85 – 94.

16. Jain P and Nafis G. Antifungal Activity and Phytochemical Analysis of Aqueous Extracts

of Ricinus communis and Punica granatum. Journal of Pharmacy Research, 2011-a; 4(1):

128.

17. Aladag H, Ercisli S, Yesil D Z, Gormez A, Yesil M. Antifungal activity of green tea leaves

(Camellia sinensis L.) sampled in different harvest time. Pharmacognosy Magazine, 2009;

5(20): 437.

18. Abu-Mejdad N M. Antifungal Activity of Some Plant Extracts Against Two Yeasts

Isolates In Vitro. Research journal of pharmaceutical, biological and chemical sciences,

2014; 5(2).

19. Ashidi Joseph Senu E M O, Odunbaku O A, Biliaminu S A. The in-vitro effect of

Corchorus olitorius (L.) on the antibacterial activities of five antibiotics Journal of

Microbiology, 2012; 2(2): 113-117.

20. Voravuthikunchai S P and Suwalak S. Antibacterial activities of semipurified fractions of

Quercus infectoria against enterohemorrhagic Escherichia coli O157: H7 and its

verocytotoxin production. Journal of Food Protection, 2008; 71(6): 1223-1227.

21. Darogha S N. Antibacterial activity of Quercus infectoria extracts against bacterial

isolated from wound infection. Journal of Kirkuk University – Scientific Studies, 2009;

4(1), 20-30.

22. Supuran C T. Carbonic anhydrase inhibitors. Bioorganic and Medicinal Chemistry Letters,

2010a; 20(12): 3467-3474.

23. Krungkrai J and Supuran C T. The alpha-carbonic anhydrase from the malaria parasite and

its inhibition. Current pharmaceutical design, 2008; 14(7): 631-640.

24. Mogensen E G, Janbon G, Chaloupka J, Steegborn C, Fu M S, Moyrand F d r, et al.

Cryptococcus neoformans senses CO2 through the carbonic anhydrase and the adenylyl

cyclase Cac1. Eukaryot Cell, 2006; 5(1): 103-111.

25. Supuran C T, Vullo D, Leewattanapasuk W, Mühlschlegel F A, Mastrolorenzo A, Capasso

C. Carbonic anhydrase inhibitors: Inhibition of the β-class enzyme from the pathogenic

yeast Candida glabrata with sulfonamides, sulfamates and sulfamides. Bioorg Med Chem

Lett., 2013; 23(9): 2647-2652.


1739


26. Supuran C. Inhibition of bacterial carbonic anhydrases and zinc proteases: from orphan

targets to innovative new antibiotic drugs. Current medicinal chemistry, 2012; 19(6): 831-

844.

27. Supuran C T, Innocenti A, Sarıkaya B Ö, Gülçin I. Carbonic anhydrase inhibitors.

Inhibition of mammalian isoforms I-XIV with a series of natural product polyphenols and

phenolic acids. Bioorganic and Medicinal Chemistry, 2010-b; 18(6): 2159-2164.

28. Satomi H, Ueno A, Hatano T, Okuda T, Noro T. Carbonic anhydrase inhibitors from the

pericarps of Punica granatum L. Biol. Pharm. Bull., 1993; 16 (8): 787-790.

29. Azuma K, Nakayama M, Koshioka M, Ippoushi K, Yamaguchi Y, Kohata K, et al.

Phenolic Antioxidants from the Leaves of Corchorus olitorius L. Journal of agricultural

and food chemistry, 1999; 47(10): 3963-3966.

30. Khan M S Y, Bano S, Javed K, Mueed M A. A comprehensive review on the chemistry

and pharmacology of Corchorus species-A source of cardiac glycosides, triterpenoids,

ionones, flavonoids, coumarins, steroids and some other compounds. Journal of the

Chemical Society, Perkin Transactions, 2006; 65(4): 283.

31. Ahmad S, Ali M, Ansari S H, Ahmed F. Phytoconstituents from the rhizomes of Curcuma

aromatica Salisb. Journal of Saudi Chemical Society, 2011; 15(3): 287-290.

32. Nassar M I, Abdel-Razik A F, El-Khrisy E D, Dawidar A M, Bystrom A, Mabry T J. A

benzoquinone and flavonoids from Cyperus alopecuroides. Phytochemistry, 2002; 60(4):

385-387.

33. Bhandari P R and Kamdod M A. Emblica officinalis (Amla): A review of potential

therapeutic applications. International Journal of Green Pharmacy, 2012; 6(4): 257.

34. Okuda T, Hatano T, Yazaki K. Guavin B, and ellagitannin of novel type. Chemical and

pharmaceutical bulletin, 1984; 32(9): 3787-3788.

35. Oliver-Bever B E P. Medicinal plants in tropical West Africa: Cambridge University

Press, 1986.

36. Jain V, Murugananthan G, Deepak M, Viswanatha G L, Manohar D. Isolation and

Standardization of Various Phytochemical Constituents from Methanolic Extracts of Fruit

Rinds of Punica granatum. Chinese Journal of Natural Medicines, 2011-b; 9(6): 414-420.

37. Jena J and Gupta A K. Ricinus communis Linn: a phytopharmacological review.

International Journal of Pharmacy and Pharmaceutical Sciences, 2012; 4(4): 25-29.

38. Okon J E and Umoh N S. Influence of Sesamum indicum L. ethanolic leaf extract on

haematological paramaters on albino rats in akwa ibom state, Nigeria. Indian Journal of

Fundamental and Applied Life Sciences, 2013; 3(1): 106-110.


1740


39. Subhash G P, Virbhadrappa S R, Vasant O K. Spinacia oleracea LINN: a

pharmacognostic and pharmacological overview. IJRAP, 2010; 1(1): 78-84.

40. Abouzid S F, Ali S A, Choudhary M I. A new ferulic acid ester and other constituents

from Tamarix nilotica leaves. Chemical and Pharmaceutical Bulletin, 2009; 57(7): 740-

742.

Documents

ANTIPROTOZOAL AND ANTIMICROBIAL ACTIVITY OF SELECTED