biochemical studies on the metacercaria of … · 2018. 1. 4. · thair •atabcilie...
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BIOCHEMICAL STUDIES ON THE METACERCARIA OF CLINQSTOMUM COMPLANATUM
rTREMAT0DA r DIGENEA)
A DISSERTAI^OM S^BNflrrED IN 1>i?i;BltiAt FULFILMENT
FOR J H E fc^GRBE OF ;'
BY
Afzal A. Siddiqui
SECTION OF PARASITOLOGY
DEPARTMENT OF ZOOLOGY
ALIGARH MUSLIM UNIVERSITY
ALIGARH
December, 1980
CLi>io<rgituff cfygCftifftTQff (rnffttkrmit t ntep»ir»)
ro»» TMr occ^rr
or
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By
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9rCTI0«l j r PMft lTOtOCY
orPMTBtuT or fOOLoev
09ec»b»r i^sn
DS2822
Aoc No.
^ligarl) iilusdim ALIGARH. U. P. 20Z001
®nitjer£(it|>
Director : Ather H. Siddiqi Ph.D. (Alig.); Ph.D. (Purdue)
PARASITOLOGY RESEARCH LABORATORY DEPARTMENT OF ZOOLOGY
t7 n(»cf»«h»r I'*"''
This i s to c e r t i f y that the uork
nrsspnted In t h i s d l s ser tntIon Nca bpen
csrr led out by Afzel ft. "^iddiaui sn«< !s
s u i t a b l e for the auard of the Wester of
Phllasoohy degree In Zoology of the ftllgarh
f^usllra Unlveraltyt ^llgarh*
Profeasor of '^eraaltolooy
nfofCATfO
TO
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(LMf ) p*». it3»*L * . ^inr^imiT
in
cwftTiTurtr
COWTCItTg
inr or Tftaus (iw>
Li^T or riGuwrs fw)
ACKNOWLCOCCHCIITS (vl)
!IIT«»QOUCTIO«I 1
or w^08L?« ^
HC8ULTS 14
0I5CUS?!0«I 19
Table*? ^«»« »»%«belie •nd-ermfuet* 37
T»iil«*3 tffvet of eH«Mle«i» en aeld «fid alkalln* ohosoHatatet 38
<tl>
LT<?T r>r ftrj^nff;,
npop
1« ^ qlycoqpn Hpolpt ion l*
d p n l p t i o n i'y
4 , «5ho«nh tsBP ?»ct!wltiP8 f t
*«. Ph«5«nhr t i a s p P C t t v I t I » » f » I n r e l f t l n n t o t tm* 4"
6 . fhoanhr'tniip p>ct f vltl*»ft In rpl.ptInn t n tt>iiiQprp>tuTp 44
7. "ho«ohFt''8i» p c t l v l t i p w In rf»l>»tlon t o hnwoqi»npte conc«»ntrntl f n 4*?
8 . "hosohrt''8«» f^ctfvltf«»« In r p l ^ t i o n t o s u b s t m t p cnncpntrnt I on 46
•5. f i p c t r o n h o r p t l c w o h i l l t y of -" cld f»nH p l k e l i n p ohowohptr-ses 6f
T cpnnot #xoT«»(»? In uorr^a thp rff»«>o
s p n s e of o r - t l t u i i p T OUP t o r»f« «tH*»r •<• • i H H t o l ,
Pro fp t sor of '^!<^r«?»ltolocy, for klnr* hplfi ?nr*
in«Dirlnc! quldancf th^^t T T'-CPlwfd frrm him r'uffnn
th*» or#«i»r>t work. T «» P 1 « O Indebted tn hlw •'or
c r l t i c p l l y rpvlpulnn thp w p n u s c r l o t .
T fjlf^cp nn rpcord ""V indphtnpsip t o
r r . I'ajlh *!• *'i?pf«i, for h i s l o t p r p e t pnr* w«k;ioo
pvpl lr iblp no povlronmwit for r»B»srcH«
'^locpre t,h?»nk« PTtf due t o t h e Moiwpr«!ty
d m l n l » t r » t l o o ' o r i r o v i d l n n th# nPCP««Bry lf»horf»tory
f C f l l t l # 8 »
I ftlao u!»h t o ex->rp«ff thpoksi t o my
f e l l o u rp»p«^rchprs for t h p i r Wind hplo ^id to
f r "f teb f f r l d l for t y o l o o t h p a c r l o t .
/»:r7f"' L f • ''T'* ' T^MT
ywT'^opMCTyo^
The tr«natod*« eons t i tu t * an iwportant groufi of
p e r a t l t l c worntf the •orohelogy and U f a eyolaa of which hava
b&wn oh la f ly atudlad in tha paatt nalnly baeauaa aavaral apaolaa
ara of • ad ios l or aconoalc i»pertanea* Within tha leat two
rfaeadaBf howavar» appl icat ion of taehnimtas tfavalopad froai
hioehaniatrvy t»iophyaiea» phyaioloQy »nt!l iinmunaloQy have v i r t u a l l y
r«wolutionlrad our phyalolof^tcal Vnowltdga nf t h ! a ^roup*
Hewevart phyalologiecl end hlonhanie«t infornat ien
oonearnin^ traaiatodaa hava r<Naain»d eemfif%ad »nra or laaa to the
aanually naturo adulta ( l i t o r d t u r a In von !lrandt 19731 5aiytht
1976}* I t uoulef ha of oonsidaracble in teraa t ie inv<»atiQata tha
phyaiology snd bloohaalatry of I n r v t l trawetodaa, baoauaa tha
anvironnantal ehanQun durlnq tha lifti eyela fttm a fraa<»llvinp to
a t r u l y paraal^^io atano »tvi tha t r a n a i t i o n froo cold to vntm
bloodad vartabreta nay ba aeeonoanlad by dreat io adapttitiena on
the part gf parea i te* Tha e r l t i o s l atudy fm theaa aaoaeta of tha
l a r v a l atagaa w i l l Qlva a clear p ioture of tha hoat-pareai ta r a *
la t ionahlp* 2n aiost of tha caaaa the paraa i t ta hava b»>an atudiad
aa indlv iduala and vary few workara have t rea ted thaw am wn
inaariarable part of « aytta»f ! • # • • tha hnat -paraai te ayatam* Tt
l a Quite inportant now that for a bet ter underatending of the
p a t a a i t e phyaioiaqy and bloehcsmiotry %m B»i»t have a good knowledge
of the habi ta t aa wa l l t in whic^ the parnai te Uvea*
AlthmigH atudl»« en bloGH(M»lo«l oompositlon end
cerbohydrat* nt>t8boll»M of adult dl^^naant hi^vi* bfK»n nu«prou«
but tuoh t tudlas on t h » i r mataevrearise Btm «or« or leaa
fra9*entsry and o r i a s r i l y d«»«crlDtiv«« ^ in i l t t r l y * th» oHo«obatna#
•yatiwi of adult trait«ted»a Hav» bti«n •xtanaiveiy invaatl^atad in
a nuiMbar of d i f f a r a n t adult apaciae but ra ra ly in l a rva l ata^aa
(lltarufcuro In von 3rand, 1*»73; r « y t h , 1976) ,
A. niMiber of workara Hava undartakon hi»toehaii ic«l Bnti
hl8tol '>^lc*i l r»8®?r«3h nn t iataoaroari ' i l oyata found In tha tiaauaa
of varioua f iahaa* Kuntar A Huiitar (194<1) ernti Aegitah (1^7)
o^rforwad hiatologioajL mn4 hiateoHatnieal atudloa on tha cyat of
Poathadioloatoiiust ainJamn and Huntar * Oalton (1939) raoertad tha
hlatolttfiy of tha cyat of qlinoatoituw waroinat^iW* Lynch A Wooltah
(1962) I d a n t i f l a d 14 aaiino acida In addit ion to aiic d l f fa rant
augara in the matacarearial cyat of £ • niniwuii* i»i«}ht ( l 9 7 7 ) ,
Raaa ( l93?) and Maatfno»rg<»lund (1934) deacplbad tha eyat wall of
IBTM&I raee io la haoatiea aa conaieting of an inner and an outer
u « l l » but Dixon ( i % 5 ) ualng an alaotton Riieroaoopa» found that
the outer well conaiatrd of two layerat «n outer tanned orete in
and an inner Muoofyolyaaocharide layer* The innrr eyat wall alee
oonalated of two l»yarc» en outer and an inner onet the forner
being aubdivided in to three l(»yere* the ehainieel eonooaition of
whidi wee mainly carbohydrate containing pretf^ln eOMolmea*
Aaanji A y i l l i aa ie (1973) etudied the metaeerearial eyata of f i v e
tftffftr«nt s p M i M naMcly P«goreM« »o«nthtff« P>«thodlolo«twiuw
l f t * t PyftHCfllff^ef^awi^^tfty i i o Q I Q l l t f P«>ff^Ho^f|ft|ot^a»^ n^x^ and
CXifteatoituw t l l a a J M end found that both • t ruet inral ly and
ohaMlcally th»y er« d i f fe ren t sntS th» nunbcr of lay era on tha
cyst wal l var iad frcNt apaelaa to aoaeiaa mii tha fvaa aneyatin^
f o m a had mora l@y»ra than thoaa uhle^ panatrata bafora ancyatlnei*
Thay ( l * e « ) alao obaaw«4 that tha aingle eyat wai l of £ • t t l a o l a a
i a produead by the hoat» ainea I t a ehaiRioal eenoetl t lon la tha aiiPa
aa that of i t a aurtetmdinQ t laauat i ^ l l a tha ainola eyat wall of
£ * »gwu» waa probably produead by tha pavaaita alona* ainea i t a
a t ruetura and ohanleal eoiipoaition d i f f a r a froM thnt of tha
auvroundinQ t iaauaa* f t i a d j^ j i * ( t9?8) ropertad that the
nataoeropt ia l oyat of 2vpoeotyla luneta i a ms^ up of two wa l la ,
tha outer wal l eontaina Mainly acid nuoopolyaaepharidaa and inner
wal l l a Mainly of protainaeaua nature* Chenp f l9§3) raviawad tha
poaaibla l i t a r e t u r a of l a r v a l tranatodva but deal t very l i t t l e
with tha netaeeroaria and eonfinwd hie atudiea on other l i f e eyelr
ategea of traoatodea*
The elinoatoiRatid Mataoeree>riae alao preduea narked
pathOQenio e f fee ta on t h e i r in ternedia te hoata m9 reported by ««!
(1966) and nay even eauaa a t e r l i t y in tha internediate hoeta aa
ahown by Naaai (1976)* Sin^h h Virnani (I97a> obaerved the d la -
lodQing of the aealea »nd tha deereaae of haenoglobln content*
paoked e e l l voluae (PCV) and inereaae in erytroeyte aadlnentntien
r a t e (t9n) in the f iahaa uhiph ware infaeted with the netacerop-
r i a a of £ • piaeidiuw# howavert Maren * fach (197«) noticed that
nellfi»r th» body eofidltlon nov length of td* b«««t Wlcro»t»ru«
aalwoidaa w«r« r»latffd to th* Infvetion p«re0nt»f|» eir n«tiie»re»riiil
density of X, auEaiasilft*
HtiAtor ft Vornbor^ (l9S5) found th»t tho iiotoe»ro«ri«
•^ Syy^iffffo^ylt JiaOtl w i b l t O uptakv Thnao A Callieehto
(I9f7t a) rofiortod th»t tho n»t«eore8rl«o of £ • oow»lanirtmi* i |i 14 wit»c« aboorbod tiero C (jlueoto imd ifioofnerfttod lioro rsdioAotivity
into Qlyoe^on th»n did Ja wltyo forwo* ^ot«ooreert»«t In vtwo pnd
JUl JlAlUt* ineoroevtttod f|lueo»«*(l»n! esvbon into alonlno* olutMle
eeldf 9luta«ln«t prolinop ttnti oc • kotOQlutiirlet e i tr ie» fuMnriev
lootlof byruvle taui oueeinlo ooldtt ond iUQQootod tho ^ooaneo of
« funotlonal TCA oyelo* Tho»«o & Rcll leeblo (iHTt ^^ ineub«tod
the poraoitoo In NaN^ COj to otvdy tfio eO^ flMstion In th»
notaoffreoria of £• aawilawatuif tm^ tbo lAbollinQ wao roeovorad
in alanlntf Qlutaaie aeldt Qlutafliinat orelinof eltr<ita» oyrtiv»t»,
aueelnatat a^ • katOfltttaratef fuMarata mnii l»et»tot and confirm
tbair oarllar findinQo (Tbeiiaa * GalXieebiOv IM?* e) that tha
nataoareariaa ooaaaaa tha naeaoa«ry oeMoonanta of tha TCA eyela*
Harria k Chant ( i f tS) m6 frlod * fhaolro fl97S)
atudiad tha ooeMiulatloii imd axeratlon of natitral l iolda In tha
nataeavcariao of LatiooehlOrldioworoha eonatantlao by hiatoehaaileat
and thin»layat ehromato^raohio taohniouao tm€ found Hold
aeetMiulatian in tha Intaatint of tha ineitbatad nataoarearlaa*
OwtInQ ineubation in non«nytriM«t toeka*a aolutlonf nataearoariaa
aeeUMulata triQlyeovldao tinii to a laaaar axtant eholaatarol aatara
and • n e w t * ft— s t a t a l In to tH» wmliuit* rvlact ft f t l oM is ft<978>
twpottm^ tha ta f t ts i lva irfanti f l o a t ion of oholaatarol aa a
eh«R«a%traeiant for tha mataearearial oairinc) in j^* conatantifa*
f r i a d ft Biit lar (1977) loen l i t a d neutral lififefa by HfatooHaMleal
taeHnlduaa in tha axetatovy ayatam of tha nataeareaviaa of
Cotylyrya fff»« »n«l TIC an^lyaia cfataotaef f raa a t a r e l , fr#f» fp t ty
aeida @ncf a t a r a l a a t o r a b u t t r i f l y e a r l d a a yare undetectable In tHe>
•a taoarear iaa* SUlar ft f r i e d ( l977) fotmd thi»t tha aiteyatad
Meteeareariaa of Cehinoat<la ygyolu^mi eulturad | n v i t r o in *»nrr
135 ahowad nautraX fa t i ia inly in tha axeratory ayataii» in^ v i t r o
eulturad worma ahowad neutra l fi^t in itddition to axeratory ayatan
i n in taa t ina mnti paranehy»a* Aa dataetad by T l r tha nautrpl l i o i d
f raet iona ware f ree f e t t y aoida for axoyatad nat»eerc»rl»p pn6
they exoreta f ree fwtty aeida in tha non-nut r i t iva incobption
wadiym*
'Taoentlyt A*la« ft Niraflii f l989) carr ied out h ie to *
chanieal sn^ hiatoanryiiolonioal atudiaa mnti d ia t r ihu t ion of
^lyeoQont orotainat l i p i d a Bnd nuela ie aoidat wntS a<xia atruetursl
orotoina* l i k e o o l l a 9 ^ »na a y a l i n ware tS^tmet^iS in t h * vnrioua
or^an oyataiia of tha leataoereariaa of £ • eeniolafiatuit« Thay f l . e * ^
elao loaal ixad sold end a lka l ine oheaohimtaaaat non-aoaelfio
eataraaea mii l iffaaa in tha froron aoetiona* 1 * f^iddimii ft ^ i i M i i
(1980) atudiad tha l i o i d f raot iona of tha a»iia wataearearln i»nd
found H io l t r iq lyeor idae mntS ohoaohelioida aeoount for tha najor
part of l i p i d t whoraaa* eholaeterol and frao f n t t y aelda n?ika uo
the Minor l i p i d eoi^ionanta* mnm phospholipid f raet ionat
6
|il«o»ph«tiiiyX«lioilfi» «fi«l plie»|>li«ticlyJUit>i«iioJta«i4fi» MMioiifit tor
th» Mijvr part of plhmpholipim^ utw^Mi ly»0|>hosp)i«UffyictM}iifit»
iY»9f»wphmtiaflm^nmml9miim «fid itniitofitifi«fi |iho»|)hoil|iict aMmml
for th* Kiner j^MmpholSpiti ctonpononlo •
Thl» |ir»li«ifi«ry ifivootioatieii HOB earrioil out on
Uio pi»tt»n«tie notiicprouriJi of ^JUOSSJtliBUi ttpjaplanotua^ Ihlft
•etooorcoria |»ooo«Bii«tt o y«i4 cfovoAopod organ ayataa ineiudiiifi
raprodiMtiva omafu and i» i>erpt)oloi»ieaiXy vary auoh oiiKUar ta
i t a aoiat wHioti ia fbtmd i n tlw ooaophague of uaiar aurataOf
tiytiiilmiB I fa ia *
Aa faf aa ia known, thia i i %ha f i i a t a t ta in t ta
obtain infaraatian on aojor biooliaiiioai oe«pono»tti« Bom mf
thair •atabcilie aml ififodMOtfi and kinetics of non^apaaific
phoaftHoiKineaotaraoaa in tha aotaaareariaa* Ca^piataiy nofiiaatad
aofiaata of oartMihyitrata mtaboiioa i ika {iluaoaa iiptaka, fiiwoM*
iaakafia and a ffoot of atarvation on giyeeean «n%aiit Mara aioa
atudJad* Tha praoant atudy yat undartakan te aaoartain i#lia^ar
a a in i i a r p a t t i f n af liioahaiiieai co^paoition end nan««paaifio
plioophoaonaootaraitat a i raporiifd fk>r di f farant adult divanaam
aiOa tad in tha eataaorc^aria*
7
tiMi Imtfy cayity sf Urn ftmli twlwf fi»iit It&iJilSt&Ilt ftfBtlltIt i
•ml tMii»ti«ii In fi»h •aUn* (fofatttr A TaMaytt 19i0)t «»fitaifiiiii
mfA too arit mi ZmS «n, Caf^ij ^•s «^t ' V ^ i«<> i^* HuH fo^
B*S an and itoHCii, ftrftf fiM 7«0 at 2iP ;t ^* ( ^ ^ ''••^ ( ^^^ ^
lAiatatfi f i l t a t fiaj^aia baltota bains waifiliad m4 f inally matf Iter
liiaoNaMieal analyaaa* fmt non^apaeifia pHoefihoaMfioattaiaia* aaaay
tha aataaaiearia* mm iaalatad in iaa ealii aalina*
rtifriAliM
Ca)i2nt.
Dry waif^t was datafninacf by talein« afififepviata
aaounta cf ttia fraah aataaiiaariaa in pfa«waiflfwiS «laaa vialat
which hati alraatiy t^an aub^etad to dyynaaa at tOO** C until
eonvtant waight wa* ot)taina«l«
Tatal palyaaaatiatidai aa glyeoyan wara anttaatad
l»y tha utiH aiftaatian aatliad af tii>nte«oaary {\9m) mvi aititabla
aai^laa waia a»tinatatf by ttoa anthrona taafioht ecntaining 0»e#>
anthtonat ^ thiauM^ ami m H^&li aa daacciliad toy flaa 6 &ailay
(19M)» blyeooan eontant waa datanUnad fron tha atandatd plat
af ahall fiah tAyaagan ^iwna Chaniaal Co*f U&A>»
a
Free v lucsosfi i n t i e e u e extracts yaet tiaterinined
enzyiaotical ly by tho nethod o f Carxoll ^% a\» (I970)t ueiny
yiucotie {iOHf AnaiHj as otatfidardi the incubaticm eiixtuce cone i s ts
o f A B 0 « 0 2 imoi, NAUP 0«004 sMaol, Vm^* 0«01 onol , hexokinasa
(yea&tj 16 unite and U«b*4>U (yoaet) 1G unite*
I&olation and est imation o f protoine uore carried
out by uain^ the procauuroti of iiaolnnir^ i^97U) ano Loury SX aX*
(I^SI) rai'i ioctiveiy* The prote in oancentratiun uaa read en a
p r e v i t u s i y eatabliBhed utandard curve of bovine aerunt aibumin
(Koch--4.iyht Labaratoriee^ UK).
Ine i ipl iK uare axtractud u i th ct»ioroforisi-
fisethanoi (2«1, v/vy by tho method o f foic*^ fiij a i . (lUS7i and
uaehau u i t h nor i^i s a i i n e aa debcriLuU Ly Miera (196(u/» The
e x t r a c t uaa ev^orat«dd t o drynees ^ MMJ^Mf ^ ^ ^ ^ r ^ u i t a n t
ra&idue uao raade tt( vrlumfj u i th Ghlorr.forjn, Total iipit'S uere
determirwd c o i o r i m a t r i c a i l y by the v a n i l l i n phosphoric acid
feasant Ojntaininy pheuphiirio aclo I4ii ««r»d v a n i l l i n ISrrfU
Uiuluaterul 23.2 un in Id', ethanoi UHB used ae etanoardi as
daicribwii by tlf5l*m3r ft Kirs oh (1%2J.
(--'. 'kiclBi'i aclcla
Nucloic acida uara i s o l a t e d by thu ncithod o f
liutchini un %%, %4* ^^ *^^^ ^ ' ^''* ^^ added tti tif'^ue homcibonatu.
3
pr«pttrvtf in tfittlllMf wsiat* I t t#M tNm wnfttlfntttf antf th«
•iipam«i«fi% u»« tfiMi«v#«d» TNi p t M l p i t a t * u— i»MlMd t i f i M yith
10^ cold TCAi* Lipiitt tf«r» wmmm» by tiMHItig %li« rMitfu* uitb
•thanoliHHtv (3t1» v /v ) t •tH«fioli«hlorttf«v« (v/v)t •hlarofom
and tthct* e«fi%rifii««tioii w«t 9«tfffiNi«tf « f t t t M « H ttati* Th*
r«M2t«fit wmi^n M M tiHnibtivd at 37* ^ l * * C in fit fi«QH« Aftat
17 h iiteubatiwi* tfl HCl MHI tsC TCA IMMT* tMvd Mid ••ntvifUQedt
i l l * •ii|i«rfi«k«fit w«« ••fiarstad antf tM«d f«r MA* Te tH« (ir««
• i p i t a t * 0*8 N fiaroHierio aQid umm addail and H«a%«d te tO*C»
af t«r ««fitrlfy9atiaf» th« aupamatafit waa itaad fat ONJi* WNii waa
dataminvd ee ler i» r t f iea l ly by th« Mathetf of nufivo i riaek (i9ff7l
uaifiQ S3 mg raCljtiMjOt 109 • ! HCi and 3*1 • ! of «{ oreanel In
athanol aa tHa aalorin^ r«af«nt* DMA was dat«ff«inad by tb«
Method of Ciloa i nyava (l90B) uaino 1»S Q dielHmylaninat l^n nl
Olaoiai aaotie aoidf 1*1 «1 N SO^ 9na 9*1 ml Of 1»8< aeot«l«
dabyda par 29 • ! of tHo eelo»inQ taaQont* CalibYAtion euwaa
ware pTmf»r94 uitN yoaat tHA and eal f tHyiKit DMA (!li«n« Chanieal
Co«« USA)*
Uolgbad »ataoor«ariaa wava inoubatod aarobieally at
37* ± 1^ C in olueosa (10 MR) and glueoao frao Nank«a aslina for
varylnQ pavioda of t i»a froai 3*18 hf t e etudy tba effeet of
atarvatien and eyntbeeie 9$ glyeoQen reeoaetiyely* Aftet
ineubation the glyeoQen content of netaeerenpiae wa« eatieatad
aa deeeribed above and the inetibatee neve tfiatyeed for olueeea
10
uptake ftfid «luoo«» lMk«Q« • • I N * • • • • aay b»* with ••oh
•xfi«ti»t<vt • •Qfitrel y tm tiftultaiHieiitiy tM mmtlh mwvfimmt
we» rcpMtsd «t i M t t f l v« %!•• • •
U e l i e aeitf waa tfataminaif by tlm italltoif ef cutiiafifi i
Uahlafald ( l f?4) it«tfi9 9*8 n %vt«tHaiiol«»lna titfff»»» /? •HWM
fflfl sfMl IDN •U9»«fi«lan 2*7S iinlt«/«2 »fid th« Mtinetioiia aftar
12i 2O9 aatf IS «ifi Mar» raeertfvil «I 340 M I end tha e«%u«l ouantity
was tfatavBiinad by nultiplytfiQ tha aKtifistltM ooaffioant with tba
•Ktlnotlon diff»tmmm*
Pyruvie aeiif waa dataminatf by ttia •alflod «f Ctatc ft
LaMnvaot (1974) waino 0*8 PI hydraiina-^lyaitia buff«tt ^ ««AO
49 an» and IDH ayapafition 19 un i ta /a l * a f lar 30 «ii i of Ineubatlan
at 3?* ^ 1 * C tha axtifMitlofi dlffavatioa yaa fi«tad at S4!l nm and
aetyal valtia waa dataf«lfiad with rafaraiw* t« tha axtinetlon
aoaffiaafit*
liiff!*tffwlfU g^y^i^fffff^^iltyii^li
(•) Pffigffrrtlffi ff »w^r»flw
Tha hoaogafiataa (fO and IVCt w/v) wara i»aoarad
in a Pettar*rlvahja« glaaa heiiaeanizar in iea oald 0«?8 1^ •uor—
and oantrifUQad at 18^1 9 ^or 10 ain* Tha |ir«eipitota waa
diaaardad end tha auparnatant f lu id waa atorad imdar toluana
ieyav in fraatar (md a l l tha <mfia»i»anta ywitpi «av»iad out within
48 h of aniy»a axtraetion*
PHo«|ih«tM« aotlvi ty WM d«t«t«ifiMl by %H« ••th«d
• f B»V9««y»r j | jJL* (l9T4) ucin^ fi«nttroiihMiyl photpHat* tfltodluii
• • i t (Sig»« Ch«tieel CP»t USA) M Miti«tr«t«* Tli* ••••y«iilKttir«
oflfitaiiitd 1 Hi of buff or (iiN •ultabiy stfluttml) with S wm •ubHrtt i
«fitf 9*2 Ml of 10^ HoBOooneto for ooicT oboophatoto at oN S antf % t
Ml of buffer with t «l oubatrata* to wbleb <3«3 • ! of i K bo»8Qan«ti
wat addod for alkaline pboofrtiataoo at p^ 11* Tba roaetion uao
•toppotf aftor 39 aiii of inoubation at 37* jt 1 * C by tho additlopi
of 9«t)2N NaOH* Tho iiboratod p^nitropbonel wao Maaourod on m
£r«a opootr^HotOMotpr at 419 n«« Cniyno ootivity uao dotorwtnod
wltb roforonoo to a praviouoly oalibratod ourvo of knoun
•oiMontratioiio of p*nltrootionoi (SlQUia CHoaioal Co** Uti)«
Spool f i e anryiae aot lvi ty bao been exnreooed In tatiio of ;«iel
p*nitropbanol l iberated per «Q protein per win*
(e) f f feat of oW on ontwie eetiwitiep
Tbe effeet of pN on ontytto aot lv i ty uao obeerved
by uoinQ tbe followinQ bufforo of 3.8S n otren^tbt pM ?«4 (aodlun
aoetate<-NCl)» pN t«f (aeetate)f pH 7*9 (borate) t PM 19-11
(«lyoine-4leQK} and pM I f (Kei«Na6K)« the pMo were adjueted »nd
obeeked elootro*etr ieal ly with a 9laoe oloetrode* All other
eonditione uere ae deeeribed above*
fd> r f feet of inoubation t i »e en owrywe eot iv i t i tp
for tbe dotemlnation of tbe effeet of Inoubation
t ine on onryao f .et ivi t iee* tbe reaetien ntxture une Ineubated for
1^
varyifiQ 9«riocf« of t i »» ftcm lf)«99 • i n In aiitia«i sandltion*
To •iiMiy the «ff«et of toimorfituroi tho roootion
nixlyro woo ollowod to oi oeootf »t ififforoftl lonooroturoo ronging
fvoM f3«70° C with o i l tho otNor eoniftttono foMoini«f tho OMIO OO
rfoooribod obovo*
Oifforont oonoontvotlono of Hawoqanotoo tongtnQ
froii S-fOC for cioltS tmtf S<*?SiC fov oltc«lino oHoopHotoooo yoro uood
in tho rooetion Mixturo Miif ootlvity woo aoooinrod oo litoooTihod
oliovo*
ftgli^vlllii
Voriouo eoneontrotiono (^ outiotrotOf nnmlno
ftm %S*1? mm U9T9 yootf in tho bufforo »t oH 8 ontf 11« for
•old ond olkolino ohooohotoooo roooootlvoly» tho roootlono woro
oorriod out in tho oondltlono 00 dooorihod eliovo* Coletilotiano
of hiehaolio* eenotonto (K^) yoro aodo ftooi tho rooioroool oloto
of deto 00 doootihod by Linoyoovor ft Suvh (1934)«
l o
(H) rff>ot of ehtiito«l» on «f t»w settirttlM
Thp effvot of emttmin ehMiiefils wi vriTyiDi* sctlvlfetwi
u«f«> VKMilnflid* fediun fluoride* todtum urocfiittcit laQnoslum
ehlorld* end eob«lt ohlorldo ymrm U9»d in thm ootiR^l renetion
nixturo in diffvvont eoneontrationo fit S «»nd 1* •#*>•
In al l »xo«ri»»ntt» auititbla blank* utrff uaiNf
enrtt»'lninQ tha feaetion wiiitura axetRit hoiioQvmsitay w icH uaa
e€ili9ti aftat tha addition of flaRH* All the »xn»tiiRanta wara
rasanted (at laaat f iv t iinaa*
Tha orotain content of aaeh haaocienate wa* detar«inad
eolorinatrical ly by the wethod of lowry j j | j ^ * (l9!!i> uatnn
bovine aerup albunin fKeoh«»tioht Labor(»toriea» UK> ae « at' ndmrd*
(i^ rleotreohofetic etudlte
for the electrotshoretfc atudfetf T* (3oly*epyl!*»lda
gala y9f oreoared aa deacribed hy r»*«f» fl«»! 4 « Tor i»cld
phoeohataee^ Tria-eitrnte buffer (^•1'^ W " oM 6*3 pnd c\ »
naothyl acid ohoaohate a* eubatrate trnd ^^f-at blue »• aa trwcWinn
dye were uaed aa d«tcrlbrd by Sur jf^ j||.» f l ^ ? U for aSkiillna
^hoaphataaot l.'tW* «)lyeine-WaOM oH 11»<1 wee uaed af thp runnino
buffer ttftdi tha at eininci waa oarforaied by the method of oy««r
(1961) uainQ ^ •neothyl ohoephate aa »ubetr«*te and fwat blue
nn aa dye*
eBybehvdrat« •»tabeli«w
( • ) pyy »fftff I' tf yffi^f? ffiontf»»t
R««ult8 rcvvaled thet th» ««tirotre>»ri0» pot8»«a
24t dry nxvttvr and about 7if€ uatar*
(^) Total fti^rtyehy^y^^^ 9^^m\
f r e t gltieoaa ecmatltutaa abmit ^•46*%56f of
of wat wt with wn avaraga^ !1»5lfi< Jt 'l.'JJS Sf and 1»91-?«3S<
of dry wt with »n ttvtB^t of 7*14!C ^ 1.1 f!5 ?r«
Clycetien oentant In tha laataoarearla of
£ • oowplanatuwi l a nodarataly hl^h e o n t l t u t l n f about
4«0->4«69l of uat wt with an avaraga of 4*391? ± %11 ^r tm6
16*74-I9»3e< of dry wt with an averaga of 1B»y€ ^ '1,S'^^ «»r
(Table l ) *
( e l To^fi PfoW^ 90*^%¥^%
Tha orotaln eentant In tha wataearo^ria undar
atudy l a about 8»27-i»9Wg of uat wt with an avaraQf of 8»8f ±
0»133 SC mnd 34«44-37«37< of dry wt with an avara^c of 36.?3<
• a.587 SC (Tabla l ) .
.1.
Lip id t in th» Metaevre-^rla under atudy
const i tu ta about 8«42*8»8S< of w»t wt with nn averaqv of
§•6^ 4; 1.181 Sr and !l5.t««3S»S4K of dry y t y i t h » mwmn of
S S . 9 9 < ^ %631 Sf (T«bl« l ) .
f?|itA nakffo (^ eboyt %^7?*9«^8€ of u«t ut with
»n Av»rfi{}» of l»n9i j^ 1.102 Sf end %3fJ3»f5.366< of dry wt with
Ml svoraQo of 1»'5< *_ 1.119 * r , wH»roii«» OH* eonotitut»» about
0 .12-1 .1*< df wot wt with a «i»«n of 1.13< j ^ 1.1SS «fr nmd 1.19-
1.1?< of dry wt with «n svaraQt of 0.14S< ^ 1.1?9 sr (Tabla 1>.
t^) Carbohydrato i»otaboli«P
Inoubation of metaearo^riaa In alueoao fri»« end
glucena containing oalineo r«»ul ted In to ths* doo^etldn of
^lytsoQim* In 3 h incubation liataoorepriao l»e»» thv i r olycoQan
raaarvo about l7 .8rC and 14«6K in gluooao f r^a and glucoaa
containinc) aal inaa* wharoaa* in 6 h 31*f lK and ?4.89< daolation
takoa oisea in QIUOOOO f rea Bn4 Qlueoaa eontaininf oalinaa
raaoaot iva ly . Inoubation for 9 h orodueod about 41.8<K »nd
29.3QC of daolet ion of glyooQan in Qluooao f r t o and olucoaa
containing aalinao raapaot iva ly* Nouovart 1? b incubation
raaultad in tha daeraaao of glyeooon t^ ich i « about 5 2 . l K
t^tl 41 •2H i n Qluceao ft— Bnti glueoaa containing aalinaa
1 ;
rMovet lv f f ly * 15 h of Inoubtt ion of notaeoreeriee r t t u l t o d In
f7«339E »nii Sf)«8n( doplotlon i n glueooo f ree ^nd ijlueeeo eontninlnt;
ea l ln« t and 18 h ineubstlen orotStiood «i«rkod d(iol#tlon of Qlyooo^n
anotmtlnf to ebotit 84*30^ «nd 78«S^ In igluGOiK* f r o * mniS eontsln-
Ing oellneo rotooet lve ly* Af ter 18 h the pereeitee do not survive
in ei ther eel ine* The reeul te ere eunnerired in f i q ( l « ?>•
Incubation o^ oereei tee in eelinee ecmteininci
19 fffl 9lueoee reeulted in the uptake ef eXueMe fren the
IneubetinQ fliediuai* 2 * 7 , S«6t end 7 juaiol glueeee/Q vet ti^ w«s
taken (^ from the nediuM by the pareeitee in 3» ^t f^*^^ ^ ^
rtOEseetively* In 12» 15 and 18 h ineuAtation the uptake from th?
Krdiuai wee 7*4, 4*1 »niA 2«8 jmol/q wet wt Cfiig* 3» #>•
r f f e o t of etarvet ion on Qlyoogen eontent wee
marked in glueoee free eel ine* In ea l ine without ^iuooee
the aetaeereeriae leaked glueeee into the nedium* 0*Sf 1*6 and
2*1 >j»ol 9luePae/g wet wt of peraeitee wee leaked into the
Aedium if? 3 | 8 and 9 h of inoubatien« 12» 15 end 18 H ineubntion
reeulted In the leak^^e of 2«9» 4*1 9nii 5*1 ;u«ol Qluooee/c wet
wt of oarael te (fifli 3» § ) •
The analyeie of iiiHiiuii» in which the pareeitee were
incubated* revef^led that Metaeercariee produced l a c t i c and
pyruvic acide which were excreted into the ineubetinQ «ediuM
at the rs te of 1»A78 and % n i 9 /UMOI/Q wet wtA» reeoectlvely
(Teble 2 ) .
IV
(•) trrmot of PH
Tyo olmmt pffaka of •nzyme netlvltifft yer»
ob«fi>rv»dt one «t pM 4«S-S«^ end Bnother «t pM ll**^ frla 4>,
IndleetlnQ thp prvspnop of both Mild wmti alkallnff phpophntasM*
Spoeific act iv i ty at pH S waa raeordad aa l.'^l? }mol o-nltroohannt
m^ ptotaln/ailn and %!3049 fjmol o^nitrophanol/n^ orotain/wln mt
o>i t1 with the rat io of acid to alWalina phoaphataae activity
bein§ 3*5 t 1*
(^) f f foet of inmibation tiwe
rnzyNia act ivi ty in relation to incubation time
ravelled that^ after the f l re t 19 win of ineubati'^ tha activity
iif both the enxyiaee roee eharply yhleh continued up to S' mln
tiHvt which i t becMie linear with no diatinet riae in their
aot iv i t iea (fiq S)*
(c) £tgjMai.stJi«RPtrf.^,Mxi
Different ohengea in the tenperature on enzyMe
act iv i ty ehowed a audden Increaae in the acid phosph^taaa at
so" C (ri« 6, M) Bnii of alkaline phoephataae at 40° C (ri^ 6t ''^t
below or above thie tewpernture eharp decline totsk place in the
e c t i v i t i e e of both the enzyiiea*
Ki
(^"^ .^fffgt of h^ogynn^y
!nor9cit«d »et iv l t i«« M«r« ctottrvvd with thv
incrvesing eonevntrationa of hoit99«n«t»t et both pH 5 «nd 11
(r is 7 ) .
(•) „firfgff,|, fff ffMbp^rnlf foggfOiljjtio!?
The ttffocil of c^lff»r»nt •ubttreta concantr«tlon»
frcM f »5->t? iiHn at oH S and It wara naaaitrael &n anvyiiip aettvlti»a*
The i'^lchealla* otmatants dat»7«inad fro* Llnawativar ft Aurk olot
(rig 8) ywrm fesund to ha 1«1 w^ with a V..^ of %^18 unlta/»e
orotaln end 1«6 wff with a ^n-^ of O.fl lS? unlta/wij orotaln for
acid and alkalln# ohosohataaot ri»ao»otivaly«
ffaaulta of tha affaot of certain chaaileela on
anzyma act lv i t laa ara aumarlratf in Tabia fS)*
'Q) CAtfftroffht^yti ff .fltf^^fi
One band aach of cethodie netura w«a obaarvvd
for both the envyisea in poly aery iamida gal eleetroohoreaia* Tn
addition a wia aotlvity waa alao obaarved at tha origin of the
gal in add ohoaphataaa only (fig 9)*
The r e s u l t s of biocheiRicf>l cowooaltinn (Ti»hls Of
o lucose optsks (rig 3 , * ) , Qlucose l»flkeo» (flti "?, "^, pffpct
of atarvptlofi on qlycoQsn content frig 1 * ? ) , oroductlon of
l a c t i c Fod oyruvlc ?«ciria (Table 7) pn6 k l n p t l c s of nc»>-«ipclftc
oh o»oh OBI on opBt erases (flQ A»9>, pre cjenerrlly In Rccordpnce
u l th thosB recorded for aexuplly mnture trewetodee ( l i t e r a t u r e
In von f'r^'nd, 1973? «wyth, 1976) .
The cjlycoqeo content In the metacercfrlj" of £ •
comolpnatuw l« hloh, eonat i tut ino nbout d»3'»^ of wet wt r>ntf
1??,T^ of dry wt, vhich i e !n nrcordpnce with the findfnne In
.-^dult tremstode»» namely, raec i e lp qtopnttcp ^rol l , 1*^61 ,
"oiethooylahe renre , nioIndieeuB jMjhcl,>»V".tufli, rpgefot^' *^f>n<*tlcp
^M'ltin, 1"67, a \ niycroeopliuffi dendrlttcug ^irM^tt * Le*^ni»r,
1^71 r-nd Wghroprchle rpn^ruet ^'^prypkf^rte _£t PI»> 1'^7P\ T S ! «
hlrh l eve l of olycocjen mpy be because the w t a c e r c p r i e invp^tfp'
tpd inhabits p se«Bir>neerobic environment, pnd PISO hpino P
t rans i t IOOPI st«»oe io the dewelooment, reoulres wore pnprov for
nrowth ^nti metabolic reoulre«ents»
nororinrlson if 1th the d tp pvpil«hle for adult
trewptodee, ind icate thrt the orote in content in the wetpcpr-
cpria i s s l o n i f i c a n t l y l e s s (S.Tt of wet \jt ?nd 36.73^ rf dry
u t ) . Moijever, Pckert ft Lrhner (1^71) reoorted P.6*^ of yet ut
as the orote in content in adult picroeoelium dendrit icua. Thp
oresence of surh e low> pwount of orote in *n t h i s we^pcerc^rip
ff>ry bp dup t o tH» nb9»nc9 of «cl«»rotlrtlrinf? o ro t^ fn« , i^hfcH
pre UBUplly lnvr<lv»t^ In #c?f| inroductien of t:r»iw»t"ffff» (*«vt*> *
ClpnPt 1* S< >. «uch n ro t# ln» for m^ct oroffuctJf-n would nr«»*"hlv
bp synithpBlzpd only £n th» f«cfult« Mownvprt I t rf«»»l.nfi
•oecul '^ t lvp u n t i l wor© df»t>»llt.d inform^tton !B mr»rfi> »»vi»fl«»h1p
on thip Tro te ln »rft'=»bol!8W«
Li-^tr^s cootlfcut?» ««br>ut R.S** of utft u t sod " «?,<»* o*>
dry titf uHlch in Hloh whpo co«»oiir#»rt with th© mpjor l ty of )»Hult
tre^t^todpa ao Per lnvi»«tfai?>tprf fllter>»t'jr© fn voo "p«»ort, i«»7' »
*mvth, 1^76^# How(?vprf t h p or#«ii»nt f t n d ' n r e «*•«• !n porp#*(«#»nt
with l»olf»(* Pf< s tudf^a on i"dult dfoiwfs^na, *^ehlat090«ip wfm^onl
( T i l t h * "rorrkp, 1«16'5>, Cotylnahoron cotyl«»nhr>rum« ^for>»>*;oootvlf'
gxolf»optum> fchineatowr^ ma.lgy??ntw» find rg j ic io loppla hua><l ^^upu^i
* *^!ddloi, 1976K Th# hlnh H o l d contpnt In t h i s nrooi»n»-tlc
mpt rcp rc^ r l a eonf l rna thi» theory of rnf^i»lHri»rht * ' »1s» M'''»4>,
uHr» r#nor t fd tH-»t th*» Icmr aurvfvine l^rvi^l «tpor» n^ H^lmfntH^
»tor#» IlolH*?,
"•"A fBi-k'^S UO n b O U t '^•'1f*< o f U F t W»t ''Od '^.•^"^ O^ H r v u t
yHrtrpnu, pwn con«tltut*»« •'T '?^ or ua t wt wnd 1.14'* o'' drv ut
t ' f tb 3 r? ' t io of 7«"5 J 1» Thff occurri»nc? of n^f .f>nd «»*»' fod!cptp«
t h e or»«npnc» of a c t i v e o r o t » l n «pti»belisw to t h i s »pt!»cPT'C"rl»».
Th«^9» r e e u l t t a ra raore op li»aa !n POTi»ew©nt with th«» f indlooa no
?»dult t ramatodeB, n l e i l d o o h o r a wtTiftnc?! ^^?»#, 1 '»'=>> j»nd £ •
h f o n t i e e 'Lpajat £ ^ . , 1'57«^>, Hoi»fwart f o r t h r r atud!*** PT-P
r eou i r ed t o i>acartpln tha r o l a lyf nuc l a fc mrlda fn th^i syn^-H^nlfi
o** ondon«»nouB o r o t a l n * In t h i s «ptsc»re»pl '*«
2.
Pfnrf p f p c t of ««tprv?*tl"n f»n nlvcofjtwi cf»nt»nt ^rio i t -?> ^^nycr*
more or IPSA «lmilor Of^ttfrna »• r«»nort»f< f** *!«»wu>»!1y mj»fur#
tte»m?»tod#9 ^von "r?'ntf, 1'?7^; Ttlnm, I'J^^t ' prvp'<^rt»» j»j j»T,,
1*^76; •'myth, 1^76^, Trom the rptultn It Is clpf>T tH;.t from 7Prf<
to ^7 h, t'^ijrr te nronouncffd Uf3ts»ki' n f t f t wHlch thp untnke Is
»loi,f ?nf* npsrly COWPB t o en #nri pt 10 " h. Th© nnfipiMp r«»p»»r>n
^r>t t h i s i^Jf^prpncp In thp rp*e af cjlucowp uatuWe mry hp thf»t •i ip'
anrpt i teo rewpln - r t l v p In thr f i r s t 1? H nftpr uHlch thplr
•c t lv l ty r!r??duelly dper#i»pp» in th!p wpdiura. Tt u?»ii notfcpri
th-'t thff ouantlty of c^luronp lppk«d hy t h i s yorm !nrrp»»»pd In
rplf^tlon to incubptlon t!mp« Thp oattprn r^f olueopp lp?»kpop
(riq 1 , ^) in th i« m(»tf»cprcprlp indirpct ly puoopptp th^t the
tpnuwpnt of the worm 1« involved in the s e t l v e tr«nenort
mpchf»niefB» ftccorrflno to von "rpnd fl*'?*^^ the e i o n f f f c n r p o^
cr-rhohyHrptp Ippkpoe eeeins t o he oheeure "OH i t I« mnpt Hkply
••; "»othnlor?icr'1l r»rocP8« in re«non«e t o exr»Pt'l»e>ot!»l ir\ v i t r o
cr»ntiltl''n8» Thp exnct mechsniein pnd the rppfi' n "f nlurosp
IppUfsne 1» not oosnible to be pxolpinedt but the nresent etudy
«ee»9 te Indic^'te th?»t pt Ippet two fpctore pre Invnlvpd, nnp
1B thp s tructure and nature of the tec?uiiipnt p>,r>f^ the othrr e
thp unoFtur?"! condit i<^ of the incubptloo mpdiuw* Mou«>ver, thf
r*pr,ii!»tlni ff olycoqen in olucoer contplnlno sp l ine frin 1 » ?^
*unc>pst8 thf t qlucose elone CPO not comolptp ^he nutr i t i ve
reoulre««nt» of ofirfeitps in exoerimentpl in v i t r o culturp
ptodfpp pnd attempts should be mror^^ to Imnrove the innic
9 '
comooslt lon of t h e Incu^ptfno wpdlum bpfwrp p^y lr\ v i t r o »tuf*v
i » un<^ertpkpn»
THF orpit»nc» of l?>ctfc ?»nd oyruvfc weir's fn f-hr
lncub;»t£na mwditmff nnd thpit rj»tlo fT»bl«» ^ > confirm t*'** »«?rtfpr
0UFlitatlvi> f lnr i lno t on tH»» s^mp w«»tsc«»rc*»t't"» 'Thnnsjji *
l ? e t l c «r»d a i n n l f I c s n t l y l«»»«» !nt#n»ip nrjotn i f ' jyruvfc prr^'** »n
f^ftB f)r«»ti»nti»d In t h f orpvtnun cH«'»t»»r clt»p>'"rly fHov
t*iAt jncirf 'jHniftoh^tnsiP 1» »iucH «ofi» Aotivff ••* pn p!Wpl!n(» 'jHnjjnHp-
tF?8i? in thi» w«»t«ci!»rC'TJp ondpr stucly «nH !« In i»«"cnrHp>nrp vfftH
thi» f lnHinns ?n r«lffir?'i»nt i»efult fcr»«ptnd(»i«tt rnf.ohrfpll p Mjrrn-
g,ln««t, ?!al i e n t y i e krnyfir t t '^nlyatfw^ f nf-.gripf ylmfitw, £_f r>1 nr ' nn
'"'jaratioxtif t f l'^c*yrntyl_# »?ntt t^t ,g> plellr'.e^h^r^^ wprI?'not,
cxlii?i?li?c5£» .'"'3lstHogylyr3hf fpn^p, ^^Ipiod.t^yeuw gubcl-'^vf'tup,
'^otopcftrt.n* _yitgllll,,flfe;fi rorpOf!ere eynnnlr' g» g>, £ , VTft tc .^ nnd
i » ESilfSHi ('^^Ifcnn, l%7b> , £ • nio?mt ICP ^"robprt ^ £ l» t i '^ '? ' ' \
£ • bf-'OPttcf f"robi?''t ^ Lwin, 1^74^, ,|f".^,g.t?.t.ebj_y hynwplnHrnr!
''«rlv'^pt5>v?=' -« f u o t s , 1'^7S^, f r^nttor^iscu.f> ,".ffl.'?,^j •"'•''Jf ' -unf r ^
!*uots», i9?tO ;-nr* .^^lagtrotHvlgx frutwffnl.^py ff"uot« *• " n s r c r l , i'^?"'^
Cr»ntrpry t o thpsFf "^ufitw * f*uotp fl'577> rpf>ot"tf»f* tbr^t ! " fsr«ult
f'ang'n t l r r l nuWt tbp ^lkf»llnp ibn^oH-^taup t s mucb wntp •^ctivp
t^sn «c!cf •jbosnihptf»8r>»
Thp recorded oH aotim^ In thm c»ri?t#n* study f ^rtr
"cld oHofliphr-t)Bt# BTw In !;H» »is»irr# r«»nop gi* A.* - "!,i^ ^n rmnnrf-p^
by W?>P1' UII workin'ii for Pdult tr#W)«todP8 (Hr-lton, l ^ 7 b t '^rnb^rt
^ Ji»» l^''^? ""nil, 1P71? ^robPft » Lwtn» 1''74j ^rfvPfttpv?* *
ruot«», 1f»75 -od ruotp * '•tmf.s*, 1«»7'' * in7R>, nhflp •'ftf ». k?»1fnp
o' «5«i9h-tp»f>t f:H«»««» mnp In pnr«>flM)t«*nt wJth Mfypinl r^ j*!^. i' *'*: n«
three tri«>?»tc»di»a nf»«i^ly, fpjiciolBQgl^y* hugkj« f .j yt rod! went dp*
hnwlnig ?nt _f » wgtlf?y* ni»Bt Inh^^bltlnr thp Rpetrolntentin?! tr»ct
o f !3lq«
After ST »ln of Incubation (rlq» S)$ tbp pcttwitif**
of both «»niypp« of thi» mptw!crTe?>Tip Cf»«» to R upturiptt'^ nnfnf
ouitp d i s t i n c t l y . Tb«»#» r»»ult« urr not In i»or«»f«»i»nt ifftb thi»
only r»0O?t In t h i s r>rf«» by "^rob^rt * twio f i ^ 7 4 \ i.h« rF Pn?ywf»
rpwinp 'ctiv«» uo to ? h In £» bgo?'t,te.?*> HowfV«>p» l ln^prHy
bptwf^en r t l y l t y r-nti increaiilno ho«no»n??tf» 0?mc«»nttntl'>n«( wns
obtprvpd In both th» onTymstit which !»» In potppmt^nt with '^rohsrt
* twin (i^7&).
It e n bi» fir«n froB! the fffsults f f i o . 6> th - t thp
Frid ^bn•lh^tp»e Is mors thrrwostpblc thr>n rlkpllni* ohn»«r»hnt^s»!
the s«»# ^hf>no«pnon uss obpsrvpd in £• h.gf><»tfp.w '' rnb*»rt * fu!'' , o 1f'7d). The mtlifn»l f c t i v l t y of plkrl lnp flhoftoh "t P«S »«t 4'» ' ' I *
in 'rresmi»nt with thet of *«iTtmi jgt ^ » ^1''75>, no slnht unpcfps
of c*io(pnefeir trww-todft from both hn«Beothsr«ic i*nr^ ontkllnthsrwlc
hopts . Thp ocjtlwfl p c t l y l t y of ^ . ct^ol-^Hptuw mstrcerc'Tl* pcid
nh08oh='tP8s *»t 51 C w^y be pn Pdaotstlon on the nprt nf th i s
') i
D^raslte t o mstcH thp "wptfibollc teinn»rsturp co«if»n)»ptinn''
(uhltwore * roldb»rp, 1*»7?) of i t « oo lk l lo thfrwic Hn t nri to
3r(?pf*aot I t s p l f for ths chnng«» in tpwo^rnturpf rturlno t t«
ttsneformration Into Bn ;»dult In A honiipothwmic hn^t fblrdl .
Thi* F<ctlvltl#A of both th# pniyaips OPVP P tvn!c«l
byoppTaollc curve u l tb tb# incr«it8ino • u b s t f t p concpntrpt! f .
Thp !^icb»?»ll8» conptflotp f l , l «'" pt oH 5 mr? ^.g «!* ^t r>M 11^
<'or o-nitronhpnyl obonphatp ar© In thp PP»BP ranop r^,''-i,«? »*• *
p rsoortpd for aowp otbpr ^jprspltppt ^» wrngonj^ ''«l»mn-cmftb *
^tf^ndpn, 1963) , Jj£2lJLS JiiiE '''^uttsrwortb * "r^bp't, I'^Tn),
£ • ^'p^yotippua, £ . bugtkl fr-oH, i<i7i> rnr* £ • bponttcp ^"rf'bprt »-
Luln, i^74>, Thp IOWPT K_ VBIUP *'or pclr? obopohp*J»»P fnHlcptap
p orpi^tpr pf^'lnity fof thp 9ub»tr?»t» whpn cnmopr«»d tilth plkrllnp
fjho*!nhr' ppr»
'^n «hfn>»n in Tsblp 1 , V5»riou« wptebo' ic inh£b£torp
; nd stiwuli^torp havp dlffp'-pnt pffpctt on both thp ffnyywps.
'^tliBulption or Inhibi t ion "onperp to bp conopntrptfon ripopnr'pot .
•^I'^prpnt concpotrptlonp of *l»r pnd •'s-'^pH oronort!onp.tplv
inh ib i t br»t*i thp pnirympp* Thpte flndlnop prp In prcorf^focp yit^
thopp of r o l l ( i « 7 3 ) , opobprt * l.wln (i«>74>, «!lypml Pt ^ , ^l^e
??nd •"rfyppt^v?' * ^uoti* (l^75)» '^eth •'o** >»nrf ro*"^ ntlmulftf th?
pnjyiBp ' 'ctlvlt ip® but thr dporpe "f ptlwul'^tion Is dl'fp^pnt In
thp tvn pnjywrs. Thr stlmulr^tory pffpct of »o** on ^cfd
ohosoh'tape i s thp «»«« ?»» rpoortpd in £ • tiorinuw 'ruotr> *
rui->t= , 1077> but dlffp'-pnt ^rnm thet found In J2« ginpnata ''f»,
1964), £ • buaki 'od £ . rppyotlfcua Tnoll, f 711 md J» hyogplo-
bf^r.rl f'^rlwrptpv?* * noot.'^ 1''75)» Hur flnd^nna o^ o*"* on
alks l ine i aho«sr»h?»tp»(» pnri Co on bntH thp pnfywp* sjfp cwstpra^l «>
with thosP nf m i l (i«»e6 « l ' ^ ? ! ) , vfypmi ^ ^ , ^1^71;^ '-.j^fp »
*"li»ctronhorptlc «tudl«?>« on oo ly r r ry l rwlHe (5r»l«t
shower' on?* bsnd ppcH at tHp n s o p t i v e OOIP of t h e fipl» •^unnpuf-.inr
t hp abne>nc» of i«ozymfi>a In b n t s thi» mntymfB- Thpup rpwutt-e f r
In aqr#»fi»pnt with thosi* of C P » T ! (1'57A^ nn ^ , ^enw^^nt« '»ntfc»v/pr,
In edul t t r ewa todes 7 t n 1 lao^yfneff hpve bppn renor tpd bv ^p
fl«if4> In £ • i i lneg is :^nd J^robert ^ Lwlo f n 7 f > !n £• b p ^ - t i r g .
' ^ iml lp r ly , two isozymps of s l k p l l n » obo«obrta««» t ' f P i»lm^
obaprvpd In t h e c e r c f f l a nf £» w g>ngonl ^"Ino jpt ^ » » 1' P*' . "r
r*»^ort i s p w l l f b l p 00 t h e o l r c t r o o h n r p t i e w o M l l t y nf then*
po^yrap*! in tbp R»?»tPcrTc*»rif»« Tb? pc t fvf ty r t tbp n r lo ln ' •* f.**c
fjpl cfprp 'oonrls t o tbp mpwbprsoous corooonpnt of tb«» "cfr!
obnpnb^tppp end Ind tcp tps tbp t ecie* obofob^f^s'" bp« fu'^ ^ r r t s ,
OOP •npmbfnnup In np turp pn6 t hp otbpr polublp* H'sypvpr,
?»lkpllnp oHnsob»t«9P i s rtiffprpnt es only nnp •^fod yn* o^sprvrH,
with no ' c t l v i t y s t tbp o r i o l n of tbp q p l .
Tbp rolp of ohosobatRspp in tbp "TTtf^boHsm bpp bp«»n
wpll d i scussed In s nuwbpr nf D^n^rs on Hfffpi rnt oprosf tp
!<opcips« They ocrur in rpolonp ubprp p c t i v p t rpneonr t o^
m p t p b o l l t t s tf 'kps O1PC» sucb es tbp tpouwpnt rod t* tb r t{RPU<^9
of tbp r»r>roducttvp system CMlmffio-*^mltb ft "^tendpn, i % l t
Malton, i % 7 b ; * o r r l 8 * Tbrepdeold, lOfiR: "p^dpr , 1'»7i! -"psprl,
1^74; ^ p n - l s ^ j ^ . , l Q 7 4 t ^ r i v p s t e v s & '"uo*r, 1^75t '^H»»r»p,
1*^76). Tbp f lnd lnos nf t bpse putbors surj^pst P funct ion for
'^'
(I'JTe^ riKiorted thpt they i»s»y R1»O be involvfri In fSp u'lfpWp j-nH
trantoort of nutr lwtt t In thp vft^llj^rli*.
Thp r»8ult« of 9rff«»nt f!nrtlnc» did not ehnw pny
k invt lc dlffpr#nc»» b»tw«»»n t h i s orootnet 1 n w#tpc»»rc!»rl»» -»nf«
rflfft»r'-nt sdol t tretBptodPn. from th«ip rpitultff I t nmy hp
surwljpd th=»t tbp9(» fnjywfir h^vp nn Of»rt tr* oli»y !n thp »»ctlw«»
r»3rr»ductlon of trffB».«tod»«» bt>e8Ui!t« In tbfj* m p t p c T c r f p ;«!! fh*
rparoductlve OTRPOB »r« u f l l d«iwi»loo8d but only ?no ornductlr«n f.
l-'cklnot while In th» vc>rt»bre>t«>i» thi* r o l e of th»««> •»nry»»»i» <n
Tiioroductlon It uiill B«tBbll«h»d (Wooof 1*346 » ?»f»a»lblv, thu-nm
iB?y h?»we «>ow«» involv^we-nt In th» corbohydrfttw m»tpbnH»w of
nonpd* 9r\6 in th» mffiotic d i w l t l o n . rurthr-ritorff h{i»toch«»iHlei«it
r(>fiultf> (f^lfifi Ht ''^ittmlf in or»ff«) oo thf «««» w»ti»c»rc*«rl8
Indicftp thr»t thp>ae eo;yiBf>s ar» loc^l iz^d in thp r^oroduct tvp
oropos pod !»l80 in th«» ir»oiont uhtre oo«BtMy »cttwv» trwn«'jO'»f
nf nutripntp tfiknw ol»c«»« MoutfVffr, th«»lr rol«» rf>«8lo(i »o»culi—
tcry u n t i l «or» d«»t«»lli>d Informstl'iii I s nindP ewnllnblp on th i s
aeapct.
Thf» Dtes^nt Invffstlcjetlon eiL^oi?«t« thi^t tbl»
««ptPc»rcf»rlR r)ot«p«*«»» mnre or !•»•« iiii»Ili»r o»tt»ro of bfo-
che«lns»l cowDOsltlon i»nd kjn«»t!c« «f non-«o»ci f lc ohoi»oho»ono-
f»tp>rpi9P9! 8» found In dlf f#r#nt »c>KUftl1y mt^turc diepni^pnn fvnr^
"'r-'Od, ^^7'it «wyth, 1'976)» Thi« *.« indio^^tivv thist bi>eld*»8 thpsi
constitupntw sowp oth^r f?»ctor8f l l k » t»«f3i»r«turpt wprtpbrptr
host enxywpB Bnd hormonps i»tc, »8y »l80 lnflu»nc8 thp wjc
oroductlon*
The Dr»8»nt diaswrtat l fm d9»i9 witH th« bloohMiiciil
eo«po«iti(Mi} toM* atovcts of oerbohydrat* metabolltm ^nd k i n » t i e t
of non*«p»clf ie pho»iihcMono»st*rci««« of th» oroQvnvtlc itotaoorcgrii
of S^iiQy^o»Mf fio»p^ir^B^^>
Tho bioohMileel eoaooti t lon of tbo ii»tiO»ref>rlc
consists of f r ss gluoos«» 0.516< ef wst wt (2.14^ of dry w t ) ,
glyco^sn 4*39^ of i#»t wt (18»3'T^ of dry wt>, o ro t t in B»7< of
ust ut Ofi.ZJ'C of dry wt ) t H o l d s S*64^ of wst wt (3S«96< of
dry u t ) f Wft %08K! of w«t wt (O.SSSt of dry wt) »nd fWft %13S<
of wot wt (CI*14S?C of dry wt)« Ths oattsrn of QIUOOSS uotnks»
gluQOss Isskags end s f fsc t of stervnt lon on the i r glycogen
content I s sl is l lar to that rsported for adult dlgeneans* Lactic
6f\fi pyruvic acids (l#478 m6 % ' ) t9 ^ o l / g wet v t / h resoectlvely>
were foxusti In the Ineubatest suggesting the presence of an
act ive g lyco ly t i c carbohydrate net^ol lcHi* Protein pnii H o l d
metabolleflis are also discussed b r i e f l y *
<\cld and a lka l ine phosphatases obtained froM the
Rietacercarla showed optLnal a c t i v i t i e s at pH 4 . 5 - S . l and 11.1
respect ively* Speci f ic a c t i v i t y at pH 5*1 yes recorded to be
^•017 fjmol p«>nltrophenol/iig proteln/mln mfi '5*'1C)49 A*"»O1
p«nltroohenol/«g prote ln /n ln at pM t l . ' l «t 37 C* The rp t lo of
acid to a lka l ine phosphatase was found to be 3*5 t t* f^axlMal
a c t i v i t i e s were observed at S^°C and 40*'c In acid and alkal ine
phosphatases respect ively* <;eturatlon In a c t i v i t i e s of both thr
anrymes was observed a f ter the f l r e t 5!7 nln of lncubj»tlon.
2a
l ln»arity In wixyns «ctlvtt i»» w»r» notievi! with the incrv«crinc;
hemiofivnstv cenovntratlofit in both th» wntyme9* K for
0-nitroph#nyX pho«oh«to waa found to bo t«1 •!*» ulth » V.. ^ of
^ H S unlto/iiQ orotoin for acid phoaohetooo Bnti 1*6 nn with «
W_ ^ of a fJIS? unito/wg orotoin for alkalln* ohoaohataoa.
Olfforant conoontrftiono of aoditMi idrsonata and todlua fluorida
inhibit and ff©** md C9** otinulato both tha onzyaaa* Polyacry-
lemida QOI alactroohoraaio ahaw«d ono eathodle band aaoh in both
tha onzyoiaa* Tha poaaibla rola of thaoo onrymao in tha natabolit
of tha nateearceria haa alae boon diaeuaaad*
Hjrm^<;^,^
Mem, <;.r>!» & Ni7a)iil» <.'•!}• (1980) Hlatochcmicr.l •tudlvs on tH» mmtmencBTlm of CUnottowuii cowolawtyi (Jtmntotin i niQ»nffR). ££0£. lUilai* ilsaat' pBfgf ^Jl* MiaaiSI t 11.
/ir»», C,(l97S) Chi«lc«l eo»oo»ition of OlolldeoHerii wtrlartoi (Wc«OQ«n»fi)« j « Para«lt»nk> 47 i 211-?15«
Mtanjit W« r. ft Willii$ii8t ff.C. (1973) Tht ttruetur* «nd hi«tooh«Mi8try of to** trMi«todo nffttMSPtonriiil eystt* 2* HglmintHol* 47 I 3S3»3fie#
Ber^»y*r» H*U»» Cewv ny K« ft Hrsotlt n* (l974)^rnxy««» AS bioehmiioai r«i>Q»ntii» Inn n»thoc!8 In iffizyiiitie nnnlytls* 7n(* Cd« (Horgmoyor, H.U., •d) Vol* I , no* 42S-S?2» J»c*rt<e«»ic Pr»»»t N»w york#
Sogltshf B«3« (1962) Tho chvMicfil nptur* of motsevroprial cyats* !• Hlttologloal and HltteehMileal obovrvation* on th« oyst of Po»thodiplo«towu« f»iqi»^* ! • Efi£jBliiSi« M < ^ - ' *
Boy*r« S«M. ( l96l ) fllkelinv ohoaphstes* In th» himm nmrp md
Butlf?r, ri»S« ft rriedi B* (1977) HistochoKlc^l end thin layer ehroMBtograoHle analyalt nf neutral l io lda in Cehinoato«» ravolutuw ii»at«e»rearlB» oul tut ad }jx JtHia* !• EJeEiSIliE^
I 1041-1! 6a I 1041-1045 •
F)uttarworth» 3« ft Probartf A*3« (1970) Non-spacifle phoaohoMonoaatarataa in ftaearfa auut !• Cffact of InHibitorsi sctiwptora mti chalatora. FKD« Parasltel* 28 j 557-565.
Carroll, 3«3«, «5inlth, N« ft Babaon, ft.L. (l970) A eolorl«atrlc aariM gluoota datarninntion using haxokinaaa and Clucoaa-6. phoaphata dehydrog»naa«« Sioobaw. Wctf* jft t 171.
Caap-ri, !•«• (1974) SebiBtoaoaiff aienaoni t niistribution pnd cbaractariatica of altcfilina and acid phoapbati»aa. Exp.
Chang, T.C. (1963) Bioehamical r«ouir«i»»nt« of larval tram^todaa. « nn. H. y. Acad. ^cU 113 t 289.S21*
a .
Cxok, n* k Lanprectt iu ( l974) Pyruvattt pHosphovnoloyruv^t* ana n-9lyc«r8t9-?-oNo8phet«* Jnt Wethodt of ynzyoiAtlc enalysi t * 2nd Td (vd* H»U» 0«rg«ty«r ) , 90 . 1446-1451. AcadfftRic Press* Nsw York*
Davis , B«3« (1964) Disc slsctroohorssis* I t * nsthods and appl lcet ien to Humtm ssrum protslns* tmn* JL»X* IfiJUi* Sei» j y ^ I 4CM-427.
Osnnlst E«^*t SHerDt fl« A DauQlasCf >?• (1974) CsrboHydrats rsssrvs end oHosgiHetass a c t i v i t y in ths •o i lute- t rsMstods
(Loos, 19^1) Odhnsr. 1911. 3« HsUintho)^* 48 t l - H .
Oixon, K,r» ( I 9 i 5 ) Ths structure end h istocks* is t ry of ths cyst well of the «etacerenriee of f gsctole iiSSLfiiifiJB* .ffiLS&L&SlSUL* 5S I 215-??«.
r c k e r t , 3 . 4 Lehner, ?• ( I 9 7 l ) Sauerstoffverbrauch, Substrata-ufnahiae und tac t at ausechei dun^ van Dleroeoeliuat dendriticuw (Tre»atcde)# i « P»raeitenk» 37 t 288«5'>?.
Engeibrecht, H» ft Palm, u» ( l964) Der endo^ene Clykopen-und Fettetoffu^eheel in seiner Bedeutung fur die r i f ferenzierunn, Lebens-und inf6>ktionsfahiqkeit der fntuieklungastadien parasi t iseher iiunaer* X» Paraettenk* 74 t 88«n4«
r o l c h , 3» , t eas , I*'. * <ilone-stenley» G#M« ( l957) « eianle wathod for the i so la t ion md puv i f ioat lon of t a t s l l i p i d s from animal t iesues* 2« M S i * g^—»• ??6 1 497«5')9,
Foratsr* *?• * Taggart, ?• (1951) Use of iso lated renal tubules for the exaMlnation of metabolic processes associated with act ive c e i l u l « r t ransport . 2* Ce l l * f oatp* Physiol . V} ^ 751-270.
F r i e d ; a . & Glosoia, «?•«• ( l976) Tentative i d e n t i f i c a t i o n of ehnlestrol BB a chenoattraotant for metacercarial pair ing 1 " Utucoehloridomortyhff ponstantJaf (Trematoda) J . Pgyftf i tol .
F r i e d , 9 . ft *;h«oiro, I . L . (1975) Accumulation and excretion of neutral l i p i d s in the mrtaeercaria of teueochloridiomorphp constr t i t i je (Trematoria) mg>lr»tninad ^ yftroir 4* P^resitpl»
ii^
rrivdt B. ft 3utl»r» n*S» (l<?77) Hlfttooh««le«l and tNin loy*r chroRiJ^togrciphle enalyelt of ntytral l ip ld t In th? •fftac«re«rlal and gdulfe Cotylurm ffp» (Tr««iftodp t Strla©ld»e)» 2» Par^«itel« 63 i 831»834»
rri»d» e*,«?obbin8t S.M. & »«l«on« P,o» (1978) In ytvo and l a JdlXfi axoyatatlon of gYgOffotyU JLH!lBi/TT7i*»toda> nvtaeorcf^ria* and hiatoonlMiefti obaevvatlont on tH* eyat* ! • Pffffft4lo|* M « 395.397.
Cll(»at K* k nyarsf A, (1965) An iaprovtd dipSanylanilna nathod for t^a aatifliatlon of daoxyrlbonuclale reid* datura* London. 206 I 93.
Goll, ff.ff. ( l96 l ) Physiological •tudi«a on IraMatodaa-r f e l o I | oioseiilsjli oarbohydrata •etabollaw, Pg?gffi^olaBy,,
r^oily n.n, (t966) Physlologtoal atudlaa on tramatodaa t phoaphfttaaa ayatawa In IJ.iff^fO^Hylfsy ||y^«yf^|ffff* "ifin4U^ogYf ^ t 115-110.
Goilt w.w. (1973) Phoaphritfeiaa ayataw In faactolopala buakl Lankattar, 1897 mti £|ff|ro<;itffVCt .fifgyptlf^^f Cebbold, 1876. ParaalteloQV* | 7 t 197-204*
Gupta, V. ft AQerwalt S*K» (1979) Phoaphataae nctlvlty In G est rothy I ax cruiPenifar (Tr3»*toda). Tnd. 2* ^wfwttol . 2 *
Cupts!, S.P» & Gupta, R«C« (l977) Phoi^hataaa sct lv l ty in into tiorinup f?o« nam^ tlc^tin** x* Pareaitank, ^ t
Gupte, S.P. ft Gupta, *?«C« (1978) Phoaphi^teea ©etlvlty in Caatrodipeug ffiaavotJaeuf (Tfa?m«,todi»). Tnd« 2» EfLSftU^*
rutm»nn, I . ft Ufihlafpld, r»t't (1974) L-(4)-Lsct*!ta datarwin^tion with lr>etiit« dahydroganeaa and HftO. In 1 Hathoda ''f fnzywa-t l c ^t>nslyaia, 2nd fd. (td. H»U. Sargmayar), op. 1464-1468. i^csdamie Praaa, Naw York*
Moltpn, P.f. (i9«7?(> f?tuf?i?'a nn ^lycogan deposition In tranatodii. CowD. Blochaw. Phyaiol. 21 * 113-12%
Heltont O.U. (1967b) Studlvt on ohoaphetaav a c t i v i t y In tr««f;todff«. 2* P a r e a l t o l . ^ « 46*54*
Harr is , K.R, ft Cheng, T,C« Cl973) Hi«toeh«aiie^l dp«onttrittion of f e t t assoc iated with in t«« t in« l enccr of Ltueoehlorldlo-«0££t5£ sofistpntiflf* Jt^ff, Ja» Wterose. ^2S* 2 2 * *'^-''^?«
Hejten, T»C» A ITsch, C.u» ( l978) Obtervations on th» veoloqy of Clinoatoffup If^r^tn^lw i" *^» '*»'9»» «Pufr' h**** (rjcroottruf tel<8ofd(?fj)7 J . r l 8 h ~ i o l . i 2 * 4iVA?f!
Hunter, U»H» & Dslton, H*C« ( l939) The cyst cf the yellow qrub of f leh {£Hnof%pm:^m SMSU^SLa^iB.) £lLS£* M £ 4 a * 12£* ,VEta* 6 ; 73-76.
Hunter, C«C« III i Hunter, u*<$* (1940) ntudiee on the development of the )iiet8cerci"rl8e end the nature of the oyet well ''f Poftho<jlfflof1^o«ia mln^miM (r^^CAlm, 1'5?l2 (Tr^m.todr t Str lqeada) . J f , a t * i a * >?;i,smffi* iSft* 12 » 5?.6"!.
Hunter, u.S» ft Vernberg, M.B. (1955) Studies on oxygen consunotlon In d lgene t i c tr^wawtods-s* ! ! • e f f e c t s ^f two extrem9e in oxygan tension* Exa* £jj£.«atilSi.» 4. * 4J7»434«
Hutchinson, U«C«, Oounie, r«D» ft funro, H»Ne (l9S7> r>(po>fyrlbon-ucli»lu .acli and r ibonucle ic acid extrt^sblnn froci rnlit^l t i s s u e s ond the ir e s t i w a t i o n . t!loch|w» ^i.ophyg. .sct^> 2§2 *
Islam, f . U . (1974^ Carbohydrate weinboUsfli of soer dlgenet ic trewttoties . Ph#Cf» t h e s i s , ftllt/«rh nuelim Mnlvcrfflty, Indie*
Keryakprte, »»•»., Ch >i.id, D»n«, & «ii i»nt, ?«C. (1v76) c»rbohydrj»tP metabolls^i of Wehrsorchls ranertp ^rlvp^tsv^, 1943 ^Tre»ptod» nl«5en«R)., y;&gi thwua.. * i'nj.M* 1* ?,^i* i S * 69*92.
t e e , S.H., <«wg, C.Y., Lee, K.B. It Lee, H.S. (197P) " tudy on the ehro«ioso«»l prote ins of r e s c l o l g h<^pqtl< a> Koreen 2*
tinewe8V#>r, H. ft Bur^, n. (1934) The determlniitlon of enryme dlseoclstlpr* constants* J » j ^ * Chest* ^co« f£ j 338-66fi,
Lowry, C.?j., 'insebrough, n. , f a r r , fl. A Winndall, 9 . (1951) Protein measurement with the f o l i n phenol rewgent. 2» 9 i o l . Chem. f93 t 765-?75.
:i.;
LyncH, o*U i Bogl t th , 8«3- (1967) Th» oht«ioal nsturt of th# m^taovreerl*! eytt»« I I * 8iDche«ie«l Invvvtli^fftlont on thv cyet of PothodlpXoitowuw wlniwuii. ^« P a r ^ a l t o l . j ^ : 24t -243 .
PjtiBlifeol. 1 2 J 235.240. J u u * - * * *
l^aclnnls, fl,3, (l97f3) I t D l a t l o n of orotvin f t notions* In i rNp»riii»nta enti Tffohniouvt In Parsoltoiogy* ( fd* Plrpclnnla, A . a . a Vog«, n*) op* 173, y«H« rrff«i»«n «ncf Ce*9 ^«n r r o n d t o e *
f^lsrsf U*K* (1968) Llvor H o l d s of ra ts sdmlnlstsrsd sxcssslvs «iMounts of r s t l n o l * SJCL* ! • l i S l t U S * i « » 697.731.
nontoQoiisry» R* (l9S7) Ostsrwinetion of ^lyeoQsn* arch* 91ochs«« Bloohvs* 67 I 378*386»
nooc;» r* (1946) Ths pHyslolocileal s ignl f iesncs of ths pho«9hQMonosstsres«s« B io l * Rsv* 21 • 41-59*
Worr ls , r.»P« » Thr#»d§olri, L»T. (1968) U l t rest ruoturs of ths tsguwsnt nf edult Sohlstosows EJBHSSSL* 1* P f l f f l f l W * JSii < 15-27.
)<!unro, HmHm & r l s o k i A. ( l967) Ths dstsrwlnation of nuclslc Aclds. Wsth. MSfitilft* MJBL' lit * 133-178.
N»s(Blt H. (1976) Trsmstods s tsg l l l sen t BlowaHalarii^s o labr f tq •n Cuadsloups. B u l l , soc. 7eol# ISJBQJI* i M » 1162 -1^3 .
Nimmo-Snith, R.H. ft St«ndsn» O.D. (1963) PhosoHomonosstsrases in gchlstosowf reensoni. rxp. P s r s s i t o l . jy[ i 3n5«32?.
Nizami, M.A., Siddioi» A.M. ft Yusufif A.N.K. ( l975) i}on-so#ciflc a lke l ins ohospbcMionesstsrssss of sl^ht spseiss of diprnstic trsi iatodss. 2* Hslwinthol . §9 t 281-287.
Pino, E.C»» Annsxy-J*lsrtln»T, fl.w., Psrsjr -Vl lpr , W. ft Clntron-Rivsrst A.A. (1968) Studios in Sehlstosena wansonl. I I . Isozyms pattsrns for a lka l ine phosphatess» i s o e i t r i c dshydr»9snesst 9 lu ta» ie oxaloaostlc ttt/fmin— and qlucosa 6*phasphets dshydrogsnass of adult uorns and esroprias* JLM* P a r e o i t o l . 1 2 « 288-294.
Probertf A.3. , Coll , « .«• k Sherwe, R.K. (l972) ??loehi«ilc»l and hl9toeh««laal ttudlM on th» non<»ft9«clfie 9Ho0oh(Miono»tt»rA-• • • o£,iJttSiaiJl fl<ftff»t^sB cobbold, t885. Pgrtfltologv. 64 ? 347«>3S3«
Prob«rt, A»3,, A Lvln, T. (1974) Klnttlc ftv(mmttlp9 md locmtlon of non*i^#elfle pho«oheMiono»at«rRt»« In «ube«llulj»r fraction! of raoclo^a b»Petlcg> rxo* Pafpaltol* 2SL « 753-.?6l.
Probart, A»3# * Lwln» T. (l976) rafcloli^ hapatlca i Tba oraaanea of oartlelaoaaaoolctiad snii solubla non-«ooetlfle acid phoaohataaaa. J j ^ . EalMiiSi* 12 « 2a««?11,
Ral» P* (l988) On tha cllnoatonetld matacarearia In a«ia of our adlbla flahaa end reiiarka on tha oatbolOQleal algnlfloanca* Ism* 1* JiHifi* lSi« 12 « 189»198.
naadar* T#ft«3« (1971) Hlatocbawloal obaarvatlona on oarbohydrataat l lplda md anzynaa In dlQcnaan oaraaltaa and hoot tlaauaa of Blthywla < {mtfgffi" ffi ,B* Eirttt^^Omyt 63 t 12S.136.
n999t G« (1932) 4n Invaatlgatlon Into tha ooeurraneot atructio-a and Hf9 hlatory of tha tratiatoda paratltaa of four apaclaa of jjt«ftaM I U Iftfflffgt^^e ( » ' « l l . ) J > JLISISlIt (ffull.)t
jl (WuTuTandX* ft won a l i i (CTnea) and Hvdrob fi la (wull.) and ^» ftwonalle (Llnea) end Hvdrob fi ^wyth) In ClaMor fln md f anwouth* Proc. 7ool» oc>
Lond> 1-32» f?oe, 3«H. & Dallayt D*C» (l966) Oatarwlnatlon of glyoOQan with
tha anthrono raagant* ftnal. 91ochailt» IS l 245»25')»
Sharmat P»N« (l97fi) Hlatoehaiileal atudlaa on tha dlatrlbutlon of alkallna phoaphataaat aeld phoaphataaa 5»nuclaotldaaa tanii ATPaav In varloua raproduetlva tlaauta of oaftaln dlqanatlc traoiatodaa* X« Pafaoitank* 49 t 223-23%
Slddloulf 3« 4 i^lj-anif t** * (l98rj) Studlaa on tha l ip id frecticma of tha Matacarearia of Cliwoatoauii ffoiiiolBnatuw (Tra««tod»). Sisa* I I I ItJii' S3Qst* *'a?ffii^ot« Hifffay* £52 « «5-6fi.
Singh, A*N« ft Vlmenl, C«K« (l978) Blood 9io*llt« Of CoUee faaelwtua (Parelforiila i Anabantldac) infmBtmd by tha natacarearlaa of CUnoatoivti olaeldluw (Tra«atoda t Clln09to«atldaa)» Ind* ^* Paraal pJ^* 2 I 131-132.
9«>lth, 3«n» ft Brooka, T*3* (l969) Hold frectlona In adult ^phlatoaogffl ffifinfoni,. Pffrgfltole^Y* IE < ?''3-298.
; ^ v .
«j»yth, 3.O. {i<?76) Introduction t o liniwel P#r«»ltoloQy, ?nci ffdltlon* London i Hnatiwv »««} ftouc|Hton«
5»yth, a.O. A Cl»gqt 3.A. (1959) r g g - t h s l l forwptlon In tr»iiifitodf>fi •nd eett9d»«« £jBa» ^ALl l i iS l* 1 t ?a6«3?3«
Srlv8«tava« « , i Cuptst S.P. (1975) «>Ho8phBtp8« uctlwlty In
35-47• Ifooarorehla hypaglobaogi (Trniiitods)* ?ool . jf^j^. ^tng.
«5ur, B,K., P a s s , O.U. & King, E,3« ( l962) «t*rch-g»l p l»ctro-o h o r w l * of o r o t t a t l c »cld ohoaohetss** £IX2&» Jit» iilill* exoc>*PiHgtp» 2* ^t-''3«
Thoi»B», 9»r. & C e l l l e e h i o t V. (I9§7a) f»»tffboll»» of *C 9luco»f» by the M(>t«»e«resirt«« of CHnoifto»u» conialBniiituii (Tr»«»toe««), ! • Peraaitol» §2. ' 9B1-984,
TbCHRsSt R»r. t C v l l i e e h i o , V. (ll!67b) Cftrbon dioxide f ixat ion by the metecorcsriM of ClinostoBuni coKoIanBtuai (Trewstode).
laaa* SiasJua* E&MISI* 21 t sh-iai. von Brand, T. (1973) Biochwalttry of PArf!«lt»ft>7nd ed i t ion ,
<^e«de«lo Pr»«o* Now York and London*
Ue«cnber9-Lund, C* (1934> Contributions to t h t devoloomitnt of the Trwiatodo Dlgono«« Pnrt ! ! • Tho bl^tsi^y of fr#»h unter corcsr lao In Oanlsh froah ystar* JJ,* pmmHwm Vldanak* ^alak« Skr* 1 J ?23.
imitnore , D#H» ft Coldberg, r» (1972) Trout lntf>otln0l fak«Une ohoaph»t«oaa« 11 • Tha e f f e c t of tetaner store uoon enxywptlc a c t i v i t y J^ v i t r o ma l a HkiSL* 2» LMS,* 2SSk» 1, ? » 59-68,
tfrlght, v«' * (1977) f^tudlea on larvtitl treiietodea fro« Worth MRI F-C* Part I« nhaervistlona sn the r e d i e e , e»rei*rla«« t^d cy«t of raacloltf heagtlec^ *nn« Trep, fieri, " a f f lto^« 2 1 ' 47-56.
Yuaufl, A«N*K* A S ldd lq l , A«H» ( l976) COMHSfiratlve aturilea on th# H o l d emiooalt lcn of 8C«e d lgenet lc trenatodea* | n t . ^. Parowittel* JR ! 5-8•
Zbl lner , »(• it Kiraoh, K« ( l96? ) Uber dlw atinntltetivfr neatlmmiino von Lloolden (nikroMthode) ml t te la der v l e l e n npturiichen Lljjolde (a l i en b«ksnnten Pleaaiallolden) gamelnammffn ^ulohoohoaphovenlllln-Reaktlon* j ; * SiSJt* £S&.* !l8Jl« 135 t 545-561.
T.i>i» CD
BlooHcMlcel ooiKDOtitlon of thff nvtecvronrie of CllnostowuBi cowplanBtuw
:A
Conttitu«nt« n
% U»t ut t Dry ut
frvo niueos* % 5 1 « * % W 5 2 . 14i%1?J5
Polytacchar (Clyeogvn)
Protoln
Lipids
f»NA
DMA
Idoa 4
4
4
6
6
4. 39*0.in
8, 7q*a.t33
8. 64^0.151
0. a8*a.ii?
0,W5i%^S5
18. 3-^1.509
38. Mji%S87
35. 9S^1.631
0«335i%009
3.145j;1. ?«7
:i
Table (2)
SiM« metebolle •nd-»produet» of th» m»teetref«rie of CLliwe«toiiw oewoXartfttuw
netebol i tvt 8xor«t»i! }mol/Q u«t Mt/h
X • !?C
Lactic acid
Pyruvic acid
4
4
1.47S*fl.36l
%?519i0,a33
i 4 / . « ( • ;
,^jr .Z..82.2_ • \
' » ' • /
:^>i
TebXt (3)
rff»et of vf^rleuft chMilefll* on acid and elkeltr.p pHo»phpte«M of th» •»t«c»rc«»rl«# of Clinottowuw co«plan8ttifit
t lnhlbition/iitifliul(»tion
CHenioele
Hon 9
Sodium arsenate
Sodlun f luor ide
F^sgneeiuR chloride
Cobalt chloride
Concentration
«»
n 01
10 15 01
OS 01
10 05 01
Acid phof^hataee
100.00
•
-
85,?9 47.05 3?.35
9?. 00 65.33 44.00
45.33 20.00 09.33
44.70 28.16 11.62
^li( aline phospH«>t|iee
100.00
-100.00 - 86.31 . 68.4?
-100.00 - 70.17 « 47.36
<• 31.57 -*• 10.57 • 04.38
• 57.74 • 35.02 4 16.05
r lq* 1« P»rcant glycogen r}«pl«tlon by the m®tactrcpri«» "^^ Clinogtoai^w ^oiwolnnafeuM in v r ry lng p e r i o d s of tn v i t r o incubat ion*
3.i
toor
s o o
O o u
• 1 WITHOUT •LUGOSC
CZJ WITH •LUCOtt
20
ID TIME (hours)
Fiq» ?• Rat» of »ndog»nou8 glycOQcn depletion In the metpcorcrrla of Clinoatowuw cowplg^Rtuw during j,n vitro Incubation. C^#—*^ j In ppeeence of nlucoee, (-,..„.) in pbeence of Qlueone*
9 12 TIME (hours)
r ig* 3 ** n.luco©<? uptnko b / t he RetBC«rc,^°"iB of Qllnoetowuw QcmnXmRUiai^ in t h e o r t s ence of n w'! giu cost*
B. niuc<5«« Itukftoe in t h e absence of f^lucoee*
PH
Fiq, 5. fniymp ctlwltir® in rpl-^klon to t i m e
'. -^cic^ aho<?«nH-t'-8«' ft rjM 5,
' . rlkrllnp nhnenhpt^?sp ' t T M 11,
r l c j . 5 . rmyme . 'ctfvifelp? in r f l ^ t i o n t o t imp .
^» ^'cld oho»oh''tr-sp ft nH 5 ,
1 '
3 0 50 7 0 TIME (min.)
r i a » 6 . f"nijrymp s 'c t lv l t lp* ' tn r e l p t l n n to tftnnpT^tarp*
' • Pclf* n h o n o h n t f S e ^t '•M c;
\
10 30 50 70
TEMPERATURE (°C )
rla« 8* '"nzvmp ^ctlyitlps In rpl^tl'^n to 8ubstrpt«» cPncpntrrtinn.
'» rcld nh09oHnt«»p ''t nM S,
45
o o
> 6
O ex < ^ UJ £
5:
2 8
z.o
0.8
n
r—•"——
A
1
•/
1 1 1
— - ' " • ' • ' — • —
1
10 15 20
5 10 15 2 0 25
HOMOGENATE CONCENTRATION ( % )
Fin . B* rnzvmp ^ c t l v l t i r s In r e l p t l " " t o 9ubstrpt«» e e n c p n t r p t l n n .
' . pcir< rthoi»o»^«tMSP r<t r»H S,
>-
>
O <
UJ
>-
Z UJ
O o o
o. cr
Z I
a.
o E
8
£. 4 6 8 10 12
SUBSTRATE CONCE NTRATION (m M )
