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BIOSYNTHESIS OF PROTEOLYTrc ENZYMES
BY PENICILLIUM EXPANSUM
USltrG RICE l-ll.JSK AS A CARBON SOURCE
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INSTITUTE OF CHEMISTRY
1 UNIVERSITY OF SINDH
JAMSHORO SINDH
PAKISTAN
MAY 1989
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1. ao.d yleld,
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6e1ect
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th. $let d stFrn Gy b. t
p.oeed by
96.
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Sbtu&li.ously UF organls t3 nBtltatned as
. pu3 Etrain bder contloll.d
condltton!, In order to optlalz ploducttd ol d.!i.€d .r.the, tn!
psEEterr *h as ttre pedod, pfi, t.[p€rat@e
tnJlunco €nzy* stDthests and olsbor6tton,undd study .dcrudes that th.
opttru condltlmB my Ery
mere dre txo
t.e seit-salId
orgFntsn
to olgFnts l.! th. .yith..lE ol
Et r.d. pe.€rtly ued for nl.!6bt.1 ploductlN
6nd subne.EEd
liquld cultue ieclDlq@ .
1,4.1 Serl-s1td fehhlatld,
In s.nl-solld t€'nfrbiio prote..€ ale
!U4l!6
rhen
produc€d by
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^e.Cllu
&y sb6trnce
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ql-solld femcntitld
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io 6ubMg.d cult@6 aF report d ln
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5r
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ln rFl€p.ci tlDt ln tl.t. p@es.
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1
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p.odsls
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ard
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fd
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ftu ttF
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p!.d*rro of a 6.rcry of .nzres\*,.
the
thl .@rai.l hyd&lytlc hzyrcs
are subEtldt t.dEtble and
thelr proddtld ts .tlnl.t d by tn addttlon of eb.tste b tlE Uqutd
1.4., the .ffet oldrr@rent I lacto.s d prot !e 6'ith.s1!r
lrE
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tnc,rrtlot
It ts gfr.Flly .cc.Dbbl. that .xt&c.Uut,. .nrytu. aE tomd
by
nlcioorsants 1. rater
pha6.6
of S.oetn . For i.Et nc. th. Baculu!
!9c.tarm(1@) uder :ep6.t.d cddrtt@ p.odu.. Depudae ac vty .nir
It ,6s obered tiEt
dynth€8ls
of
€nz.yde
$der trrst hatf 6f l.lrlthntc ph.&
.o*esp.nd! rttn the groxth
6hd then rrcFa6ed r.p1dly
5t lt of gtltl.narytdE4 . T'i. out of tlrle. rntnop€ptld,qa
.vntneolzad bt BactlluE Eubttltr !n
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shile the ocitvlty of
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c tnd.fG E& to cl$tfy tll. lasstbl. dlfiefu.g t!
.rbbgFph.tc patts of p!6te*., sql.s
th€
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d( ?cz 3ol.r.li,h
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by
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by
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orn
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trE rdtral Cl ol tlE EdrE h.d ! tEotddr eft ct d Fota* lvntlF.I.
E fi&! tgllll-u :Jadl {d trr r:t'! d"t* qn1rr.& Es .tbere.d tn
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4rr$Esis or Dlot
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5.0-?.o ldtrally s.G !r!h yL1d or
Fot--
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a
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p&fo6d effedt of lb. t rp@tle d tb. gett]} Df !t@
d th. F.datlo
ot E rB de!.od! m tb FtR
cgElts. !6 lFt s. tlF prdstlo
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6t Fot !{
6, rl€&
ty Dw bct d! .rd
aD 6ptilu tr@@i@ t! Fp@ted !€ld
'
go and .sh &1& .F kEE D!.DhlUle tn FtlE*. -d!u p!.drito
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t-tco br P.nrcurlu D$!(16), trr:sg!::,
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r
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a o9i1@
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trr tr* ot cqlt !rEtro(r@), ptotar uroryotuars iy Prr|flrir
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9a',1!a ol criltE.
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tLtsldsld .td dljrg 6f Edtu cotd.trt€. rU& tt @lr€1r tf,
ie'!.Et4 .dl &tubtli? ot s.!.!
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It 6@ttsrs .h.ktt8 !8...
pl.detrd
ard
by
!s .ryEay'obd dldld.) rt
of tlE .ultu.. fd 9lotag.
(r0a-111)
lt1cs6c.6 4. t. obr&rta, .! lepdt d !y !d rdtE
rtattgrry .. c1I a lileditnttt
.r.Lar ol t.tt c!tn!.. fdll.d to
{rtlt rI4 tn 6ryE !t sisnrtlqnl1d1. fid€c! 4!!g4E!4r
t!od@! hlghe Fot
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0n
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(112)
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Pqftflub !$edt x -1o14\'-l .'d $EE!!!!!!!E_ 9p.'' '"rt'w -'rFodatl6 of Fot6e at rri.@ cdna. cddlitd.
1
n.4 Eaf..t o! Nblhta:
1.4,4.1 Effe! 6f €rt n @..!
C$bohtdrat 6
a.
mEt lrDortant
cl.s ot 6rb@c€ou.
nubtht !o! t.t@t!t16 F6.r@. u.d !n ildeds.lrd,
.ltrroudr
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arn oDE
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.16 utllt4 .rt& ..t<1., hy&edbd!
codpohd..6. c.!bd !dc.. R.qutr.@t of c.lbon
blr
dlfl@nt
sp.cie6
of olcrcorsantln Ery td c2 m1@t.r ( lc.tic acrdj €uEnot) to
.oplq mred.! (poryscchltd.6, le.t tn!) ud .m Mttc
rR
utt112.d
E.furly\'-l . Th. urdt4tl6 ot lrdu.t tat brs!.duct6 6!
agrtcultlEr sat. rhich 1. rot oth@tse u.€fu1ly pr€@ar
@cblel In tlE laodatld of fffitattE
.u€6,tll.
.Inple
cd4o6d.
c.-"od'
Th. us. or
could b€
product!, !au!d uEn i.tE
tr€ 6l ddt4E \ r"//.
drflent tF..r
@nohydla+r.ldts1n t6 leFrt€d rD
t,|. FodEtl@ of tne Drot€olytic €r4,tu. by tlr. itcledganttu. ad tn t *€
a 3!od
r&ld.6t D!6t?'*! srroE6r-
by
@
!t!:!g! rn a culr4 d€dtr
dpplffit€drtur !6 potlto st lch ,r*.oyb..n @r.nd 1d r.rry !.D.r.t !t
1.Eortdhy
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hd XlEn .t61\"// dtun.L4t d hrsh.! yt.ldB
6.. Fld*trd !y rn-d !tg!E
rh6 dlt|N
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dih lf odEt€.D ltqu.td!s {ttn t( s1lft4 a6. €lbd .o@.. ig::g!!!]g
9:Ig9 lhor.d st@ prote& plodetrd rh.n crdrw mdtqrrd 4D91.6t d
_27_
rrth rtei bEn
(tt4). otrotb *.
*t-r
r€poltcd by Abd€l-Fattlh
epplffit
d
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FoaEtld of !&tee h.!
.1(115){hn basl au1t6. n dluo
l.
ti+n .uga! 6o mfaa4. lnst€d
1.4.4.2 Eff.ct of nttroaa .oac€t
tt. !€qdldai of ntt4o
nicr.-or@ni.c ts wlsttl. .nd
as
cdn gt@
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ntd.iat fd
tne
ertn of
{.rI tn ,t6t.v.r b' lllt ortaln of
rlt!6ss E@e' t€E i1n.a.s t 'e bio!v.th€.i! of a pglttolt! !!od@t
.@tit b€ udc in4a.€ ot dlffeqt ni6oa.n 3dN.! t. Godided td
.nzyre produ.ttd
rhlI. alllMlm saltr oe
f.uDd an
.tc.llerf nitr'ss
I'nl\
.oec td trl. sldtn or tt.r orsant#'"". tn ss6l
to uttltz. botn rn"rs.ntc( nlt€t.€'rml6 d a$d16 E.lt.)
a..
caDabl.
6nd
orcsrlo ( ariro a.tds,popttdeE, rtlt€in hvdlo1tst
coorr.d
ce..l.6d rtlk rhq) nltiosd li)lfte.
Varlow niteg€n
.dcer ced
gldtll of fqgt fd trE rrodEtro
r6Dorted
dF rd
.11 ntdo
rn ut
bv
or Dsotas
!'
rr
3ovbdn'ft3h
!ol'
g,o,tr,(105).
duforat tdk*' ln tb'
/rra\
sh a. ChopE 't ar\"-/,
tlEt the dlflereni i.olat s ot $PgEqLE I!g:!
Ep6ct€
obtrt!*l
frd dtff.rdt .dccs ha€ dtffa.nt EqutEmt of nltlosn ..@€t tn
prct66e blo€ynt6€t.'f@ tctaF. lslate q .nit Ar( kolttttl tM
d.@.ed
Enbuca
otflctnalld ca.!!n.nd cltru. n€di, fDtt. F!!.ctrFly)
-2eDroduc..
rrld
!6oast!6 6f Fot{.. tre .tun18 nltEt tE t!.d ..
. nitr.ao d's rtw .. r&r'i. t (rFl,i.d fld !5gg dd.:Hdl)
Foduo.! opillu !fota!. 6ryE {tEn asprEEtn.
15
qd .. . nltroC.!
o, anUnn .t a1(117) haw r.Do't.d rrlt hlC!.! Fdstrd yl.ld
ot
C.nt. futu |16 -dlr
Fottr s. ..hlcatt .i6dlrttt
{d
.upp1dt3l f,lt! €*tn !s!ad ol e*h hvdtolt.t .! . nttos-t &@..
1t!.
lndsild .ff.ct 6f tlt dmtc.d F.t tn ot .a! $tt!int-t*
ctu €t .r\' '"/ b uF F.dEtld
nrdulan.nd
oa
dt !c.rlul4 Fot6t
tt ss !.pdt d tlEt t{o to t}E& ro.ra Erater
tn
t0'
tt
lgggEl:
rnomt or
.rEa..Uul!r .ctdlc,rqrt-1'ri.lb1t6 Fota&. B qDllr.dad |tn
cultd idt6 n!
!.f
e8a
4Ep16.nt€d
,tth h6i lflctt{t
rhlt . th. cdFantlE .b.tl.6 ot tE
of *tE .rn ,!t ..110L! Fot64 dqby
d .ga
rhlt I! !1&. .l
Dlra.apn .oE
.
tn tlr.
F.dtilo
tt &.i.|!. qltndld9@ !.Ddt
d
atd.l falt h.t.l(115) !uss!!t.il mdM lcodq.tl@ rn o@ ot.d..i..!
1tq6 ,h.r 6.d .! n.ttr€ad !trd. ln co.Fd@ Itd!-t rtr.t ..d Da64
.1a€ rlt ye.t dt!.ct
.
r..hi6to .r ar(1'e) hv.
6,F- o. ."r*"p,1ftilo**.-
Fota* t' adaanLtr rhpontt f,-toi4 ty ualng dtftc.rt
d€anlo .1ir6ss
or
lndc.nto .td
.Mc.. .{rt!t:tdnt.d iltl| rtFt t6n dltc. r.<Itu .td
thg'obld.n
tL! .'dls arts ru4It rNH4tFo4'
(xrl4)z!?oa
'd
Etp, r!
tlF !.rtF dt o.l t 1,q( t@*..8 tlF 4ntt6d! of 'ot'l !tot6*'rb$'
tL o!.tlttttat oi €.dlr nltnt d@-d ilt rynudtt ot tlt d'
otr
tl! ots
h'td Gsrnro ( gorvlct'td" test
qt!t't'ct)
nlth'd
.6!@ r.d D.l!'|lflant ?ff.st d Ot rytll|t'rt ol Fot'*' Sl_dtDlt
th€ sbraty
d
!1o6vntrl..t! of Drot
.!.6
bv
s!r@' @g !'!'o1" 3f@
und'!
ac.. ( c!E!' 'td rda'!lrc)' t4tF t'd uEt tn 'D
o!.! Fpld. .nit lvBtn rd tt b.ttd nltttlo tdo" tLn tndgttl'
d1ff6i
.rt os-
nlhorpn
"t -.(120).
-to-
1.5 PEIJi6i!6 of Ntd4.
1.5.1 n6!!qr-e!-l!ts!99llg!3!-!!91!9!ei
F.m.nt d b!ot$
."
o@91d aqu'ouE
ntxts' 6l otlltrsrubl' trit
aid M odw+lttr'
ln.orulrc €:ir.c.llula! lcodwtt 'cohvdtlbL
tre s!a6t'd fd
to
Eleult!
o'11rn!!
ftd
Pa!tt.l.!
..
.ub.ht
tlEtt dlff@c" In t'!t! 6t dv5t41td crdtel
st4
iltffGitrb
'tc'
rao!4ii.. dh 6. !14, d6.Itt'&1ul'l1ttt'
or 6't b lq' Fi&r'c
ant d.GtE @ uF f4tEe! $toh a1ldt 4Fttlto
$lqtld t ..d
tr
uslna
@
filt!.t16 aid c'itir'stu-(9)
'
Dlft.i.lt E.!6FtId !f@'!-tE!"d
{t€p.!4na upon
th.
Daod@ts
W
drfr'rht r@t<'!'
deltrtd' !o! Intbnc€ tn tn€ '€D'raiid of
rot6!6 trd l6thr d.tclto$rl's f6s1t ''' l(4!Pd4!1I' d!!:'
E!9!q!UE !!:g9, EP!:AU!' lqutslo '44!!@ -l*ierl 'P!'Eelll'!-r
clqrng!' E494rrgi g4p6essg''
pacc oDF.!
E4'ull-r {l'qcd!g,|
turgt4)r -o!p"."r:r* r,',qg'('4"16) "d
lnd
P9!!9!111!
trrouCr ntttn
.tr"---(rzz) wcelt fM fMt d blotn !v tl1t'tto
!v@1la ftd
tc1 lilie F!et, flor.la rx.r nort- "t "l(1t2) *FEt'<t
mtt4 uE@8tr '
cu1t8. both dtatnlng !l.lt 'lotttq atd !G6t'o1vtt'
s.d.ck oi
"t(121)
@d oetri. -5r2 r,t, th. 1.E6!€d b!ou! of
-r1-
P.nlctlllu Jsthrsl1r to ficl trt' ttF filb'tt@ F@'86 fe l,}r
s.Fdtld
oI {c.11r, uEoulh
&!.ntd frd
,Br
brotb urough
by dtrtflaatt@ ai
.r
i1(11
llltd
4ooo
pr.B6.
frllttto
F@rtr I9gq!&!q4c'1ta
bv
BucrlA
/o\
t g td , d!ut'3\'/ '
)rsokied Fotes. e..dwti@
on
flN'l'
loudd
th' 6tn'! h'nd tdf'r
E!!crlr+]! Iqry!q+! rrd cdtE
bv
.}!.1tb thlo!6h c'ntrlfugallon !t 10oof fo! 20 'ttut"
614 !fl1tp6r' ttlt'r'
fo11o{ed bv tlltratton @lns 1'o dtfa'nt !6e
broth by !cp6!.ttna
,nd
€eh as o.Eu .nd 0.4th.
plod@r
s.f.dl dthod. .t' @d t6 obtaln t$e e'k'cellul'!
tn a c@c€ntaied fon pr.stnt
!t
t'tte
llquld pr€:' 'I
3eh as plectptbtion' e6p@ttdn s'd
prcctp.lt€tl.n d.th.d
tt 6'sttv
1n6
cult@'
rcd6'
ultrsftlt'at16' Ho*thr s'
enplov€d
fo' eatEc'llubr t[ot'45! ts'lalld
l12r,l
|un !vcroPr!'! ;
stntrtcitv. For tr't'nc' p'_tas'E frob Pentctll
dL '.
pellculru Janl4bel]u(2u)']!9.*-rr* -'*lt4(t')
!gi!9-!(t) re'
ol
5o-3oi{
adonlu
and
!!4bE4!
+r'' addttron
prectDltlted rrob c€1r rree cult@ ttoih bv
6urphtt€ 6nd prot€Baes
ftd Psl"lr-lq
arcc 2/r56(12t) 6nd !:!9l9roqE9_'
iglg,rrdlu lie!-cor'{t
,.F ltatpit tld ft*'11 ite cult@
$i*u. r.s l.ft td
one
d_lPoil {-rola\
c'r'vr'ra' \''_'
broth uslDs cold etbnol 'nd
hou at Aco' Th' precittbtes obt'I!'d te'
tlp
''
4FEt il by .6tdfua!tt6, trE ooll*t
d
F..tptt t ! dt..o1r€d t
lpDiopd.t bdfa6 !r. dt.ty..d .g!ir.t eG hufte .t
n!!ht . slood
c@sul6ilDg Droi.ad trod
i- 4*r.s €s lcerprt
t€d
tt!. qoe€ts b1E of cult|E
b.f@ tEuE pultt@itd,
th.
9€t !.dwtton tn o€rt
courd b. chl€!.d
ueina
tr..
4Co
td oE!
c.tt &6 culiE€ brotn ot
rltrr .6ton (r4).
tcotn
e &ds.d t Iru btd.
ult .ttltdtl@ prca*!. .c. rn!ta*.
curte€ bloth of Srb$tfacnu gg1!Igls!!g
usolran. Diono
tlrbau@ sy.ti adt coc.hd
Fts.n.
Ur_10 uErng
tJ6 antcd
TC
-s I. dlalyr.d oE ,ight i!.trst
P.€clplbttd xlth tnorglnlc ert! o. u.rr8 orSsr. &r@t.
uttiattltiattd
t€clbrq@!
FlEll,
dir.
od.@.<t ,rth ,6et€t!
and
,hcttdttd
.r ad ,tti tt.tt d Nc.s tc 6?- DEttlo{d,
PElfl€tld ot dllnlc Dl.@r.! & cftlr, .chLrtd b, @rrS @
d t}l o6nblBtlo of tlE torrdrrrs .lEdto8r.ph1. !.ubi|! .uch .! (A) C.1
f!ltEu@,
(B) t@
.rctj.,. .nd (c) Aaqdk chfoFros.dv
d.p.,runa ulon
tl, EtR .nd .t4 ot tll FodEt _l@ul.r.
h gd drdto8Rph, irE GsaDt .ub!t E . @ er.ht€d m flr
dilfdenccs tn
b66ts 6r
th.t! ..}*ut.? .!rc ty
Disstn€
th6 doh
thlough
I
col@ contrlntnA sioller s.t !€tttcte6 ot apFopalate BtEe & uEt tt
r6.Ae
Dl.cule.E eldt d 116 set befoF tl,c m1r.! &r.cule..
heE6nily tlEee tyDos ot
s.t!
a.e @d In Sel ch!@!og!aphl, seh
.!
..!hro@ (!3!o$), .epb6d€, (co.. tr*ed d.xrEn) .nd Bres€1 (pory€clyt_
a'ld.),
TrE
r$1att6 or tho p.orltn botcd.s
slna ft of the
eel! ts tuc.Bsfutty a.h1.Ed
abov€
For Instan.. uF err6cetlu1..
lfores€s of
lurftert
20ttl,
t_Fowh Aer
by 3.1 cb.mtos!.p1,Jl
sodl6 bdat€ buf€r of
SEpbJ,
ana rcport66(12?).
phr.rltr@ cJglll{,ee
slng !eFd.:
pH
chldt
C_?,
.ollb, €qrdrtblEi.d rttll
s,5, contatnlns rroid ilacl lnd o.o2* s.dt6
ur. &acrton. xe.. etut.d rrtn th. se luffFg4)
IBI Ion dcharc ch.ffir.caolsr
a21d. and
'
I4 ton
exchqnse
bo6d to goups
ol
chlomtognlhJ th6 blologtet mcr6Doteule. a!.
the oppo.it€ ch.l8e ol
t6 dchrn3. mtrk. tnd th.
seDaEtton ot bowns!b6t!rc.. ts .6hieed
,h6, tn6
.tntl*
.ha.a6s
{tth
ozyre p@lft€rlo,
strona tortc .oncentElton
an
aDprop.tat pt
4.. rcc@d.d to Frnt€t,
8nd on ttie ton
acLrvtty or
*clEnse!
t,},€ hlAh
dDd
and
.r&Eolyre! .l uE
sr. erced .
For
$ttsf6ctdy
al6. 1dl.tc stlhgth ot .r@t
cha.s. bofl! or he
.lryrrc ,.ot€tn
thi6 iould allo retrh rh. gotubtllty
en4m\''ol. td rDsbnc.
tJF ?@o
and
tord pulrtotl@ of tJ!
qt!...uul.r Drot.!. of C.ptatolDdl@ .!*t.B h! rpct
6t.r'- i ulne
DEIE
uE Daift@tl@ ot p&teo r
(o
.
by 5.t
rr
seCEdd 4-50 f6rto&d bt sDhlder C-rOO, ,r,.-
cdblDtto of trE t{ d!.rr.t!!.nd 3d tlltsitd
5.0 .nd 1.5 f61d rn
rt
tulrp€cttEly. st
IlEt,
Ds.ies rr "t &@9:!9gr 9qe!49Sff
!.tna DEA! sepad., t-50 *ttn na! nacl
and
31n3!. srdg
t oJ6fi) cmhtrattd sladta! rn o.eh dblrrD€ bdta of dr 7.o
.l t6 d.lE gE. d cormEt tly .Eit.bl. bn
.f tlm (a) 6t1u1o- t@ .rctb,ae ..d
(b)
oqt
eEh rd aor.nsa rF iod
nfr 6dtrbl. fd tlt putflcetro ot azycE(126).
td qohnse dtftst clFle gtdD€ or dnldlc d
cletel
elffi a!. .tta.h.d itib c.-tutoe bv/rdctrd ud+ corroried oodttt.n.
of ddt€tret|d .tn td lftt (t..6 trEn r !€9/s) e o'@h b.@lE
ID @1ru1@
(, rdsh
l€El Ey &!16!r. us @u!:.oe xd
6r J tr'.
ld tftt atlds
and.lutt6 of polt6r&o&tcs in nlld c..dlil@. In c.llul@ lon
elchisF .y.t6 r@t€rE (n-{Hl+) .E absb€d d to or{.tlul@ 6trd1c
brndtne
€:clanse. ( oeu'tlo*-4oo-)
!t I tt
.Dov. uE
ro&r6btc por.t (,h.e
plot.in har. legltrF crE!a.).p.ot rD (R-4OO-) .r.b@h.d
c.Udo* .ntd16 etEDs.r.
on
t Dm, -
16 4.hisr rrr.. e Fi4 l@tuble DortE l9c@ It Fbt
bst rynthetl.
Id
dcrEn3.
r..tN (poly.qr.!d. .ynthslzld ,ltr
tb€
cdblFtld of ltyde ed rlwlbqr.rb .6 portfttbcryllc tb cdniaitor
l€a
6t vtwlba'd.ra Ftb6yuc acrd) rttn fr SFta @laclb
d*lop.d
irnl
btoto3t€1
Th€
.*h.!
dd. cffi!.uy
'tE
aGtLbl. l@ tlF b..t E&lutld 6l iL
&l.cul6.
do.t
-OH
Unked hlEh
potrid!
| -g),, -aO- 6 ||'.
@
contqtn anidl,c a'|d @ttonlc Eroup!
dblr dcr.nlp rttr blml*rd.l *'.
(C) Eftnlty ohloetos$Fhyr
c.l liltFtlo
6 Id achru. crEdbsnpl!, lr t. wr Fobd.
to! ..!oEtld .! bloloAtcal Femt.od6s, *h !! ,mt6tn,ro1y*eh.dd.!
.ttd tuletc 6otrt!. Ho,€q itE to ngtislbl. d.rtt t!@ tn udt dvstce
ch.trt@l prcp.ltte€, ihe.. EtlbdE
pulit€tld:*ttl
hydlolysla
16
r@t",
ar
ombelg@ and trk6 nuch
r!re
tD
tlF .4u1t d6 to d.[tEtt@, o1o6gF, drvnilc
et , ! 6st.i._nbL .eut of .cdvrly d6t!S l.ot.t]d prd.t@
To
ov€!3om
tlts rttlftculty o @...Is,6D.! rfftnity
chmt gF*V tr deEloFd tb G6t ya.. f6! uE l|Etttotto of
btologtel Gdrh16u1€!
on
tlE b!Ei. ot tn 1r drEcie.tsu,o plopett..
to b&n dtn otlr! Dteul.. sFciftdUy anr tlEltbly L, t6rnS a
->t
spdi!!. dl.&cbbl.
duE .ub€trre. d lrtdttltd.
!@h
.. dztlra
!ub6b!t d.nryE trlh.tbltd oo.dq.. d otbc tul.qil.B
tdn.64u
rhicn
co.p1q
e !6ci .!.clr[y dth @t!tn rl.cdG. Fd
tDat
r
acttct d !Ft64 of torln. ht tr
.Dd
Ebbli 6t ltt l rel.
putfl.d
.nd
,lf1nliy crrdbg!.tv
by
MroFhobt!
trtddr@
tlt 6tctr
iq.
usrrS
drlvtl.d.ee cLl6 a*t --rn sCE&s(1!). srrrrsrr.cla lcoerlu
l'@ r.belruE EjEOI ad !qa!E !Det.€ t@ ?6ulcit
.raodt e4plv uer!|l rrk Dtrt@ ,rA aril
l s.4
C@
by
.ffblty
(1v,1?\
l-E.Crr@ e4.ctiEly
fi@ntbt
fi@ghlty ol . btoDolt@,. r*drt
FF-td to ..t b1t!h.<t
pEl @tlon & d@n.trat d by a .hgr. b.r(r m 6!r.c t .lntqEs of
.sFFtrd
.rp1ot€<l .rah
6.
Sl|5
.1@tr.drdr.rs,
urt drbttwAttd,
tg@t*trtc l@d.!,thln Lta rrn p.F! clEdt r!.dry $toh rdrlo+rr tn
tl6eE.. tbt laot tn 1! !!e tln rtrp6rtr.s.nt r! a: .ccprrd drt dd.
F@ rEt.d trE rr-s.n!t' or Fdo'€ or
!9!9:!!!!s rr,u,rnorr-(26),
a.r
P..louur !ggt!\"/ and @!rtd!g!I!r rut!!nr!4@r"/
I!r. odrtd
by rhdlng..lrut. t ttd ulrra ${,.t.dboprsGEt., tn d.t!.rpi.i
uF
t:!&lr.!!.l! !.qerdu prot-e F!.rtt6rrd Fdd., iodt6 .t 'r(,i)
dtuoct t d flE lpotB 6 d.ddtn3 F!.r ohldtodhdv sws.6t1ns
hFrti.r r
.h
'w
tctlr lbFd
hr
d6..t!
iu.1ya!r lnd
td
-r'
ex.h.nsE chlomto3Epw
rllileu u!
"yntrr.u"
to .cht€vo the ftnal honoaenlty of th. p.oduc!.
hmsenrly o?sr.Clnocec6 6E@
poptra"6(1',4),"" dorchstFted
llys crir@t e€thy.
lE*ty"ln(rrr)
aB Btnsl€ spot by uBtns
",d
thln
Penlcllliu
by
expamui
llnt . tll.m6phiti. fuaE
ard
. Bft
p!op@..t
1t.k ln 1409, bolongs to the ktn3dd of TlEttophvt3, .las fusi
Irp.rf*Lr
o d rh*edln6e\"'/, lt tsd Dtlnt p.Lhoscn rbgs, los€.
&!tt 1n tlanEr! ad sto6s€(lr6). penicrtl u gllEnu t.
and
so-9d applo
dlstltbuted rn a *Ide Eng. ot
$tl
oa
dtfferot rtur€
rhoush out ti'e
rorldr ert ndtng trd luIace .nd depth dd to 42 rnche6,3..tt. esron
to
+^he
subt oplcs an.l
Thde are naeroa
fbn
Eea 1ere1
to hlah €lpt.. 6o!(1',?- 1'9),
tsldts ilEt tt t. i$lat€d l!0r s!$ l.El ,$1i
s!3h,s11ne b€ch,the root reston 6f ash, c1orers, $a mtetdanpttsy,
nests ard
tdtlEs of fr.€ ltytns blrds,stded 6ppl..,grains of {bst, cdn
b.rly,.asto. b.ar, to6to, pod. of :rolnd nurs,
stored Feai,
net lroducb,
exFngh
frozen
rrutt, eke
doad
and
forc Fttdly ardtng
flurt
adult
.Aar and Capek agalr att€intns a d16@ter of 6.5 cn tn A-14
qutcEy
tlEoush
cold
iuicoB(117).
coloDies on
6!oD at 2tco, The colonle6 are Sser€rty yhtte at !rr*
tes,
dlt 4rract
d,y.
Hho
nu!.hadrlg
yelld to ble AE.nr Dd. c€l.ndtn. srea or *a. gre.
xlth rrpenlng of th€ c6ndra. colontog s.n stno.Ely nouldy,llke ol rottd
appr" or
ardt& r!urty(14o), Tl;" s"o*th
tnblbit€d tn
vlt
o by
dpeclos and a net
or
c@hllodlelr an anttbtoNtc rct roltte of ClEeton1u
bollre 6btatned troh potclUtd pgMl9g9ls .nd
-9pcntquu roo(1
1.6.1
Product€
17)
ot P.ntcllltw ednlu!
f6!cll1t6 dp.@ t! ted rrdelv
rrv
dtrl46t Y6**t rt tlt
dtlcild o! btosyntlE.tg ot hydlolvtrc 6t aE
oi toxlc/ .nttbtotlo
tl'e nlo6y.tn
816
p.oroFctr'..(1t' 6r+crr.rerc(151
Ari.!rh.
rdt r
1n
and
th. dganlc copo'tna
Etwe.
ot hydrorytio .na,ft. sEh .! '4+ovr...(141 '142),
146),
tn* ta..(rr?'14s),,r"*(t*),
rr*prllsl,
), erruorytt arym(t?), p +r*o"ra.*(151),
pho.Dhat!&(154) lnd
p6rydrta.tcoE*.(1'3)
reportea uy
ertour
P.'tdlrtr @9!!g o Et@1 .rd .tnth.tt. n€dlt 6dd
@ dl-&ltd .ultc cordtt.tdr. Fe tn.t @ ..,- aw] e
by u.lDg
subclsed
btoElntlF6is ny P€nlot11i6 6:pan$ Btlatn r-1045 (tRiL 54r) usrns
cd ste.! l&E .rn gtsoe Fdts wde etnFA,.d cdrt@ cornrtr@
h"E ben lit nt"d ln urlt d stat€(141).
1.6.1,2 Tdtc com6d.
.nd anilblollcBt
;c€iyrct\r@brD{1t9),2,1 !uty1.B s1r@r(l60),p.turrnG6r-15',)
ana crtrrnrn(1&) @e .1& rynth.dr.n by P.ntctlltu elplnEu ullna
app:,.
.y.!
.rtc.., .!pl. lut@ rtt! rtffil
b.oth(r@6t
r.dtrr lotlto datbe
nroth
ltd
.rt .ct .t!t s@o&) oF Et'm1 .rn 6dt!1.L1 !.dt.
r'ATENIA!5
TIID IIETHODS
-40_
?4c--.848$!E-!glq@!gr
?,!_-!c4:s
Ir Drcdi ddt rlc. r[rk ot Id-P.k 6 colr.ct
Dll!!lc! L!!kE 6,.trd,P.*t.t ! c. u!.d
tlEouah out
oFrrel .iusl. of rt6 bult E. @t.d
d froi
ric. nttl lrllaF
th1. .t!dJ.
\16'
oul by !€portal mtLd! '166)
It .o.l.t6 ot h.rta!1u16i eUuloo .tit tlt^b .loia rttt! tlabt.
c@ponotr
llke
63Fnlo nltro8E!,
d.s.l! etl Itdd..
3J,€-sE4s:P€ntc11lt6
g3!g!' ltnl
C-t(l
,9|61 Es obi.ln rl
of ttloroblolog, Uniw.ltty .f cla.a., UJ. Th.
2?to
and
d
asa!
st!nt!, .dt tnins ?l b.to.s.! ,
orgF
lta
trd tr D.Fltdit
.ls sa tutntlln d !t
F t F !r'd A d.ttro-
RmEd .!ra sry ultE -nth6,
2.1.f
Ch.rl@1st
All .heotdl. u..d sF o! .dlyttel gad. rluut f6tll*
2,2, iE$oDst
2r,1 k?6frtl6 of in@dE
Th6
.p@. @e e!.p.d ty Gtns lodp and ghtle ltF|.in8 of tlF tub.!
tr{ tlE rto.k colt@ F$rctted rit$ !t€!11. dlltilld rate.
re!. cotrtfqed
fd ]o rtrutr. .t
dtotiUed
on
splttntc ootlttuas ( Hea.E J-150O fi.C.mrv )
lz,(DOrDr.
iats.nd
Th. allq@ts
th€
tlr. !F.e reslds E!
EsEFnded
.orcot@ttd of inocutu 6! Gtnt tn
2,2.2 Biodnth..lg of Boi@..
In tn st€ltt.
d jO
i
106
hrw.!
'1 ot b.$1 Edrr(167)-r.or* "t r, e.o (reFdrt r) cdt tF
tns 0.t a or rtc. husk fiF pdd.! (40 mEh) sE t!r.n !,2, !! cont@l
Frrqy
fr.*!
,!tn colto
pluss.d
r@1 .rd 6ut@bEd at
lJ xrlcj2 fe 20.r^*..,
Clso& .olution of a lodn 6tr..gth autdtav€d s.prlatlty E. rdd.d .dtl|
6t riur.d ptp.!t€.. TIF flasts sE t.aulrt€d,Irn 0., d., rdurr
contalnt.s 50
r
106
Bpee./ol arn
incrbotd (cltt.nranp) ai
220
rd6
tnclbat€d
rn !. oo6t..l .lErria o,lrtol
.ev tr-1 61 21co,
Th. coEa of blosynut.El. of ptot€olyuc .n.vhe @6 fol1d.d by
r.tuahs co.lel flGt. atta b€nt, fo@ ho6.lrt€Mlr TIE cultw tcoth
,r!
*paEt€<l
xyceltutcshld
fld iyc.I16 !y ttltrtld
irE6,reh
,rEttur,to-l fttt ! !.0d,
,ltn dlE rlcd Ere to rtuE &lur,t. s,ym.d
a2rej.ct€d dE to p@r htta..1luLat p!6tea36 6'!"iiv'
rhere as
'ult@
blol.rl
,ss studted fd €ttFtto! of 4t!6.eUu16r p.ot6se,
2.2.
lff€ct ol
The
tlft E.
optlom 3rdth
6f ircuhotlo
'aE
leub€t-Ion tt@ edlo<t 9n
ch€ck d
aa
stnd.t€d by
deslb.d tn t$€ ectlon 2.2.2,
.i dtffe.eni tlre
The
Ensi4 tror
24
tlE stand.rd 4tnod
D.otelvttc acttvtw
16 to
120
l6,6tM
acttvtty ras obt trod tr tlE D.ltod 6f r'3 houa gror.th ar !h.m tn
22.4 Elf€ct of dlffddt Ebd
orotae enzret
The
and
effect of
srEoe ilth
sJntr€EI3
s.
and
cornEteep
'tubut
Bouced
o the lwtlElls of extecelluld
Uquo. r nd16s!€.,ratftno4, €Eros6, F1tos
11.6 busk ftDo
pdd.!(
40
studled. Ctucoa, se.ose, m1to6e and
6epaEt€1y aDd added
DtagF.D
dsh) d plor2oe
latftnoer@ .terltt..d
agept!€uy th. cuttue Fdih b.fore !tuu16tt@ .
Ihe t.@ulat d fl6Etrs
*e.
t..ub6ted tn c.oted
zj'cofor 5 days ond r6.ur!s are
2.2.t lffeql6t rtc€ hGk tIF
dbtt t *Etlns l*ubatd at
lhoh rn-Iabt+6,
ndd c.nc€ntrattoD ot
rhe -rrect of rrce h)Ek ftne po{der (ao resh) rn rh. cdcatlatron
Ersr.€ f.oo o2t ro 2.os p€r too nr cdr@ mdta ar a @bs ssc€ d
pfoia6e synihe.tE
h. Et!dt.d.
The ln@urat€d
lfast. rd€ r*lbated rc
4>
4S
ho6! .t
z?Co
.s d.&rib.d t, sttd
2r.2. Th E&rt. 6F dft ,,
2.2.6 lffe.t ot rdull rg d Eolio su@r5,
ara
optlu di of F6t€E rj,iflE.f. sr stdl.<t
by
nrnt hfr|a
oultu€ edta at dltfe.nt gI, E.atnS Iron lr, t! gror Th. asbr.rt tt E.
.djqtld rtth lf,1 d
r.B
srrd
our
b.td. 6t!!lUaild,
!t ott
Th.
ltrurltld
b' .t€lnad Btl|d a. .icslb.d ln etl6
.nd lE,rbrUd
2.2.2t
n
mrlM dyrtlElls of petqe n6 obslElt.t llt Etu.! of about 6.0.€
2-2.7 Effet
?ne
oa
.ultlft edta
.ontatning dce hultt
.xpatur.pc€ !GD*to,5a
5o
r
t€r!.8n4! Ersl.s f6
to
55 co
E. ob*Ed to b€ @
25
n@
lhcd.t€6 srtr p.nrotltl@
to6 !pd.s/i1 rcre
td
4s
rEubt <t.r diar€qt
hoE., rrF prot6* EDt 6t.
a! rrco as.hM i, llsFr,
2.2-A Eftet of dtffeht nlborh .@.
d tr€ m6dEtrd .f, .rErdrrrrs
tt6ts* slEr
rn er.c. or (mt4)2 Sot
ln Ui r.mnd r.'ffit tio Ddlu.
tg,IO_
$ uF lim .@.nbatld .! a rlt oa6n loee q. studtod
.ccddin6 to tlr .t n.t t.l EtlD.t.! d€lelb.d tn
Git@ 2.2.2r Th. $€u1ts
lXO3 and laXoJ
.E FldE€d tr a.!16 ?.
2.2.9 kbaotton o! Ip'tF4tlular Eotad frd Palcllltu .r'tjo
I bdn retst't ol t!.e,
tb1.d
q'ell! s.
8loutd ln E
roull
rctat tn F.rmc.
6l sl.B Frde ,ItI lc. cold lrb.dEt burr+ dr ?r, TlF dto.t Gt
@ntrlfusF{t
rt
12,0m
Fot6e .ctlrtty
!p[ le
accdding to
2.2,10 h6t@s asa ct
Itote.se
.t
!
ntdtd
end
th. &4.@bnt eB t!.t€d to!
Drot6t .*t
-tlbd a. -polt€rl
lodr
.ctivlty tn tl.. cu1tG. tr.th snpl.. Er d.tdnrmd
or petjE,,ra
by
tL
rut"r(&) t odbrdtie,tih Lh.! of L@r et r1(3').
ro 1.0 dl ot cult@ blotrr, ,.0 nr of tr.-+tct
ll'id dt 7,
of ca&tn &lutld (0.1t
a'd rrcubat€d ai bco
?o 2.0
actd
15 otnut
r&
pH
2.5)
{e
d .t lrbE lsctld drt6.,
6d,C.d ard
k€pt
td
20
.ddcd
2,0 !L of 1tg
ard
blchtdEc.trc
dhut . ai rod t np€!at6., Th. p!..tprtrt€
F. rtuval tt cffttflgdtr@.t 4oooa tq to d.ute..
In
an 1.0
d
6r&@t ol th. abo€
of o.tx.od1u hJ'dtdld.
add€dr Tho
.U6tt11€d
!d
1,0
!r
of
qt
Fo:r1,r
t6tal bttn E. tlFn cd. npi.
Et.,
A
rtueai.d
or.!@t!rt,4,0 it
trEtur !-S€nt
1O,O
tr!
by
( 1!r
V/V)
r4
ldiU|€ 4.0 dt ii6!b1€
bl* c.1@ de!.to!€d s€.lot rotEd afte 5,INt.
at
FotB.. .cttvtty sli s.
th€
rit Et
d l rla of ty&!In.
deflt€d .E tlE
.tu6t ot drv- ttri
add tl!. 6tlndad drdiitd of .!E!y,
2.2.11 !.t€rddtt6 of dot3inl
P.ot tn cdteni of the botn 6nd twc.lia1
by t E EUhod of Ldry €t
al'-".rttl bdlF .€u albrln .6 6 .tatrdald.
2,2.12 lt.IltrldF siudl.a
on
tn. cnd. d,re:
p.ot€& act&lty E€ fo1ldrd .t dltf4qt ttr p.dod! tt
Tlf,
t
dtEct Es detlidrd
tlng 1.0 trr of cultwe brotlr, 1.o
ticl butf.!
dt ?,5)
.r
a
tub.tlat
tapt r ur qctt@ .triEr rt
rt
and
of 0r1X c.etn (dt.&1e€d tn
l.o !I of Tlldlcl lultc Cl 7,5
R.adtE a!. .hft In
,'Co.
Ii ya! ob#!!d ilEt th. r6te of Rcttd ko6a.
TrE
p.ot€& .ctltrv rach.d pLt@u q,to
remlred c@Btentr tn.r€toE ln $b4quert
petod
E. s.l.ct d 6e
2.2.122
h.@
trt&
90
up to
@ h.6.
.tNt . .rd then tu1y
dDelmts, tlr ,@ctt6
td pf.tB.. aottvtty d.teDiflttor.
EIf6t ot 6
T1'.
9roted !ctt?!ty E. aftnstftt
6t tlF dzrft lanAlng fft! 0,25 to
Z,O
trI
d try ultns
cultq.
dttt &t co@nbatta!
blotn
{ttl!
1.0 Dl of
-t$ o.l,( @srn as
Ii s.
,ith
r !ub6b!te. T1t {.urts a !c!4nt
.tec!..Eln8 In
dde
ln rta€.
tb als-' tlEt i,r* Ete ol toctid
ob!.F!d
dzym ooitnt
<l
ltlortpt 1.0il
and
Gs
lesar
th4 Dro+€s actlvltv sB
r
a,43t--34tcs!d-s&!tsci!-!@s!Ee!&4r
tb. .ff@t oi 4!.b.t
c6 folldrd
r'y ustra
c@hti€tt@ d u. Et ot Fot ory3r.
qeln .5 a lub5tFt ot dtffd.ttt cdEttt ttort
6.slns t!@ o.ot to 4.q6. ah. acttvrtq st.ttlFL<l
ir ett@ 22.10.
T1E
bv
u!. dtlr.d
r€P@r,<t
fsdt. 5n €l'd lb Ft&5.
ri n: obsEd to llE i..ult€ ttEt th. opttou actrrlw
@cG.
concntr.ttd.nd !h6 ttotas.oildtv E! d@..d.t hiah*
4bdcat c@..!!ltd, c6uld t d@ to OE .fiei of 6?m _ob.E!t
ai
O,1X
(1$.r6D)
.
22-12.4
.ctlvltr.
tftet o! Dlt on fttea
TtE-.af6t ot
r,s
{ plot 64.ct&!ty tor.rds H..tucrobh.nd caetn
d6 doted@d at dl Fngln! fid
alrclDal]cl butlo, Th. rolurts
!i Es
.b..rtd .t tr
ldl.n
ob.ev€d
5,o
tld
2.0 to 11.0 !.rna a
.r.
Fts-7
6tudl
blfte
.nd
.horn !n !ta-7
uei oDttru Fota4 etrtvrty Fs
teidc tlrtugl.btn {hu. tltE d oDtIE
@!r s. u&il .r a dLetl t tt .z.rt 7.o..n 95 .
orEt€it
ItF plot4e .ctlvtty ot cult@ taotb a''Dl.s*F dldt.d a!
to t@6. Tl!. F@1& @ lrbh in
dtffd
t rDqlbds ttalnA fr-
FIa-a.Dd
in oDtlfl +rrDeat|@ of pFtd& acttvitJ E. tut d
6trd
25
55c6
fitu lcota4 acttvttt eop!.d .taqt1v.
22-12-6!ff€ct of hat tEUott
crud€
cdtE. lroth dID]€ hs tr.ircubt d fd
10
trtlut c !t
El@ t !per.iE.! En3frg fld to to 9&o. And uF F.!.ortt!.
.ottr!t!6. of
€nztm tn
.ultr lfotn reF e.ttt td .ft t
lod teop€Ftu. a. lepeted @Uer Ir 4ottd 2.2J0
.rG
cooltng
anil +J'.
.t
re&lts
c.rplfd tn Fta'-g.
2.2-12t
E
r.ol of Elou r@t.
d E.t €4 actlvttt!
th. rebtt€ actrdibs of tne DEotas i.F d.td.rned tr F.@c€
of 6!toq RA6t .f trl, @Mtc.tr@. ft. F.lttr aF tabulat d .6
_44_
22.1t P.*lal dlltatto
of dtrlc.Uqld sot€*r
P..ttal putltottd of labta* s. 6ai.d oui by fEcttdiio
rlth .@tde, 3.r ohlodt.8Ffiy .nd td @llna. clEcto8ladv.
of co!.Idceton. @.
sdtt
d
2,5 lord
t t0 d oulta. t(.tir.rd tbts dtrrlal e. Lfi
for o€r nlsht tn !.fll8eratd bd odtrtlw.d. TL collect d F.ctttta!.!
r.!e iLtB.oltql tn 1o nn h1!-r€l bujtd
Ectlvtty of
i
DH
FirElt Fot€ae E. olend)
?,5 ( In {h1ch tlF Mxtns
6nd dt.1y4d to! 24 hoc.
aFln.t tlE sF butfe,
22.1t.1 C.l clldtohollt
\e lu.ly8.rl 6!plc aE th.n rpplt.d to +n. S.thadd
(trt r
lo)cn enurubeEt d
.1ut!d
rttn
.qqr.i^'64 o 'rm Na(L,
B tld !.t€ or 26 !l D.r
Ftt m 16 shorn t!
oolm
rttlj t!r.-Jc1 burf* di 7.5. rt. cole -!
: 0.111Tttan|c] !!ftd
$l ?.'Lqt
rN cou.ct€d FnErly
G-100
hor.!d tlF ancttdtol4,0
and oh.o*ed
Flg-10. Th.
fd .ctlvtty!
T}!. typtc.r
euird leot .s-I acttvtit
Dl Ech
.lutts
rao ilet ct€at
fEotlos , to 9, lh@ .s uE -&r .ctlrl{4 &ctd,ntutr.1-II lrd
rD611r Fote- E. lorld tn ftutto. 1o t6 ,o, l.iiE fEctld of
1n
actd,
@tr.l-Il .rd .ltaliB !cot6*s, ur
ras p@1.d aid dr!ly!.d. TrD
EtE@toaEglEd
@nt
it of tub.s 10 to )o
dr.rrd.of !@r.d fFcttda sd.ul,Jet
d IflE g.plr&r l-50 co1@ toc fctxe
@t
d
Etto udd
-t19-
dl .rd tdlc gldrat cd*6!.itd!
0.2l|
usrlS
4
O'll T!t!{lCI b|!t!d 'dt'I
l6cl elutloi.
a;-!.:33q-s!SEt93ser: jes!!!q!
tll
aboE pooLd .dr
s€prEd.: r-50 col6n(2.t
*
dl.ly..d *tPl. Fr tE a!'Dlt.d to Etl
14
otr)
€qdltb.Ft
d
_
*tin Td.Jcr butt4 dl
7.o:oE 6rle G. .tqt d rt0r dl-ar.drent udns Tti.'l€l oJt brftd
coDt€lng o,zx
ard t.O
r6cl !.nrtns trd
7.0
t
9.0
tl .t s floi Ft' of
'o
n\/h@
rl fcctloE rer @rl$t!d cMllt. The !Ectt@. Eldtn€ t.r!'
db6..!.rc. at
23o
m
@e.E6t.d rd.ctiUct tutlal 4d.llt1tc FotaF
rctlvttloE o6.hfr !n Flg-11. Th. elzrm|lellv actis tt..ttoN of
att6lt* trotas fld
!!ot€E* fton
26
t
2 to 16' .ctdtc
41
p.ot6!. ftd
fEcilda i@ lool.d .
T1)e
17
to
otr
25 ard
.11
n6t"1
lalfioitd
9'
'* ar lhdn in
Tne pool.d f€ctt@! Ere aBtted t' stdv tLrt ch'$otet!tt"'i dlltffit
Es 6lDror.tdt lv
20
told dut
. rocEv ol
,n
1abL-
pdmt*s of AH' t !p.rats., h..t.bbuttv.nd eft€ct ot dlffe4nt
th. F.!6$tt6 of col{it .ia stPl.. .llrtoitd
r! llt rldd Iv atd v,
aF d€*rtb'd
- 50'
22,14 ch.@cietEttcs of tn. @tft.d d,rEr
22.14.1 H@itv:
lEctl@ of.ctdlc, mutE1and.lta1l* !aote*! e;
P6l6d
crEk€d
d
tl'Iok .tlte
t.}lln 16vq
se1 G-6o
crEo6 s.!ht. fd hoeent uy(15),1!4). o--
pbt€. ( -.
re!.rdlr -yi ) r.R E. Et
on hou ai l1co In hot o@ .rxt .pott d {Ith
10
d lor
rrl ol 6oh coUacllon
ty Elns but r6lac€ttc.cld- btrr (4r1!t v^) as. sl€nt. A&.!
rLEloplngr plate6
|@
aFry€d
,Itlr rlnhydrt., 6llldl.
6pot
!nd]4tt.a tht
.U !oLd factidE @ l@C.|t@ Fot tE.
r6trE tms.dE ol tl|r fFctl@ Fr corttrEd
tV
sD€r
Fl
.l@trolh@sts ( *. lpr.ndrr-v ) cln3/t6nuFoM bufte! Ey.t r !!
(17)
..po!ted bJ D.vl.\
'/
and
rbd.
&
Rt.tdo.d :
Ih€
!.6uli! as lhffi tn
2.214.2 chu6cie!&i,tc d@dtt ! of hoe6l|@ de!!
ltl fEctt@ cf actdlc, tuEll I
&
II,6!d alblle pbtaF! |6
!t'ni.d l@ dr optlE, t€4.nbN optid, rEt
.td .otlEUd eff4t of @lN rg.nt
.nry@
!t uE
Et3nddn 6tJrod.
|u.
6tab111t'. 'rh. lnhabtito
studfed
.! d.@tb.n tn *tr,o
d tN -4L. or
2,2,12,
RESItLTS l!D
DtSCUSSI0T
-tlr,9_Eg4!l@.![!G@:
th. s of F.nictUtr lrF'ro tu!! fG tE Fodalto ol
cofficit11t tiDoltant rlodmi! lE! . ld..@d 6ttd1t oE uE F.t
hlf qtsy' Itr -ln !@pd of od! .hrdJ E. f@r*d o i.rr d.tosI ot
lsltcdbrl s.t .dr ar !16 htt tlrloh dnd !. 4ariltt utilt*d 4 .
@td
and
.E6| loE€ fd th Fodatto
.ubst!ee!, In tht. dl.fecitqr, tt
of
b! ot.md
en
V-. .rd ollFr cIFl4I
rn p!.l!@rry
hBsilsaitd
tlEt UE raodutlo ot hy&oryuo ddd. l! 3!|g!r .d Fota& tn
Frttcur.r n! .a ft.t El rEqtlrrr thn ooE dty-.. It E. trgt @,
t* opttlraiio.f Fot6s btoqDu'.ltr hB strdt.tl tD.Lt rl, DF
patfi€t!6 .id.tf.ct€rtEtt.n .f trot6e! r4 t6tlE c.!!t€d out ftd
!E!!g! .t!.t! cxltl6tl/lte'atc.
sEr€y !€gl.d ort !@h ty!. ol Et 6 F!t6* ta.dEtro tv !g49!l]!5
€f!!gla! udd trE rqrtF-t ol alo|.th cdn1tld ri. Ft b.a d@ e
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and 43
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and
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tn
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REFEREf,CES
NEFEREiCES:
1.
XolwlltE. cottt ot t'. lnt@Eilc.l lhtcr of Bt@rFd.try ,
En.r* I:.|FrcLt@, l@d.crc DFs ( Xd to!*, lordd) F tt-156
(1934).
2.
L&@v, Proces€ Broch.n.
,.
rl.xJosartyr P.J.crlffir lhd l.tl,J.Vo.t P!oc..! BI@IM. 9!27(1tf4)
4.
G.l6rlar
5,
5.
?,
1.3@i (.1@ .td
3.
Y.K@4t n.G.tt
-.11,1?
(19?1)
tr@it l S.Oldl1y dxl C,ooa1.rt
1j4.. 2o5 (1*t)
tJo.Er
E!r,
FEas
lgtte6
Jtt@ld,:3' t6t (1961)
x.E&lksq.dr B.&tc6l EE,r,Bt@lE, 1?!,6t l19e)
t{J,Pr&\tt ltn 6J.co.rblr JG.lrclbror.
!&' 2rn
(19d1)
f,.laEt l.YadiL.rd l.ti.nr Agrio,llol.Cra
2a, 216 (19dt\
9.
tlI.rokcz*d€lBt |tJl.o?d.k.nd l.PolMlttt E6J31@h€n
72,
557 (1eat)
10.
R.ldskvlsrr
11.
!Ji@y
12.
'1t.
J.rede ei a1, Bl@h.d
],.Rsy ei a1 r
1t!.
JJtlr.t .t alr Elot chool er@N,. 1!r 12t
15.
s.r.r6aairr !r'd P.J.cltffrDr &9t.r{t@bto1,_jl!i, 135 (19a1)
and
Act€ ,Ch.tr !S@^.
3L
1121 (1963)
B.s.{t1dlr stot olhor.liee. 13. l?9 (t9tu}
.t
Brdphq!.Act
,_4,
Blelrr .t Dl@hr.. r.t ,
7r Gyr)
1?9, 329 (19n)
(19F9,
-101-
16,
&.t{dtlEE.t.tr rtdr.Bl@iE9ro*v..
1?.
l.ltar.hdrt.!td tl.tdgtLt rmJi-t. Agtc.crtFoo.J.Fr
Abgt6oi FIOO
16.
?U
21
,
(
(1956)
anit X.Eb.t€,
J.l!o!.clE!.
(196r)
19, I.tidtlE.Dd iJs@ttr
20.
5?
(1965)
r.fidthaE, tl.T6utuklt ?,obt x.tnN
2\O,
132,
Jll'dst 8.lch&rn-
At
b.Bt@l-.Bldvs. 126' 620 (1969)
.nd F.tocnd.t Aalt..Blol.ctn.
:zgr paa (195a) !d_:9, r (1e6t)
K.rio.tlrar T.ota and t1.Tr@Ut Bl@tst.BlothvA rot ,lltrl€2(195?)
22.
l.t(lEGtor
2t.
n.fitubrDi.r 1.I@s aid Trtdrot!&r rDplJ|t6ltol.
24.
g,ltaEhiroto,
E,
J,Bro.h.!iq1' 160
Y&FtorT.Is.,
(1967)
4'
.nd LYokot$rdl lpDl'll1@blo1.
5& (1en\
Br6l6r.ht::'
25,
s.Ebtr D.V.W.!E ant c.^tao6btFdt
26.
l.sof.m ltd R.sbrr B1@Lr.Bt.prvsrct!. !grt4,
27.
rrld&t xo. 12?,215 (!{1@!to1, Ab"tr.
26.
N.s.Ye.dov.t al, Prlkl.BbkhtlJ{1bobbr.9, 654 (192)
29.
n.Rq1a.ut@En!.n ard
\.
c.zoE@
t1.
,m(191t,
442 (1C16)
(1964)
?.7411 1972)
u,s.r{.@L r can.J.ltt@blo1, z, 2a1(19f)
.t !t, Bl(.,jFn.t 2t lktllClt)
HJ,tdt*r J.R.3rlm6 qnd c.u.stin r J.'at!y *L.27,52a
(1974)
P.
YJdz.t lt.colt {.t4*t r,t dd..Dil TrtllEl &rtcptol.chh.
4,
tt,
216 (1964)
lt.o.omnt l.F.Abdul-fatt€h .nd s,S.lFtroukt J.C.tttclo!1o1.
t2, 1t1 (1969)
a. K.Nak6'{Ehr' rJr6h.r! 5, 125) (1919) .nd I'r1 (1960)
t . T.olo 6r'd x.t dth... r J.rsrt..ch.!.s@,J.Fn !?, X7 (19r)
16. N.s.PaLEl lnboblolorL , :9, 10@ (1969)
37. F,Iashrda et a1f Aerr.ALol.Cl.. 4, ,\1 &rr, (1*t)
aa. FrYaEbld. .t .1r Bl@tE .i Btcdn.. rcb. l2r ,60 (t96t)
s.raw .t .r,, asdcptdr,ch.:9, 6a4 (192)
t.
40. !.st€@r St*h.rlol so..TnFc!t@, L:,23t (19&t)
41. sJ..e arit f.oh.st rsrlolroltha. :' €29 & 3N (191)
\2. fi.s€klfr, iadclBtot.ch... Ji96 42ot \19t2,
4t.
JJ.s.!dlF6r !ppl. &6.b1o1. j':i,]g (lt2)
A\.
ti,z.stenb*a, J.D.by.S.t.:!, 1'9
t15. fi.R.rhni
Pli,D
Tt
ds
(1915)
(1971)
!nlM!t!r
of skrtrFlydc sr!!ao, u.R.
46.
c.A.sdkutt.nd r.J.B.bul, J.B.atcrorr 12,1407 (196€)
47.
Ir.rrrD,
J.Ju and
s.IEBklr
fiErxoDs
m E zrxoLoc] (G.E.F.d@
.rxt L.r,@nd Ed..l v61. iq p446 A6d.trt. p!6!s Nq
4€.
lolk (190)
E,R.rEI1., J.o.ti@ .Dd S.E.l.lErdtmllt t R.v.L.t{tu lllcbblol.
27, 1@ (19€l'
-tt5tlg.
J.rtn(lmb
!.:1, 710 G96?)
c.D. J.It@!!.l. !1'
arr ASlrc.Bror.Cn
,o.
"t
Bt,tara.rn cJ.ti.eltiFt
51.
H.L,lrors
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,r.
n. . .!cyr T,c.Ena.lhldi .nd J,t{.Upadhy€y, ttycoFth.ro8r.'4't7(198i)
54.
T.sat h, T.Beppu
tt.
r.!amb.!t and s,P,J.h1.o1.r c.h.J.nlooblol,:q, 1463 (1934)
t6.
c,J.BFdyt Br@h.r.J.39, 6rl (1t61)
,7.
r!.s€leti l.llDt
,s.
s,r.alrri l|.ra6s .rd s,sultrr ht.Jllr,rid.n.r.:!1, 2t6 (r9o)
19,
I.I1LF.i
727 (r96t)
ad c.s.t|lslttftr Bl@t'... st aftdv€. tot , 14,4r9(Elo,
and
and
LArtnr l6rtuAtor.chetr. !1, 29, (1977)
P.Pr@l4ltt, J.P1.nt.P1vdor.:ll!, 1r,
c.siclEffcld,
(19€4)
C.s.hl!tFe.*t t.zEtgr c.cdatert
E,E6ottol c.Iapis and s.o'ndUyr atot@!m1,L.!t. Z' 559
(1965)
60.
n. l6hlrat3r J.Btoob6.9:,
61.
J.n.l1r A.Pln.ky
62.
u.lha! Daboir s.A.alt
6r.
gr.
s.sht@
P.raliigr
PJ.Ld.l
65.
xI.LyE
ard tl.A.c Sadforit C.n.J.at@lE!.c.11 3101.62' 1o9r(19€t)
66.
6?,
L.Tudemn and t.J.Pr@kopt 9E.J$@h€o.
and
E rl
1169 (19a4)
s.Cro..Ent c.r..1Jlcr,32, ,92
end
a.n.r6drr J.Pbh,Pb.lhlr. Ltr. g'1 (r98r)
r.n*ulhtFr Asl.Jr.l.ch.r. rl,
!r<l
(1966)
1?16 (1969)
s.P.ltrldtr t.AstcJ66d.Ch.r.r3,r22(19q4)
Rl.d€llaren; IEs.l.tt, jgl129
(1990)
j3:'
545 (1932)
-106_
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(19f9)
6s.
o.lt.D€tl.*tb .nd t.L.coldbcat J.lror'chfr'
69.
{.S.sas .td
m,
J.!; ana, J&vurr, J.H.Ro ry P'coliln ttd A' ldtt'rrat
ll.@Ie.J. l9:, ,rq (1919)
7t.
s.I.htElt
72,
a.Po!€ and
7t,
T'l|rddt
n.tolhlMat lJrhlrit l.s'dt1i A.{litls6t l|'ust'dt"ti
(Jeb!..hlt
.satut
J.Er@h.r. q,144,
r&-t@
aid
RJ.rttont N.Eoc|.
r.srol.clM. ?!9, ss6l
H'slatt't Fos Ltt'!!A'119(196')
ridl n'!'
T.!r@t!ttt tl.U'd!t T'tttEkubo'rd
t9l
?4.
E.{.Ret*!d.€
t'hr'htr
n.!.s.1$q&t1'
t.A'rory J.os.PnYalol'
o.Forln .rd
9r
(19€A)
(1965)
74.
7r.
76,
n,
c,F.B.Sohdcher .ntl
(ly6)
J'Bi@har'191'163(19€8)
B1@lEr'
!9' 9 (1912)
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v'ctoeli,ut rsbr!"r' A'@7 (q7\
6 H,f,lo€t'ltv+l I t'tr'6dt tn !tr''#lo"r td'
YonL Vol' xlv
s.P,c1o'Ict md Ii'o&p14' Ae'Ltt6 ?rF N4
Dt
P- 2613 (rEf6).
A'z'Etst tttol'ch6'?'g'4562
79.
J.!.S.1iat S.t.Adttnt C'I'Gt. t
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c.n.E@ lri J.D.Rtd€[ai adr.BrodE' !g'
al,
s.rbt€Ehlt€t
@.
t4Jsltz,
at.
o.!.LoEv, N.J.Ros tsoultht A'L'F$
@' 26, (9'1)
t.lBd
6nl
ard
(193!
)
411 (1934)
t.Ntt'r aFlp!o'r6' 16'X5
(1966)
J.co.Pr\YRto1.19' 459 (19t)
!Y'd
R'J'&rldtll J'Blol'cYan'
34.
D.P@r qrd F.tt.A.tonr Plant F?ry.rol4zr 719
a>.
c.E.fblt DLt ard L.!.char ,ld1.B1@1q. 19,1?5 (1965)
86.
i.soa.E &d xiTar.r!.htr J.Br@hd,
67.
Y.Icblh!!6t f,.s.s.*
aa.
s.s.lrrnlnar aElpt@re.1!2,
49,
u,TtElld
90.
r.D.c,|qt rE1. Br6rs. jAt Bt (1%)
91
,
6nd
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9t.
H.oulttup, t TrE u. of
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l
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(1t8)
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I
(1932)
o9d.)
aBl.Bt6h.tr. 1j4,9
h@€B! Bl,@lEfr. 2t t9
9,
17al
K,tar.h..h1! J.Bl,@he.9gr
and H.tr.D€nkelt
r.coUtdt
!r,
(1967)
(19a6)
(ll'l
tulltat. , 2,
2r8
G.*l
mtrtld h €nr,ft l.rgtaitdr
A/s thpubltlh€d mnlJ@tpi
lrdo
ctt d tn 'Btot€.rologer
App1tstld of &rt tn rd Eh4dr Ed. !t zldrt lnd ta.gElo
I€d.nto pF6s lrd Yor* p-41 (1916)
94,
n.Davl.., ctba In ' lldlErtrtry of Indqt tlr |Itc@!g.n-ts'
ld. l" C.Fllrlbd .rxl I.l|.Rd , A6d€.tc F.!s {d lo.t p-64(r9$)
,,
A.*.r@du and
96.
A.shhyor r.(*aakl .rd c.T*utr J.Feffii.telrbl , 16 n1 l19f€,
97,
J.E.Ballry and D.F.ouiat, 8t6h6161 frgtr.Ftna iurdaMt3lr,
2.d..dlllm
96.
S.Dqr !!p1.r't1c!ob!.1. 19, r99 (190)
corFrv, v255 tt\d 74 (196)
o.P.i.rrtt clt€d t. t l&ebtal Edtd .!d lloterhobgt td.tt
rE crar-8111 Boot
t.l,i,Fo8.!iy, Appl. Scl. lur)lllhosrld Ydlt pJ32 ( 195r)
-1oa-
99.
l.shldv.
aDd
c.Tdulr J.F.ddt,T..tno1, !9, 51t
1OO. I.R.PoU.clt I fh. lact r!!r Ed. by I.C.GsdlB
Aadqlc P!.s., nd Ydk' Yo1.4 F12l (1964)
l('1. T,t.Dsttrr J.Ftut.
r@.
Teclbr. !!,r1,
(lto)
R!t'St3t16!t
and
(1966)
c.A.s.ikutt .rd F.r.Bat 1r tDplJlc&bt61.
1or. J.nt{@l ' Ind6trbl l9p1letr6. of
j:, 19
(967,
tt@bto1o6ti Ed.
t{.o,}io!..nd J.E.s'1t}, sEly Utti. P!.s U.lC.'
b9
(lyfl)
104. I.nJl|nt R.Bashl! sd K.z.tsull.hr J.ttat.s.rlbtl'r. a,
'lot. l(Jundrupt Bliitsh P.t rt' 1'r06'2a| (1958)
59(1904)
106. S.t&y!-t l.OobF*llt O.T.dtrt S.$ta6Fl tJdE.td E.Iohllhl-i
rr'Dl. !t@btor. eZ'9, (194)
107.
E.rchr.l'tEt l{€t}od! in riryelogyrlg,rtl
10€.
n.Pra4d, n.(.ualtk atxl D.xrltEthEl
1o9. I.tluatn arn I.r.nc Dmldr
r.J,rcDodldr can.J.itcf.rbror. gr
111.
c.tt!11!
112,
S.I.jlu.1-+lohsnt s.A.EL
11r.
I
lt.R.Ktht J.!,BlaIn
!'2t
e5 (1934)
('1953)
251(1960)
c.n.JJlleoliol.@'d(r9a4)
J,N.caeb€11r
a9rc.{a.!e.,19,
A.ls t.Darlt R..,:'
c..,JJtc!6!161,
1r0.
t
(19rao)
At!*r
.nd
l.F.Abdcl-llttlhl
(19€4)
and
J.D.E,P.ite&.t
tp91
JnvtonJlleoblol.
27, 719 119t9)
11(. B.rJbplsr D.lr,Ja6d
2!, 56 (1e3)
lnd R.li.try{t BlocrEGet aloprry.lqlct .
'115.
A.F.Ibde1-i!tt€h'
17,12'
115,
(1*)
s.choFa
s.r.a!d.l-rd6
and
sJJr{aEl lattcJl.tt€tl
lid P.tlchtlr Fou. tli@blol.:9'1r?
G9q,)
117. JOrSul1t6 ard c.x.lhtll1sr J.cJrdobtol. 4Lrt9 (lgall
116. P?,c!Il and v.l.$odtt rolta l@blol' ?g'76 (1931)
11g. H.Halhletot l'ri.a€ ond T'Yototsdtt a!p1't{ictubbr?!'946(1t12)
12o.
n.s.th.nt tlJ.arht"'& ltJ.D.chusht't, P't'Jitocha'!3'63(t979)
121. O,P.St!d .d lt.D'srldr ndn6t.Br@hn.
'122. ||.1i.! |brbtr s.l.J.s.t,Id.R...:9'
iz'2o9
(1900)
19L (1937)
canJ.!torq{9, €5 (lylo)
121.
J.sod.k 6rd T.lto!*ni
124,
J.D.yr sJ@rel.tt D'c..clo and a't4cPh'r'dl JJto:LhtD'3!1'19?(1966)
1zr, K.odat H.Tdl.htd and s'tlGor asdc'Btol'ch'Df0' 651 G96)
126. J.E.L&.r.id J.r'Ih1b.ot r..h.B!G'tt'p!'dv!' 139' Aa (196?)
12?. l3!.rllmrt.nd t.J.$tl,tt'crlldtosddrro ll'th'dll Pdb_u'lF
clE Ftr ard fi,rU, londo 6.1t.* rdt p-93 (1935)
'
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Slr.t 0n1r. PE3!r Pl''l (l9ar)
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cooDt.\Y, oxfoRt
1to,
-
Net York
F1 (1m)
R.coDrlaldt.lE.nd J.F.ltodt rB5
r'tt.
!91'2r'6 (1935)
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1:l2. tt.s,itck*t .ri PJ!.8!td.-{Gat can.Jpr6t'!€!'l2o (196)
11
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