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Antimicrobial activity of some

ethnomedicinal plants used by Paliyar


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"
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