tribe from Tamil Nadu, India Veeramuthu Duraipandiyan , Muniappan Ayyanar and Savarimuthu Ignacimuthu Entomology Research Institute, Loyola College, Chennai 600 034, India author email corresponding author email BMC Complementary and Alternative Medicine 2006, !35doi!0"!!#6$!4%2& 6##2&6&35 'he electronic (ersion o) this article is the complete one and can *e )ound online at http$$+++"*iomedcentral"com$!4%2&6##2$6$35 Recei(ed # ,une 2006 -ccepted !% .cto*er 2006 /u*lished!% .cto*er 2006 0 2006 1uraipandiyan et al2 licensee 3io4ed Central Ltd" 'his is an .pen -ccess article distri*uted under the terms o) the Creati(e Commons -ttri*ution License 5http$$creati(ecommons"org$licenses$*y$2"06, +hich permits unrestricted use, distri*ution, and reproduction in any medium, pro(ided the original +or7 is properly cited" Abstract "ac#ground -ntimicro*ial acti(ity o) !# ethnomedicinal plant e8tracts +ere e(aluated against nine *acterial strains 5Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Ervinia sp, Proteus vulgaris6 and one )ungal strain 5Candida albicans6" 'he collected ethnomedicinal plants +ere used in )ol7 medicine in the treatment o) s7in diseases, (enereal diseases, respiratory pro*lems and ner(ous disorders" Methods /lants +ere collected )rom /alni hills o) 9outhern :estern ;hats and the ethno*otanical data +ere gathered )rom traditional healers +ho inha*it the study area" 'he he8ane and methanol e8tracts +ere o*tained *y cold percolation method and the antimicro*ial acti(ity +as )ound using paper disc di))usion method" -ll microorganisms +ere o*tained )rom Christian 4edical College, <ellore, 'amil =adu, India" $esults 'he results indicated that out o) !# plants, !0 plants e8hi*ited antimicro*ial acti(ity against one or more o) the tested microorganisms at three di))erent concentrations o) !"25, 2"5 and 5 mg$disc" -mong the plants tested, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Punica granatum and Syygium lineare +ere most acti(e" 'he highest anti)ungal acti(ity +as e8hi*ited *y methanol e8tract o) Peltophorum pterocarpum and Punica granatum against Candida albicans" %onclusion 'his study e(aluated the antimicro*ial acti(ity o) the some ethnomedicinal plants used in )ol7loric medicine" Compared to he8ane e8tract, methanol e8tract sho+ed signi)icant acti(ity against tested organisms" 'his study also sho+ed that Toddalia asiatica, Syygium lineare, Acalypha fruticosa and Peltophorum pterocarpum could *e potential sources o) ne+ antimicro*ial agents" "ac#ground -ccording to :orld >ealth .rgani?ation 5:>.6 more than #0@ o) the +orldAs population relies on traditional medicine )or their primary healthcare needs" Bse o) her*al medicines in -sia represents a long history o) human interactions +ith the en(ironment" /lants used )or traditional medicine contain a +ide range o) su*stances that can *e used to treat chronic as +ell as in)ectious diseases" - (ast 7no+ledge o) ho+ to use the plants against di))erent illnesses may *e e8pected to ha(e accumulated in areas +here the use o) plants is still o) great importance C!D" 'he medicinal (alue o) plants lies in some chemical su*stances that produce a de)inite physiological action on the human *ody" 'he most important o) these *ioacti(e compounds o) plants are al7aloids, )la(anoids, tannins and phenolic compounds C2D" Rural communities, in particular paliyar tri*es, depend on plant resources mainly )or her*al medicines, )ood, )orage, construction o) d+ellings, ma7ing household implements, sleeping mats, and )or )ire and shade" 'he use o) medicinal plants as traditional medicines is +ell 7no+n in rural areas o) many de(eloping countries C3,4D" 'raditional healers claim that their medicine is cheaper and more e))ecti(e than modern medicine" In de(eloping countries, lo+&income people such as )armers, people o) small isolate (illages and nati(e communities use )ol7 medicine )or the treatment o) common in)ections C5D" :e chose eighteen plant species used in )ol7 medicine to determine their antimicro*ial acti(ity 5'a*le !6" In general, these plants are used in )ol7 medicine in the treatment o) s7in diseases, (enereal diseases, respiratory pro*lems and ner(ous disorders" /roperties o) the collected plants are also pro(ided in same ta*le" E(idently, there are not many scienti)ic studies that con)irm the antimicro*ial properties )or most o) the plants collected )or this study" 'he phytochemical research *ased on ethnopharmacological in)ormations is generally considered an e))ecti(e approach in the disco(ery o) ne+ anti&in)ecti(e agents )rom higher plants C6D" Table &' Bses and properties o) ethnomedicinal plants collected )or antimicro*ial screening 'here are a )e+ reports on the use o) plants in traditional healing *y either tri*al people or indigenous communities o) 'amil =adu C%&!!D" 'he de(elopment o) drug resistance in human pathogens against commonly used anti*iotics has necessitated a search )or ne+ antimicro*ial su*stances )rom other sources including plants C!2D" 9creening o) medicinal plants )or antimicro*ial acti(ities and phytochemicals is important )or )inding potential ne+ compounds )or therapeutic use" 'his paper reports the results o) a sur(ey that +as done *ased on )ol7 uses *y traditional practitioners in /alni hills o) 'amil =adu along +ith *ioassay test )or antimicro*ial acti(ity" Methods (thnobotanical survey /lants +ere selected )or this study *ased on their medicinal use" Eresh plant parts +ere collected )rom the tri*al (illages 5/aliyar tri*e6 in /alni hills o) 'amil =adu, India in ,an -pril 2005" 'he tri*al (illages +ere appro8imately lie *et+een !0F !2A !0F !5A = longitudes and %%F 26A %%F 33A latitude" 'he ethno*otanical data 5local name, mode o) preparation, medicinal uses6 +ere collected through Guestionnaire, inter(ie+s and discussions among the tri*al practitioners in their local language 5'amil6" 'he (oucher specimens in duplicate +ere deposited in the her*arium o) Entomology Research Institute, Loyola College, Chennai 5India6" Preparation of he)ane and methanol e)tract /lant e8tracts +ere prepared *y cold percolation method" 'he plant materials +ere dried under shade and ground into )ine po+der using electric *lender" 50 g o) dried po+der +as soa7ed in 300 ml he8ane )or 4# hours +ith intermittent sha7ing" 'he plant e8tracts +ere )iltered through :hatman =o" ! )ilter paper into pill (ials" 'he )iltrates +ere dried until a constant dry +eight o) each e8tract +as o*tained" 'he residues +ere stored at 4FC )or )urther use" 'he remaining plant residue +as dried and soa7ed in 300 ml o) methanol as a*o(e and the e8tract +as collected as descri*ed earlier" Antimicrobial screening 'he he8ane and methanol e8tracts o) !# plants +ere screened against a total o) H *acterial strains and one )ungal strain" 'he test organisms +ere Bacillus subtilis 5-'CC 44!6, Staphylococcus aureus 5-'CC 25H236, Staphylococcus epidermidis 54'CC 36!56, Enterococcus faecalis 5-'CC 2H2!26, Escherichia coli 5-'CC 25H226, Klebsiella pneumonia 5-'CC !53#06,Pseudomonas aeruginosa 5-'CC 2%#536, Ervinia sp 54'CC 2%606, Proteus vulgaris 54'CC !%%!6 and one )ungal strain Candida albicans 54'CC 22%6 o*tained )rom the Christian 4edical College, <ellore, 'amil =adu" Preparation of inoculum 9toc7 cultures +ere maintained at 4FC on slopes o) nutrient agar" -cti(e cultures )or e8periments +ere prepared *y trans)erring a loop)ul o) cells )rom the stoc7 cultures to test tu*es o) 4ueller&>inton *roth 54>36 )or *acteria and 9a*ouraud de8trose *roth 59136 )or )ungi that +ere incu*ated +ithout agitation )or 24 hrs at 3%FC and 25FC respecti(ely" 'he cultures +ere diluted +ith )resh 4ueller&>inton and 9a*ouraud de8trose *roth to achie(e optical densities corresponding to 2"0I!0 6 colony )orming units 5CEB$ml6 )or *acteria and 2"0I!0 5 spore$ml )or )ungal strains" Antimicrobial susceptibility test 'he disc di))usion method C!3D +as used to screen the antimicro*ial acti(ity" In (itro antimicro*ial acti(ity +as screened *y using 4ueller >inton -gar 54>-6 o*tained )rom >imedia 54um*ai6" 'he 4>- plates +ere prepared *y pouring !5 ml o) molten media into sterile petriplates" 'he plates +ere allo+ed to solidi)y )or 5 minutes and 0"! @ inoculum suspension +as s+a**ed uni)ormly and the inoculum +as allo+ed to dry )or 5 minutes" 'he di))erent concentrations o) e8tracts 5!"25, 2"5 and 5 mg$disc6 +ere loaded on 6 mm sterile disc" 'he loaded disc +as placed on the sur)ace o) medium and the compound +as allo+ed to di))use )or 5 minutes and the plates +ere 7ept )or incu*ation at 3%FC )or 24 hrs" -t the end o) incu*ation, inhi*ition ?ones )ormed around the disc +ere measured +ith transparent ruler in millimeter" 'he same procedure +as )ollo+ed )or the )ungus also" 'hese studies +ere per)ormed in triplicate" $esults and discussions 'a*le ! pro(ides the *otanical name, )amily, local name, plant parts used together +ith their traditional therapeutic uses and properties )or the !# ethnomedicinal plants collected )rom /alni hills o) 'amil =adu" .ut o) the !# plants tested )or antimicro*ial acti(ity, !0 plant species sho+ed antimicro*ial acti(ity *y inhi*iting one or more microorganisms" 'he results o) the antimicro*ial screening o) the crude e8tracts o) all species o) plants are sho+n in 'a*le 2" -mong the plants screened, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Syygium cumini and Syygium lineare sho+ed promising acti(ity against tested microorganisms" 'he tested plant e8tracts +ere most acti(e against gram& positi(e microorganisms than gram&negati(e microorganisms" 'his is in agreement +ith pre(ious reports *y the se(eral +or7ers C!4&!HD" Table *' -ntimicro*ial acti(ity o) the he8ane and methanol e8tracts o) collected ethnomedicinal plants 4ethanol e8tracts e8hi*ited a higher degree o) antimicro*ial acti(ity as compared +ith he8ane e8tracts" 4ethanol e8tracts o) Albiia procera,Cassia auriculata 5lea(es and )lo+ers6,Peltophorum pterocarpum, Punica granatum and Syygium cumini sho+ed acti(ity" Punica granatum possesed 25@ o) tannin C20D and the anti*acterial acti(ity may *e indicati(e o) the presence o) some meta*olic to8ins or *road&spectrum anti*iotic compounds" -lso ethanolic e8tract o) Punica granatum +as most acti(e against E! coli" /rasanth et al C2!D reported that, di))erent e8tracts o) Punica granatum )ruit sho+ed some anti*acterial acti(ity against P!vulgaris and B!subtilis" RaJa7aruna et al C22D reported that Syygium cumini sho+ed good acti(ity against Staphylococcus aureus and Bacillus subtilis" 3oth he8ane and methanol e8tracts o) Syygium lineare and Toddalia asiatica sho+ed antimicro*ial acti(ity" 'he essential oils )rom the lea(es o) Toddalia asiatica +ere most acti(e against E! coli, K! pneumoniae, P! aeruginosa and S! aureus C23D" Peltophorum pterocarpum and Syigium lineare had the highest inhi*itory acti(ity against *oth gram positi(e and gram negati(e *acteria" .n the other hand, Cassia alata sho+ed only slight acti(ity against *acteria such as S! aureus and B" subtilis" In the pre(ious )indings lea(es, )lo+ers, root and stem *ar7s o) Cassia alata sho+ed a range o) acti(ity against se(eral *acteria and proto?oa C24D" In this study methanol e8tract o) lea(es o) Cassia alata sho+ed anti*acterial acti(ity against Bacillus subtilis andStaphylococcus aureus" 9omchit et al C25D also tested the +hole plant parts o) Cassia alata and sho+ed acti(ity in the lea(es against Staphylococcus aureus" Syygium lineare, Punica granatum, Syygium cumini and Toddalia asiatica produced the largest ?ones o) inhi*ition against Bacillus subtilis, Staphylococcus aureus and Staphylococcus epidermidis" <ora(uthi7unchai et al C20D reported good anti*acterial acti(ity in P! pterocarpum and P! granatum against Escherichia coli using aGueous and methanol e8tracts" 4ethanol e8tracts o) Peltophorum pterocarpum and Punica granatum sho+ed acti(ity against Candida albicans" In general, among the tested micro*ial strains, *acteria +ere )ound to *e more sensiti(e to many o) the test agents than )ungi" 'he most sensiti(e *acterium +as Bacillus subtilis, +hich +as inhi*ited *y methanol or he8ane o) !0 plants" .n the other hand, no inhi*ition +as o*ser(ed in the Eruvinia sps" 9ome organisms e8hi*ited only slight suscepti*ility" E! coli +as inhi*ited *y methanol e8tract o) )lo+ers o) Cassia auriculata and he8ane e8tract o) Punica granatum" Proteus vulgaris +as inhi*ited *y methanol e8tract o) Peltophorum pterocarpum and Syigium lineare" Klebsiella pneumonia +as inhi*ited *y he8ane e8tract o) "la# scandens, methanol e8tracts o) Peltophorum pterocarpum andSyigium cumini" %onclusion 'he processing o) the plants per)ormed in this study +as not compara*le to the traditional approach +hen the /aliyar tri*e used +ater )or e8tracts +hereas +e ha(e used he8ane and methanol )or e8traction" In this sense it is not an e8act replication o) the traditional 7no+ledge" -ll the same, gi(en that methanol e8tracts +ere more e))ecti(e then he8ane e8tract, it is li7ely that +ater e8tracts +ere +ill *e e))ecti(e as +ell and possi*ly mort so" 'he anti*acterial acti(ity o) Syigium lineare, "la# scandens, Albiia procera and Acalypha fruticosa are reported )or the )irst time" =o pre(ious report on the anti*acterial acti(ity o) these species could *e )ound in the literature" -mong the medicinal plants tested in this +or7, Peltophorum pterocarpum and Punica granatum sho+ed the most promising antimicro*ial properties indicating the potential )or disco(ery o) anti*acterial principles" Eurther phytochemical studies are reGuired to determine the types o) compounds responsi*le )or the anti*acterial e))ects o) these species" 'he results also indicate that scienti)ic studies carried out on medicinal plants ha(ing traditional claims o) e))ecti(eness might +arrant )ruit)ul results" 9e(eral plants used *y /aliyar tri*e e8hi*it some degree o) anti*acterial acti(ity to+ards gram&positi(e *acteria such as, Bacillus subtilis, Staphylococcus aureus and Staphylococcus epidermidis" 'hese plants could ser(e as use)ul sources )or ne+ antimicro*ial agents" %ompeting interests 'he author5s6 declare that they ha(e no competing interests" Authors+ contributions <1 has carried out the e8perimental part such as e8traction, inoculum preparation and antimicro*ial e(aluation" 4- collected the ethnomedicinal plants )rom the /alni hills o) 'amil =adu +ith the help o) tri*al practitioners" 9I super(ised the +or7, e(aluated the results and corrected the manuscript )or pu*lication" -ll authors read and appro(ed the )inal manuscript" Ac#no,ledgements 'he authors are than7)ul to the tri*al people )or sharing their 7no+ledge on plants" $eferences !" 1iallo 1, >(eem 3, 4ahmoud 4-, 3etge ;, /aulsen 39, 4aiga - An ethnobotanical survey of herbal drugs of -ourma district, Mali' Pharmaceutical Biology !HHH, ./!#0&H!" 2" Edeoga >., .7+u 1E, 4*ae*ie 3. Phytochemical constituents of some Nigerian medicinal plants' African $ournal of Biotechnology 2005, 0!6#5&6##" 3" 9andhu 19, >einrich 4 The use of health foods, spices and other botanicals in the Si#h community in 1ondon' Phytotherapy %esearch 2005, &2!633&42" /u*4ed -*stract K /u*lisher Eull 'e8t 4" ;upta 4/, 9olis /=, Calderon -I, ;uionneau&9inclair E, Correa 4, ;aldames C, ;uerra C, Espinosa -, -l(enda ;I, Ro*les ;, .campo R Medical ethnobotany of the Teribes of "ocas del Toro, Panama' $ournal of Ethnopharmacology 2005, 2!3#H&40!" /u*4ed -*stract K /u*lisher Eull 'e8t 5" RoJas ,,, .choa <,, .campo 9-, 4uLo? ,E Screening for antimicrobial activity of ten medicinal plants used in %olombian fol#loric medicine! A possible alternative in the treatment of non3 nosocomial infections' Chttp$$+++"*iomedcentral"com$!4%2& 6##2$6$2D +e*cite BMC Compelmentary and Alternative Medicine 2006, !2" 3io4ed Central Eull 'e8t 6" Mlouce7 /, /olesny N, 9(o*odo(a 3, <l7o(a E, Mo7os7a L Antibacterial screening of some Peruvian medicinal plants used in %aller4a District' $ournal of Ethnopharmacology 2005, 22!30H&3!2" /u*4ed -*stract K /u*lisher Eull 'e8t %" 9andhya 3, 'homas 9, Isa*el :, 9hen*agarathai R (thnomedicinal plants used by the valaiyan community of Piranmalai hills 5$eserved forest6, Tamil Nadu, India 7 A pilot study' African &ournal of Traditional Complementary and Alternative Medicines 2006, .!!0!&!!4" #" -yyanar 4, Ignacimuthu 9 Traditional 8no,ledge of 8ani tribals in 8outhalai of Tirunelveli hills, Tamil Nadu, India' $ournal of Ethnopharmacology 2005, &9*!246&255" /u*4ed -*stract K /u*lisher Eull 'e8t H" RaJan 9, 9ethuraman 4, 4u7herJee /M (thnobiology of the Nilgiri :ills, India' Phytotherapy ' %esearch 2002, &!H#&!!6" /u*lisher Eull 'e8t !0" =ataraJan 3, /aulsen 39, /ushpangadan / An (thnopharmacological Study from the %oimbatore District, Tamil Nadu, India! Traditional 8no,ledge %ompared ;ith Modern "iological Science' Pharmaceutical Biology !HHH, ./!3%#&3H0" !!" Ignacimuthu 9, 9an7arasi(araman M, Mesa(an L Medico3 ethnobotanical survey among 8ani#ar tribals of Mundanthurai Sanctuary' (itoterapia !HH#, 2!40H&4!4" !2" Erdogrul .' Antibacterial activities of some plant e)tracts used in fol# medicine' Pharmaceutical Biology 2002, 09!26H&2%3" !3" 3auer R:, Mir*y 41M, 9herris ,C, 'urc7 4 Antibiotic susceptibility testing by standard single disc diffusion method' American $ournal of Clinical Pathology !H66, 0<!4H3&4H6" /u*4ed -*stract !4" 3u+a L<, 9taden ,< Antibacterial and antifungal activity of traditional medicinal plants used against venereal diseases in South Africa' $ournal of Ethnopharmacology 2006, &9.!!3H&!42" /u*4ed -*stract K /u*lisher Eull 'e8t !5" <alsaraJ R, /ushpangadan /, 9mitt B:, -dsersen -, =yman B Antimicrobial screening of selected medicinal plants from India' $ournal of Ethnopharmacology !HH%, <=!%5" /u*4ed -*stract K /u*lisher Eull 'e8t !6" 9rini(asan 1, =athan 9, 9uresh ', La7shmanaperumalsamy / Antimicrobial activity of certain Indian medicinal plants used in fol#loric medicine' $ournal of Ethnopharmacology 200!, /0!2!%&20" /u*4ed -*stract K /u*lisher Eull 'e8t !%" 9amy R/, Ignacimuthu 9 Antibacterial activity of some fol#lore medicinal plants used by tribals in ;estern -hats of India' $ournal of Ethnopharmacology 2000, 2!63&%!" /u*4ed -*stract K /u*lisher Eull 'e8t !#" /erumalsamy R, Ignacimuthu 9, RaJa 1/ Preliminary screening of ethnomedicinal plants from India' $ournal of Ethnopharmacology !HHH, !235&40" /u*4ed -*stract K /u*lisher Eull 'e8t !H" /are7h ,, ,adeJa 9, Chanda 9 (fficacy of A>ueous and Methanol ()tracts of Some Medicinal Plants for Potential Antibacterial Activity' Tur)ish $ournal of Biology 2005, *2!203&2!0" 20" <ora(uthi7unchai 9, Lortheeranu+at -, ,eeJu :, 9ririra7 ', /hongpaichit 9, 9upa+ita ' (ffective medicinal plants against (nterohaemorrhagic (scherichia coli ?&</! :/' $ournal of Ethnopharmacology 2004, 20!4H&54" /u*4ed -*stract K /u*lisher Eull 'e8t 2!" /rashanth 1, -sha 4M, -mit - Antibacterial activity of Punica granatum' (itoterapia 200!, /*!!%!&!%3" /u*4ed -*stract K /u*lisher Eull 'e8t 22" RaJa7aruna =, >arris C9, 'o+ers ;>= Antimicrobial activity of plants from Serpentine outcrops in Sri 1an#a' Pharmaceutical Biology 2002, 09!235&244" 23" 9a8ena <M, 9harma R= Antimicrobial activity of the essential oil of Toddalia asiatica' (itoterapia !HHH, /9!64&66" /u*lisher Eull 'e8t 24" Mhan 4R, Mihara 4, .moloso -1 Antimicrobial activity of Cassia alata' (itoterapia 200!, /*!56!&564" /u*4ed -*stract K /u*lisher Eull 'e8t 25" 9omchit 4=, Ree?al I, =ur IE, 4utali* -R In vitro antimicrobial activity of ethanol and ,ater e)tracts of Cassia alata' $ournal of Ethnopharmacology 2003, =0!!&4" /u*4ed -*stract K /u*lisher Eull 'e8t 26" Ooha =arasimhan 9= Medicinal plants of India' Tamil *adu Cyber Media Bangalore +ndia 2003", II! 2%" =ad7arni M4 Indian Materia Medica' Popular Pra)ashan Bombay +ndia !H%6" 2#" Rastogi R/, 4ehrotra 3= %ompendium of Indian Medicinal Plants' Central ,rug %esearch +nstitute -uc)no. and *ational +nstitute of Science Communication *e. ,elhi +ndia /001'/002 I3V!