Indian Journal of Experimental Biology Vol. 6, (4) 1968, pp 232·247 Screening of Indian Plants for Biological Activity: Part I" K. 1.. DHAJt.. x. lot DHAR. B t:. DHAWA..'\. B. No VLHIlOTRA .. C. 1lA\"' W:~ P\.t..at. ProJeCt:. Cz;ctrU DnI.c J4. . ~", ,.ut..u JI.nmrc.b Jubart.£.,. l.uc.~ .t ).lr 1961 T_ ~ oaa.d aJabty-O," bota.llkaU, lIkDd:6.d pl.iuIl --.u.rt&1Io a.a ...........u--.d with bt pu -=-r .tha.a.ol aa.d lb. e.rtT&.ttIo pot Or'OCllb • wid, blo&otlcaJ ~ _ 6J ...ta. ~ todGd .. ...". .... -tJ.bacttri&l. _tlh.rfta&1.. &.rIdrlral,. _tlbdJn1alhk. _d~hOW ' _~. _tUtrUUQ'. 1IJnooI1~ -.d a .. 'de ralItt 01 ~lo.l.caI acth1t1-. PROGRAlO!E: tor the screening of plant utracu for a wide ra.n«:e of biologic.al aCUVlI ) ..u $unc-d .1 this in~titate about four vean &1;0. So b.r 0\"'0" ODe thousa.nd plants uve -~n coUeettd and e1rli: hundred have ~ Mn:Cned for tbeir biologteal aairily. 1t is mteoded to publah tbe d ata collected UI this programme from tune: to time In thli . the 6m of tbl s 5el'ies of papt.n. lite data OD two hundad and eigbty-five plant material compniinc 1""0 boudnd a.nd tbirty-seven ~p«:ies are reponed AhhouCh it is customary to prepare trtr.l.cU of plant materials with a variety of solvenu (non -polar and po4.r) ""b(:D investigatiog possible biologic.al awvity. It wu decided at tbe onset of 'this proI"a.rum.e d U I oaty one extract of eub pl~ t mate:fi4.I. a 50 per aot ethanol txlr.l.ct. would ~ uanuned for blolocical acLJ\'1t}" Tlus dttis:iop ...-as ~ t o c.nahlt t.hc: ProrrUDme to cover a w«t number of plant au.t.aU1s. The botanic:al idcdity of each pbnt was established before the- at.ra.ct wu pn~:pud1. Almost all t.besc utncu hut: been tested for anti~Clcrial . antifu.npl. utiwiraJ. Ultica.nccr and a ,,;de RJlge of pb.umaco1ocic:al activitiC5. A fair number of Ulncts lu\'e bec:a lested for antiamoebic: a.nd bypvfiycumic actint)' a.nd :I. feW' for activity !,¥ainst bdminlhs and .pirocha.e:le! and for inhi blOOD 01 sperm.uoceoes.iJ and prevention of implantation. A W.le.rial. u.4 Mel:bocla CcJ.kdic. _/ ~ ~aJ.s - Plant malerials bavoe been colicdcd from various pa.ru of lodia tbcit itkotity aubli5hed witb tbe- aid of tr~ on rqtoa.al Sora and compariso:t ..ntb h· ~um .beeu of the ;autbmuc speri~ Tlk d d e a.nd IOca.uOD 01 c:oDection b..a~ ~n rcr-ordt'd w 232 PTep.rcJUM oj uUatU . J fr~ - The: allcined plant m.atc:tWs haw been powdced m.ec.bA nica.lh- and e..xt.nct.ed i.Q. 300 , . lou by t.m-. oold pe:n:::ob.tions with SO pu CtDt dlwd_ Tbt cambined utraaJ. have thm been c:oaoentra.ted mlW =loco! J""SS'U' (bath '=p. SO"C.) aDd 6ully dried in V&CQUZO desiccatOR.. Prelimioan' fra.ctionation has i.Dvolved atractiOID of t.besc mat'Uws ....1tll bezu-.c:be foU~ b)' partition of the raidue in .-b;a:tzDoI aJ)d ...to' 01' chlorofDml·:nethanol-v....tCl'. 'Tbc thre.t fncbcms 50 obuiotd have been subjected to c::onfiJmatOf)' ,em. A. ArJilHJ.::taial lull"l- The foUowiD&: two Gram-positive st.runs. thr~ Gnm-Dqat:iw atn.1nJ aod ooc: acid -fast My~... str.ull haft been used: BuiUtu $u1Kilu (AJ. S:.p~ luano.s (A.'. S.I~ tYPAl (-'.1. ~.Gl",:: cotl (.". Alnbat:kn"m , .. .u.facicu (A..' aDd JJ>~tI .. Iwlwr~$ H17Rv (A.l . Orrzumu A} to ~ ..en m.a.int~d Oll Zlutri~l apr lhnu1. T tstJ.D.c was don< in ptptom broth. A.ft.c:r inoculatioa with • loopfw · of mhon fnm the tl&Dt. the I«4ed. broths wen. iocab&ted at 3r ± 1·c.. for 24 hr. Tht. t~fold aerial diloticm t.eclWqut' WU D5ed The plant utraet ",1.) db"olft:d In alcohol or acetone to obtain .. 1 q .Jm1. IOlutioti . TllU .a.olution (0·2 ml) was ..dded to 1·1 ml. of tht seeded broth and tanned the fi.rst dilutioc One ml . of tbis was diluted with. fartbu l~ ml. of tbe loCeCitd broth to produCt the ICCClI:Jd dilatioZl a.od so on tiD six such diluLJODl WI:ft: obui:Ded. A I>I!t of tubes containJ:n.r only inoculated broth ...-as lup' ., coauol. Ahu incubatiQll far SI 'hI.11w: lut. tube .ntb ao r0wtb of tbe miaoorpaiaD was uka tf' ..-e.pn:scZlI the minimum I.llhilatc:wy coaccc.u.o!%. (MIC erpreucd 10 v-: /ml) of the d.."\C'd utntt A plant. lnb1bhin.c the growtl!: u. a c:o::a.ntnuor. of 25 )it./m! W'U considend attt\"(: B.Icttria A. (M . 1"q..,(;..Mms HJiR\ ..-a.! rro~ 0:1 JUT (l ntemauonal UW OZl a.pm1t Tt:~os.aJ medJum F o! t e5olUlg. 2 ml. of tbt m~~ containinf lbf. dru, were taLen io ten-tubes a.r.d Lnspw.at~ Into s.4.nt, a t 7S-C. for 3(. tr..tr. The- planl exuaC1 v.-u L')corpouled in the a.~U!D prior to kspisu,· t IO:. Z! con(~ntratl0:1' of 10. 4(' and 20 t'{!ml Fer. co:r: jl.iTUOr. Uon lCOUnlC iI:lC bydnzidt lII"&S wd a: co:;,cc:ntntloM of O·S. !'-25. 0-12. 0-06.0-03. HI ud 0-007 ~ Iml D ru,-!rec co:mn! s!u.u -.rere also oset fot co::npatuon Th~ ltIocnitl!!l ..-u p:e-pueC from a pr\mU) cultan by n5pe:1o~ a.nd DH,\li " aJ ' SCREEN ING OJ" IN'J>lAS PL.\:..iS r O I( fJI OLOG 1CAL ACT IYIT\, m c:c.hanicallr sh.aking ) · 2 mG. of b3 c tcria In 0' 4 ml. of t.lcnle- chst illcd ..... ttl . A foopfu l of tilt baClclial 5o~pensioD ...s tbm s treOlkrd on eac h $lan1. Lab.:,·j · line 01 t br growth \A'a5 dune "'Iter a JlC-liod of 28 da y, accordins t o UK Medical R~.a'ch COUDell honiuid Tria.) Rtpon }\o. 3 (rd. 2). A It/KOloa)' tnd poinl \A'U la.ktn for purpClSC'S 01 !.C.1'e cnin,. B. A.Jlf_t~I tali", - Thl' loll o "';n~ h.·10 str,ajn" of yun.lJl:c l nngi. dC'nn;uo p hytes ~d .a pall I p~thocenk IUllfUs ha~"C: bttn clllployC'd: CcrediiQ. "lbiulfu (BI ). Cryp1OCO«W$ ",ojrn lfl.Qn.J (B, ), Tn·' It.... pA.\1fna fAnJ.:r ~Jsylu (B,l . It.1 iaO$par v,,. (."iJ (B l .and AJprTtww1 "ItfT (B. \. • All thesr su .a..ins \o\~C' mainlOlioed IOn Sabouraud '5 apr sb.n u 1 , Testing wa.s don!' in Saboun ud '~ broth . l..oopfuls of Bl .and BI cultures ltom $.~u W~ lt inocul~ted into broth Ilnd tbe respec.tin inoculated broth1 u~d {or l esLinJ; a h er incub-Ollio n for 24 hr.at 2S-± I·C, I n t he c.a.sc.s uf the l emain· i n~ thm:. sma..ll piea:s 01 mycelia WCH' jntrodu t~ int o coniQ.} Buks c:ont.:lining SO ml. of the broth. The ft...ub ""ue then incu bated ",; tb shakinc for 24 hr ill tbt cue of B. and for 18 hr in tbr ~ o f B. and B. . After iDcubation. tm: clear broths wen· taka! out of th~ fl uu and serial dilUtiOll5 o f test DUlteriab. made. u in tbr c.a..~ of Ik baauia AI-A., Subsequent incuba Lion prtiods wele 24 hI 10: BI and 8", -4 8 hI for B. aJld 96 br for H. and H •. E\'aluaticm of anti fungal activit), W iU t hen done u in the Ultibotcterial t est s AI ·A. C A,Jiproto:tHIl 1.t11'1t1: - T tli> " ' II.!o dont on ".'0 E ..J.~ AilloJyt iu $traln STA (Cl l . T' yp"n 'JJOTI't " n '/3"si {c.l. Tryp"" o.JCJm" c' u:i (C.. L PJ...l~iu'" be>"l:ki (CJ llnd n"~.J'i,, , hod" i ,ci lC. 1. • The pr~ure tor a.ntiamDt: bic t est ing h as been published"'. Testing ",,-u initiaJly d Olle in n O t,,,. E:rtnd.S havin, am~bicidal activ:i t.y at II. concen t n.tion of 125 p.«,/m L 01 less wrrr thc!n lCSIC·d ire wf!'O. Suiubk d(lSasc for tht 'PI vitlo t csting ,, ~ d el('j o mmcd iu a preliminary l oxicity Icst , An aq\l"ol,;':o !>oiulioD 07 suspension """U ;admi.ni.sle,ed o,alh· to I:loups oj t wo wea.nllnJ: rollS (3 w ed:..s, 1&-20 p. body ...qbl) a l Uu'et arbi t rary dosr ievd5 uf 900, 300 and )00 n )g,lkg, body weight . T ......o c.ont. ol ntl welt "ven only the vehicle , Thl' animaJ.~ " -ere observed 10T 7 da~'5 for any morta..lit y . If t hue vas no mort:llit~· the i rJ vil o t est ing " ':l.S done :ilt 300 mg.fkg. If t.here "Wen" d eaths,. l csti ng ",'U dOnc .at 1/1 0 ~ lolera lrd cos;e. TcsUn£ ~;un st tbr CI~hel 1.,c1<.oz.....a wa...~ done by a loxiat}<\lIn·tllc r _i-"t' u\l c l et.l i n mi cl Jl.!. u(':.O'il)Cd In an caIlitl commu.ni c... tHm' D A~·i~!m ir.J;'i{ IIcJ il"fJY- T)u!- .... :l.S d om· on I.lllnC O~es Ulrinji (011. Ch::mU"e1I t:: ;'.c:..-JN " f'l (OIL :'I·lppGu ,Cff.f:l1w, ~1I. S'illt:nJ, js fD. I, llYTT'J t: ok pa ,...:: r.Q (D41 ani! Al,,,r;a',,, r.;;111 (0, ). Tht techniqur" 0: sercenlnf! af'u.n~: cctlOn·r;;.l fihnl ( Ll'N" ~ l lid(J (.:I"''':'; ) a .. d esc:-:h--rl by 1h.,,· ~.:111'" .. nd 5c,,·e!! 1 a.... d St\l:clJ and H :& ", 1.:.:'0(:' wa .. cmylo!'l'~ Sc.r«T'j llf a.f&.inst D J W-.c..~ done In cott o:, r:i!.I~ .. ~ 1/5 .r.e 1lIUl.'1lUU, t olcult°o d~ lo! nucc. . All e~IW-~t e of t his fipJTe ,,'Ol! obt;uoed dunnf tbt- l~ t:r.f .. fai n!! C, SueenlOfg a~ain ' l D I ba~ not bt-tn dont in the p!e5enl s~ries, . PA Rl ,·u,,, I" .and i" ..-it-·o teslin#: OIg~ln~ t D, ...~~ COM as d ucribt-d earlier'. A d ost 01 250 IIIJ: j/..t ....;u. employed. A ~oup of J inft'ctt'd anlffl;J/,. ....a.." ~d fo r uch uttOlct whik anof her f!r oup 01 3 inlrtl('d oaninuls 1i.Cn.'ed as control. /ft ntro telollo,£ ~n~t D. "'..., al!>o don~ by the $.arnt 1lK1hod. t he pH 01 tM medium hein£ adju Sl~ t o 6' S in th l5 cue . For i" ""(10 t aung , a modibcOitiOl1. of Whit· lock's method' "''"&.$ u~d . }\e"'I\' "" uncd a.l bino uu (25· )0 &,) wtTe moculilled ;", th SOO brvac . On th t 8th dlY afttr infec ti on . faK.a.i pt'lieh ",' ('fC e.umined 1 0 establish prde:nce of O\'~ lOr Uli' mals "~ rt' s l af'VC'd o \-emit:ht ~d tbe on t d~y Oil &roup of 3 inlC<lC:d ;ulImah w~ j!I\'Cl liO I1I~ I/...~ of t hle I DI e.xt~CI by Ca \"al;t. and anot her l"oup of 3 was kepI U COlltrOI. Food . 'U 2I.{:ain . 1th hc]d tbt ncrt day and On the m urnlng thc , e...he, ti le aninlall "" Cft' ).:illtd and the WO nta n-cnvcTr d ;lila counted, I" l oS"" !>cl('cninl) ag.ainSl D, .~ ~Iw dOIle' to, lDodil~inG t he tnet bod employed to! D. Tht pH of I b~ medJum "'.... adjusted t o 7,2. tilt !>uJtu'f; dilution of t hl! t est d rug in t he- meWum ", ;u. 1: 100 w1 / vo1 . and the period o f incuLaticm WOl! <4 8 h r . hi r i"", KI'«"ni n,; . 'a... dont' on clue/..s infectffi tv. o monthi earliel ' , Tbf birds wert 51aJ"\Td for H h: and ... £TOll)) 01 t W'O d ~d ",ilb liO tng.fkl; 01 11)(0 eA'1I'Ict. T ..'O birds ""U~ kcpt as u nu-eate-d coDlroh . 11K lac<cs 01 tbot: birds " 'ele uaminl'd OVCI I S da\~ lor ,"'onn ~ , Thl' bi ld ~ wc.I'C u crifu~..t'd 00 t he Jblh d Ol.,)' and the worm IOOlid o f thl' trr'OII I t'd IJld eooHol r;ro up" cUIn,~ed. E. A rcJi~'I,"1 ""iul,,· - T h.il> ...~ dc-t,.lUllnf"d as::unst N.c t:l k1t..t: ilJ t~lt vi rus ( £..1 " lid l' .II CCHII ~ ViruS (F"'I \. T ht:y "" ele- Ctown o n cborlo-ali;ul toic. membl'1Jlt (CAM ) cuhura or IOn m onola \'~ 101 chick embryo nbJo b lut.s. (CEF). The mClIi(.d follo..·cd for ptcparuion of CAlJ cuhu, ~ h l.S bc..m descri~ prc\'1ou.sJ\"u , ld onoJayels of CEF wert f11tJ».Icd hom 10 da)$ ol d chid: cmbry" by a mcodibc..atio n of (.oore", ffit- tbod " . The- ~'Irus and lobe plan l l'X1r.r. CI . ·Clt UlI).cG and add ~c to CAM CUltUll"to 01 eJ::F mOfl o~\"c:.n and lou.:l... trd 411 lj -C Tile bn.J COnCenlralitm o! the ulU =t was 0-05 m,; .(ml. TIlt: cun«ntnllOll III 1:.. ",. ~ 0 ,0(,4 haemaulu'tination (HA) u nit tmi and I h.l.l 01 t hl! Ea SO po:x-lorming IPF) units/m!. ~ Irst wa s done- 1b rio: usinG 6 reJ)ljcat C'S In CoiIch l est. TIle ~·ir ;J..I proccny \l'as meiLSUlt"d by ootm/? 1~ HA titrl' of t llf' c:nh u rc fluid ... her -i s hr JnCIJb;!.lIan in tht' CUt' o f E I a.nd after 72 hJ In t he Cll~ o f E. The inhibllOf\ dif'c i of Ihe U:1raC"1 ...·u ml'uu:rd ' a ~ Ihc d(,(,fU ~:e- III I).e- (:on Cel.tUlion of "Iul p'Ofeny produced 10 the CAM cultllr",~ CI CEF m('no~q!~ compal("(! I n the- cor:1roi!. 1:- f"': ! cent lal1 l n \11.1 prOf!C'ny in CCIoC of E, and J O\r pe r CCliI 1.. 11 In ~ of E.. v.c::re 13k t'n;u SJr'lInc;n t. Fo~ lill'eC: ;;Enln IIi.! aeli "lI Y, in L·' -I, O l e.c t l!1C \I." ;'~ dent t,~· mlnt( It,· extr.ct {O·S m,:.jrn!. j and ' 'ltu!' (1)- 192 H,~ enll ::.: ~ and 150 PF u nit S/nil. E..l a nd trlC'\l~tl.1"r a: 3;-"C for ~ !::. Ttl~ CA~ or CEr t-uIUHr"" .... rt( micCt n! a\ 1 : IC';. dilution of t he male'C' a:'lG "'.,raJ prOfcny product .: .... u meuuTed Oi.\o c!t:S.Crikc .:....H. Tht~ teS\ " 'a' G(Jnt only in c....s.t of Uolt .eu !>bc,,:in.& !>Ipif.c.an l a cti vlIY io t h ~ pre"iON rest n,~ U· \l leH l>hu\\.;nt ~l-"i6canl vi ral i nhibito ry p !"e>p<:n y 23j n .d po... ~~i n& no dif ec: "':1 " '. i l .. 1 .i " uv, t y II, 1.1'1111 .. ~r c: d«med 10 h.;l ve :lnll v ' T:'] a(! [ \'l l'.· F. A1l1ilpiuJ( 4:u! JI .1(':1;":.'- - Thl; ... .. " IC) I.·d ~Ii:J I:1, t SpirocJuuf.:l r aU rf c-,, :a 1I : !f ~ ~' (!'I :11 mk<. 1hc mtlhod used has bo: ~n d C':i cri l"~<:! e:lr !l.' I( G .ifl1 l[rrr i luy lelll r1g - Tha \O~ CJr:j"d (j lll tol : ~o!ony br\·d ma ture male ~"l,j lem .. lc J,Jbllla l:lb Two typo.~ 01 tesu, \·il. . an tispcr.n alo ~cnc: :m IGJ .llId anll.impiUlt.illon IG l l . ..... eft u ~td. The plan t eX If'2ct was macera ted 111 111'1 an c<!u., 1 qU llzuity or gum auci:l , s u:s pcnded in distilled ....·a ler md ad m rn i~ l o!~ onJI), in :1 d~ of 100 mc .fkg . daily for ~rioru mentioned bc'low lO groups of 5·6 raI l.. For G 1 " s u:;pt:n ~io n of the extr3 c t was admin istered dlih' to m a l un: m a le ra ts fo r 50 d 3.\"S. n~ .. niml h "';Clt' sacrin~ 1" hr J iter t he Ia.~ t dCN Spt nnatol Oa wert colle-et l'd fr om the mid· sectO! 01 ti n: V;t5 defe rl"ns an d thei r SfthS morpho· logy no ted. The we ights o f the testes, the semina l ve..lcle~ lnd t he ve n t ral prOState g la.nds ""("Ie It(oldcJ and the: testes .... c re exami ned hjSlolo~ i · alh'. For G 1. m.l lure m;lie: a nd fema le rats 0 1 prO\'eu (wilil)' ~·e re agcd together in t he: e ... rni ng in the r:lll io of t ..... o km.l le;; t o OIlC m:tlc. In t he m u m:n!;. v;agUlill sm c<U":'> of the female r3 ts wac t a:ke n and c,umllled under :II ml cru:scope fo r t he pre~~" III:C: o f 5pcrma \01o;l. The day on w h.ich ~ p('. rtn .. ta J.oa w .. ru found I n the s mean; W:\$ cohsidC:lcd Ihl:' [u- s t d ay of prernanc}·. A suspc"nsion of t he cxtr act wa.s aJ· , mlll isiered to pregnan t fc.males fr om 1 to 5 da}':$ b post coil u m . The a nimals .w.,re l.a pal'O l oln ued on I the 10lh day of pt'ltgnancy to noll;: the pres.ence o f ) imvl.lnt:l,lion sites ami corpora lutea. Complete absence of tm plan ta lion was I:l ken t o indic;'l te anl ikrlillty actlVil)·. H. Hy/,oglyC4L" ..·C 4ct j""1·'Y - Al bin o nts (ilboul 1 00~ . body 'AT-ight) were fast ed for 18 hr bu t watt r ..- Y1.5 allo ....·ed ;ul libiJ wm • B lood 'A :.&..~ Col!.:-c ted at t he I beginning of lIu.· t xperiruen: (0 h Qu r s..mpJe) and ;a s1l5~nsio n 01 lhc extnct w;u ad zmnis ter¢d orally ,to il group 01 3 OlI1imills. :\ single: d o se 0 1 250 mg., q . Wl.S used.. Blood was collec ted 2 and i he late r (or estima tion of s ugllr by the m ethod o f Nel!ion ~d SomOf::yi l l • n le m:U: Unum per cen t loweri ng of blood sug a r was calcula tt!d in tach case. T he n:sulu wt' re compa.~ Wllh blood s ugar levds of a IJOUp of unt rr at ed rats:lnJ a third group of sul pha. n~~uru (250 m!\:. /kg .) t reated ra ls. Lowering 01 blood sug;lI b}· JO ~ r cent or morl" w3.S \;llcen as 'Signul ca.nt. J. .-t r.!lllp,,<mlc ~1I,! jty _ Th¢ lest h.1.s no t been ~ rformcd wi t h the present :.cries of plan t ex t racts. J. £/«.1 OIl ,up,.,4!icns. c;o,Ji/'J1J"-Sfwla, l ySU,," iI,. d ~d iLaJ z ,,& ~m.b,",, ( - ." dult mongrel d ogs or cats of t i l ~r SC.t. anusthetiz.ed. wit h pentobarbitone sodium (30 mg,{kg. i.v .) w,:: N: e'm p loYl"d. Thl" ·tnche;!. was cannulated and conntc tc-d to a :!duc y's tm'l bour . one nl Ille common cilIolid U1 erics ..... a.s Unn ubttd ilJ\d connected to a m ercu tv manometer. :Records we re made on a slow m oving k n n ogr-aph. J1ht dru/:j we ~ 3.dmin isle:-ed In , ouKh il can nulated ', m"'J wi, ~ u ch extr;!.cl was ini tia.Jly O\dm ; n~ t tred in a. dost: '01 25 rn g Jkg. i.\·. If iln ~6ecl w:u o t.i HVed . snl.l llcr 23. d,,~.. \o\ o:':r · ~""I cd and the mlnH~ .•7 l"!!~'.~; .. ( ': .• d ct~ !'~. ··Ie-(! !1 25 fm; i"g. pr w .. ~.' :::~:~ ~ ~ ~ .. :npon.c, :::'e e:i t"<.. \ v! ::0 O'lg .. kg .......( j. :~ d..~ •.. ilt 1i1l~ level Wu :I;,: :n\C;~ .•.. ;<,! I!'x :raCI IIllCtl·.'C' R~!)r i rd t i" [l U ll' T:I'! c:1c:c : .l.nd arnpilt uck, "";U ~ ...:J. c:- : .. :~ Hlood 'Hes;'!; : " ';1:' rh '!' ~: !e<:: ... ~ ",UC:r : :0 the n j"lrm :1I blood p : e iru ~ t 10;\,.. 1 and ~ so on prbJ.0r r~poMC' to ;Adr en :Jlr~ \0 ·5. 1 ;q i.;. i.\· .J Uld d eprC'l>sor rc.ponSt"s to ~ celylch oh::e L"ld ruH:ur.ll:e tboth O<!.5 ..()·5 ""go/ kg. L"·.I. :\icUl au ng mt.m bnne (JI): :\iC"'.Ju,Ii.,ng ::ltr.:l . ~ r:lnc: prep:u 'l.l ions wen: not u~d Ul the p:-ocrH , .. lh:5. reG EIe<.:trocardiogra rn U J : Sta.n d.ard Lud II was reCOr·J ...'" ....i th a Hdligt' EK ·i5 Simpli$~r:.ptor ciKl l ocardi ut,'>'J"a ph . K. Effah 0" "l"{rU~ti tw W(S - Guinca, plg Leu;'11 (KJ : l ".r~ to .6 \·t cen timetres long tc ~tllln:U por · I ,OIlS of ileum were taken fr om ~ fretbJy ~.,.d guinea-viii: weighlug 300-500 g. and s~pt:Ddec. .n ac:ral l"d Tyrode:: !>Olut ioll maint ained at 3+-35'(. :n illl isobted orgilJ'l ba t h of 20 mJ . Qpadty . Aftu 30 min. r~t. 2....f. dl>.>e5 o f spumogen wue addt-d to Ihe bat h 10 obt:un uniform amplit ude of CCI:lL"-· l.ion whicb ..... as recorded 0 11 a smCllri.eti kymog: 'oph by '" frontal writ ing le' ·cr wit h 5·6-fold rO'ogr.t:i.:a. lion " nd 3 -+ g, Il'ilJolon on t be: tis5!!!!: , The ~ pa..."Ul()ge n 5. :lcet vlc.holint' ehlor:de 15:.< I ~ ti. ) .lntl histamine acid phosphate p X 1Ct" g.). v.!'le a llowed 10 ac t for 20 s.c:-c.. In some (xprri.r::c:::..s nicoti ne sulphate, S. hydroxyt.rrptanunt CUlln!' $ulp ba t e and barium chloride were:: ~ to PfoCU CC appruX-imill cly tile' sa me beigh t of COntu,ellO!!. The aq ueous ~I u tj ons o f t he exl ncu were adcffi to t lU! ba th 1 min . before tbe addiuoQ of the: $p.umo.gen. ERect 0 1 gradtd doses of the: utract on the: s~mogen. i nd u ced contTict lon ..... as SCtD a.r.rl J,,,:,r cl!nt I'Nuctio)1 of coul.r.l.ction me llun:d . Thl! t~u e WilS wu hed t wice a t 1 m.in . inttr.-4.1 iltlG 3 min. rcst W~ given to it. II Ihl' test compound i tself cauJoCd cont r.l.ct:on, d letts of a trop;nc s.u1 phil te {I x \(t't g.) ~d m~pyr· ami ne: m:Ueal e (5 X 10"' g. ) on t his coolr:::l.ctior. wel e st odil!d 10 de termine t be natu:-e of ~p.aslOoget.ic :u:tion . Guinn.• pig hca.n (K J: Guineil pi~ "" eJ~hin g 400·500 g. .....ere stunned ;and bled 10 duth . The he.ut ....";1.5 removed \\itb .a. small por1t!)n of t!le asct nding a.ort a.. The ilon il was ~"lUblcd 4:\d t be heart was pcrfu ~ wit h ox y~atcd R~~~~r· Loc ke solut ion .it 37 "C. Wlder constat pr~Ufe by tile Lang1! ndoril mtthod.. The fate :u1d OlfIIplitude of con tr~ctio n of lbe heart were rrgisl~red by a Starl ing's htart levu OD a smoked Icymo~~:;;~b. Tht plau t e:o:tract was i D j ec-t ~ d $lo..l), in:o t he perfusion fluid in il volu tne of 0·5 mI. Uid perce,lil~ clung!: ill the amplilude o f con t :action :c;d 11.5 dur'H iDn mea!>ured. Ra t ute rot.!. (K 1): f em..a..le albi no rau .....1!ft t:t;n,·d wi th stilbocstero! 11 00 1-16./100 g .) ~llbcuta.n(.c~~ J}' one dar bdort tht e xperi me n t. The riU "'ti e killed by stun ni nl; and b leeding, une horn of : rle utenls w ;u diss.ec tcd out and suspended :n ...... orgllllJ bath of 20 m l. cpac.II Y CO:lt:w"lZng i!: ;lt~d IHIAk " /I I ::.cRU:..." I !'C OJ I ~ I)I.-\.!\ l 'U :':l > 1'0 )( d l: J alon', SUld OI l 30·-C.. T)" rrspon5>C 01 Ihl: r;o l ut er us 10 d lft rlC'nl d U!o t'5> of l h,. C' ),lIi CI w:!\ IC'c ord t"d b \' An isolonl c fr on l al wn t m h I ~ \ ' ", UI, • !>moke'd k~iTlo~ta}!h L C,ou Iffu /) "'i d tffub. tm et r.I " , 1 nnl/OUJ Jys:t rl1 - T ho.c- \io UI'. \O t u(Ju·d in mll:( of ci tbe: ~ ,.. w el~ bi.ng \)(: hort"fi I ~ and 20 Ii Gc-nr:ali;• .5 anim als were Wlplo ~r d :n uch dox Ic \'d I ni t i;ol l eMin!: In .'' usu;a.Uy llecn done a t 1 14 . 1/ 2 LD ..e If aCt i,;t)' ¥.4S detf'cted. s maller d ()$t·J. were- s u ~ qut:' nt lr t e-<-II:'d, G, ~ d Tt'C1.s ( La): The ani mals w~ r ~ Obien 'rod for 2 hr alter dru~ adm ini ..t ral ion fo r any (',;de-nce 01 c.:nlnJ nt t\·OU" system s l imu la u o n. or depTr"SSJon or any a ut onomiC effect. T his in " ol"ltd n ot i n~ t he efi .-ct on (i) sP'»nun~u ~ mOlor a c t i"ty. (ii i certain n8!:xes.. 1'.£ tlE;hunC. cornC'.ii.l. PU1D ;s.1 . (-.ii) muscle t one- (rrduccd a.nd c.atideP;YJ. (I\'} pos tere and ~a)! ( .. taxia alld w,Tl thmgl. (\.) ... y~ (pupil and pur.» ..). ( \;) l oplrOl tion . 4U1d (vti) observin& Ihun fo, occur· renet: of trc.m ~n.. coo\<'Ulsions. St~ub' s tail. piloelection. C)"anosls.. lacbrymation ur diarrhea . Eac.h e Bf'Ct wu graded as mild (+ I or strO;1& (+ + ). Hypo th("flllia (L,.I : The" skin or rt"Cta1 t t'm l"en t ure" "'''U measured -v.; t b a t h ... nnome"t ~ before end i. 1 and 2 hi" a lter drug administration. Thlt f<l.U in letnpo:ra lurc in il1l the- 5 animal$. of ~ puur .:It any pnucuia: l ime ""'a.!. t ot aU~d Ollld a \'3.luC' bo-lo ....· 5- F. lJ:nou·d .'I.nalgC'ilil IL,I : T his was t est ed U,\' HaHner 's tail c lip JDltthoc:l u . If 2 pos itn'~ Jesuit W~ oblainit'd furt her connnnat io n was d o n r by tht- hfl t pl:tte m ethod of Eddy and U.imb3dI U , Supram:u:imal electroshock sc:iture p:a tl em t est {L,! : Current (.. 8 m a..) ,",'U deliverr-d IhTOUl!b nr electrodes lor 0·2 5e. C. using a T echno electro-con\'utsoOle tcr . E ffec tive druSj; ;lholis}wd the t Onic \"X lensor componen t 01 the S('izure. \! elr<iwl !I ' :'urf' lhr ~h old t c.<. t (L,.\: Dru ~ "",etc tested fur tllt'll :.uili t \· 10 prol l·c.t fr mn t11(· cun\'ulSlon ~ ptoduc..-d b y ' 80 ml! ./I:s. mtt r:t:ol Ili" cll s:Jbcu lanruu!lI). t he a ni m Olls IJein b o~ :~".l' d for th .. n l:' )''1. 60 nun. Stn 'chninc loCiz:ure t hresllold l est (1-.) : This WOlS simila.r t o tilt roe.trawl seiz.u r~ threshold l est ex«pt lor tilt: U!oC'. of strychni.ne (1 ·5 mc./kg ,) in pJoI C(: of metr:u.ol. B;1r bitu rOllr pol e nd atio n t (S'! (L..,): P t'nlobarbit orlt /4 30 m g Ikg ) ""'as udminisl erL-d te' :0 cUlll rul and :1 drug treOltrd puup and t ht' ume lot w h ich nEllI in~ trfiex rC'.malll~ at)!>!:t1t wa ~ n oltd l\ compan!OOn o! t hr dat a. o! I ....·'. cr o up~ ill.:ilta l t'{! Ii tlierr \104$ :t!l\' pole.ntl atlo:-, .a.mphf.: otmm. hyperacu \; t y t t'$ ! (~ : H \l>e: OIrt :'"In- ....~ .. mduCt'd b\' 2·5· 5 m:: ./kt!. :nr.pilt'tamUlr p,:e-;! inlr.llpenumul)y. T : •. uti,'''), (i ~ t he: chU; ,,".;a.I!"d ~rour W1S co m p4i t('d ""lth a control poup p--:'ln;' tlm:"h"::l minr a Jon ' 15. 30. 6{1 9(1. 12(' 1:-, ;Inc I ~ " r.::r.. altc: tl; .. 30m::-:'lt.1!.tlUfi 01 •• :l. '1 ,. , .".:- ;-"' 11:-'" l lC" ~ iu t\ i;: .: ;::c-~ .. ;n· rmet' (L, " ,". ::';--,":dI::;:, ;;J nlt; I ~c I i ~ .. ' J:"\t'f. 1'-' tho C!.I;:::,,! OIf.1! G:l.:!: u eat e.·d :4:,!ma.· H"O\:?~ 01 l fl :r. ,'~ n,:c ud,' 1"'''0 gt'ou p:. "" l"fr p!::~fC In rquOl! • :~: Cl!culll COI.f'~~ a nd rno:-: .h: \ nOI .. ahr: .5 a ne :?' l':: c IH lI UJt..I ~ .\L .\ t 11 \ I..... J'.ltoNl I M. AJ11iu"Ut lClltr.Ir.I - Ant IC;u'lUI Kr rt'mn~ was d<'nt a l Ih ~ (..:I nC'r r (1lr 01nlhrr:orY h:tlltlllal Sr.rvice (cnl el Screenin,: LaboJ.. t on !' ~ undr' 11'11' UOIcl 10 t N- 1\ :l t ion:!o1 C;a~ ('.., JII,w..::r Ii· Ihf'-.I~Ol. M ~}'I;an d . USA. The folkm'inE: t Oo t s~"J.lcrn .. "'l ilt' 1.:)(,:: H uman ~PJdt'nnai d cuo.')o:ru of Ibt' ril~J b:u\1l:\ in It!l.!lue- cuhUI f' (MIL \ \' .. 11.("1 .. IC:.v...l:COll ... 256 (ult umuKlJlar ) In Ibr Tat Ilr l l. L 1:210 hn,pl,uld Ie-ukautu<l In 11ft' mou~ n,! , \. ~ .. rc ... ~~ 1M' II lilt rnoUSC' lM .t hl:'pa:o!D<I 11\1 II, tl,t mouo.c t': . Friend 'n ru! Jeukaernia (!oObd) In t hr mou -.c t~: • . lr""" lun, c.an:;moma m t he mow< (ll,' a..nt5 H~l bumUl s.arcomOl in t h ~ emb~'011a' rd cu. r~. \ Tblt pr oloool o f methods has. b«n publhlled I :'. d daiJ u . T oncity- Toxicl,· ""~ oe-tc nnmK b\- tl.~ 0 : ... 1 or t he: Ultnpu1toneaJ routl:' In 2 0: j OIdul' .. Iblrt, mla 01 either W'x and uru~!I\' 1.5 t o III f In "'I:'I{:~'! The: dntd plant el.tr.llC'u,' ,,' ('J(' S.U\PC:'\~ct! n. (j~ per 0".01 ag~ or in 1 ~: ct::lt p:m .. ~a~ In dl~ulkC ~.ter. Coocutraduns .. erlt 100 ad )uot\ t'd I b:l t .. 20 , . mouse l K.eived a volume- 01 02 n11 T h~' initial doSlt' ",,-as at a )C" \~l of ..00 a~ 300 rn/;!Lt . f oint up or do-...'Jl by • 100etOl of 2 Ocu,u:mall~ an iDtltn-al of 1'.5 ""AS ~ for c1 ~r "ppll':L}m<l' t lon . Dos.es w, 1~ J lb ~ 1000 m t I I.( '" rn not ,cnenlly \lSItd . Control a ....lllDaJ!. "" fOrt :..omlnl"ttrrd on l\, Hiit' Vl:bicil', The- an una.h \Io rlt' (Ji'7>rr\ rC Iv: .5 1 ~ 6 ht <lilt'f dos.:lg~ lo~ 101.ic ~\iTlpIC:b ib d .. :_I·rC u nder 1;'1 Cb) cfltcu 11 dea th QCcurrM! dUll:,!: Inl:> time, t he. U US(' of du tb \Io~ fe-coroe c MOrl.lht) d Urinc the nt'rt 24 hr \Io.;tl rl'C.ordt'd Th":l.J rHlXI' male. LD .. \oil..... estim.a.teG :iLIld tht Dl.l.Um Um lOll" rated dose \io U also recoded for w.t m lioU\J:5,(:t.Ul'Ot inves ti''' li on ~ Ruults In T~blc: 1 Iht' pb ,nu, OU' (' luted alph.:i\odlully \Io'l tll t heir local ' ·ernacu l&.!' nanl ~ :o.n:t! t ht pl ... nl 0-1r1 (')( t racted The: m O:lth and loc.:;.!.Jt \· 01 n.lIt(· t lOn , t be blOl~iQ l t est:, urnC'd (".i:. 3-0\1\\ o~(' n' C' d iUld maxim um t l.lh; r;l1 eO oo~" \~q D 0 : LV" .m· allOO f!h·en .in thr o t her co1u:;:n~ co! li'.I.~ l abl!:. Son~ 01 tht' aCllVC p lant utnct! n.. \'t' lx-en hacu gnattd furthel atld the fractiOn!> \(' ~Ird . These pb.n~ a5 ".,.e11 as t heir PerlormallCr' In c"n' bnnOlIory t(" 5~ :lit' listed i n T.:abl~ 2, Diacuu ioD A coml'lI:'h~' I\~: \,!' hioioFiQ.\ scree-nlr f 01 2t ~ )!OInl l .ul!:na l~ occunmr In India hOl ~ L-("r unl·a lIul !to !.. n \' u l li,t"t' p !.. nt.. ;I.: .. Il'put t"d I e i. .. \ ! r. , · d:~n .. ! l 'lof ~nir~ .. 110 sow !..H It Lorn. 1:'\\ t' .. t1f"ud t'Olri:n h \' o tl." r \oII OI kt'r ~ al!'>t.o lr, ~~,·.raJ Ir, >:",H{t~ tnt nnrunf' o! 0:;;(": wo :).c: ~ ! ..;.\~ 0."';. COrTul.o:"lrd Il It' . r . A scir!nrH {""us.:: : . .:" BtI,:~c; r f'n:. U ... hilt' 10 oih!! , C;o ~t'~ 45. '::-:':' I ~"; ,. han b--~r. U1.o'.lIIt'O 1t'1:' Atfi( ",~'r-.( . : 11 . .'\.t:.~ .. ~ ... ~ ( ,IO . T ~ ... .I",.J.u I r.i. ,:: ~, Tr.:- hCl"''"'\r' . ' n',: ur,' r xpe£lN! ~ n cC' ph ~'t O('~~ulucnt~ ;::'- •. 10 ..... ' ((' loLl'"~ oc:pc-ndlnr 0:: e-colt·p':2: !~~1('" ~:..t .. ~ I:mf c-I cotl,ruull. chm.l c. . h..lbl: ": c t; .. 1~.:~:~-t.;J: ;Ie:" " I\, hu a ~ Lren ob:a.u:.lrc 1 ~ ""'me i ;.: B ... l.~r}_ hl.\'C' not I...--l":n i:' \'rSl lplltd eJ.rhc~ Te.• < i :,t.:t,CI· ;011 the- p!U11 s ho ....; r.£ anIlCa.IlCCI 4."'.: . . :.tl\':';; ::-;OI",S j . E!\ I' TU Lr I - Lin Q,. Pl. .... ~a ikll t.l ~ ':l, a:'1- ': 0 1- 6. ocr- ' nER 1965 P,lI; f lo Ln.~" ~': ~ c r . ", ;,0 C <> l~ . (l'O 'C T:\t J: 0 ' hr s :~l> OIH " : " l D : .... nl/;. LC'(;: CII.L ,':l' -:.·· ... u :, T h l"l l>" ( 1"Un ! 1I~ :::'Cl 1-. , .r. 5Nl ~ ' ~':~ bot a :;;.,. :" .. . ~~! ; ...·:-;;:.. _lu :' S;lC :.:-• .! fl.:':,', j -:J..~ ' ,.Ior..J kI! ~_'':o !;;~ =~ "' !"- c t ..,i\nreo.crll t " (FL). /I'\U' , FR,. lea! Ln. r , t " . ;',v l Pt.:. ;:' 1::1 ~~ 'l.': '!:.; : 00; \ V X . .... : !I:.:k R2. I""..:.,,-::.r R.H J. rool ( Rn . "l.C ID bull. I S8 1. ~' 5 :&>, . Itrl"".; ~T, . ... ~ of : "..." I"' R ,. "':JOo.l 0 1 . I t ll'! \\''51, D on!(>,,~~: :" 1.3. q~-_ CJl. A. U).uboltt.ni.&l : 11. an nhlnp..l: C. ~upr~ lo)l t' ~ 1. O . .a il r 'IUIDtkll:. E . a:: tI\'1n.l : F . • t; t:)r-lI~.IoC'U'. G. = -:=.:.:-~ :'\' . N. bypocl)'Q-Cn:It : K . L. p~ oto~ : &110 ll • • :l. C::CICot"~. .\ 11 r x tn.. :J. !ar t OllJ.C ! Y u~d y .. 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OCTOBER 1%8 P ....TS USC». PLAcE "':-;'D T IlrI.• or COLLJ!,Cl'IOI" Rut: L'l' ~ Oa T.OJN r-D IK BI O L OCJ~"'" T L<rT1S 0-"",U SC.EE :-;' ~D. PlAnt d:r.u, Pm .k' T Ime 0 1 D15tr1cl coU~on WooJ/«tli.tJt;:iu.J . Kun.. PL ltucb x............. ,:,..-sll.• Aug-...,t 71 RT ~d POOIlIl. lf~l71 Drhn. Dv.ll. UP ST October T ...au 2 - L1sT o r Ae-:::z""f) obkf"'ed '·!Tli £,. R •• 1..,.. 100 H 100 LD.... S l<lU (Dhuri) ( ythrv;dC) l.I.o.Q. (Oott.-rokll Nl tCoclpoutUJ Z..l b.,.I.,. aJ.rv-o Ro& b. j'Tltagf) (Ru t1.C:Q.C) BaoklriuJ ksts ca.nitd ou t ,.SD 5 "'·11:.1 ..... , O!' AlmO'l1l.. UP n.-.. Z...... DL,.........•• ~ ... • Hr. A.-.. B ...... c". D •• E.... . ·'I~.' F.H. J" .. Jl. ... Kl. .. 1., .•. •••. ::!dl. ,.... ........ Br... Ct. 0 •. ~ . •• H. J.... R .. ... Lr-..... lJ' . ...... ~ FJU,C'noK ATap '-0. TiI.&I. BIOLOGlc:..u. AcnVI TT 1".... II .~ Spa.smolytic Spumotyrl.c. a.rdJott:l~C do H YJlOIly~mic UypotCI..-,g:,'e ADdan~r A~~e potal~~Oil Ul~~ kb('>h..D e Hypote<lSlYe. - " Spumol)"tie AntivirtJ do do Spumolytie do Anti vial All. timc:n.to l Spumolyti.c Hypot.casivc. spa.s:uolyuc ADtica.Doa H;rpo(!yea.e.mie J.Zltiampbcu.min.Q Sedative Spumolytie /UJticQn'~t .u t.bdmi.o~ A.D ti~ A dn:Ju ,llnc pou.on...b.o:l ADtia.crtykbliDlI: AntihistuniAic. ..pou- .sc.:. .... ,.~• ..-pw.a ~p,u (FR) .KM."l..._ (FR) T.-.anaJ.tU •.J"u eFL) Tri.w.JAn.. """OU' •• eS"I) Vo..,.._ ..,.,.b.tJU1ll U'lu",. iUOea {BUl 246 IPU .....= H ypotc.Q.J;i yc M tLlCc Z,lc.holillc An th 'iraJ A; t:..lI.::J pbc tamiz:! II: AntivinJ DHAR II .1.: SCREE' Sl~C Of lS,D1A?\ PLA!I-"TS FOn BJOLOCICAL ACT IVln'~ J'ART J ani \1' ) (Tablf' 2) 1n a fc\4' ca!te'5 ~imil;u phOlnn3 ro· )o~u! aC l ivuy h as b<"tn repon f'u by other workt'pilom clO"C:I.\· reiatt'd planh (e . ~ . llrpu l Cn.~ '·~ OI ell· ,;t~· in RJuxitukntlror. mo..nmJlm iCOlV('lou. and spa:.mo· l;enic aClh·,t)' in SympJoc.OJ r/Z.UfJ't ~u). A perusal of Ta bJe 2 v.-ou,l d indicatl' lh;;,;, a nu mbc-t of plan'U sbo..-ed ..act.ivi ty ll; th the crude cAtr.'lct which could Dot be confirm<d on fnction. tion. J'hi.s could be in some: 'c asrs due t o dimin3tion of tht inorguic consthuc:nu du ring fracti onation. In othtr ases t bt act ivl ty observed with tbe- crudt' utnct could be due: to s \'JlC'T,:r1nic d ircl 0.1 two Ot more: constituent!. .....hicJl mi~h1. ha ve:- ,ot conrrn· tnle'd in diCerent fract jons thenby leading t o J~5 oj activity in t h e: h actions. Finally the possibilil)' (If some labile constit uen ts getting lost C\"'rn durin,; tbis mild chemical treAtment c.annot be' complettly TUled out. A number of pbnts, "" bere- activit \. could be Ul'D6....mcd afttr fra c,t iona.tioD. have ~ wlecttd i., • d:t>ilod chemi<.al and ph&nn.colog;w Sludy. urther .-ork on t hese ) Jants "" ould be- reponed htu. AckDowledgrment A Kheme sponsored by the Nation al lnstitutC& ct Health. USA, for t.he d evelopment of anti a.actr ag<D1S of plan t origin (Grant No. PL480 131304) has CODrribul<d s ubstantially to..... ds tho cost of plan t collecUon and, as men tioned in the text, the a.nticanctr SCfttniDg dati have beco obuin.c irom the Cancer Cb<mo,ber.py National SeniCt Centu, Kctional Cancu Institute, Bethes.da, Us:."", The follo\O\-1n1; pmvided t echn.iC6l assistance:.: 1(. C. Ar)">. (Mrs) S. Bajpai. v . N . Bhall• . D. K. ~."1\. Bh.tt.chary•• S. C. Bhl.l. H . B ora! . H. l!. ChUnv.rti . s. K. Chili.vorty. T. K . Chouilhari. (Mn) B. Du. A . Dulla. P . S. Gupta. . A. Hai . A. ] .J"m. (Miss) K K_ul. W. 11 . wq. ]!.~ lb,urya , R. S. Ah!t,a , A . R. P a..rm a.nik. C. PawaI. Ramanand. T , 1\. . Ro)'choudhnn. Illt .... ) P. 5.t;nani. G. Sh::ml.:;u , Sh~hikOlnl . G C. ~'('n4 .:.nn ){ . C. S I r\'ut :"va R dtnocCi 1. c...,,~. F •• cd •• A .. t)"fi(. r _~.., ( A.~ckwuc: 1K- Jo,:.c:.- Y Of k L J %3. 110 WcdicaJ RCIIQ..I'dJ eo&utcil., L ..u t. 1 0911). 11) 5.uu". l!_ D.u. S. R. a: S..GK. B. K ....... b ... &.r.. • ." . M .....- ~ 11963). 231 . BJu.n.. au,ay.... B. J\ . A Svr. A.. 13.. ArrL ,..,/. P ..~· rAn .... JJ1 (1962). 61liAw&JMG. F . Sno"&.U.. P_ B, . } . P"~ .. .1 (l9fI). liS . ~~u.a.. P • • H.AWKJ'u" T ... /J,. ). Pl.~ .. . 0950). 239. }"me 2. 1. ... S. 6. c,oJ,... '* i . S ..... A. B . " H.,.r KJlfc. "'6. r-_ Ii,. J. Pl., •• : .. U (1960). I . WJIIIln.ocJt. J. H •• J . P.r&nl_ Z9 (19-4) 1. <42' . .Aa.aa. K • .8. & WA1.1).&-, A . W .. £11'. P~u ul .. , U9S6). 560. 10. L ........ O . p •• ] . U'ii'Wl • • u . RtI.. »C (1961). III 11 . Coon.. . r. "'lI"~ . 1 (l9S~1. 112. }\.-.uo... K .. SoWOCin. )I.) . 1".1 C . .... ~ UJ 0 9+4). 375. ».. n . ~CIl. c.. 4: T.LAJr CUQtDo'1 . J.. B, ) P .. ~ .. • (t9S<). 210. 14. EDDY. N. B. A 41w .... ar . D .. j. F •• n ..t . • rl TAn ... 101 (l9Sl ). liS . U . ProtocDla lor K7U'UillC chrmJo l a,aH.a u t ll.1~ral produC'tl .,amst animal ,·omen. ~d othn ~ . }..uau. c.-..u,. CL'~ oIjt'T. Rrp .• l.! U9tU ). I. 16. JUt L . YUJIi. S. C_ J:tn:c. H .. CalVI . K W . • T ... H . Y .• Vu . F/ ...d . H nul . P.tJ. JJ 11964 ),10 h £. a. PlUMYU. J. P.. J . P--- . P"~ .. J" (1964 ). tIS . U . A uuJC. )t •• JaJ"_j . Pw-.. 13 (1961). JI ~ J 9. C OJ UL. III . L I!c S _nlll". P. N .• ) . / J .... .,J A.l.I ... 2S (l95S). ,,,. 2:0. FUG. P . c.. fuYJC'U. L J .. lt~ J\.. E... P1.cwvu.. J. Jl. " S W",M·n . H . So. A •• } . pu.--, P _ u. JC (1961). 556. 21. HQrI.DJlt. A.. ll .. )=' .... tlllll . ) ~ H~. A. it. }J •• IT-... 1'1.,.,.. s-.... E.o~. &1 (195.5 1. 101. 22. )d o ~ , N . c.. I'tu:KJI. 'P. .E .. l"o..IO.)'&. W. E. A JUCII.t..Uof,OJC. A. P •• ] . P~ . .sp. Tlt'l' .• 111 (19!.f ) • -US . ll. Sast. JL. 1a.1..- J. P l - . . :l4 (1964). 129. Ji . T1»ro. P . c.. H"yt' u. L J .. aLt.")'.,,.., 247