Anti-diarrheal activities of medicinal plants of Similipal ...openaccessscience.com/pdf-files/vol2_1_mar2012/IJMAP_2_1_17... · different species of Shigella, S. dysentriae is known

Int. J. Med. Arom. Plants, ISSN 2249 – 4340RESEARCH ARTICLE

Vol. 2, No. 1, pp. 123-134, March 2012

*Corresponding author: (E-mail) sk_dutta<A.T.>yahoo.com http://www.openaccessscience.com©2012 Open Access Science Research Publisher [email protected]

Anti-diarrheal activities of medicinal plants of Similipal Biosphere Re-

serve, Odisha, India

Sujogya Kumar PANDA1, Niranjan PATRA1, Gunanidhi SAHOO2, Akshya Kumar BASTIA3,

Sushil Kumar DUTTA2*

1Department of Biotechnology, North Orissa University; Baripada, India-757003

2Department of Zoology; North Orissa University; Baripada, India-757003

3Department of Botany; North Orissa University; Baripada, India-757003

*Corresponding Author, Fax: + Tel: +91-9437143598, Fax: +91-6792253908

Article History: Received 17th September 2011, Revised 23rd January 2012, Accepted 23rd January 2012.

Abstract: This paper deals with ethnomedicinal uses and anti-diarrheal properties of medicinal plants used by the tribalpeople of District Mayurbhanj, Odisha, India. Aqueous and methanol extracts of 72 plants were tested for antibacterialactivity using agar well diffusion (sample concentration of 100 mg/ml) against eight pathogenic bacteria responsible fordiarrheal diseases. The results indicated that out of 77 plants species, 47 species exhibited antibacterial activity againstone or more test organisms. Out of 168 extracts, 54 methanolic and 43 aqueous extracts expressed antibacterial proper-ties. Nineteen plants are newly reported to have ethnomedicinal uses to treat diarrheal diseases. Among these, Bombaxceiba, Buchanania lanzan, Butea superba, Coccinia grandis, Curculigo orchioides, Eleutherine bulbosa, Ficus racemo-sa, Flemingia nana, Helicteres isora, Lannea coromandelica, Mesua ferrea, Semecarpus anacardium and Smilax zeyla-nica experimentally proved to inhibit the diarrhea causing bacteria. Salmonella typhi, Escherichia coli and Vibrio cholerawere the most sensitive strain. Shigella dysentriae showed least activity compared to all other test strains.

Keywords: Anti-diarrheal; Similipal Biosphere Reserve; Preliminary screening; Plant extracts; Ethnomedicinal uses.

Introduction

Diarrhea, particularly infectious diarrhea isthe second leading cause of mortality and mor-bidity throughout the world in children less than5 yrs of age. This is especially true in develop-ing countries like India where there is poor sani-tation and overcrowding. Estimation of crudedeath rate due to diarrhea in India was 9.3 per1000 population (WHO 2005). If this continues,then the predicted case of burden will rise to126.35 cores during 2016 (WHO 2005). Amongthe leading causes of infectious diarrhea, Sal-monella and Shigella contributes highest num-ber. The current chemotherapeutical treatmentof Salmonellosis and Shigellosis is complicatedas a result of drug resistance. Moreover, majori-ty of the people in these developing countrieshave no access for modern health care facilities.This necessitated the search for alternative ther-apies such as, the use of medicinal plants. Shi-

gellosis is an important cause of diarrhealdeaths. It has been reported that not less than140 million cases of Shigellosis occur world-wide with 6,00,000 deaths annually, 60% ofsuch deaths are of under 5 years old children(Peirano et al. 2006; Sur et al. 2004). Among thedifferent species of Shigella, S. dysentriae isknown for its fatality, and life threatening situa-tion. The emergence of multiple drug resistantstrains of diarrheagenic pathogens has made thetreatment of dysentery more difficult hencethere is increasing interest in plants as source ofantimicrobial agents for the treatment of suchdiseases (Munshi et al. 1987; Monroe and Polk2000). Salmonellosis, another type of diarrhealdiseases is caused by a group of bacteria calledSalmonella. It is primarily transmitted throughingestion of contaminated food by infectedfaeces from man or animal, through fecal oralroute. Active cases of Salmonella in man are

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124Int. J. Med. Arom. Plants Anti-diarrheal plants of Similipal Biosphere Reserve

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source of contamination and transmission toother human beings and to lower animals.Strains of Salmonella species with resistance toantimicrobial drugs are now widespread in bothdeveloped and developing countries. In devel-oped countries, it is now increasingly acceptedthat for the most part such strains are zoonoticin origin and acquire their resistance in the food-animal host before onward transmission to hu-mans through the food chain. On the other hand,indiscriminate use of commercial antimicrobialdrugs commonly employed in the treatment ofinfectious diseases lead to high resistanceamong the strains. This has forced researchers tosearch for new antimicrobial substances fromvarious sources like the medicinal plants. Searchfor new antibacterial agents should be continuedby screening many plant families. Several stu-dies have identified that the compounds withinherbal plants are effective antibiotics (Basile etal. 2000). Traditional healing systems aroundthe world that utilize herbal remedies are an im-portant source for the discovery of new antibio-tics (Okpekon et al. 2004); some traditional re-medies have already been produced compoundsthat are effective against antibiotic-resistantstrains of bacteria (Kone et al. 2004).

Diarrheal disease continues to be a majorcause of morbidity and mortality throughout theworld, particularly among children in develop-ing countries, often as a result of infection bybacteria, viruses, and protozoan parasites. Giventhe increasing resistance in many common pa-thogens to currently used chemotherapeuticagents, there is renewed interest in the discoveryof novel compounds that can be used to fightinfectious diseases. There have been numerousstudies that have served to validate the tradi-tional use of medicinal plants used to treat orprevent diarrhea. Several methods can be usedfor selecting plants of potential therapeutic in-terest (Vlietinck and Vanden-Berghe 1991;Farnsworth 1993). The search can follow threemain routes: random, ethno (including ethnobo-tanical, ethnomedical and ethnopharmacologi-cal) and ecological search (Fabricant andFarnsworth 2001). The ethnobotanical and eth-nopharmacological approach uses informationobtained from traditional medical practitionersand other people such as village elders and localwomen who are traditional users of medicinal

plants. Ethnomedicinal reports are available onthe use of plants for the treatment of diarrheaand dysentery by the tribals from districtMayurbhanj (Rout and Pandey 2002; Pandeyand Rout 2002, 2006; Rout and Panda 2010;Panda et al. 2011a,b,c). There is limited infor-mation on the safety of traditional plant extracts,although some clinical trials have evaluated thesafety and tolerability of herbal medicine prepa-rations used to treat diarrhea and generally indi-cate that minimal side effects are observed.Based on earlier reports and data collected fromthe present ethnomedicinal uses, plants wereselected to find out the experimental validationof tribal knowledge for the curing of infectiousdiarrhea.

Materials and methods

Study area

The Similipal Biosphere Reserve (Figure 1),one of the mega biodiversity zones of the coun-try, is situated in the central part of the Mayurb-hanj district of Odisha (20o17' - 22o34'N and85o40' - 87o10'E) and covers an area of 5569Km2. The reserve has been divided into threezones i.e. Core Zone, Buffer Zone and Transi-tion Zone. The core zone (845Km2) is reservedfor wildlife habitat development and no exploi-tation activities are entertained in this area. Thebuffer zone (around 2,129 Km2) is partially pro-hibited and is allowed for some activities likeresearch, education and tourism. On the otherhand the transition zone, which lies in the peri-pheral region covering 2,595 Km2, is allowedfor research, tribal settlement, tourism and otherenvironment friendly activities.

Ethnomedicinal documentation

In Mayurbhanj district, phytotherapy (treat-ment with medicines from plant and their de-rived products) forms an integral part of the lo-cal culture, and the information about plants andtheir uses are passed from generation to genera-tion through oral folk-lore, primarily amongstthe elderly; the natural retainers of traditionalknowledge in their respective communities. Thefield study was carried out from September2006 to November 2008, and the information on





the use of medicinal plants for treatment of di-arrhea was obtained through structured ques-tionnaires, complemented by free interviews andinformal conversations (Huntington 2000). Theinterviews were individually carried out duringthe first contacts with the local population, na-tive specialists were identified, i.e. people con-sidered by the community as having exceptionalknowledge about the use of plants.

Figure 1: Map showing Similipal BiosphereReserve

Collection of medicinal plants

The present work is based on the explora-tions made in Similipal Biosphere Reserve dur-ing 2006-10. Field trips to Similipal BiosphereReserve were undertaken for collection of medi-cinal plants. Identification of these medicinalplants was done at the Post Graduate Depart-ment of Botany, North Orissa University, Bari-pada.

Processing

Stems, leaves, bulb, barks, roots, rhizomes,seeds etc. of plants have separately been col-

lected during field trips to different parts of Si-milipal Biosphere Reserve. The roots are dugout from the soil and the adhering soils wereremoved by shaking and washing. The leaveswere plucked from the trees, washed properlyand infected leaves were discarded. After col-lection, the healthy leaves were dried at lowtemperature to maintain their green color andvolatile oils, if present. The materials werecompletely shed dried so long it does not allowfor the growth of any type of fungi, molds, bac-teria and other microorganisms. The driedleaves, roots and stems were powdered separate-ly using mortar and pestle and then were passedthrough sieve to get the uniform powdered sam-ple.

Preparation of plant extracts

Twenty grams of each powdered sampleswere dissolved in 100ml of sterile distilled wa-ter and 80% methanol separately in wide mouthbottles. The aqueous samples were prepared byadding distilled water (steam for 30 minutes)and stored overnight. Similarly, methanol sam-ples were incubated at room temperature for 48hrs. The suspension was then filtered (WhatmanNo. 40) separately and made up to 100ml withrespective solvent and utilized for studying theirantimicrobial properties.

Test bacteria

Pathogenic bacteria under study wereEscherichia coli, Escherichia coli O157:H7,Salmonella typhimurium, Salmonella typhi, Shi-gella dysentriae, Vibrio alginolyticus, Vibriocholerae and Vibrio cholerae O139.

Maintenance of bacteria

The bacterial cultures were maintained onnutrient agar (NA) slants and stored at 4 оC. Ac-tivation of the bacteria was carried out bystreaking culture from the slants on to a MullerHinton agar (MHA) plate and incubating over-night at 37 оC. Single colony was picked upfrom each plate and transferred to nutrient broth,incubated for 1 day at 37 оC prior to the test.





Antibiotics

The following antibiotics disc (Hi mediaPvt. Ltd., Mumbai, India) at the given concen-tration were used to determine antibiotic sensi-tivity profile of reference bacteria: Amikacin-Ak (30μg); Amoxicillin-Aug (10μg); Amphote-ricin-Ap (100unit); Ampicillin-A (10μg); Baci-tracin-B (10units); Cefoxitin-Ctn (10μg); Cef-triaxone-Cez (10μg); Cephotaxime-Ce (30μg);Chloroamphinecol-Ch (10μg); Ciprofloxacin-C(10μg); Erythromycin-E (15μg); Gatifloxacin-Gf (30μg); Gentamycin-G (10μg); Levofloxa-cin-Lvx (5μg) ; Naladixic acid-Nal (30μg);Ofloxacin-Ofl (5μg); Polymyxin-B-Pb (300unit); Streptomycin-St (10μg); Tetracycline-Te(10μg) and Vancomycin-Vn (30 μg).

Sensitivity testAntibiogram was done by disc diffusion me-

thod (Bauer et al. 1966) with commonly used

antibiotics. Antibiotic sensitivity was tested inMHA plates. The test microbes were removedfrom the slant aseptically with inoculating loopsand transferred to separate test tubes containing5.0ml of sterile distilled water. Inoculums addeduntil the turbidity equaled 0.5 McFarland (108

CFU/ml). For each of the bacteria, one milliliterof the test tube suspension was added to 15-20ml of nutrient agar and transferred to the agarplate (90 mm diameter). After cooling the in-oculated agars at room temperature for 25 min,antibiotic sensitivity test discs were placed onthe surface of solid agar. The plates were incu-bated at 37°C. The plates were examined the-reafter, clear zones of inhibition formed aroundthe discs were measured and antibiotic sensitivi-ty was assayed from the diameter of the clearinhibition zones (in mm) (Figure 2).

Figure 2: Antibiogram among strains

Screening of antimicrobial properties by agarcup method

The agar cup method was followed to studyantibacterial activity of the extracts. Overnight

Muller Hinton Broth culture of the test organ-isms were firmly seeded over the MHA plates.Wells of 6 mm diameter was punched over theagar plates using a sterile borer. The bottoms of





the wells were sealed by pouring 50-100 µl ofmolten MHA into the scooped out wells. 100 µlof extracts were poured into the wells. The wa-ter was allowed to evaporate and the plates wereincubated at 37 °C for 18-24 hours. Presence ofzone of clearance around the wells, confirmedthe antibacterial activity of the extracts.

Results

The antidiarrheal screening of 77 plants col-lected from Similipal Biosphere Reserve, Odi-

sha, India was carried out by agar cup method.The selection of plants was based on ethnome-dicinal uses reported earlier as well as freshlyrecorded data (Table 1). Eight (8) different spe-cies of bacteria that causes infectious diarrheawere selected for screening. Two different sol-vents viz. methanolic (80%) and aqueous usedfor screening the crude extracts of different spe-cies (Table 2). The plants have been listed inalphabetical order of their families. The botani-cal names of the plants were given along withparts used in the screening investigation.

Table 1: Ethnomedicinal uses of medicinal plants from district Mayurbhanj, Odisha, India

Botanical name Family Partsused

Cure for diseases References

Acacia leucophloea (Roxb.)Willd. Mimosaceae Bk Diarrhea Panda et al. (2011a)Achyranthes aspera L. Amaranthaceae Wp Dysentery Panda et al. (2011a)

Rout and Panda (2010)Mohanta et al. (2006)

Acorus calamus L. Araceae Rh Diarrhea Panda et al. (2011a)Adhatoda vasica Nees Acanthaceae Lf Diarrhea Rout and Panda (2010)Aegle marmelos L. Rutaceae Lf, Fr Diarrhea Rout and Panda (2010)Allium cepa L. Liliaceae Bl Diarrhea, Dysen-

teryRout and Panda (2010)

Andrographis paniculata(Burm. f.) Wall. ex Nees

Acanthaceae Lf, St Dysentery Rout and Panda (2010)

Angiopteris evecta Forst. Hoff Angiopteridaceae Lf Dysentery Panda et al. (2011a)Annona reticulata L. Annonaceae Lf Diarrhea Padhi et al. (2011)Annona squamosa L. Annonaceae Lf Diarrhea Padhi et al. (2011)Anogeissus latifolia (Roxb. ex DC.) Wall. exBedd

Combretaceae Lf Diarrhea Panda et al. (2011a)Mohanta et al. (2006)

Bauhinia vahlii W. & A. Caesalpiniaceae Bk Dysentery Panda et al. (2011a)Bombax ceiba L. Bombacaceae Gum Diarrhea New reportBryophyllum calycinum Salis. Crassulaceae Lf Dysentry New reportBuchanania lanzan Spreng. Anacardiaceae Lf Diarrhea New reportButea monosperma (Lam.) Taub. Fabaceae Rt, Sd Dysentery Rout and Panda (2010)Butea superba Roxb. Fabaceae Rt Dysentery New reportCalotropis gigantean L. Asclepiadaceae Rt Dysentery New reportCareya arborea Roxb. Lecythidaceae Bk Blood Dysentery Mohanta et al. (2006)

Rout and Panda (2010)Cassia fistula L. Caesalpinaceae Lf Dysentery Panda et al. (2011c)Catharanthus roseus (L.) G. Don Apocynaceae Lf Dysentery Rout and Panda (2010)Cissampelos pareira L. Menispermaceae Rt Diarrhea Rout and Panda (2010)Citrus limon L. Rutaceae Fr Diarrhea Panda et al. (2011a); Rout and

Panda (2010)Clausena excavata Burm. Rutaceae Rt, Lf Diarrhea, Dysen-

teryPanda et al. (2011a)Mohanta et al. (2006)

Coccinia grandis (L.) Voigt Cucurbitaceae Lf Diarrhea, Dysen-tery

New report

Crotalaria spectabilis Roth Fabaceae Rt Dysentery Rout and Thatoi (2009)Croton roxburghii Balak. Euphorbiaceae Lf Dysentery Panda et al. (2010)Curculigo orchioides Gaertn. Hypoxidaceae RT Dysentery New reportCurcuma angustifolia Roxb. Zingiberaceae Rh Dysentery Panda et al. (2011a)

Mohanta et al. (2006)Cynodon dactylon (L.) Pers. Poaceae Wp Diarrhea Panda et al. (2011a)Diospyros malabarica (Desr.) Kostel Ebenaceae Bk Diarrhea Panda et al. (2011a)





Diospyros melanoxylon Roxb. Ebenaceae Lf, Bk Diarrhea, Dysen-tery

Panda et al. (2009)

Eleutherine bulbosa (Miller) Urban Iridaceae Bl Dysentery New reportFicus racemosa L. Moraceae Bk Dysentery New reportFlemingia nana Roxb. Fabaceae Rt Diarrhea, Dysen-

teryNew report

Glycyrrhiza glabra L. Fabaceae Bk Diarrhea Rout and Panda (2010)Grewia hirsute Vahl. Tiliaceae Lf Dysentery New reportHelicteres isora L. Sterculiaceae Rt Diarrhea, Dysen-

teryNew report

Hibiscus rosasinensis L. Malvaceae Tw Dysentery Panda et al. (2011a)Mohanta et al. (2006)

Holarrhena pubescens (Buch.-Ham.) Wallex. G.

Apocyanaceae Lf Dysentery Panda et al. (2011a)Mohanta et al. (2006)

Indigofera cassioides Rottl. ex DC. Fabaceae Rt, Fl Dysentery Panda et al. (2011a)Mohanta et al. (2006)

Kalanchoe pinnata (Lam.) Pers. Crassulaceae Lf Dysentery Panda et al. (2011a)Mohanta et al. (2006)

Lannea coromandelica Houtt. Anacardiaceae Lf Diarrhea, Dysen-tery

New report

Lygodium flexuosum L. Lygodiaceae Wp Blood Dysentery Panda et al. (2011a)Lygodium microphyllum (cav.)R.Br. Lygodiaceae Lf Dysentery Panda et al. (2011a)Mangifera indica L. Anacardiaceae Bk Blood Dysentery Panda et al. (2011a)

Rout and Panda (2010)Mesua ferrea L. Clusiaceae Lf Diarrhea Dysen-

teryNew report

Momordica charantia L. Dipterocarpaceae Bk Diarrhea Dysen-tery

Mohanta et al. (2006)

Morinda citrifolia L. Rubiaceae Rt Dysentery Panda et al. (2011a)Moringa oleifera Lam. Moringaceae Lf Diarrhea Mohanta et al. (2006)Musa paradisica L. Musaceae Fr Dysentery Mohanta et al. (2006)

Panda (2010)Nicotiana tabacum L. Solanaceae Wp Diarrhea Rout and Panda (2010)Nyctanthes arbortristis L. Oleaceae Bk Dysentery Panda et al. (2011b)

Rout and Panda (2010)Oxalis corniculata L. Oxalidaceae Wp Diarrhea, Dysen-

teryNew report

Paederia foetida L. Rubiaceae Lf Diarrhea Panda et al. (2011b)Phyllanthus fraternus Webster Euphorbiaceae Wp Diarrhea New reportPiper nigrum L. Piperaceae Sd, Fr Diarrhea Rout and Panda (2010)Plumbago zeylanica L. Plumbaginaceae Rt Diarrhea Rout and Panda (2010)Psidium guava L. Myrtaceae Lf, Fr Diarrhea Rout and Panda (2010)Pterocarpus marsupium Roxb. Dipterocarpaceae Bk Blood Dysentery Panda et al. (2011)

Mohanta et al. (2006)Rout and Panda (2010)

Pterospermum acerifolium (L.) Willd. Sterculiaceae Bk Diarrhea Dysen-tery

Panda and Dutta (2011)

Punica granatum L. Punicaceae Lf Chronic Dysen-tery

Pandey and Rout (2002)

Randia uliginosa (Retz.) DC Rubiaceae Fr Dysentery New reportSchleichera oleosa (Lour.) Sapendaceae Wp Diarrhea Panda et al. (2011a)Semecarpus anacardium L.f. Anacardiaceae Fr Diarrhea New reportShorea robusta Gaertn. F. Dipterocarpaceae Bk Dysentery Mohanta et al. (2006)Smilax zeylanica L. Smilacaceae Rt Diarrhea New reportSoymida febrifuga (Roxb.) A. Juss. Melastomataceae Bk Dysentery Rout and Panda (2010)Spondias pinnata (L.f.) Kurz Anacardiaceae Lf, Fr, Bk Dysentery Panda et al. (2011a)

Mohanta et al. (2006)Syzygium cumuni L. Myrtaceae Bk Dysentery Mohanta et al. (2006)Tamarindus indica L. Caesalpiniaceae Lf, Sd Diarrhea Rout and Panda (2010)Terminalia alata Heyne ex. Roth Combretaceae Bk Diarrhea Mohanta et al. (2006)Terminalia arjuna (Roxb. ex DC.) W. & A. Combretaceae Bk Chronic Dysen- Pandey and Rout (2002); Rout





tery and Panda (2010)Terminalia chebula Retz. Combretaceae Bk Diarrhea Rout and Panda (2010)Tragia involucrate L. Euphorbiaceae Wp Diarrhea Rout et al. (2009)Vitex negundo L. Verbenaceae Lf Diarrhea Mohanta et al. (2006)Woodfordia fruticosa (L.) Kurz Lythraceae Rt Dysentery Panda et al. (2011a)

Rout and Thatoi (2009)

A total of 168 plant extracts belonging to 43families were tested for anti-diarrheal activity(Table 2). Among the selected 77 plants, 47species showed anti-bacterial activity against atleast two or more test organisms. Twelve fami-lies did not record any activity against any ofthe test organism. Two families with highernumber of plants didn’t show any activity be-longed to Crasulaceae and Euphorbiaceae.Acanthaceae, Anacardiaceae, Ceasalpiniaceae,Clusiaceae, Combretaceae, Moringaceae, Olea-ceae, Punicaceae and Verbenaceae were someof the important families which showed anti-diarrheal activity against pathogens. Screeningresults showed antibacterial activity with 54 me-thanol and 43 aqueous extracts against one ormore test strains.

Table 2: Preliminary screening of plantsagainst diarrhea causing bacteria

Plant description PU E S1 S2 S3 S4 S5 S6 S7 S8AcanthaceaeAdhatoda vasica Nees Lf A 12 - 11 - - 12 12 12

M 11 12 10 12 - 11 10 10Andrographis paniculata Lf A 11 - - 11 12 12 12 10(Burm. f.) Wall. ex Nees M 14 - - - - - - 13

St A 12 12 - - - - - 12M 13 12 10 12 12 12 10 -

AmaranthaceaeAchyranthes aspera L. Wp A - - 9 - - - - 11

M - 10 8 - - - - 12AnacardiaceaeBuchanania lanzan Lf A - - - - - - - -Spreng. M 10 - - 12 12 14 - 13Lannea coromandelica Lf A 12 - - 10 - - 10 9(Houtt.) Merr. M 14 12 - 14 - 12 10 14

Mangifera indica L. Lf A - - - - - - - -M - - - - - - - -

Semecarpus anacardium Fr A - 13 - - - 12 - 12L.f. M 12 11 - - 14 12 - 13Spondias pinnata Lf A - - - - - - - -(L.f.) Kurz M 12 - 13 - - - - 12

Fr A - - - - - - - -M - - - - - - - -

AngiopteridaceaeAngiopteris evecta Forst.Hoff

Lf A - - - - - - - -

M - - - - - - - -AnnonaceaeAnnona reticulata L. Lf A - - 12 - - - - -

M 12 - 12 - - - 12 -Annona squamosa L. Lf A 12 - - - - - - -

M 11 - - - - 11 12 -ApocynaceaeCatharanthus roseus Lf A - - - - - - - -(L.) G. Don M - - - - - - - -Holarrhena pubescens Lf A - 11 - 11 12 11 - 14(Buch.-Ham.) Wall ex. G. M - 10 12 12 - - - 14

AraceaeAcorus calamus L. Rh A - - 09 - - - 12 -

M - - 12 12 - - 14 11AsclepiadaceaeCalotropis gigantean L. Lf A - - - - - - - -

M - - - - - - - -BombacaceaeBombax ceiba L. Gu

mA - - - - - - - -

M 10 - - - - - 12 -CaesalpiniaceaeBauhinia vahlii W. & A. Lf A - - - - - - - -

M - - - - - - - -Cassia fistula L. Lf A 9 12 12 11 12 12 11 9

M 13 12 13 12 12 12 10 12Tamarindus indica L. Lf A - - 12 - - - - 10

M 12 - 14 - - - - 12ClusiaceaeMesua ferrea L. Lf A 12 10 - - 12 - 12 12

M 10 12 - - 14 - 12 10CombretaceaeAnogeissus latifolia(Roxb.

Lf A 10 - - - 12 12 12 12

ex DC.) Wall. ex Bedd M 10 14 11 12 15 14 12 12Terminalia alata Lf A - - - - - - - -Heyne ex. Roth M - - - - - - - -Terminalia arjuna Bk A 10 12 12 12 12 - 14 12(Roxb. ex DC.) W. & A. M 12 15 14 12 12 12 15 12Terminalia chebula Retz. Bk A - - - 14 12 - - -

M - - - 12 11 - - -CrassulaceaeBryophyllum calycinum Lf A - - - - - - - -Salis. M - - - - - - - -Kalanchoe pinnata Lf A - - - - - - - -(Lam.) Pers. M 12 - - - - - - -CucurbitaceaeCoccinia grandis Lf A 12 11 - - - - - 11(L.) Voigt M - 14 - - - - - 12Momordica charantia L. Lf A - 12 - - - - 12 -

M - 14 - - - - - -DipterocarpaceaeShorea robusta Gaertn. f. Lf A - - 11 - 12 - - -

M 11 - 13 - 9 - - 12EbenaceaeDiospyros malabarica Bk A - - - - - - - -(Desr.) Kostel M 10 - - - - - - 12Diospyros melanoxylon Lf A - - - - - - - 11Roxb. M 14 - - - - - - 12

Bk A - - 11 11 10 - - 12M 12 15 10 10 10 - - 16

EuphorbiaceaeCroton roxburghii Lf A 17 - 13 - - - 12 -Balak. M 14 10 10 12 14 - 15 15

Bk A - - - 10 - - - 14M - - - - - - - 15

Phyllanthus fraternus Lf A - - - - - - - -Webster M - - - - - - - -Tragia involucrate L. Wp A - - - - - - - -

M - - - - - - - -FabaceaeButea monosperma Lf A - - - - - - - -(Lam.) Taub. M - - - - - - - -

Fl A 10 - 11 - - - - 12M 12 - 12 10 - - - 14

Butea superba Roxb. Lf A 10 10 - - - - - 10M 10 12 - - - - - -

Crotalaria spectabilisRoth

Lf A - - - - - - - -

M - - - - - - - -Flemingia nana Roxb. Rt A - - 10 - - - - -

M 10 - 12 - - - - 10Glycyrrhiza glabra (L.) Bk A - - 10 - - - - -

M - - 18 - - - - -Indigofera cassioides Lf A - - - - - - - -Rottl. ex DC. M - 10 10 8 - - - 10





Pterocarpus marsupium Bk A - 10 - - - - - 10Roxb. M 10 12 - - 9 - - 12HypoxidaceaeCurculigo orchioides Rt A - - - - - - - -Gaertn. M - - - - - - - -IridaceaeEleutherine bulbosa Bl A - 14 - - - - - 17(Miller) Urban M 12 15 - - - - 11 15LecythidaceaeCareya arborea Roxb. Lf A - - - 10 - - - -

M 12 - 12 12 - 15 - 14LiliaceaeAllium cepa L. Bl A - - - - - - - -

M - - - - - - - -LygodiaceaeLygodium flexuosum L. Wp A - - - - - - - -

M - - - - - - - -Lygodium microphyllum Lf A - - - - - - - -(cav.)R.Br. M - - - - - - - -LythraceaeWoodfordia fruticosa Lf A - - - - - - - -(L.) Kurz M - - - - - - - -MalvaceaeHibiscus rosasinensis L. Lf A - - - - - - - -

M - - - - - - - -MelastomataceaeSoymida febrifuga Lf A - - - - - - - -(Roxb.) A. Juss. M - - - - - - - -MenispermaceaeCissampelos pareira L. Rt A - - - - - - - 12

M 12 - - 14 - 10 10 14MimosaceaeAcacia leucophloea Lf A - 10 - - - - - 9(Roxb.)Willd. M 10 12 10 - - - 12 14MoraceaeFicus racemosa L. Lf A 12 - - - - - - -

M - - 14 - - - - 12MoringaceaeMoringa oleifera Lam. Lf A 12 10 8 - - - - -

M 14 11 12 - - - 10 12MusaceaeMusa paradisiaca L. Fl A - - - - - - - -

M - - - - - - - -MyrtaceaePsidium guajava L. Lf A - - - - 10 - - 12

M - - - - 13 - - 14Syzygium cumunis L. Bk A - - - - - - - -

M - - - - - - - -OleaceaeNyctanthes arbortristis L. Lf A 12 - 10 - - 10 - 12

M 20 - 13 10 - 12 - 10Bk A 12 10 10 12 - 12 - 14

M 23 10 18 12 12 11OxalidaceaeOxalis corniculata L. Wp A - - - - - - - -

M - - - - - - - -PiperaceaePiper nigrum L. Lf A - - - - - - - -

M - - - - - - - -PlumbaginaceaePlumbago zeylanica L. Lf A - - - - - - - -

M - - - - - - - -Cynodon dactylon Wp A - - - - - - - -(L.) Pers. M - - - - - - - -PunicaceaePunica granatum L. Lf A 10 10 14 - 10 - - 12

M - - 12 - 12 - - 10RubiaceaeMorinda citrifolia L. Lf A - - - - - - - -

M - - - - - - - -Paederia foetida L. Lf A - - - - - - - -

M - - - - - - - -Randia uliginosa Lf A - - - - - - - -(Retz.) DC M - - - - - - - -RutaceaeAegle marmelos L. Lf A - - - - - - - -

M - - - - - - - -Fr A - - - - - - - -

M - - 12 - 10 - - 12Citrus limon L. Fr A - - - - - - - -

M - - - - - - - -Clausena excavata Burm. Lf A - - - - - - - 14

M 14 - - - - - - 12

SapendaceaeScehleichera oleosa Lour. Wp A - - - - - - - -

M - - - - - - - -SmilacaceaeSmilax zeylanica L. Lf A - - - - - - - -

M - 12 - - 11 - 10 12SolanaceaeNicotiana tabacum L. Lf A - - - - - - - -

M - - - - - - - -SterculiaceaeHelicteres isora L. Lf A 11 - - 10 - - - 12

M - - - 8 - - - 10Fr A - - - - - - - -

M 11 - - - - - - 12Rt A 12 12 - 12 - - - 14

M 14 10 - 10 - 10 - 12Pterospermum acerifolium Lf A 10 10 - - - 10 - 12(L.) Willd. M 10 12 - 19 - 18 - 15TiliaceaeGrewia hirsute Vahl. Lf A - - - - - - - -

M - - - - - - - -VerbenaceaeVitex negundo L. Lf A 10 10 - - - - - -

M 12 10 14 10 - - - 10Bk A 10 - 10 - - - - 12

M 18 - 16 - - - - 17ZingiberaceaeCurcuma angustifolia Lf A - - - - - - - -Roxb. M - - 8 - - - - 10

PU-Parts used, E-Extract, A-Aqueous, M-Methanol, Bl-bulb; Bk-bark; Fl-flower; Fr-fruit; Lf-leaf; Rt-root; Rh-rhizome; Sd-seed; Tw-twig; Wp-whole plant, S1- Esche-richia coli, S2- Salmonella typhimurium, S3- Vibrio cho-lerae, S4- Vibrio alginolyticus, S5- Vibrio cholerae 0139,S6- Escherichia coli O157:H7, S7- Shigella dysentriae,S8- Salmonella typhi

Some of the active species have alreadybeen reported earlier to have antibacterial activi-ty. The effects of few medicinal plants viz. Jus-ticia adhatoda, Achyranthes aspera, Androgra-phis paniculata, Holarrhena antidysenterica,Cassia fistula, Carica papaya, Punica grana-tum, Moringa oleafera, Vitex negundo, Hemi-desmus Spondias pinnata were previously de-scribed (Valsaraj et al. 1997) and was confirmedin this work. Similar results were described byseveral authors by adding plants such as Achy-ranthes aspera, Annona reticulate, Annona squ-amosa, Cassia fistula, Diospyros melanoxylon,Holarrhena pubescens, Moringa oleifera, Me-sua ferrea, Punica grantum, Semecarpus ana-cardium, Tamarindus indica, Terminalia arjunaetc. (Ahmad et al. 1998; PerumalSamy et al.1998,1999; Jeevan Ram et al. 2004; Prashant-kumar et al. 2006; Parekh and Chanda 2008).





Figure 3: Agar cup method showing zone of inhibition against V. cholerae and S. typhi

Discussion

Diarrhea is known to be caused by severalfactors including the non infectious type of di-arrhea such as metabolic diseases, food allergyand other organic causes. On the contrary, infec-tious diarrhea is a type of diarrhea which iscaused by an infectious agent (bacteria, fungus,parasites and viruses) due to the invasion andcolonization of the host tissue. Infectious diarr-hea is characterized by an alteration in a normalbowel movement, an increase in the volume ofwater content, or frequency of stools, nausea,vomiting, cramps and/or abdominal discomfort(Sood and Pacheco 2002). Studies on pathogensresponsible for acute diarrhea in developingcountries, revealed that the contribution of Ro-tavirus (15-25%), entrerotoxigenic E. coli (10-20%), Shigella species (5-15%) Salmonella spe-cies (1-5%), C. jejuni (10-15%) and entropatho-genic E. coli (1-5%) (EHNRI 2002). E. coli(EIEC) enterinvassive and enterohemorragic(EHEC), Salmonella, Shigella,V. cholerae arethe major bacterial pathogens most often re-sponsible for causing pandemic and epidemicinfectious diarrheal disease in developing coun-tries (Black et al. 1984).

In the present study 168 extracts (both me-thanol and aqueous) such as 54x2 (leaf) extracts,4×2 (whole plant), 9×2 (bark), 1 (rhizome), 4×2(root), 5×2 (fruit), 1×2 (gum), 1×2 (stem), 2x2

(flower) and 1×2 (twig) extracts were testedagainst eight infectious bacteria. Among the 77plants, 47 species showed anti-bacterial activityagainst at least two or more test organisms.Twelve families did not show any activityagainst any of the test organism. Some of im-portant families with anti-diarrheal activityagainst pathogens are Acanthaceae, Anacardia-ceae, Ceasalpiniaceae, Clusiaceae, Combreta-ceae, Moringaceae, Oleaceae, Punicaceae andVerbenaceae. Screening results showed antibac-terial activity with 54 methanol and 43 aqueousextracts against one or more test strains. Solventor the extraction agent used in the preparation ofphytopharmaceuticals must be suitable for dis-solving the important therapeutic drug constitu-ents. In addition, solvents used should be easy toremove, inert, nontoxic, and not easily flamma-ble. The aqueous extracts have commonly beenused in preliminary studies. Methanol efficientlypenetrates cell membranes, permitting the ex-traction of high amounts of endocellular com-ponents in contrast to the solvents with lowerpolarity. Such solvents are limited for extractingmostly extracellular material. Hence, methanolchiefly dissolves polar constituents togetherwith medium and low polarity compounds ex-tracted by co-solubilzation. So, the aqueous andmethanolic (80%) extracts of different plant be-longing to a wide range of families are selectedfor screening based on random sampling and





observed ethnomedicinal uses. Plants viz. Ano-geissus latifolia, Buchanania lanzan, Eleuthe-rine bulbosa, Helicteres isora, Lannea coro-mandelica, Moringa oleifera, Nyctanthes arbor-tristis, Pterospermum acerifolium are newrecords for antimicrobial activity from Indianscenario. In the present study, 19 plants arenewly reported to have ethnomedicinal uses forthe treatment of diarrheal diseases. Amongthese, 13 plants viz. Bombax ceiba, Buchananialanzan, Butea superba, Coccinia grandi, Curcu-ligo orchioides, Eleutherine bulbosa, Ficus ra-cemosa, Flemingia nana, Helicteres isora, Lan-nea coromandelica, Mesua ferrea, Semecarpusanacardium and Smilax zeylanica were able toinhibit diarrhea causing bacteria. Among the teststrains the most sensitive strains were recordedin decreasing order were S. typhi followed by E.coli, Vibrio cholerae, S. typhimurium, V. algino-lyticus, E. coli O157:H7 and V. cholerae O139.However, the strain S. dysentriae showed leastactivity in compared to above referred strains.

Nigella satavia (Ranunculaceae), used totreat numerous ailments including diarrhea andits essential oil has been shown to exhibit activi-ty against Staphylococcus aureus, Salmonella,Shigella, V. cholerae, and E. coli (Ali et al.2003). Swertia corymbose shows antibacterialactivity against a wide range of microorganisms,(E. coli, Salmonella sp., V. cholerae and Staphy-lococcus aureus) that cause diarrhea. Vijaya etal. (1997) reported 10 Indian traditional plants(Allium sativum, Bauhinia racemosa, Camelliasinensis, Euphorbia hirta, Cissampelos pareira,Acorus calamus, Psidium guajava and Sphae-ranthus indicus) used to treat dysentery and di-arrhea with soaring antibacterial activity. Me-thanol and water extracts of a number of medi-cinal plants used to treat dysentery and diarrheain the Democratic Republic of Congo showedactivity against one or more enteropathogens,including Shigella, Salmonella, E. coli, Vibrioand Campylobacter species (Longanga-Otshudiet al. 1999). Diehl et al. (2004) evaluated 60 tra-ditionally used plants in human or veterinarymedicine to treat worm infections (worms ingeneral, round worms, Guinea worms, or flat-worms), diarrhea, dysentery and abdominalpain. Our present study correlates all theseabove cited research.

Conclusion

The present research highlights the scientificinvestigation of use of plants by tribe of Simi-lipal to treat diarrheal diseases. With respect totraditional medicines used to treat diarrheal dis-eases, such plants will continue to be used aslong as there are communities with limitedaccess to modern therapies. In future, it may bepossible to supplement conventional ORStreatment with plant extracts resulting in com-plementary treatments that may lead to a reduc-tion in the length of disease symptoms. Undoub-tedly, the evidence provided in this paper, en-courages further investigation in the study ofisolation of active component with in vivo tocure diarrheal diseases.

Acknowledgements: The authors are grateful tothe authorities of North Orissa University forproviding necessary facilities to carry out thisresearch. We are thankful to Dr. Anil K. Biswalfor identifying the plants and Mr. Bikash C. Be-hera, Mr. Santanu Ku. Jena, Mr. Yugal KishoreMohanta (all from North Orissa University) forhelp in collecting plant samples.

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Anti-diarrheal activities of medicinal plants of Similipal ...openaccessscience.com/pdf-files/vol2_1_mar2012/IJMAP_2_1_17... · different species of Shigella, S. dysentriae is known