Group Title: Eimeria tenella: comparative pathology and lesions of experimental infections in bacteria-free specific pathogen-free and conventional chickens /
Title: Eimeria tenella: comparative pathology and lesions of experimental infections in bacteria-free specific pathogen-free and conventional chickens
Full Citation
Permanent Link:
 Material Information
Title: Eimeria tenella: comparative pathology and lesions of experimental infections in bacteria-free specific pathogen-free and conventional chickens
Physical Description: 145 leaves : ill. ; 28 cm.
Language: English
Creator: Radhakrishnan, Chittur Venkitasubhan, 1937-
Publication Date: 1971
Copyright Date: 1971
Subject: Poultry -- Diseases -- Research   ( lcsh )
Animal Science thesis Ph. D
Dissertations, Academic -- Animal Science -- UF
Genre: bibliography   ( marcgt )
non-fiction   ( marcgt )
Thesis: Thesis (Ph. D.)--University of Florida, 1971.
Bibliography: Includes bibliographical references (leaves 124-143).
Additional Physical Form: Also available on World Wide Web
General Note: Typescript.
General Note: Vita.
Statement of Responsibility: by Chittur Venkitasubhan Radhakrishnan.
 Record Information
Bibliographic ID: UF00097686
Volume ID: VID00001
Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
Resource Identifier: alephbibnum - 000407591
oclc - 37704197
notis - ACF3896


This item has the following downloads:

PDF ( 5 MBs ) ( PDF )

Full Text

IITI;Eih: TI 'I,L'.: L"iiV PAJil.TI',E FATHO!'..'G"Y AUID
LL: i': li'S OF E-'FtI.R IlEii'TAL. iJREC'i'TI IS l
C.ITE,'; I A--T.F:Ul., SP '.?l 'ILU F.',TH' Elil-
REE AI1D '; tOViEWrTIJ, iAL ',f I CI'.I!( 1 I


C'1i m. 'i t:cs "'. ;- ACt'ubi;in IT I.hAR.' i ?lii-h]_

A Diss. t' tL i'-,i Ptr.S-: n.-I .'l t h' e Glji..,l.-,te C ourj i.:.l of
T:he U',v- 1 L .' :. I : Fl: 'L. In Fai t ijal iPu f illcriint of
ihe i LI .I t- 'i : O: '-.' of Do, r ;-,: Phiil ':zop'i

i I i.*,' '~4 riT4 'I

r,' I p :.'.':.: ml.i il "1 : -i ." .1,1 .'111"i t L211i ,i -'l

' Ij,:, i .l'.- _. L t ,:' i '," I _' ii-r C i i La .:-': r'.

A I: ;: L.EDGi [C!ITS

'Phic tholr e::prTe-::: hi; ?ppre.i-i-tion ind deep Zc-nle

of grat-iPtd tro Di Richr rd E. Byad :,, Sr., chr-iirian or

his : .i ,up-rv izcry Co mitttee, Cor l: I pful ii ,g!: .tio'l, c'oujis.el,

ad3i enco uriI '3 :.C t tnrlrci hiioi t t i i 7 lii. a 3ri d ring preprta-

tion :iof thic di'. rrt.atio H.:e i oc. irid:-bted to Drz.

I-. T. L .i:, E. i. Hoffiri' n G3. C. Sruart, Jr., S. G. lam,

anid Hr. P. E. Loc'vin:, r:'iber'- of i1n SuoerviLory ot Committee,

for advice. e ard -Uid ce. I.eci :i tn: n' -: for help i-i r,-in,

Ja:.'. io tI: i;r:. W,'h PFre;t.s e i .1 a i r-iloi .-adut1- e .t1

rir. David J. 'Jei r. [I'. Louis I!. Crtle -ain [Ir. Jame: E.

Dic!'ini-s on, Jr., proi- idel teclL iie.ic l help for iliicn the

aut. ior its iindebred .to tr. m. 'r3at:--'ful th'ankc 2 ,c to i. H 2:

.Jean E- irr 'rfy r fp:-cial t-c.uiic:l I l.p. T'''ioo fa' ulty and

'iP-adli.uate stiurint: in thl- D:p--.trc: nt of' V'.t ri3tar" 'S ien',ce

ha-e b.--:n hcipfui i ,. ,' 3 :.77.

T"hi .autth .r t;. i .;i :- o a .* . tu i- -:t:':o sd-o.:. ti :

finaricial i nppFort 'of 1I D':. part 'i it :1 V'aetitriy- Sc icienc

in p'ro.idii:"' - -,-- 'l !lp i; suip-

por't d thir throuhoi: ti: :tud, I'l, r :r .--. 1 : c7rrie

ouLt : a '1 prr of ,- q*ajh ;- p o... ci ..-e C '.-10 O ),

ent titled "Biolog cIal:. [ietnid or i '.,ntroi for I 'a li

Pirasi i i:es of Livertoc:l:. He i:- greatly indebted to his

'.;ier, Jaya, for her help, encouragement, and understanding

throughout the entire cour.'. of thick work. Thlanl: :Also

ar extended to [lrs. Jo.vce' Jarvis for typing tie manuscript.

L I I'S L D,1 1 I'S ......... T........ . ................ '

Li i '; .,F TA E. : . . . . . . . . . . . . . . . . .'

LI 'T OF F I-jRiLES ....................................

. 'i i . . . . . . . . . . . . . . . . . . . . .

Ii fl 'D ..i;r . . . . . . . .. . . . . . . . . . . 1

LITE1L T1.1T E iiCE I fI ..... ............................

Ei- i ri rr -, : L i -.':l- : i l,.'orp o .....

Sh:: : :f E. r -i Ini ction .......... .

P I .:' ..:.f '.., 1 C 1:. l 1 .... ..... .. ... 2 1

_crf ,,.lo -ri H, t .. ........... ... .. -
i G.etr-Fre C, ic: ....... ......... ...... 1

fP Licir; .T' IAt' .- inril F recite: and
F. .: i i i .- i . i :f : : t -t . . .. .. . .. . .. 'M

i'R[l'L Ii .CROF- I L F L.,F ." F C'HIC L.'. .......... ...... 0

I TLERI,'L3 ;' r ;P'iri'.,D ........................... 44
Sl .. '" ..! .. . 44
i. ,,] t, on ,..l nt 'jl o'.. C_,,.r .4....... 4

II. r i ion of i P L l: L-- ....... ........ 'i

III. F ,ri.Jo ct.' .r, : t j ..] 1 'iick '-t n; ...... .

IH. 3.uri' .r ..-- 11-.. ....... ... 47

V. F h l ':, .;l . : .... .... ... 4

V I. : .l- i ................ 4]

A t. h e,:7 r i -. i j ] ,- f -
rth e c" u,-,', in ,. *-*:n t :l .:. ,i : li'.. i' '1 . . 4-

B. D.-i.i.- in-t l fi n of nic.riobial floor 'of
thi. c ucr-,i Un -Ic i-f'ree SPF, and
co I'.'.: it I cl. cll C :e,'-. ... ....... .. 51

VII. P-.c cd'. Ccll 'olume ............ ........... 51

VIII. Total Sc-rum Protein ...................... 51

IX Sr ru.i lec tr ph i ................... 53

E :posur;- to Ba' terma and Funl;]i ....... ... 54

RES ULTS ......................... .. ............. 5

D,.cve oop -.nt or 1licrobi. l Flora in the Cecu'n of
Co v:,', int io' -,i l' 1 ,icL:!- Inoculat-d i ili C. t rc.- 11 a
and Uil inroculatc.dl Con Itrol. ................... .... 55

BA.t.-ri I Isolates in 3-'ic l:-C-1-I SPF -,nd Con-
Sc 1 Clii lc.i In'cilatcd t.i th E. t.7n r.i
cntio ,l l Ci hii et :i C c.c lt'n o d ith t... : .

I C'J U.IoIl.t. . . . ................ ... 6. .

DI CU PFL:: DI : .................. ............ ..... ... . .
S'.JR:u Ft Y '.,D CPCLU' IO3 ]S ............................ ,9

A PFL i: DIX A ........................... ..............

APP2i DIX' B ................................... 12'j

DI LIC 'G:. P IHY .. .... . . . ........... i..... i

EIC'. \PHI.;.A L Si'K TCH . . . . . . . . . . . . . . ... L:


'Ta.ule 'sPce

I Fr ,in r;, I ,l':-it ion r.:din iU.e7 d ifc.r the
Be:c:'v r' and Id:-n ific-.tion of ab'ctcris
n-id Fi.mi._i frm C:-c ofr SPF and Co:n-
*.;-int ii n Chicli:iel.n ..................... 52

II Cecal Flora of Y':c'.nr ?c'.et ional
C hi.':,:.:l E:.:too.-, t.a : i -er, il ten lla ..... .I.r

III Pr-c .orl'i nant eacte 1' 1 "lndj Fua.ral 3rp..cier-.
fr r Cnhic Ie .3 F .n c h i'inF Lc c-.c!l ,s f' Cc'C l
Ccccil :.i 2 icr1 t'r:'ri i'oiexpc.sdc C n-
vciz. ic n l 1 Chicl :ens ...................... 4

IV Fat~hlC, o,i,' DI.1- to Eiii.r'i tm-nell, in
act :lr Ia-, FtIirl- :iLd l' i.- pl i,.u oni -
Li\', Or.c'ni-,ii p -Fr.: -c Chic'.1r ns i ai.'-ed in
Plastic Film Ic1-.l tor: .................. .0

V F?.t:i ilo- .' Due toi E-, r i t':ill lia -:Ind a
in,-le Sp cC. I- -C f Finr i.i- 1n t-._acteri -
Fr'e Chiicl:.:is a3is.ed in Pl: I tiC Fili
Is c l t r. :, .................. .......... ... 5

'VI tf'; i'- loi ', Due t, Linr,-lr ta r. :lls ai' a
,Si le Species n'-f c teri : in Chi.celns
fi'-. P.i in Pla'tic Film Isclators ........ 0

VII Pa- thol.ilcr. Di.i .e- tO. ir.F- i- fill in
Chi,.:en[ R'i:e. in fla r -.c Film
Icc _l: r- P' -lyc': 3r-i ta 1nin t..-- u it
.c ri an 31 : r Fi. . . . . . . . . 4

VIII P-'?.h .lo '/, D t.:- 'o Ei,,r-ir i t.r, -!1 in
Ii :: or 'i-Fr' :- C ~c ......... 1

C'I.r lnt icoiil C ItC!. c... ... ................ ... 1'

occ, r trperc c. l C1 ] t 'i. .i 1 , . ri,.'r-', . 1

F r: :i.:i ':., rL*'.-. i :n:-11. C iici :' !. r. r,,' ;.ed
to EC r t 1] . . . . . . . . . ..... 110

Table rsa.{

XI llean PacLed C-11 V'ol-.e (PC'/) of
E.F ct ri- -, FLn 1-'i-, air-id PF'LO-Frce,
Spi-':: ifc PatLhogs-n-Fr-cc (SPPF arnd
Ccjnve; icon l Cf ii:kllcrn E::rofed to
i 1t f '.- ] ....................... 112

XII T'otnl Serum Frotcrins -in- Seruri Protrein
Fra~ctinc in Infct:.d and Hionitifected
Ci :,'crentionlal, Srec- if iO Path 1:W'-.n-Free
(CPF) and BE.ctr-iia-, Frunci- andi PPLO-
Free Chicl:en ....................... .. 11

*. 1 1 L

LI;T i:.*i, fli,].li

f .- -,7 . ...-- r. : .-I' .. I --
il "+ + + I. . l, . r. : .' . i -' . .

I, -,- ,, r ...1 .- ,, r , : ;i. .:. .

H+ T . . . . . . .. . . . . . . 1
l, I _- L", ;," : tl7 i r t r,,-'I ] .i- ,_,1, I

S. . . . . . .
..i... . .. -I .
in +++. ........ .. ............ ......... 1-
I- - I
-_..__-: ._. 1_

,__ ,- _____ _. -

.: -,,_ ,. , .' ,- ". .. ',.'
.. - - ,

I '
I j I ~ I- L' -, : 1 r, 1

+ ]'--'L. ... ..... .... .... ... ...... .. 1

A atr act of Dizsert -ti on Pre-ent,'d to the Graduate Council
of the Universit' of Florida in Partial Fulfillment
1o the REquiremints for trt:- Degree of Doctor of Fhilosophy



Chiltur Venl'itasubhan Radhakrishnan

Dc-emober, 1'i71

C}-iirman: Richard E. Er:.ily, Sr.
MIajor Department: Animal Science

In order to determine the role, if any,, of the

irn.lijgenc-uc cecal microfl-ora of chickens in influencing the

development of and disease due to Eim-r-ia ten'-lla, the

patholo.-;y and leionsn fo.llacuin, exi':rimrntal inoculatirnn

uith a standard dose of E. tenella infective oc.c.c-ts i.

b.actoria-, fungi-, plut.lropneumolnia-li:e organiirss-free

(PPLO-free), specific .athogcn-free (SPF) and conventional

chickens were tudied.

The dominant nicroflcra of apparently. healthy con-

ventional chickens a ,nd changes in the indigenourZ flora

following infection uith E. tenell were studied in chickens

aged 1, 7, 1t, "1, 2., arnd da:.':, u.-inc standard micro-

biological procedures. :peri:,-entLl carried

out by oral inocult icrn ith 100,000 surface-ct cr1ilized _.

teneila oocvyt:t ?lonic or cou;.if.ncid 'itn .in-le cor multiple

ope.:-c of tbLctcr>-, -r'.'-r fun -i.

1il clinic.:il symptom:, micrtt.allity or gross lesions

n .:rer ,,:.i i~ i rictal of 2' b1r.~it ri fur..Li rriL FFLO-

i'''.e c. l 'C.L i: irinc.: il-.ted i: h :lrL-. ten-ril 1 l,:'I 1 i'I: i. I ihe.e

rn ost- ret fiardat Io.n :.f trhs d'v:. lopieritt of t, endoceqlI

st.ajz. of -. t.-i:' :ii: evi.?ert. Th.: r:. n c'.: of a pure

:tvr-i o E'-'I- P.'IL'-'r -. C, l '-t' r i, r.,-, r-f ri' .en ,

L..:l i'iIi -i .c011i, L-~ ,: i:t : 'illi? p. Or 1 tr [ rl'-,:l ci:: ll

:.1 or sinLr -l '- 7'CC C Oi f tih fru-. i C i' -i i l1

o:r i.:or i .p. resul:te-. in mild 'e'al c:icc diosic: 'llo.ini

iitc1l, :At ion fit E. tene]la. The typical 0 :31l

.is E.'yr:iiori.:s dev'.'lop.d in iic':>.T : .l:'..orinr 2 or nor-

z Fe'ie. 1 of imicrofior',v ., C. r-'rfr i .,i: arid S. i0;

. c.- i and S. Cfe,-ii ; E.,-ter i:.-le .p. C. r,1rfm ri'-ns,

E. colt ar-i 'w 'c Ol is. In CFF :hicl:-..-n typical cecal

co.c'idi,:.Eiz .-livElop .i foll,.ini :.:pe-? ,1r r- t.11 :i -f.c ion,

jith a mortalit: r ati of 3.O and 3 -.'n :m.oss lem C, ore

of ... In conventio:ar. l *Zhici':n., t m:.'rt:llity r,-ite w'--

' id :j th mejai ,: i ao.s lSiic, score? ,i.1. C. -

-'_i_'__ ,,-__ : ic.:1l .1 t- I m, orn'c fr aie, ntly front,!f ct,-:.- CSPF

c ic.i:.:c i tiisri fr'.r) noi ninll-'cct.ed c.fln,'-ntiontals chickr,-:. A

t- imulat ion of LL'u'.th of C. .-, fr'i r"-' and coliform;

oc .l'rr d lt'ith a ,'o L' i t r*. : ri'i in tah"- o'I routi of

L-,rt.. i l us sr. in SFF rnd .:*Wn,'Ert .n-il c:-h l::cin CIiOf F-

inrt, i. ,' ypi'?.l C:c:-il c.0c:,. 1 :,: .-.

Th-] I.2 iniiiat,: th'"it ,r-rt'Lirn ;p cies of thL

ilio ii .?i' icr :i ': "r-I :t',. ':t-. i. al to rT.ojdu

the t'"pic l c:a-' ]. c,:ci.c.i .Lo: :".tdro,,:, follo'iith, in ::- rio.

ol F. tr'-:n.:11i irif ctiv., ,cc., ::tz .


Li .- ria tf 'i-ll [2Cillot and Luc-t, 1891; Prc.tocon:

Eiracrilda-] is the mort ccn-ion and pathogenic of the 9

species of Eim-i in described from the chicken (Callus

dom-., tit). All the 9 rspc ies of Eimeri. occurring in

the chicken ar- initrc:.:llular parasites of thc epithelial

cells of their intcstinai tra.-ict producing the disease I:nowin

as ccccidiociZ.

Coccidio-,i. is a of great economic importance

to the poultry industry tlhrouahcout the world. In the Un'

State alone, a total lo:z tu the poultry industry of

.3,;j:',000 was -timaL .. during the period 1951 to 1960

by the United D-p.artment of A.riculture (1965). Of

this cum, :015,123,3000 niere attributed to mortality and

,8'.19,7 1,000 to rnorbidlity. In 1966, the c::pcr iiture for

coCcidiost0-tic- ICu,7s in the United States ais estimated to

Db tetuu',11n '.'~0,00), 00- 50,00 ',0 00, for broiler and laying

f'lo-k r.:p.i c lene t rhic'',ns. ,. LuteL cocc dio- i.: nith a high

rate of inortalit" is rc ul1-1:; a-s,:ciatc I ith infection

due to [. L.1 i- l and cin,: e t:'. 1-1.; i-.:-.- -.- .onfined to

the c.:ci. th- dizc'?:"Le is oft' n ref5. red to as- acute -ecal

o- :' i..ti . Yo :. ch ic .:, : 3-: ,',-r.': cf nig.: Ii'e highly

suaceptiblI to t. his di'ea-.e i.Lh 1 real susceptibilityy at

arbiLiL ':.'-.t I (,:'if 'i:' it'1 l'fi. ': I'5), 2 r v=r :l c, trer

.auiit1hjrs 5i s zuie- "t t 'hat V'or,.i c1 ic I:';i ,: ic or': su. -

c eptible than older :rne (Ty:-'er, 1929-; K.rrann nrid Prc'li,

1.'33i Ccomn.ermely, mar;, ctn er i: :ii-,ect that th, r.v- rse

is tr.i, (T z::er et -l., l ?- i;:,-i u;, 193'T; .Jcrnis, 193;2,, 1' ). It is impFortant to diiticicuisl between

cusCeptiobilit:' C.' hiC:kenl t:o linUcal dien:'-e anid 1u.-

c Eptibilit- toC *:, a idial iif'ccr ion a, .n:: fl;il*'d ti.:: oc.:,g;t

prc.du' ti :i Based on, c .:; t Frpi; duc: f ti,:n, clder' Li rdis are

n:.r:- usce p? ible to the p'traza.t thil L::: c.un*er o.r-ne

(R sc 1 .":'l. T!i 1 is dlii t:c the hi her rate :f ::; :,c t.i-

tion Cof ti. c ocy s : ; D':Z r -In andFI P rr, 1965; :*e, l'c 7r

Older bild? are -ilsc su-:iF. title t: lini'al i'irpecticn built

tli,. rate- of mcrtalit, is isuijlly louI diue to rin :pprarerit

;-uiioipr'd irimunirtv ftr pr r vic is c.r-t ..l *..,posures.

Leviine (,1r6 soe.losJc ted tiat tne acquired immtiuit' is often

no.t bzoliute,, bu..t :enerall'. oi', a c: ni r ition f' r elative

inimt.iit',. C'c-:-dav,-oJl r -u:ce -ol. .i.*ckens .'are le s

i..: .t i ble to t ir i : i i.'::t 'i2i r B in *2crr p irl..L to c[hi.c. ens

'-5 'I+dl:S .i' 3 age ( :.e, 19? 7t). Tlu.i thle severit t orf Ilhe

inri'e'ti :..d. dI:w c:.- in cv'i:::c:n i'.lcr f,'.Ild :cnliLionc

is r-:l. -1 t to ,'-e an~ j brL:i.l 'of tli ch11'?k n1c rr-evi:'iosI ex'-

rpo nrc .fi tl .--r'c-: of :.p -' i.'- t.:.. ti infective stagc

: r tlr- p r:.:ite. ,.t the pec nt tice, co.: :li: i i1:

',i, .1l l i .; t.i riO n: ine UiC.e .C rt .i t:o:c idi'e tl i.

dr, Ii. -',: pr'act i -' l _Fo.r *.;'-': ni.: rc-, ,: tlese iru-:-' ha'Ve

to be incorporit- d in the feed or dzinkin-c water of the

chicken from the da:' of hatchinif :nd continued throughout

the life of the bird. This continued use of diiruic h1as

resulted in intcrfei'ence 'iith immunity (Davies 3nd Krndall,

19 ; Reid, 1"160), side c.ffccts. su-h as reduced fertility

(Jolmer, 19m I nd development of drug-res 1 ctant strains

([!cLoin.iilin and Gardiner, 1961a, 196'.1b, 1 62; Pcllzrdy,

19ol 19c22, 1962b; Gardiner .and IlcLoughllin, 1963; Vegh,

1963; Jo'n-r, 1970; IicLoughlin, 1970i. moreoverr, drugs

preccntl.y available do not offer effective protection

a iinct all tha species of Ci-v-ri- pnrasitic in chickens

sni nmost of the current brond spectrum coccidiostats are

not suitable for prolorned pEriodr of use in chick.-'n

intended for ihuri;- con-.umption. In spite of tn,' hiTh

efficacy of modern coccid.io3st.tE, outbreakl? of the disease

may occur (Joyner, 19',0) due to highly levels of contami-

nation in the environment, reduced upt!.:e of the drug or

development of drug-resistance, and a high degree of suc-

ceptibility (Joyner, 190, 10 71.

lUmil-ri natural c-ndi tions, cec:i] coccidiosis occurs

thiou,hl in,."r ticii of 1 -irge ni bers a,' th.e ife: ive stige

of tLr-C:e .l called spo;ulatcl ooc '- r Ti. unnl:.pori'l tcd

oc.c'.-:ts uhi'.n re forJ. I ed in t': ,nitheli-.i1 .lls of the

ceca are c:!pelled with the feces. The: develop and sporu-

latc ni the ;-rou: ,l if con:liti.. ..' '.-f o:-:.';:n te.'t ion, noise ture,

terip-rc-ture, a.nd other environe-intal factors are suitable.

Th.; si.or uilite. c.7oc'',t ?oint jitu i~ :. zri ro:cir.- 'i tlhe

infr c' ti .ve : t -,- The fir.s ste. in t -he raLiiO. *-..:- i o

th,? di-7 ':rei' iF excy:t t tion of tih' i;i :.ted .oc t- Z..e

Ii'"'.r. f'otluid rapid e.:c r.tat icn of the naijoritl.' of tlic

C..:c: tz in hii hl,' ?u. z,'epribi caiii n: 4ii; 5, oi .- :

oid, whi le:- :u:cc -z ful p::,?, t t.ioni a .d lou o :-'St pro-

le0.: :u:c ptibie to ,ecal co.?i.dic i- Sh.: :.?ritc the

r?,: for vevy iou prc.ientae f-" ::,nstc:.t:lon to in-

m 1t.rit. cf the ho, t: ( i eas. action .c. tle _;zzar.1. *'all

,Iud cutb- :pt in l conil-intra ion cf tryptic j,.i:-;. T)l,,r.:

are -i 2conbination of r .-ctor-, tiec. ?ar:' for E :.:. tsat ioC

.:'ch .:-z bile and rpincreatic i[?? : (Lxvinre, 1 -'2'; Ik!eda,

195.;o Hibb-er't et f : ., 1nQ).

Our imr..:,ldger re A:.r irng the role of iite-tinal h 'l _.

in the initiation, E:veloprient, or ev.rit:.' of .:eCal ,oci-

iosi is limit.1. Ho.;rev.:r, it ha; been r'?.or t;d t:'y

Joiian.7.7o suaid 2.arirlc Z (t1':a 4 I t ha. c durirn E. tAu- i.: inrL'ec-

t ion the .roiuth of Cl niri. i iim r ',, .--< i. i c i ,-latc-

tunlle -'Otb f L ,act. ."i' lii :p. i crir .:p .--.I The.

,eca-l L.i t:_-ri l 'lora , ti .u.:cc '0 uf Lth- tol 'l i3t'o-

initestii .l filor'. 3n.9 is of' *r.- i .: :1 i. ':aI :. t o.

th-: h:altli of *ci:,; ie', fCor :..'nlh 5:V i .' -- L.-" i. *;i :.min.,

ICo.'t: : rt j l., !'4 ; Tinmm Ir .-')

IT e c "-r 'lc t ,cf Llori.lj) l.a-c : 1.1 l i l- r; i Cu bi olo '!D :i

ani irr.nriol.';,' of the ho- i : at :n: ip Lb-ticrn

this flora and certain diseases have bea-en inve'ft ig-.tgei by."

maLny uorkei'L. Pnillips Et il. ( 19.. rising e-rm-free

guinen pig pro'.ved that pr'esnce of bacteria al flora is

Ce.ent ial Pfr c-urv'ivzol of Erit- rlno' b' hi. tol'i: ico and pathio-

jecncciC oir aornmebias i. Bared on theze findings, Wlit -ier

?-nd Rozentbnum (1970) ctudicld the role of bt.ctcria in modi-

fyiin the virulence of E. histol'.,'tic. Bradley: anid Reid

(1l966) demonstrated a dual etic'lo;:l involvi' zi a proto-oan

(Hi-t..r'on , m.!e:.'ridi ) :rLinld a r-i1ngle .pic i -es of bacteria

(Es 'cir'h'ti- c -l i, C. nr r'rini- ns, or B-ci ll'ic ubtli s

for infectious c:nteroheop-it it in ti rk:c'.s. Several other

studiec hTi'ave been inade on tne role of bacterial flora af-

fctinL the course of infc-ction in infectious entero-

hep..itit i (Doll and Fr,, 1963; Franker and Doll, 19?-,,

Reid ,t! '_.. 1969; Springer et Al., 1970). Hejde et p1.

(19o9.1 studied the patihroenicity of E. Brunc-.tti in t4actcria-

free chic!:ens and zhoued that the para!ite can develop and

produce disease in bacteria-free Chic!-ens. io substantial

studies, hc'iever, '-i-e b-En nmaie as to the rposible role of

bacteriia or other microflora in relation to patholo;ty due

to ~. tainr llr usinti-; ..outobiotic i'bact~eria-, f~uii- ?rid

i.,lcurc'pr' curonia-li!c- or :-;ii'r!ns (PPLF )-fr- ) chickens. In

pre li:-. inar1y stu lie", Clic. ct *"I. ( "..) f id no .liffer-

Cnce in the course of E. I ,n.:l -, infection in b.acte.;ria-frce

and ccinvent iona-l chicl'. t''-r'c -. hone'v':p a dela of

1' to 15 hours in tl.:- -ippearic-eie of the 'aiJ g:fnerati ion

ri : P::o .t: ii t n ? fe,? of -1.:tobi:,ti' :a L?; V 1 fc.: :nd

PL- i.t i ( I ':. qi.. .:.1 .,' He,;.i: t l.., 1'... ) t e, rt. ci 1

rclation.hipn .et ': ce hlt- micTiil.P lor'a .acn -. 1. in

t ,e l:r-dtc,:l:ioian ,:cf rlHe :,.. 1. *zc::.? ,idica i .:i -n.'l,'i"r,'c. .Ac H -rie

Lt li. (l ',':) rI t ted t i i'ct, c 0f th Lb.act-i 1 1 f lor'-

?n t e:- pat: n n :,_ ten icit of t hi :p c., i- e.- .PC 1 r ,it.' I.Tn':. 1 'v .Ji."

It i. nZILOt leA I he.-tlier t ihe b-c terial fl-ra niri.l2I's C. r

o1i, '. t.incll-_I to initi:t':s and d*.velop tnhe 'i, -e c:ntity .

Tt,,- przei-c"nt -tu, dy uw-s w.uvile rta'e-n t c. find out t the inIflueInce,

if anii, ,f t r e riCt':'Lb::es normal r .' ::ent in the ca.: o:l

chici::. in thl *e lcmnt of cecal *cccidicsai, cy

I,:-terniniii.-l- th.r l li ty r.f E t r-, .- 11 t:, pC r du 'ic t I,?

specific and legion.; of Cccil CC:-idioSi in

:'1ii,-:n, is i bl..:,'t-i no :, ,lde,:-e, able rtii .:,.r' ism :i.nd by

cc.rp.rtinrig th, pa-thc.lo of : :xperim it a':tl infection it h E.

te- 11'.: i in Act t'ia-, fu_,,i-, and FPLO-C ft e- clhickens,

p:r ci i pii t:]se In- r : e ci -1 eins arnd c :! en lti.Y i O al 1 c i.:un:.

T'I:. .pr e i;:.:l'h po.-;late ith reg rui tc co: al Cc.?ci io,'l,;.

r ct _r.ial c.1:,1 't: r ':r c ui',venition,. l ai:.-iae-frr-. c jhie ens

--nd chick--ns :.. 7hinig typical l c ion:z c:f cecal coccidi,. is

i- ,r- a- cl:, c.p:i r,',,d. .ite 1 :i fu n i-i- :n.tL F LO-f roe a.- _id

SFF chicl:cni 'icic incci.ulat?., i:ith .ta dose:-c of iF.

to 1- -:l1 l :r.: l1-it-d o. K ,c /. l.,, 13 K r, I O na !u.r ll;:.'-i.W',C. ed

ca' e:, eith .r ailoie r ..:.I: .. .L th bacter i-l ':ci- s.

Ar.- ..'t" ai L c. i o'" .. i tc: i of -i-.: .ni. : c -:i bl of pi:-

dui'iL 'e.' il occidi ii: ii, pi' tidtoI.iot i clhi,?:r.i wI ere tSn ln

isolated :-,l tihe i:. 1 ltc.- used for, further inoculIt ion of

conv;,, SPF and t--cteria-frce click lcs. The l.nou-

leaugL' of the intci'relation-hip betieeen the normal microbial

florets or the co.-ca and E. tencll b ained by the present and.

subzeqiu-nt studi.-sc ii',' ult imLatly lead to better, mean:i of

control of cecal coccidio;is.

LItiCt-iA L[Z~E RE" 1EV!

E_, li- tr-rell-i [Railliet, and Li,?I. 1L'1 -. 'a

prc, tLc.-.n t,-l, ,fin t'c the F--,Tuily Eirm,-r iidae, *Clas

Sar : c a. 'i'l'i- tern .cc.L i. is" i. g nce r : Lil sc..i t to de -

c'ri b? :pec'ie beli gin; tr. the F ril- Eimrn-ri ida' i Bc e-.r,

1'.4 *). DLurin- the i..t 10 y Jar t tLidies on I;he fine

:trltr ure cr f c.,cri ..i and related crI'o. upi i._-.r rc ':*ale:d C T

cre'' t ntr'.atcr cl,' h-, -:imil'ar' tr',r tuieU c namel;, the :,,.llicile,

the rF. Lar riii;:- tie otp.:11iiiiCLij nicr;:.tut-.ilc tlhe

I h .':-.; ie? t'e i C ) 1 Iro 'in- the i er rc '.',:- and ti' cc': ,.

Tle.'l fine : ruc tural imiiari tie re ie : c id-e-i r:i d aI

indici aticr cf a ?cloe: Ir: 1 t, icnr-rii p ad Levine I 19',.) pcc-

pI.l d a s ig hlt i ood iiica3ti.,n o :.If t his cIlas C i cicaticn iand

Si: 'l:i -'c : a7d -l (c1'-70.1 h. '.- ,i d Zi Ci ..:1 tI) pIr-r...tbl-':ms

r'f t '7:n'.,in,' c'f Sp',,i'co --Ca. In th-e d c'm es I .? chlic'en t'3 11 Li "

drer...' ti,?i ), epr:-c les f ti-h.: cenu.I. Ei'--,l ? E. :c ,rv',j i -i,

E h,. i, t l- .. i-_- P. t---" ini i 1 ti- L'. r."j -J i E- .

..I-..-.-' ir-, E. -i ---c -nd E . an-ve ree7. n d ..' ?!-i e

ra p.e.- sites of r I. -oith lial c -li. f ti -, ri,-u i.--iCn.

c.[' the iriteztinml t ract ',,,iest-. :r.J I'.-.. I. E.

n *iv:ti is tle :il' cpe ihicilh ri;,' b: f.und in v.-eral

e i ,r.. c ?f t:, i t-. 1in 1 Tr 1;..'t ,j".ir :ibcld, 1' ', .'i

T'ie '3 rpi-eci.- cai r di fcer. ti- i t -. oirpholr c;ic'1

teI-i t Z ?C :;-or lati on tim: ci th. ,';cc';a d,;,v,'.rl..nent l

featur-es, localiz -ion ini tLEL hoct, inl deg.r ce of pathlo-

*,.pnicity. Of the mirphc.logical chr' ct. r-', I-c t "ucLtur

-f the occs-t i: uaLj-Llly' u:icd to tilntify thi pecir:: at

least ithin a ri v.c-n host Lc-'.'in 1961), but oocyst

c aractcrs alone 1have only 1 ii ted va].-Il in differe'' fntia Fin,

pcc ice of Eir-i:'ir. (Horto.n-Snith iand Long, 19(3).

AI.moncg the mnii,' means of biolo;-ical diffcrcriitiation

for Eineria s icciec, the location of ;nd'.coge,:nouiI stages in

th.e pacific region c.f the initcstinal tr.ct of th hIost

and species specific i niri,.it:,' atre of r'3j-'r' importance.

Each spcci&e shoiu a mar .ed regi'-onal specificit, (Tyclr ,

1929; Ty',er e t el. 19?.; Heirick, 193c6) for the develop-

ment of cindcgcnou- ct ages. A lsco inifect lin ,ult h a cpecio2

r.-cuilt" in immunity cigainct thait s'ci,-'.: butt rnot against

others even withinn th-e .:,sre host. Hence c'ros-i:. lmni ty

t;st? can b:- used to diff-rentite the various peciez of

coccidia (Tyser, 1-9'; Ty:zCer it rl., 1932.1 but tne

specificity of acquired si.-stance may not be rigid.

Rosc (190.' I) found: crors-i irilni ty b: tx'c," E. tn n ] a and

E. n c- tr i ':h-.n -, ncirg pcr'c. Ooi c -, of E. n' .- t i t tc

induce infectio in lie c.-m,. '.. reti ui'i Lr e a .'o-nbiration

of factors is aI l":.,l u :ed f sy- i : if-.nit i icat ion.

Ei ei t enel Life C i : i i.' .-li '-i: I--:

Acute cocal cocc' i, '. in '..3u.v'J ,'li,. .n is r'e gLu-

l rly" assc>Lcizated witi E_. ,- ,.'' _,d this is- tle

:no.-,t comnion -aind r.C : patl'c.enic cf lil found in

,icl-Ini: D vie:-.s t a ., l'.i) T : r (1- 2 ': blilhed

: .1. tai e ild d.c',:.r pt i:, of :.t[ihe r-,lr'r. ,lc.,-.r Z'.i life .of

t. ten 11a: '.,i.:h ha b,:.c n confr.irc'..l in all detail by c,.b-

.i:ql'nt in:.' ti; ti; .:. (E.iint, 1911.' Lii!:c other piti: es

Io EL ri ic.::i. l s' .- c l : 1 ge-lr at i n o?:C.ur i the-

2. -e host rol ,lo 1c An 7 ;,,ce-:tin of' v.'iab :- -:p :rllat :-. -c: :yst:

t r,:i. .i- t'cd ,.'or ;:, ter-. T'hruich a pr:oce:. of e:.:' at a-

t ic.n, sp r'.:oit esC: -. f. ir oi tie E .-rc.c" It: a.nd ooc/ t' s,

libut th the: f ct r.'i co rt tI'butI i n- to :::;- stati n hi n:ot bee

d firi itely' *:-sta.rl i : i.: Stl die- b:.' Lrvine .19' ) and ,

I e 1,l'.5.5.a 1&55f;-, 1 ': ., I'.:). re'cale,1 tn.t pan:r.- tic

, i .e, in part icilP.r l.r'p:i is onr- of the fa tor: r:-

.ponriLl for ,::c: iocl. G,:odrich '. 19' .' ob -e; v d .

e cc ape- po : i thi o i lir i; I i .'.ai lab-6 f act urc in

th:- c t u-li 5-10 m nut.lll i : a. l er tC. C:, 'Tr il has: hiC n

phi .d in a 5" t r./ So Iut ion,, mat iit i e:d t .70C

H:',.I r-o ni]- 'i n con:. eri'Lt' r ic', bi l ara d buff" .wer.: als,

url.,nd to be in l ort a t factor: in a::'.?,'st tion of va riou:]

:ppC ie cocci ia 1.3S tana. 1 3.."', : Lo.t-e i1 L.- -: I -9-0;

Deran and Par., 1:b2; ;i yb- r in. d [f"li.O d, Q l':,'l Hib.eL rt

- .1. i.1 t, :tul:,- i r th. 'eci. :. H, C.f r., bile

-nld bile acl. .,i t :.: ',1' : C -c" '- ites of V 1' ri

Li n'i :p'?e inclilin .. i 1' i L 1.ind n c :.'.: l'-.tion

lih,:i l .--:- Of tLb: bill: -aci,: oi : ,,.i iT,: ..:.r ,:ci. rl:an bi 1: uar

Li'1 il 'iie i itiiout ;,r' *i P. I 1 H *r! tte c- ,:' r d F-- 1- .

t.i n C" E. f -'.' anld i. "l- '. .' iC b-V. L i.e .il ,: : -

ta i _i n a h r,-- ." p.r 1' o i o' b I:. .'i. d I'ui:-i Iilien

trypsin alone vras ius- only ,. l.ov i jocysts excy:ted, the

other 9? secies of 'im "r i- iImnluding I 2. ten, 1.11 did not

excylt. The precise iay in which bile actc in e::cy;tat ion

of coc,'ct a i nolt. kno :-n t pr'c ent Doran and' Farr (19 62)

sug':c:ted that bile acidc m Ty laterr the pr.otlein or 1lio- n surface of the zLcid.-i body in zuch a way that it

is thiicn eadily nacLed upon by p.-;cri'atic ,ncyime. inl/'or

mat facilitate entrance of onlymes into the int at oo0yst-

throu rh th.'- alt e '-ed micriopyle. Lot e anld Lek: (1969)' found

that in aduli chlicl-ken.' ab.:ut 40-,O minutEs may, be required

for F. tnic.1 a oocy sts to be : iri 'ted from the jiouth to tlhe

large inte:.tinr-. A Cper-:i-mnt .Openinr or micropyle was not

otGs, vecd in thi oohyvt ,:all tihrourh :'hiclh poro.cite.a night

eccarme. Tli-rcfor' the release of larive rinumuer: of s.;orlo-

zoites into the dige3st- iv tr-ct of chi'.l;Ena requires th'f

rwall of the oocy:L to be tbr l:.n, Iea!lened or partiially di

solved. Lotzo iniid Leck (1 .69') observed the ,.all.-; of nany

aporulati;ed ocyst e:peild thrclti fece- to be structurally

changed. D.-evelopmint oCf iimmrruity e not hind-r e::cy tn-

ticon and ion :ill occur in innriune and non-

irmituie chi .':en' u.'n.dier *:.-tinr l cci:li, lon5s ort n-Snith -;t .i.,

Li .r. -. 'poro".-' tr- .c fu ifor',-, IC n :- 1.5 in

di-ricter, tra.:iisp-rant ,iL' 'ot il:c -ch 1 -oroe ite has a

uclo -i ;ro-. ail .t .".*-.; i I .. :-I L Ile *:it the uuti-ided e nd,

anti e".libi '.'ri-'u ty'.-.. of o O'.'cr:n They r:i'idly invade

ti ." l:trfc':e .-pit lii CAR:1 Of- the c :'c i ... th.t.n. pinE:-

r t I' :l ? L i' : t n nr-:-.:--n.- to :r t r e: pI iip ri

tir,:'.i'-i i, .n th'l', --ip hi-j payr ,'.:" fr '.- ,ri i i in tl'- ffiC.,cro-

pi -::, to C.lail reac ii .he :pit l isl c ll liniii' th.

fuid':-.C tone Liabork-ihu :nQdF -mmr- :':uil in rIPx oducti kc

l -,. 'r* i.rL. -,-; '," ,: t ', r i'. .ll iO]:;, ;l hi, i.'-t I: ; i t iml .tr

.\ithin :2i.: I l z. Thi ,ni c l.c i-.i thi invr. d call t --

- n rI r t-_ hid tiLi-, l-- 3) ..l thc r: t IO L-d

C l .- :.i o ut t] ip b'.', 1-, i E" '. .'-a icivt f' crl

t i.,.it 9' '0 lr- :: oi- : r" .-z:r, t1 t"' ) ."-' p i'i 1-:t i-t 1-a

in idi,'t i. -:, '. : t.: .=i ,- t.' r- : le- .. .: i om b ', .ire le 1

, i ..-io Fc ii i tt n t- r a o Iiin.. I 11 y i r-:. p-;r-i-[

t _at-l on ii: m- i fl jlr) -te n- t, i;:b'L-:.L iitli-.:1 1 ,1 l r: f,:[ t,,.

t'. 1'.- : 1,':p a- t -, it geli1.;n it F.i .:hii:ointil:z. T''i .ii l ultr : r..' I ,.ir-

:,f .- m r-.-.c ite : lri.- i the i i ict ii- :j ( c c i.n.'c have beei

describe!i Q: [CLaren i1.1 F- m A 1 i ,.:.. 1n IcL.arc l u09).

r )1 ii of t i1-. il i K-. l' 1, ..OCl h i it 1 1I I tI a'id -. t in

- C h iir r .I rc .!I *u. O Z -:, c :ii i i-: ii : :I' O i: E- ,

c: in o. con:, : %..7-. T'he r1. - ',. :t:.,-. .:ii .-,. .? ,:f I : .

.- ",-r i -a r,' fc'J in t ,-. it .-l_ *1 ,1 .L i t-'-.-:-fr I :t

iav'e r':,,-.l fro:m th- e .- r i t:,li- -1 lI ::.-; .n o tl:- -:ui p: i :itr i -l.r

lan,' 1', Sij WOZlC 3 .1 : >0 ut --i -all j4 l- 1 .- o- the

C .A. T_ i na .rT .ion An1 :i. -: .L V L )r c I ib : o' Ca l

clnerat ion ner.--oit ez causes extensive dcttruction of the

epithelial cells an! severe hcnor'rlige occ ure into te cocal

luw:n I C ollclocd by tissue necrosis .nd thickening of tne

.cccal iall by the 4'th :ad 6tn diay. T'in- 2td ,generation

,ro.jZoitezs are considerably larger thin the Ist, 'nvcr.iging

abcut 16 p) in length, 2 p in wi-1th and 2.00 to 350 in number,

many of ihilch cntcr new host cells ajnd begin the scx:uvl

pha.e of life cycle by developing into uit.lici ncrof_'-aeto-

cytes or mricrl',jaetccytes. ;icroga.nrito'ytes .r'e smaller

both in size and nu.trbcrs thn.-n mri.lcro .icLtayt':es rind the 2

ar: iin clo-e pr'c::imity.' within colruna r epithlielial

cellE of tie ceca, belo- thc: host cell nuc.-li. Fron each

inic'rog'rietocyte nicrogd ctcs develop. Each micir orncte

lhis 3 fl-Ta ellia and is notile.

Young rmncroganetocytes are large irriegularly shaped

crlls mea-uring appro::imat.rly 5.3 pm :: 7.5 ipm. Tho"- '-e

young macnroP;n-etocyte still retaiinis L heo zhapc of the ..r',-

Soltco, iL can cbe diti1;'inguislhe-l for.! schizont or micr.c.ameto-

cytes by the pr:sr.nce of "wall forming" or nieribran:-ou-

bodies I.Scioltys eck,, 1 i' ) un..1.e thic -electron Ficroc. cope.

Latcr l: bodiic-E" d,-velop chich are t o'.Oruiht to corresponki

to the "plastic L' 'nilc s" :1. u rr'ib: 1 fri -- li -t LiicrccopL.t

inve-til:ation: (Reich, 19'13; Dofleln a'-. i. RcJ- enrio, 1953;

Chei3sin, 158). After fer!:il1-:rion these bodies nizrate'

t,., the p.:riiphl,?r o'f :y''ott -. ii li ,it mcrLran,: of t

-,', ote th'en separate fro i.n the cell to bt-conm the oute'rmost

Mei br.t-.tln- -of tli c:.-..m yz trill. F':c ,iiyIdl: l .Ic t'r o cyzt

--ill 1 : -.eicop fI,:,ro:, t "d.ani bl ,il- e an.i th "ih Ill

for 'in- b'.liE ." iv. iZ'-s tC: ti'e x -r 1i':;- ,:r -of the c -. --. t Tihus, in E. t-n--~=1li te oc,:-,..:t .Il i i" tr il:m'ante.

'lh:C- th-,: oc.,:: st .-.all is .co rnr -, tet thc ,:,:cy/st. ic :: [t .e.i

f['r-n t' ho: t tizu46- -Ind is' p. :e to tic ::teri-, ii t

thi -. :-:.- Thi.- p.-rici f f vr mA tlIh t riie of infecti c O to tie

13;. pp. of oc:, : is usci.'li: 7 da.:. T'h. C:o-:,s

product- ion th l.i:e c.rir'-.",cc:,- -O tih-: 7 'thl i 'J Ioll ii, i'- Ce::p..,s.t:- ,

-re-..:.l z a -'r-:',: --.v tlie c th i i a a.i i. r:ipidcl,y d-e :r ?- es..

,:.:'- t-. orf nr'-l1 m c0.:-i-:ll, Ecl ai. ti-:.i pari'-:cnt, withl

Su1 11 I eI'i ,-Id dOWCubli Out lii e. Tile ,, G :. r -,1:-. r c-.f t I'e

o,:,: t '' ill w; ui.1 n L- cl'-u L L rot i anr.'d th i luier r

is lipit c-.?o-t firmi:.' .:: .?iat:1 uith prorein larn-lla

I-:-lfb? .ni d ii ., 1 ': 'i. Th ic ofI- t -i c:. c O zt s vi-n-

from 1i p to 1."I :p 9.5 to '.8 ., i, oift.h .2 ti-can cf

S.':, ( O. 1 .. 1 .1,-- i, I. c.' I :i :er: 105 .:. The

optnimunm temperi-tir--: foir spc.'ul;rhiL i of thric.ooiys- is

2.0: 7 1'-' C. I.Edlia', 19". ai.] :t tl: i re-pe lra, Lure -:pomr 1 .l 3-

tio.i ,ill bi- copl: tA l -I 1. hcur: tbitt N t r:.-o teripraCt .:

it ta':: out 'i3 tour:. The porul-.e. Dc :, 2t 'contai n-i

4 -p noc-.'-t : achli :ontai inin- 2 :.. .. it Liln- in othe-r

' ioi- f: c ,f Ci ric nl.- I t i 1 : I '.. ii A n. r ..: lc" i .

p. t-'l 11 ,-.i 1- ct uit i tA.:- in' ti: 0. -.; rop n chic.:k .mbryo

I L:,n-, 1" 1 3:-.. 1 't- 1 : '~i t1 .: cilt.'ur- .:. ll

(Fa.Lon,i 1-.5 P '',i!:, 19.'. I''. ; rout N- r. a O. l i te,

1''-.; I ;, tsuolck-i et ki. 1t ''0 ; Dorn.l, 1970), using rp.-.rc,:cit.-..

obt in l by' '.t v trr, e::..yct:t ion.

Pthc-cncsi of" te"'-11L Infcction

F.-ctors -ii acting p-thlropnicity oF t i-n:. in-

clule the nutmbr of cocycsi inreAtri the nur,ber" of host

cell: dc trcvyed per ingested occy,'sh, the dieg.r of rP-

infection and the ztate of imilmuit/ in the ho.t. Thec

severit.y of the disease decenjU uoon the interplay cf tihel.

I.nou.n an.1 other lnl-::no.nri f'ctorc a.: li I'anr.' I'rc'm a impcr-

ceotible reaction to i.:ath (Gardincr, 1955). Cecal cocci-

,di-;is unier field conditions occur. pri,Jcip-:lly in yotun

chi-.;:: nz b but sell. on in those l -s than 10 or- 11 'dlys old.

The range: o1' cage of' s'usc eptibility is froicT 2 twieehk to 1

.int .h ;Ml -I of the wer t out breal:.t occur at tl 'he a e or 6

to r.jceks (Biester and Schuarte, 1'65). Herrlck et _1.

(193r.; in -i t. ,y on experimental inf: tion I-oun tlihat the

heavi.:st mortal ity anm greatest d-l,:creacse in cry t}nocyte

o'rcurr'Ad in chicl:z 1 mDnth old: heavy mortality -lso occurred

in chli!:s a.-edl 2 .ec::S anr 2 monMtlz a-lhile in Alder birds

1.3, 4, 7, 10, arnd I 1 month o h: .,c i nr' .-r alit' ... lo, cr tlhouli drop in red cell courtt t nel d ir..-m -'9' to

,'.. ':. 1 955.) e- l ,oyil .-: : ~_. 50, 000,

100,000, an i 0,.000 .. p'ul a t d C....C,'. t iitfa t. yO un

c;',ick -ns i. r .:"i':e groups or 1 4-, I :. W, .-cks.

Those iQ- the 4 -rup r !].r ffccl:-' 2n.i t'ho"e

in tihe 2 reek rl'cup i'rc 1 :-:t affected Thoi'..: bit.s 0.hich

rc_ ':-ve from nif -_ct i,:' b:ic.:'.m': iiL:- ie to r.: inc :ecr in .ith,

E. t -11 Hoev.'e', thi' is .1.t ari ab c'l, te:- iinurIit, .

.oi.i r *:' n i,.:n; ,' r- s, t he- acquire r d irmniu.nfi I. 1 r ':ti.r-

oi rdz r,-y b'real-: d.i c an- i -. ;,'mptcn, of th' di af :- to

re ipp i-r (L.:- i.n.:, 1"*'.). L-vi .;- (l' 0. in a r-tuty ;' .?ub-

cl' inical C:C:" i.i'l ii'- i i, i ull,.- : at lea t rr: m ntthi

:.1 r : t:-.l th- ne'?.:- : E h 'i 11a in ': '. In F:.-:neral,

nic:,,ver, it can 'L.,:- .un i t click ri i.::. lI lated -.i fro

iriC'.:- t i:.n remain fully and u.i.' rr'. :il, u .c .pt ibl' t hro. Ti.r j-ut

tn-ir live and. that a1 r :' se hc ':' inrl'.e'cc- on ren ist-

._rn:_. 1n..ier fi-l.l ,:,.:.n itio -.r aln.:, t alill ,:l'cn! t n ,rei- early

::pc,'cu to' at least ligc t inrf:-:tin and c: nCa-nl: all

c ide:s re,: than .:n :- a" :.l. Have c'e l 'r-: :- f r is t-

Inhcriitd reI :itEr: in C*ic r tri: tL' ch.lC'ErrVn tI

inre,'t,:n .i.ith n te ,.--13 h 5 b1.:,:-i r pr.:,.r ':- b:- riosen .-ilr-',

I Il 1. n. Ra,:,.nb'r- ., 71; Gibb-,_i l .?' u it I I ,ac-i-nIr'aI

th erc i li jI :t c-:irLei I anI" ti Zii ic:a t va' riatl 'n in

.r_. '?, ptibilit 1 t:e., Ltir.'.Ti ,ii .ffc 'cfnt bre -'r sir ,in. 1' --

hic ': i : : I H.,r- c: Er. mit r .n.1 L,.|., 1:3 i J f[r: andf

'.':I it i I I..'' reIpIrted Ih cIIr -i ti ili and i cr-

t .ality of r naie c:hic:': -mbr y.' '' ':., 1 :1 f.r nt g :- t ic

zr].ii .-:C2 to 5. t:.- --1 i. inf'ec.?ti :. Li.-:*. :.j..i H,:r, ic: (1 ''-'1

pri-Xuci? cviei r ti.c .I:c.:l t.hi -t ri.- r irezclrc *:flr act in the

di'-e:ti'-re tr:i ct oE ci rds it .-: ti-.:- lI inl C'l.i rc:-.1i::-

t1 e --everit 'l the i eis':-. Hcl -: er al. 1.'-' :'7. .i .e: .-

that inc rc e,.1 r- it i r nL nray reu) lt in chiol;ern

hi -no:r m.' n iount of -y' tcr S',Iell in th.- ratic.n. IThc niir.ibc r

of oocyC t: rei .sillt ing f '.o:i n inft'-ction i.. not n ti ue

inli'-ltio.n of the de-gree of infe, tion Tyzzer (19220

postul1 -rd that, t heore tic:rilly, infection iiti a s iingl

oocy.;st or E. t 'en l l.-' crc.ul1 gi'.0 r :i:lc to .niproximn:t.clv

1 0C, ,,',00 ooc,:st7 in a period' of -' to 5 drays. During tlhe

co* rsr of an in 'i;,: t_.n, hc.i,.'.r, t t,:r,2 ..i .,v ra,1l factor:

, ohich r,-i." cause c re lu.: ticn in this pr'-Lentill inclui ng

loss of mcrozoitcE over cr '.. .ins, irl tiscie darii.e which

results -i a los- ofc suitable cells. -r:a..':ett i0d Blinnici'.

(1949, iim5.. o ri p-rted tiht for ea.cli ocvn t of E. ten,-ll

inoculated in light inf: tioi ni pprox:ci:.atr :; 10,000 o tcysts

are~ prlr'c.l ed-, arid th.'.rr ic no direct correlr tiion be.ltuewn

the ciz.- c.f infCe.tive'.' 'e io.-o the final di :'ree of i11

tion. Hoev-r, J-nnsiii (1927) repr,.'ted tha1t thl. sE.vcrity

of cecal coccidioi is .l-pe3il on the uni -ber of zror!ilated

ocO'v:ts tLiat thi? bird t ceive. Jc :i.i:iic rid Scofield

(1 934) repor'tAi th't a dc.-: .c c.f ui. to 15,0 ar[o.lated

oc:c;,'st: rodn! ce neith-r vri '-:'s nrior' rL:'rt'i l it 150 to

.500 c'.cys.Is pirob. el : ht I,-: or.i-'e Ln no: rnrtalit,;

1,0'0 to -,00, c,.:.:.' t. foi']. hc.' de ir of ie-.oirhcge

and r' nort lit ( -d :.'r .'. : .-. r.p' eiu he-d

.ae-vcere liherorrh-;-. -'. ii h ,: cr ality. 'Iih prepritent period

in E. t ln.- in.if',:ti.o: i ," '. ',... Lih p-,tent period

v:nie : .'i h in .li iduil ij[ recti...n F'P -h t3]7 31 i r-pcrLted

ti.::t :,ccLr. s ier:i not Fpr -zent in the .\ro. iqia z. of the in-

fe:tc:d L'Lrd a't-c 1,' day,-r -ith u.i Ty z' r t 1. .

re-,,x ,i *C :' t pa t-.-' *o r ? ,. l i_ 19 da.'" ri'it-

in .cct ion. Tnc ,:rc: st-*r i'i-ib rs f c .:,cc; 7t r di.cihar-g -1

in a y'" r, 't tiy 'r,,-: r A- '1.. 1 ") t, th: t-:: rc-

13 ,a i cni bt' n'- f t I rl rp.e.1 it l-:r in ti t isz;,Si. e 3 i. n th l-

c.-.: i .,rtint c it:; ,ti rre-,lari.. r.-1. :.c i. U i -i rntur- l

many,' r.a oo: r.: Itfor ln perai ti ,e. For .-:-iMple.

Le. -in-: i '-! :) .l-i :r .:d a:o.:' t f: !' t :-L 1 il : in the drop-

pir-s of out of :0 bird; : i cich did not i:i an:;, mnrpto.:r

of irfic t :iri .

-he dii -:.:-.-: -.'i'rpti-i: i.n C: C: : i i:,c'r i .- '-: c:l,?':-ly

r":1.lted to ti: o: ur.'-: of i'f cl, io,:n a;,.d in g-era'' l tl

d**r:- .'f p.:th:'r ni:ity i rel t.:d t-. t l-: d pta o r;,ich

th.. c.- l iall ., ariti' ed. B. te -11, p-'i.:. ra.

,die pl:, jL.i i "C.:r' d,.i :.ilZ t ti t Dur in I. the dl- el.eli :n:-i it io

t-ie ,ar':_,itr-. ther-e is a mni,*pratic -i f ,ara'iti- .id cell:

i t the .u'L: .ri it.,:-h l i al -ii'n h,.er,: theya i:icr -e:-C n:lr'i,..1iisl

in c \- :.uc t.i I ..-e 1: d- tr Li d and t l i i: oir f n C.c .

o:,1cr '. ,. -: ,h: i- r,. ,:.]f r. tnirt i.aati .-i' id .er-nie i c:,i ," ii ,:riint

3: ca' 1:' n. t 'i '.tt fuijr el i .f ti.- :n -n .1 of.I'--

enr,:,r i c :.: .r du :- t o : h.i .:- 1 .. 'e t. the bl:.:od

'c :1:. Thi: l.. :--d n.i- i I lt rio:t i. : rtL ': : t f.r -the

r-.-ra 1 r t i n r. i:,ort -Uil .- i : 1 l1:.!; t .. .. I L ho r o' n:.

-i-il conrt ,ii' oizi L e jrinr,- o:c r.i s f C rrn th : lth to 'th ,t '.'.-

p t -': p o- . : ch of th."- r, '". al ." i' : c-- rn

sc-.ondary rh c i ..-l 1-l i'nfcction of r1he ..ei' ii L which the

ori h.llim 1.: l completely dJeztro;,'ed lEt i i:J I, 1 ):'1 Hem-

orrhlige is a gre Sait s r'esa oan the infected chi ckl'n and

feeding andi movement riire at L rMiAininiar luring this period,

bLt con Lumi tion Co f r'ater is iiic i: sel 2 to 3 tim e-- that

see:n i., u ini'cted birds. In a typical zeverie infection,

bloldcl' droppings x;ill occur 96 nourn pcgt--..pcsirei and

pa'age. of large quaintitics c f lood in tihe droppings con

the 5th aind day post-exposure occI.;r. The di-:e- i

at its peal; on the ?th d .,' poF .t-e-:po:i're anl W '. of the

mortality occurs withinn d-ays follo.:iig initial 'c:':pocjutre

to ocy,,; ts. Chicl:rios rlurvll'ving 9 dayv following e::po'ure

ill IusZallj recover A chronic condition', l io'w:,' r', may

occi.1r ?as tih re" ci' retention of a. core of necrotic

tissue in the ccwa uith consequent ,'c-lA dy .runction. III

the flrci: as a Whole, th' diIeasie is rrarly alaci:ys of short

duration. It is often fouid that a i-cndit ioin cf dizeeie

arizes only rhen infetion, he i vy in ,elation to th. pre-

vi'OLl. e-xperienco-% of the birds, i-. acqui red during .a period

of 72 hours or l-,s. If infectinn is picked .Ip Ino'- jl y,

thc.n the bii'dz c OILcOr int l.?for: cliiLy:.' 1 fect:

.-ppc:'. 'J?n chic:lken n re r- i .rl n ep it ttr D' s i i most

parrt.: of the icrld, the ooc. .ts ar .t :. .-. ly dl -' roved

by th.- heat of ia -r ent-iat ion, LLit I i o:0 tih u1nfavor-..ble

enr iiro.iirlnt .ire pr slmo incir'- tly v nI'-p'rinli-L 5. Cia ~t;':.-. -n the

crnv rocr.ient, notaIly nn incre.- se 'i rn icstjture .id -'r te nperat ure,


fr .:, :' hi !h nd rate .:f :i ru.litic.i l:adin:_- t..: i

:liniC: l di.e1 in thit il C!:..

'.pp i ri: ':e f i e:h lc''1 irL th:- d' l:'t.:.,,i 'V and

s.dei. ni de, t. h i r1 of di" a;--n t: ." '.'31 ,-" i n ': -i: -Il ,,:,:..:> ,ii:,i: .

Clott i :i,- f lO d L pre'- nte i .i' r i hg t :h '.i : .S$!ez t-,'

zone n kniy.:-:,i. ri ,f .ct ..,P c I'. L v i t l l':., mci d i,: ic",,:i,:,

*:" '.'i ta i5 E ir :reL e-- patEh,:.t : iic:'it, t.: -.. I.-' i ] .jri ,1 E

r;:, ,t ri:. tLD:i'ies et ,'.3.1 El:.,:d dp:,. :- t, :-::.*-

pric:': flr:nm Ph- -nt. C:, .-T-end cir :.'i > g '.i :.1 up .itlint- --, t

nco,: rl-.. ,t n c:lcp-,: bl..:.,..-filleId (..: 3 ,and r,-.:-cric- ..:.i

de, l .:Pa-e: cf: E. terll c11 : .:cnfim- ti e .1'- ..,:'-

i.- CCCC ,L. c, -,, i.1,, . i n,--rep gfre e of .C,, .. r ,

nrt indii:C i of di-e.--e .Lin,:.- .n 2. t.-- i 11 ii-f ctl,

,:,-: y ." r.: r.-o .:t ,)rdin3rili,' zee-n in a:i infectiC:,.i 1uf i, i i. l,'

e:: t_- t-:, c: .. e disCieaZC .rnd de-th iDa',ie.: et -.1 1'c.

Fc:ir :xp.-.i'[met,;al infe:ti.on,; of c:..:cci'i.] i: I:nc-.oin

nru'te-rs c-, ..pc:,-'urlated .',:,cc,'y t are a in: ni-riist:le.i:d -,iall',.

H- .:'. '-::r, Da'; ies ani .-,Yine L'- I, -'o' l .1 ,,i ..ha t' atnil F,, id .

Ei rceeded in pr-i'.i.: i C& :al iniectlions b',' iPntr:inii tint U.

t'*:- l:'i- occyst: Sub.ut.aic:,.i : .', iintr'a c--: ;,-, iintr -

p.r -itc. -nc:,l_:: or inti a-iu :i1lar-i-y in ,ic' .'i 'inct i -. i l.-c

.:pr-..: :ite- u:r; intrc liiccd b,' thre .r-n-- r-..:-t3, i: t-:d in-

r':, ,i; '. ,:' rc .,'.-*- ,I d. TP : te :d ,: '.at ic:-'- a nd

t-ran rf-ir' t:, th- oile c.if Inf i cti n f.I c f: ci,: ,. .: i.-i

i n:cc l:t7 i. p or':r t-te.-all: i- n-t ', t ',, i-:. t .- H: c.rt n- -r'it

P n.d Lo:r- ,1',. ) ;cn ir' r:, th e.:e fi.rli,.-s p:rt i l 1 ic thatit

aithou.Th they could obtain o:,c',ysts in- the coca af tr

i;-tr'a'.'enouz and intram.nuscul rT adminiit ration of o.cyc,'lt ,

t.he:; fiil c.d to IccO.ver oc,-:. Lt fr'cnm ceca ;,'h.n :hiic.:.n3

r.erC inoculated intrtaFperit .oneall', or 'subiilc.,tneously. th:y

. sUr cd thatt c:.ocyat in:culated int'to thKe blood etlreafi tnii l.i

Lc r.enove: d fron tie circulati:,n by the liver alon'7 iith

other foreign bodies. Thcy rcpor.ted the pre'elinc of di;.-

inLCegrating oocyits in the li]ei'r of chicklens inoculated

intr a:-iu.-; ularly and Z;u-rez2tcd that the sp :r,:csts and

sr.-oriozoite mi,'ht reach the inte.tin-e and cca v'ia the bile

duct. ratn.,' : (19r. !) reported tihat Jilcn oor ,t s ist rl

placed in HillliFpore .chanbers grafted -l:ithin muscles, e::- tooe: place inside th' chi'nber iith tllh help of

nri:yncs produced by the infiltrat ini Itul:.c'rcyte .. He al.:o

fc.,i1 engullfmint :.f pot'r':c oites by r;,acrophares in ide' t'he

chamber and postulated that tlhe carry,' the cp:.i

to vr.ri.'io p:rts of the body. In all these: c.a.-e3 muci

lighter ihfectiio: occur-red after parent-ral inoculation

cori;ired.. ,ith th; infection.- occurring after o'r-.l inocuLil-

tion liorton--miith a-nd Long, 196I ,).

P:1"L,1ol,r '- C, C?.---l Co '- id r i.i ?

In'vol've, re-it of the C'--oa I :, .r th-v.- of ti'e aM:ll

intLc:tin,_ i on hiari.c.e ri.tuic oCf -. t- ali a in,'ction.

HOI,, '.'- 11, ;i the ceca -re ln'.r' rci-,ov-,-., or il 'v :ry

he-.,' inflection. the ter iu: l: portion of the la3i .. in-

t,:tine Oill be par.Mziti::ed. The lesions asaociatedi with

S t-. .ii iwnf-e:rioi in the c.:c h-:.: bc- n de.-cri o-i r'

': :-.i' '1'29), y_-_cr -i. i '. nul.d irr he, 1l' 1.

T1i, ,iia-tc.:i. ;: -i t of th.- .::.,cu Vn i prirar. ily InI.ol.;led ,.- -

i. : u rtial i da: :.a iz iue to th i e l r: 'c-er". of rela-

i'- iy 1.-r.: -. d -r :ner'at ri ': .LcTio;t pr-e -eiLt ir'z rhe

, :e.-rer ll'r.,i'a jl p i 01 t I -- niu i: ,.:-. Fr':'.ducti, :r ,'f

sr.-cific' hi; not yet tIe n d.o n'O trat-. al ti'u h'

pae'i'Leral di.nit at io0A of c::tr-icts of o'c -tz il lethal

to r. bt, r : .i. .t Io chi, I .:r-n i. r'r is, hl 9 The ? it ,.i-

I .-'1: o '-ct : t ire i 0t .- r' .':' il :ont aid 1'e I' S:

of 2nd h 2 .jcn 'nat i n i roe:r ::'i-.'r. e',,; anid loui hninr

of the c-ritir-oli l 1l1 iiAi of t,,: cCuiI, rnich:l- is: c time

-trippr-d .oiin to til- ba;: of the: siL.nuccs: arid i t1 replace-

r-ient by, 3 coi e CCO .ip c ofI nCI 1: i 1Otic t iSSu Li C':

C.lood, d e:., r.. I. J .: .-:- 7. lo: p :nt ] : t ,:..:: c-f the

pa i'a it.- chief ,l ooc-,;st T'i; c.': i; at first a h.i n 'nt

to t .:-: wal -but aiter pay go-t deh- '' lied :nd lie free

iit iiri th-e l rre:. An i fjII't'. r-1, bir.i ...-.v' pa7 this cor' or-

a' bl'o clot' in the .r-O ria:7. NI't- ce:u-, .' rec.:.erinr-

bi y -i Ni it F r': : 'p-' r'aCnc-;e t r:- iri sl i:htil

t:iic:-'r:- In 11 l-hter iif'' cti cO rs r-:c cv.'ery ia oft- e co-,i-

pl1-et: rnid r:pli, .iut in i -i h v*i.i' i ,ecti'n .eco-.' er' i1

:ic.i an.l ti :- rhi.. : o. 'i't,: 't l. i- i ." i n ,coarpic tc: r- ::ir.. r ti o_. .

Tl:i co' t ini, O 1.u1 S h ',-I,:'.. l' e t 0a G I '. I; h to t th d y p.o;. t. -

':p: uL' rE ilts i' proi'c l n.:a iii,: ii of'r :- 1 1-1h.

cauLJe of ,l>l- h. ThI-. po ...1 !-iii :n.. nl i : nei .bra'e.s

betccI: pallid. Erytihroc'yte count: and herani;.o.o it deicroce

t c. hou t. 5I0 ':. of tih n:orinal on the 5th and ith d- y after

inrcc:ti'-n al itii 50,'- 00 E. t nelr ]- co; a,'c t_:, .ind the V'.slue:

retIurn to noiri-il in Toboun '3. day: iiqtt and lH'iPricl:, 1955).

[!-tt (1195) observ'cd lymrphocytopenia and h cteroI:hilia on

the 5t li day and 3n r':ai.ncphla1 on thie 10th day rollouing

infection uiL h F.. ren-ll:1. There iere no significant

chanc'ge2 in the raonoc;,.te and basophil numiiberi' duri -lg the

source of infection. A iarkcl'd lCucocytosi- began on the

7th day post-infection an,] sistel throuLih the recovery

phase of the dl-seae. FPrat (1,0lO, 1941.) obLerved an

inlcrea5. in blood su ?.' dulin1 the acute rc.3aes of the

,di*-reasce .Cith a eci'e.rise. in th= rnui cl Clycoje '.'a:'-

1'741) also foun.ld a rise ii blood -.racl on the 5th day

post-infect ion~i ani a r'ic in blood cliloride" on thl oth

3nd ?thi dayc p:o t.-irifect ion wiith :aco;ipaii.,'flnj r action

in muccla. *hlo'id Houe e .e', P .er.,an i 1 '0) found no

il:pcrelycemia or chliian'g in hepatic glycogin, bu.t lie iiotjeJ

a. -cinific:unt; rc.iuction in the prla..cml lactate concentrra-

tioen on tlice t a;ni 2ni day: ,' after:. c:'poure :nd a i'ize in

r-.rdi.-e l:'ccocen on tihe 5'.ih r:'1 y p'T t:-irnf'ction. accordingjing

to D.-:L hert.y :':l i.' ric : 1 *" : ) during th -'cute sta:e2

of :h i infection, .:u..l: -nce rpro..,c'c .l1 in thr. cecu,.: r .ciuced

t'n-e abilityv of tir brain of tl.he chic!::-n .o utilize gluco e':-

but not Ih- :c-c di h'. rhI t:. T i cecl LO Cccid i o:

S c'.eCi'e ii C: -fere ice ';i th ro', .'Al .'-p.rp''crylC ti'-c I. Lo-

'dr..te utili It. io ': ay o.-cm'. C'-ni L.-y (1 "'I ) not ld an

incr,-as in adrenal a-cojbic cid lnd a r. I]a corti-

co. ,te rone coinc.--eitration: duirinic ti- _'cte honor rha. ic

ph?-e or the inf.: tion. Bertek .]) rI. ni clear-

ancO- or uric acIid in c*nick-ns inf ecte1d uth E. tEneill'

c:r:eatezt at 2 to 4 d.jys -after irfect: .on. Thii, tu

i. ecttcd th-t death i ri.t iue ro *?-c l I i ue .~- trl'. c-

tiro r .er se nor' io it enti'el diue to c'-ci bl.eling, but

to the f Iilur' to re.?over frorn an ifiitial choeI, resulting

fro:n thre d-evelorpm,-nt of large Inum ere of1 e ienoi'U7 U .e3.

It is also reported( bv ,.Johnson:'r and eid i'197C' that in

-imi' c-. e'. the r-o s le isl in the ce in li .e birds

ill be -orze severe than tno ?se in d'.?ad bird ii iaje

anid Sunva 1970) tudiei the ,ritro. ,:n n:ta tbol im of

chic!:en inri ecte;d witl E. t-en ll.? anri noted that the total

pnId protein nitrogen decr i-a:e withliin th:- ct1eFOriies -f

all a:- on the 'rd a ni eI pec ially or th- .th day and

thii coiri ide. nit h there pF i:i 1f dev.elop.ent of eni orj:-no u

stages the pa ra.Zit Th~: as3ui. th?, t tr, e:- di. or-de r.

3re --Ocilated Wi t h many f:i tor., Ef ch di.or lerl: f1

ferment_-.tiOc' arnt. sui:.t io,0 0 -n inrilu nt e of Pnc et a b i, J. ri'o-

duct of th.? par it Tn cncli' ,rl .h:t c.n the b''a r of

iroteiD meti :3t .olir: 1 i thi liver f e. ?r.. :-.r i.h.. l", icF 1 .l hlOnFe-

occr in ,ourc hick.' ti-r.n iiin ?l:cni' brt"...: .

Lar'ly u :ori.- Lbv Lev ine :n1 H.-rI'e 1' :: 0, 11 Ci

slio:'ed thai1: tIhe ol t t.' : I r i : tel bird 'c:

iunmarle to do r or'e t ai i of .e 1,or Jo n-.e I:.y iL;Iz'l. : oif

n inf ect ed. hirric hcrin t iriulati d '.via the in :' ... : lo-v r,

Free,-,-. ( 197 0 i f n d1I that j h-ie thI 2 mus lec .air dir ,:..:tl '

t imuilatc,.t they a-re acl, to do: uorl: -ind speclaitedr thit

an itpp-irrlent of nervcCui. cosidiiction at the lneurlo-mLuculliAr

.i LrCtion :cciirr ii' cc' i i.ol .

A\ fc,' cc il] larl r'ezpcnse rPirce eft il. 1 1-?.2)

-sho'ed tl- t dur'in-; tlhe primalryl, in! ec ci n vithi E. t.:ii .: .1

hcaterophi pol.' ar ,Ccella infi] trate inrt. the.

subm lc.C a it irr'e iri- n niijmb..:r especially ron tlihe- "th1 c,'

5th iaay: po's r-c:: pos;ur e chen t he 2'n6 gener .-:rat: c', -c Ii t C t

ire ,!cv:l.piran 'ii.l maturing. Leion -corihi,; has c:-.n

frt'qu:nutl],' uedr to compare quant it atLively,' thie rxterit of

,rroz- lesion.: aid pathaoloy'. Ihr ir:': ct I. (19' I 2 first

:1e c,:ribed a netl:d of .t c:ri r E t ri 1 :1 1:n :: on. usinr a

0 to 4+ !corin~i sy,' tiolr. T'his ccO'riv,.- r.:2ter. 1; i te- .n

ColloI, .el by ri.iny uoorker: (Ri pcorim ,lnd H-.- r .ici;:, 19I 5

Grd.iL-er and lP rr, 1954; CucI'ler, 193''; Lanko : 1 i ct :il.,

lo ',;; H r.-r-to'.-- nith et .2.1., 1961; L:.'n.h, 1961; Britron

*.t :- 1., 19.7; Turk ste? hen:, 1967; Dun::1,'y, I 1. A

I p'I: 3), P:iir and '.'i.:-ir ?'"5), L i'. -. l Ci 'P r 1, r' !'),

'Pletm:!y and Hi:.he:- 1.1'. ?-'I" ") H:,L,.'_.,-l.i~h.n eC 1.

,. 1i 1) used. a Zyst,' for r i. cr -,.cL c.npi' : r' cfr l .. .1_.s

-:nd corrl 1 -tel rhi c. mi. i o.. :,pi .' i ._: f p:-' it] .1,

.eprn.i' I ; up i n the pr'.:-:,n? e o01" ?n. i',-riLi s t -'c i ol.n.Ln

and Rcid lC,0 ..,. i ._ r' : .::,st;n for ;'Lros p .hlology

iLil el: .I.t''d .. .i ti for d..vel":I ii.nitJAl st:t ,:? in'

the : *. .:,:.1 t, t:r L... iO ." '* ri]" iz. tir,e *:Di-i rIi,' .

hoiul t -l. r ,ii-et-1 -IL'o i. i Ci' L. .h u 1 .F : r-i t ivC eint rli-

ti.- -ppe:. in t i. penuz rTL.:, ..L :t.-.- t ri r ,- ni.: r .: : pic ttu. ies

n'. b, ,TircJ tlP o decide .'i.thiCp leii;[1 2 iM. ar u: I t:y

,:,: i i .:a :. ;i.r'.::'.. rni. ''' i:O ( 0"17 ::: ine.l in-

r ti al v .1 J.' front ic *: s: A Aliniral.1ly dWInALywJ..

0 1, Suz p..:. -t '- "1. L : .0 *.:C i, .::.:.i- i ti ;zt l. ;-i.:-- l l'. a 0 In-

i !.Tri: ?:'" 1.'.-: 1 I z'.I. I "-:'[ tile ':~ ': i,) )ec i ric

.atfl lr:i. 1 man i fe t ti':L L C-o l-I i, H u, -I t h1: i it ii 1-

weeothu i: *1 ti. ctier e Cerat v 1, 1:1 int'itici-

-f tl1_ i i.i.. 'l:, '.c :i .1 nici *- .:4~ opi e *:r'ia-ii. latcri cf t.

i:te -t -in ?.l t iz-:iie f". t ': ,_n f irr, iiig p -,.th,, l,*:,;-:,' u'., t."', E .

t..:.rn- 1 1 i 'fere l icn.

S.-r flr n ni H. ':, n

in nui.t.:i a.rinim 'i iv'.' in iiti-te coat.-.t A"lth

rniu; t r'l C r riC!I: ite iji .:to.r i] n: tr: t ui: foiun.i

.i in t :hi irer' i -te i i r 'Ot.t:.t, ::n It'- [-I'I' Ial

t :' ; .i :T, o t L i h. I- .-.. '.":tr i '.e- i:- -l I. t .:i" i'. n)u .

Tii .. :1 : -.. :-L- 'iat ri hIi., it nF h t Ca:- :. -": bio-

Ic ic vi.: :i.:. ',ip.. bet: .:-,; t'.: ':.-. ".1 -ri thc .m micrO-

:,r. .T i:rj ,:. ii. Lt 'r,-: h, .]Li" 1 :t L'- ,..:l-;. rn, iL Ve

* riir.;1: 1 i c C i i'u eiirY L';.cl c l-:-l c.: .x-: c ni y

their in._1 ,i oli -i :. cro-Clor:. nl I t'e r-e anir.ials ar! ..;'.n .ac

"c-..''.:"l'i .n ." :nin-is. In -i o: trast a "r i'rn-fr-':-" :iimrlr

iz on.: f 'rom iic.h it i not pF ole to e cover an' vi-ihle

orP -iiism. [a- i;: lLc'ator'Tis r"pl tects:- to detl'c t: bac-

teria, f i.--i, hel.ij rith pirn ..ite- PPLO, ardi certu-lin viruseL-

to cletermnine the ri r n-free sLtte of thi? anirim-1 Il C ljtocn,

196.).. The t-:rn "rELotoblote" Re'-iers nr -L 1., 199 ) ic

al.'o u.T.:i I r. -ferfin,; t1. the germ-f r.e anir.i'1 and ilso

niil-ns ,*:"'-yi n.'r .p'?i s ,-of o'r.anizm a. A "specific

p.-tllo;-cn-fr'-:-" ,.PrF) .ra i on.r fr ee of Epeci.ifi d micr'o-

c' .ga.i:zms .nd [1.-,r...] te ::n;n to ,:-i-c .di .: l.Sabouri'y,

1'?5). The SF'F zminrail are fur.ctionally -nd struectiurall.y

ide-nt ical i.ith tih ir convention al ccuuntclrp'irts. hut the-ir

flcr..? .Crd '?.nu.,. ar:, to oone e::tent, cont'rolled. The

i'o'h i oif g- rn'-frnee or SPF a!niminal is a problem if the

part!.:, .' -.. r:al : have > rtai.n co: ,ernital infec-

ticins .5 -1 -.- '1i ri lln.'-i in hicil:l n-.nd T:'-.':ner ni.'

in dog: R l.:-c t -1., l"6 ; Grie .;-.c.: t l., 196-, .- are

t''o inlfjt- tioi rI-,- iE : 1i .le- co--g, nit lly Careful s.elec-

tio"' ,:ind. i;c:.l3ati O of brc.- Iin- tockl free of tlie.e infec-

t lo .i .- ;. -i i 11 t ic"i o.f y -'un- niilI s o1"r li'- I':Ii .:.i,' .:ci a eiut:l iO nfec ti O- -r c n.-r -.'*-. -. r t.o in.:ire

rb'~- r;-.- -f _.L: ,i L i 1 : L-rt Cn ti "' -.i. st; cc : i R e, :.c

e-t 1., "t. ). T1, .-e elamor- -Le d-.iUrecs e.-t hlis':

an ri .-.iai n-"r..c :. ri ip..lh-o -i-fre'e -nirkalE

fcr ro 1} .i C r?- ill] -A itr i n [.t.c lit ii-,?e sity aor

usin i: e. '; :I- :i. 'iirrn-l.- cf r.;no:-rn -ii:.-' ..e exp.p sure,

a,e tr',din11 anm r o t irport.nrit env i' -onil.-nt li b-,l:-.grou.) .nd_.

The po '-Sile i rinfluerice of trhe "nncro" .anl' "micro" en'v'irrn-

ment in *discr: e pr'oc.E --i O'n be FtI1di I<-l IutIEn .t ctoc.: c f

animals v derived froir. a br-.eed ing colon: .' i r' tised

iii--e r .' di f f r n t erLi.'i r-;iLri e- t:--c-onve t io liTli, 3FF aid


A inumb,_r of ror-:erp h'.'v attnted tc o rsi-e g-iem-

free animals itr'nc F-ateur' icpecul.a!'t iCn n 1n .-5 that the

hofr t-miicrcllor'r t elati n.shi p is lbl i, te But the in-

vert itat ions .)f' S,?notteliuz- ,1 .'') Cc.hendl; ( 101- ) ,

:Col'endy: and Uocilmnan (0! ) itCter (191 ), ClimEtedt

(1036., EB lzs n I,1T37a, l V3b) Reyr'n-i rs (l;iit, I.o'4 ,

1 6 )l, -GuS tafi con ( 1~ .3) nd Iliyakl:. (1 '' pro ,e

F-P teur ' --.ri 'inal az unri tion .. on 1:.L th. .;;erm-frE

anima.-l h- become a 'cry, useful tool for Atudy"in- true

hc'ieo:-LsaEi of the i v-atobiotic hol t, the. indi'.'ir al a:'t.

and interact t iins of iTiicroor arni smsin ? tnd tiie r. pon.r,-, oe the

ho It to theei c r -~:riani mri. Th'r. e li'ter.'izctions hi-ve not ,et

ocen il511 de'- ined althouc'h a fev- ,i c-ific'a,-t i _ic of llort-

hiost rc.l:'tion.-hip? lhiav jeen ecst':li ;h,-d.

The Gerr--Pr.. Chick.

Tine embryo of health.' tbird: : n 'it:iin-d in a

.,erm-free condition iiirhiir he h; 11 until Latchiing. Thi.:

has en,-,nbled germ-fre-: ic '., ti l nnd orl-r r I to

bt obtainel r'ith relative en .-:. 'er. -rre.r chicl:c-nz are

very popular as e.:perinental asrrirals nnr h-:'.' been

sEcce'ssE'ully used- to study such research pr!'oblem-. ac t he

origin of blood grr'ojp B agrlut iniri. (Sprin er t al.

1959), the gro.,th stimulation of diectary antibiotics

(Lev and Forbes, 1959), experiments on t'.umor'igencsis

(R.eyniers and Sacl'ste.der, 150'?! iev elopmcrt of parasitic

infections i.Brallcv t 1., 1967), and for a stud', of the

%tern-free state per .se comipasred with the conventional

3ninals ( eoyniers ct al., 19- 9, 19o0).

The gern-free chiclenn: o'e less clean than con-

ventional chici:s due to the high humidity in the unit

ann- also by the frequent occurrence of -ncl locall. a'3id

loo-s: nature of the fece.T of the crm-free :hilcl:kns. The

morpholo and function of thc giatrointetLi 3al tract

are altered in the absence of viable flora in the germ-

fr'e animals IReyniers, 1960). The intcstinal raucosa

hIil le-s lymphatic development anId ess coruiective t,

mas.- in germ-i-free chicken: and the general picture w:as

that there were e rore ablorptive elements but fewer an.i

l a well developed eletricits of defen3eG (Reynier et al.,

1060). Gordon (1960) fouln aboCr.t 3 tines greater nu',ib-rs

of reticulo-eniathelial cell: in tjue ni.coa r an1 the sub-

mucosa of the ileuirn of o.Jnr' conventio,-ni l chici;:ns

in ,-er -free chici:en-. Thi i -ff-en '.:- ua lo noted

for "sc',ollen" or lobulele l'l:.cyt-s fnunl i within the

~tpith.-lii" :ci;d in tn.? rn -r 0 of r. o ll ind l.,'npho-

cytes in the Subnucoja i.ndI l1-mini nropria of the li.-;er

il utim. HCc tl pitli .:l i l c.:1 1 :' jI tLri t '::i ;:-r-t

in r:--rri-fre c;'iic :. Th: 2ni':.i,Lt '.f I:.ina prpr i i n the

;:t..:t l L st ctiudi':d 1.5 .. 0.''. in '- fre:- c hi .

anl. '.. 1.' i (.:ti' l 'hi ':c;: rE, I : n .::nd

L'D .:.c- r ,19 7-) re.F-rt that the Lei rit cf the Fs- ll

iu'testii .. I''' i'.-: t r i i .:, :.i tici tfhan i :. im-

fIe= *h 'c Enila rce ,- t f' t c ::'.ir, 11 : :. LI:

Chlli -ge in se..'cra.l 'p.-c2ies *,r ',lri|-fr., -'.i lriirrl- i(nclIdiing

the r'at, ui .:". e, rabbit a:.. 'i.i ,:3 r'it 'J,.:. t ,: l j ln -

iBr u:ic.itte -iat.rdol :, l''.9,. Ho.'ev.*-', :ec:il ,distent ,ici .1s

ni't bse ':rv ii : r -T' ree .: li?:e'n[ n d t uri eyI Lui.r''e ,

1>"'- 3 T .-: .2c.:c? .:-f t n 4; .ra- (r.'e C: li lns I.: I f .u d to

fe ci -nil' ':itl -hi.:rt.:-r in len-th tha, t ,:.-ce ch ci :.-ns

i t r r.'r.i:Al I -t r'ial li:ra I .: e 1 '-. ) Fr:.,

,r.:.: :b._r;, tion, th-: it r iint.stin, .ri -:ic-'ea f co n-

venlti a.l1 aid ,-'r r.lI e i nic :: i -e iril :r. H,:.- er ,

tb i ~r.l'ti -t lr it of tlai --e i 'L- e ti i i r, 100 ,i c e ri,:

tb:.: i.? ri .n: c', re te .r in cr ;.-. rf1 :-.:. L-:hc .'r'n cliicI:eIns.

Ti,: r ':c.s -,of rn-: 'rr'- and c.oa-vet i., l ?]hi-. : l- I I .i' ',:- 2l1

Si ilr.- ut t ,I C- r'-i c .? e e .: I i -ce t-- 1" 1 i r.l- t.

dtL cch The 1iyrrir -etic ::-:tc 1: .i : ri .L-: lor.. i-

ier i- t : a- nimi ? l .. ic. r- L :. -1 :.- --,1 -, sil of bitr--:

~ ''1 ind.:: *-f t .:. rri iy n *: 'l ":1l.-. vnt, i .-r 1 -1ii, a

,i-';t ii 'ferf:f nce y:e-,: foiuni:l iC I L Z .,.t 1., 1'-'.0o; ,Torri:ni ,

1',:.,') itr- ti en .*: rn-fr-re,- "-d cn, -,- t io i-.' ,:Ie '.cn:,- 'i'll.:

l:co--: c,-el i n.i-nc ..1: *:f : o-.nvr')' ti,: .'-,i1 d:hii.- ':fl,; 1:21 f[ jSrrer

size, full and more t.ur'id uhile il.eo-cec.l tonsils of

-er,--free birds i.cere flabby,, pale and inconspicuous. Thi

difference p-:rsist.-d till 5 :niinths of a;T.: nnd from th.en on

the diffcre.ncc br:caim less apparent. The rel. ive eight

of the trid.nt ait thr ileo-cec.l v?,l,:r ,iilcli c ont~ iin r both i

the 1e1o-cecal to1nils u Cnn cocistntly' andir .:ubstantially

-m..ll: r in :?-rn-rre cnic ;:ens I an in Conv,?,itional bin d-' .

The cornientr..tion of lymphocytrc. in the 1iiec-cecal tonsils

of the g-rmn-free bird-s was from 1,' to 1..20 of that found

in bird:; harboring bacteria.. Thorbhec:e (195Q?) found

no plasma .cells or s-econd-ary: nodule. ii' the ileo-cE ..:l-

colic junicticon of g-crui-frc chicd:: st 2, 4, 3, and 14

'i,.;e::. Thiy i 're3 founl in conventional chicks oC all age:

and in gerrm-free chick-: at 6 ueeh:s of g-,i. In ilhit.

liandotte Ban:-itn chiic.k the burz-3 of Fabi-icu; of c..n-

vent.ional chicks were larger p.'r 10" gri n: 'tody ueint Ulthan

that orf r-.rn-fr' ee chicks. In trcr-i.-fr-ee clhiic-;.r the splc-:n

:ias of smallerr rize (RE:,niers- et -i. 1 i9t0)J bit the. general

I.rucituree, color 3nd c on i enc,2 y of the Fc'r ip'.i';rl rnid cut

surf.*'Ces c ,r'e i.d-nt i,:al iiwith t!h'.t se in i'n covr',,-ttiron.l1

group.. Dis'tion I f p ,l.aia cells ,:- -i il.r in horlo I

the group.: but t :is.. cell- 'Ic r? c- less often f, und in the

thy.i i., ilceo-eoc : jlun- t.ii;, --d f' l .lic:l of tihe bur.-n a of

tie g-crm-fr'e chick:s Thcrb :, 17 T -l .- outh of

cerm.-fr'e c.hii cl:ein re i rcd c.n .. c'iliZ.?i. p l if cld lic,'t

.13s found to, be ,omp'arnbl" a ith co tr'. l chi i s .e -r.--d on

i;itural ,C'c:,Trr i1. l ict i..ut th.: .Ar'c th r nAt r-c. t- : ..

t o b :. f-- r-r in : n-fr iI.- : F :ir .- P i:, I f.' ;

Forbes E t --1., 1'5'* R -'[icri c:; 'i. 1-60' fo: -In ,r'.:iuthn

?.ri1 '-r.roi. t i.. t : e I n rlr .1 bu.t e,;T r,'C i.tCt i. i-!ri

-hat:, hibilit, poor in e. n-free .:hili:' The redi c--ell

,,T.:rph _l,.j', ,', h'l r,,n-,19 lt, i i'i "-: n -lC i ll'..n ,l .: ',!:,..i t'.' I' ei t va ^lue:l ? li

th.:. rc c.: -I Ulcel :.ciIm: r'-c ic tic l ip ;i I -r i o andg

*,:,,V it|i: n:"J l c:hi.'l:cl E, 1,.n.:I': I.,- --: IIer-I'e t,: t i s:

Ji.r i i -:-tO i i' ,:r e : i nal *,'; : l:: e1 Ci r;ulsat i ;.I ly'mph, c-

m'e le re lsO hih enr..;Lth to : ta o. that i: thie r -o-n. -& of

l ivii- *ii '-,lorg ni 0? i a :r' t h'ir pr lu't:-t? n 1. i ef- oa~ t

On tlI numir -' of 1:.' pt'rr c.'tc I.Luel;-,', 1' _" ). Th',. lo'f

i' -l ':. i: L ': .' -rc t :,it'.-- c b i li fr-cr tio i: I eC :f

t h!i ,:!i-r r-':,;-ci' t- ji:i -s *:-f t ', g.r'r,-; fr.-e .niimn l (E li::a i ::C'.l

FPlillip-, 19.:o; T" rb, -.I' e ]. l':- Tii. g:.1 :1--1 ,

f mTm-- iarl .':ta rl-I.Lti: onr :' th ic t'l.,, in fr-- t i:,i or r thi

blc. :. *,f ;r--r -fr:c-, 'c:h i e l :o 1. l ';,:.sto tiinn, 1'i i .-c

Iind tnlii:-: ii ': .'.:!iti,:nal c 'i : io h 11 ii .-

F r:,.,j ;I.r? l:nl in riC uT- in : n-f r' .*.hi., .r: th: ;,

m? .1 r'-.: .i. I i; i !o n WI it LIe 10 i M in- lot-ul 1 .,O f

the gerwn-fr!'cc clh : 2 ''ir i ste'" :'r -......i .1 o' a ren-alt

:.f a t i i!'.o '- 'i t i i,: i :tit, i.,:. ; : 1 I 1 "r.7

: t _ _..1 ,. i -l, U -:. -, :. '. -' r.: r t:,:.l -

l j- r i :n;d t t ioi of n-r.:Lil i2 i I .1 c1 fr V-f ro-

ri '. L '"'. i .I-: r :. L- r: rt c i f ii :i ..r ni i- .-1. C'

tltr'- ti-.- ,:';'.erall .r''niujl :.,,';, _p.oi.t- i".

.,:.r'e p:, Iti'--.: ol- -i.ui-t'i.uir -'c tio,: t r .tcnt -il :.-i

cec- l]. con Ltnt--.I r, prs dli rectly related to t h. a, b .enc.. cf

, .ttr ii.tnt.-Zt nal micrc.'lor '. 5salish :nd Phillips 19661.

nid Srin i er i1 19":) found. the: o -:idsia ion- -re.uc tti on poter,-

t.-ial. of bLcteri-k-free content: of the cnick'en

strron* por-itive )while those of tlit coa-iventiotnal cick-:n

: tcr:- strc.'ily negative. EBlich and Phillip? ( 19u6) ie-

Fot-'L;cd t ithat th-i pH ir.s higher in all S.rmznrrtl : of thi: h. iu

in g.i-on-,free chick: 'hl.n compared to th It ae:n in conven-

tional chicl :. Tihe above stu.die. indicate thal. the gr rm-

frree chicl:z3 horu acceptable' nori.l tr 'i thi saind r productct ion.

In n-dtlition, parocii or c pont tiunrs ar e not ct'mmon

in ge,'-fr'-.x chicl::. Th}.y survived ;:-irrai.i tio better con'.' it i.n-il chic::; ihen the dosage ;;as beloi 00 r

at the rlte of i r per hour [c:L'au hl]in at '1., 1Pc ).

lio ,Tro. piysiolo.,ical -bn,',orn-llt; has been rep r.ted in

p et'U- -frte' chic:-ks. COne cc.:ilitio;n calle:;d ".jitt.e-'s" w-a:s rc-

po-rt,'d - Gor'lur t 1. ('19 9, i, ger'm-free chickL du- to

.:: ll l-.r Crolifer'_lt-ion in tlhe br'ain..

TIT: zer' l of cr-fr 'e lnimnls hin-: been shoc.n t-o have

Slou: c-lo'.'liiL c-.tent. an '..,' fw; .nti-L':teri aa;jl -

tilnil:-, Lt ..I. ,i 'j it r.t- '. .teio e. : l tinii'i s are

pr, r-.' ''s to.:?tht-' ith -nucit of lI:ul-o.:vte andi

poor r aC oc t -po -11.- I r', .- : r.-,-fr'e :n ilc.1-

v' :.' .-,uscep lble Lo pr,.tho .-en-, -ithi.u. h, -iurpr' ingl',',

, ".-. r-e i 'er'' .-ur. -. r v .r:' Fr'. ct iv' in 1, '-

ing injc' Cl p- -t icl,-s ,r- de ." b;.:cI c: :'i ,(Lrcri;e:', 19'.3 ).

The "'-t I'ol ',' of .i n" iuf',--.-t .,., ,..;i..-c ha b .en -,zu iied

I ii ,-'. i I i i.' 1 .

P-,thc:.--n) cit i. of Int: -t in.l P:ri iet.? *: -,. F- -. 1 i-
in Crinotobi:ti.' Ho.:t::

S3 T',:ral i '-.- tii t tor- hae utii t : d it :' to t i

host: for tho e stiud"' :f c, rtairn azpFcct: of the hio. I:-p:isr:,ite

r-lat icnr3 hip. In contrast tc:, the r,:se arch on :'-:l"lic (in

'itr'o I cult i action of p.ii- itic or r nir sm:. o-hich i n:l ber',

di .-ct.ic tonir'di lr 'nia inr t ,M bi chremri :c l tr-' rd iriiTunoi- emic :'

charsc l:iicrtic: of thih or-sanri m, rn uce of zrcol.obiotIc

hlost: hIa. bien oriented t.r-:sr.i tlh in I-: '1' ctud. of thi

ei:lo .Aird pitho.'en-ris certain diz.1ase". A number of

tC.!.l:le on the: di-'. rlopf-i ent and .pith:-''Len-l:-si: of inte sf1 in?.l

pr:.t.:coa and iem inthllr.s of inman arid -nrli.Linil import ance have

t[-er done in ,no-to i otic: h rt s. i 'h :c rudi : h.a'.'c- lin

the re.liat ionr iip ,in tile n.c-t the parasite arnd the

host': inltestirn'l micr.-llorn ( 'chi.f' b1:ctcria nd fun,, -i1'.

FhillipE it Pi. (1 '95~ sho;.ed that Enranci "l- i' :I.clr i c

can produce pat holo' lc .ionr in ruine.z piE-: onl:.' v '-

cr-cie: of E bacter it tuc :25 E.- 'fnr-icmi. (C':,i and A-ert

l:.?nr are pr' :ernt. Hoi -r -riothicr p:jr~ iitiC pr.l:' :'-

: ,oai, P.-r-trii h inmon- ('TricI,:inoi:- .1 hcmini: de'.elor.,d i 1

large niuLj .ers in perm-free- pnuinca pi--: hilll ip:, 19?.'i.

F nt:' rie ho ITr ch : .: '. t -1 -l- '.en zuboutiii-

:u l, irj iri :,:. i -nt -: ri-free ,-.ui-i pf prr.luced la ,g

l-:i 10 _, iit rimi ai' ?d, iin, ati:; 'n i! c n: :n.': l' 1 i.on-iil Luiri-A

plc-s re- ulted in di .'-pp.;ar-ince ,:f the r.oto-in"' in *a f'"'.

da ( I,"ton ,?t :1., .19 ).

E:.:pe'rmentail iinr.cti-i of ,.- ct 'i:.ioti' ~.,c-? ni lth

'l __ti-.o_ r. .:i_ dur) iu- an0d int i "- r -- : r - C 111. i

h:-ve be-n produced anid -Ltuldie.d (:a::';ton et a 19'9;

'cstcott 1971 1. In general nore- plra- it.e veI- lop-e d,

infescti i cn i trere ,f lone -r du. riion, iand m.Torlc helmintih eu'2

'cr I'uc ed in tihe ccn';ent itionil thain in germn-free hc.te.:

CEO:i nophilia wa:. imalr::.d irn g'rm-f'rec mice following iena-

todlc infect in, but, no oosincrphilii, wer- seen in ccriv-etionrl

mice. iNoduile developmentt: ii t'h' intes' ina i l .iall .a: a en

in both types of ho.-it?, hojev-er the noduleF di.-app)ejred

fron, cc'nventionial ijhosts rapidly but .'?r -2lstei up to f.-

days in ger m-free m"ic?. ..'!eir tein e t 71. fl' ( '-.-2 reported

that t he larvae of !. ihi n .i'1 ill not de:ve lop to the in-

f'ctiE/. stl-ae in feces fro- gercr-free nice as they rn'rmallly

do in fece: of con.cvnticon-il mice. At le-i-.t in this c:" ,

th.i int-e tin.l micro' fl1r.- c rontr ibutol ed to the prlcin:Led

-urviival and es. production o-f the. h-Ieljintli rpec'ie.

Anrortnc: int.,reating s-tury by r[,uton ct nl. 195?,9) revealed

t hat the MiO nLr e lielmi ri in .i'-.' .arl H-'mn,=n .. r 12 n t '

which h drio not develop to r.nturity in Lhe conv',ntion'Al .l uin

pi.r, can do o': in the gerri-free animal.

.Jo;L'arcn et m ( I1' "': nd .R:-hov.' and 'Gric:rior

(19,67) found feline infectioua .Lt.:-rii ti in t'e- ,:rm-fre--

cit i s r lild, i:OnfAt.n1r:l di.:? .0-e ith s5" to : or, lc ':c.-

pen i t iyn lc it rop'. n.l :rlh .1, Ui. t .iitloI .t

noipholo-ic inte "tinal le.- ic'i ,: cliii c al Ci:'n In .2PF

c-.ts, clinic-l 1_-i.;, ultr'-- iructr .-. ter.L ion Oi' the

int.- tin- 1 n ico2.? and r luced en::y. ac a tivit icr.. Iote,-

(.JToi,sci -L 1., 19ti.7; Fo-l: and Ro1:Iovrl:y, 197ni..

I- iL' L- ica t -: In :i :-fr-', : tv urkr.i .cy Doll a it F r -iriti :er

Il'*-.. :'id Fr r- l:et :nd ['o11 1'-6.') i 1ii,1 ;hli t i: cL' c

fl' ':., i ;." 'if,-:t i, i':.oCC r e' ? cf i, nf,'. :ti- i h .t' n -.nii Ito.'

H. ,-' 11 i i ,-' ,' tile pPr :,t u' n l r,-i,--L: r-i.lIV. I. iLl, -

qui i'Ltlt EP'-dl- et al. l,1"- )I nd F,- :.l-:, -inr F i- I1 i r.i
l.. ;ie,-,,.:r- trt- ared n ,,, l c:'ti,:,l ,, inr'".- i'ving i prc'tz,' an .

o., 1r.1-- rt-i.i ') a i- 1- s ecies C' f ebt.rl ci; IL. c li,

r. r frin-i' -, or .. -,It i1 ) tf, inf ti' I't rc'-ep'-

titi ; l-uii :.,-W .r lin.z- r :t -.1. (1 ClI? rr:..crt:.1 that

thie bac tea ri l r:-q ei[ -meri t; for pr ic i ring inife ct i''i.i iz in1:cr -

I-.:pit,: it i i bacti ris-r iee CiicI er er d i 'i [' ie.t I r-rr,

tii 'cc for t i- diej i a c ii n ,onbac-.t.: ri-fro- e tu l::c,'. Tiiey

attribjt. i: t:- .:.-e role c- t.cter ia in ki.n. r1'.p ic -g n s3i or

ii-ife tio -' ei.ter-'iep:,titir tt r. .i:,- tihe ,.ec: p tir'.o Icn-

rc e t r i i: tiol for thi c iir- v 1l cf H. --n11 -ri iri -n.ce

hii rcm.:on:-. riav':-: bcen reo i. to rvi-e in b-.: te i.a-fr'e-

-. .\-. A enhancmien f viril cannot be ovc-'- r ilc.:i

tlii irnf: ; in :-. i7 i n- ti- ':'i" ]urf ,iin ir-c.i.c i:-. Ii his

S.,'l' :triie. c:nri tc: -.c -. -ba ter.: ri l:.: ti t i:,- ip i F ;illip

(l- ~.:-i -tat:rd tiat it a: Alimri t' ccrt iO n t]ha': tile nctcAri al

flc'r, r. 2c ;., t:, "p r' i'. ii ..-; :, -1i.1ti ' -.nv ir.:,- , r-m t, [,:.. ic:,l

ri.d h-lner] cal, for :- :.: .' t a cti:n a ,. bl i- i ii:L f liIn'Icn

inf-- t io unt il ir:H t1i i h t. t ;.. : m* on[ r tM ti: i E "

Ri:-c. entl y, Fhi llipz and Gori t i. I 1',:..I- ;--7 c .icmd -ion.trAte-d

ti: L lan:ous tic i r-i -I p,:,C i-2 alt --r t 'irulc, o:f f..

hii to:'-t -' rci in ar- : i:.,b -tr",--.tio 'or-:- cul. t-ifl r as

menaured b'' sutbsequcnt inoculation into animals. 'littncr

and Rosenb-ni0 1 1970) stu dying the role of bacteria in

mnodi f:.'in the virulence of F. histolytic found that th':

incrcased virul,:nce is associated only uith contact of

ameba withl live tlctoria and pcecullated transfer of anl

episonc-like v.irulence Factor from bacteria to the proto-


Reid aid Botero I1967) repo ted the Fgro'mth of the

coestdce, Raillietin-. c:cticillus, in bacteria-free chickens

and concluded tha-t nr contribution to the establit-hment of

the tapeuorn or intcrrerence from the normal bacterial

flora of the di-"stive tract occur-s. Johnson (1971) r.-

poitei the rvc':th and development of A w-ri~iia calli in

n ototbiotic chicken; -nd the data iiidicate an inhlbii l

of development of the. nienatode in the bacLteria-free


On the other' hand, ?":lizh and P)iillip: (1966) re-

ported thrit oral challenge with C,.radid : 1bic.-s resulted

in crop infection in all bacteria-frec chicks but no in-

fect iiil ocuri'red in conventional chicl:;. Layton and

Sinl:ins (1971), in their .ztudi.,s nith *'"i-.** -r a-- ."" li-

sfenTricLutn, founi nolrt ali tv '. r6 .. in ar-n-fri-e chicks

than in *sonv.ntio 'r- chic!:: (3 ). In .notori Ol ti :'.:1ne,

Lohl:r and Croso: '. 1'969, 1 1?l) 1 have described dlirrhc.-ngenic

-effects due to E at-ctablc" fil tr-L r. of 1-.oth cultur-es

and .whol cell v-3:ait.s o0 cl- r i-','liia coli and Ilever ut nl.

( 196( 1'.7, 1971) dcscrib-.d a polv,'eroaitis-like syndrome

die' to :. E. c l i n gc 'i-fr'ree pri.'z. 'T'h:- : .- i li- al l

inrdicat: that ncrrial ficrn has either a beneficial cr

i nita :roni t l,- ac tic n on .ni:i. of th-e .c patio r, c ie c an-

i-n:. AnJother i ;lt.ri -l ri n- role fotr the a~c -o iated mriicro-

flcl'o irn tlie liozt-prlri.l 1te relationship, perhaps, in

d. -te- r iriini n li..? t spe: i cf cit, I: the rI.e i tance cof th'

conriventionril gjine-. pig_ to Tr l yrnoz'-,ma cr-,. Hoiwever ,

a Tmnac: it., of the gE ri -r -e: .li ri-a p1i.:-' h1'ir. r'ed the

tr:.'rpna;omez in their blood f: llo u ing irnocul.a-

tin ( hiil ip: and H lfe, 1 ". In .con. tr-a t, tlhe

bacrteri al flori had no rol' int lt etab1 1 'li irent and

pat ho:-tn- i.::i7 o f E. ri'J- ietti it n the c1hic ,.'le intes stine

i HFe d:- eit nl., 1 ,'~ Ciar'l: et al. (l' :,2.1 l l' f d ver-'

little diff r,.~ice in the pathocrcnici t o f E. tencill: in

cnvcr, tin .cial and ba ter i -fr'ee chickenj altIhoI'.i-ih there

I-s ieia. C.i' 1 to 1. hasuro in the appearance of the

2nd ,ene r.ationr mer:roi te iIn the f'e.c g-- rCnotrlAitic

hurt t In a rIL i.l're r-e2e n nt t ud", Vi.ce rid Buirn k1\ r'c..u,

auotFd bt:, Herrde 1t: al., 1'r:l e'.l: ported rio mortality in 41

gFOtoL.OtiC *'hii:n-s n i'fecte .ith r. T-.a1 1 a : c r.p.aied

to ,77: ri-!'rtalit',' in ini'crftc-J convention ,al chicl'en- The,

conclujdc.- that a ,clo: relay t i:rn:chi'p c :::ist: L :t .e,:n the

r:ost flor-a ?nd the i;irc.l:,:o n in Cir t cn- prelI'iAt l. ioni f *.:cC l

"o'?idic ic :ndroTe. -emp t i ,1. (1971) reported

diliaye- l .ic:-elcpm,:nit of C: do ,_eCnouL. 'tt-- of 2_. .'- n-: ili

ir- g-irm-frce clic i:., :cr-ci.all:.' 2.1 genr'-r:,tion c-iii -c.nt::,

gin, to:':tes and..- oocy,'ts. There :as ia clil:nin la: cf


ret iculo-FndiothOlial cells in the laniri n p i,' pri arid

.3.bnucosi and substanrti li-: lou numbers c.f i;onooinuclear

irfinT ann t.ory cells. Thus, the effect of the llact:rrial

flora on the pat.hogenicity of this specie. ri-nain


!C'Rl[:.L [rI'COBIAL FLC'ORA F C.i!Clii[

Th.-r imo'ort-nice of inteztinr-1 nicroflor.a to thli

weilf.r,? of the hoc t In e b-e, : rcc.-cI- e rl'. in thc

hiztc.rv 0. mi: ro'oiol,: .r. A,.-' :i reF. 1lt, the nr -.tir of tihe

int.stiinzl i,icrofilora 01 nT.n:.' ninial op:.:.ic i2 r e'll

d.:':uncntd ii the lit trc .Iturc cSniriu :'nd Crabt', 9ltl1;

Ui i rii. :le a ,nd Bri g 195':..; Dut'.- ?ni Sc?,:-r, 1'9.2;

Saith :nl ,ic.nes, 1,.-t .; S , an 1 D,..b,:, '1 ,.;?; .::a.t: .a d

tic'ri rhit i'n ,'; S. e Ct n 1 u c. ?, l')"70; Ball a-t ,

17'i 1. In ;i'.i spec :' li.,e mirrofloir, ztuildiez h:ave

ueen li it cd t c t UT'-: ', rn- chi e 1:113.
Co:I: ; t A. i 19'') studied i-- effect: Oc aiti-

bi:otiC.: cn thle i"nt.a t in l iicr.:'flc'r of tur l::-.' po'u t

al Iqi er al. t 197T,':- 19t?,:, I tudied the irtestin.; l [ra micro-

flren of c.;mal health, tLuve;. s froL 1 d'Y, to : ,e!:h: of

rg and al:0 in trho: inf'IC tc1.. itl ".lJuecon ." The-se

Etudi -e: indicated I.- 't 'i turi :c- pouilt.; sho-t ''t-er

ha-.tching, the in t.-tin'i tmr.ct i ir-.' i'd L:,' -CV .i'al

-peci.s of r ,ct.-ri Thi i f-e :C.* 1. C th,:ln Piltiply

rapidi, r-e clhing- hi,:h nu; ars i I 11 A Ih: ir'.c -' to l .3

h oi~ rts of l1if Fin rin.c -'.? h?-v: z I .':, 1i'ii I r in other

i'ir, l ',':. (L. ai~~.~r -1,.- -,:, I'.t .. ", c'. 1 ,, t -.1

nith 1r.C L.) A number r of tieiri -is hivr stiille.d the

nornml baL ter i-l flora of conventional chicl:cn- (.Jchn-; :.n

pt .1., 19 ''8; Sha:piro and 5zarl' s, 194?0 Lev and Bri.,,

1956; L:v et al., 1957; Huhtanen and Penc3ck, 19c5; Smitlih,

1965:; Timnms, 1 ?.8; Barnes and Impey, 1 c6 1. Factoru such

a- age, alimrntsary Itict :tr*ucfure and function, diet,

fcedinE habits and environmental Cactore hav- teen shio.:n

to influence the bacterial flora of the normal gut

(Joihanmscon et ril., 19,4i.; :3rith, 1961; Smith, 19?65a, 1965b; in.-l Crabb, 1961). All the-e studies indicated that

the number: of bactcria of all groups: uere found to be

highest in their cec.l content: and progrcescively lower

nuImber.Z in the po-terior l-arge and anterior call inter-

tine, respectively'. The oirgani -ms constituting the maIjor

part of the flora ,,wre E. coli, (St2 rto:oc,.

fec li:-), L ctol:~,cil s -i ., B-Ct'.i.aii es Ep. and C. p r-

fri,-r- The ab:zence of R- terrnidrc cp. in the sm3ll

inte. ine, the preponderance of B- .rteroidis -p. and Lacto-

bt'-illus up. in the ceca, rand the loi levels of C. p r-

frin 'i'-nz in all Cites rei-re of panzFti.ular intere..t to the

investi-antojr.. Shapizo a.nd Sarle: 1.'-49' ) found the count

o2 aerobic and an'i-ro'lic L. Ctlori to t -imril- a in chic!:kens

of different -,gez. On the c'-n -.r','- H'-luhnnr,.n ain P isack1

l190-'5) found a prepo n-nder,..:c of -.;i,.-.c ol : after 2 :.eel:- of

age. Their re.c:uiltr .also in-iicat'd that in day-old unfed

chic:. t. flor. ccnisist.ed maill, of L. r:--Ilis. Tiese-

enteroco ,- ,si -gradua-Jly disapptearcd from the duodenunn after

6 d-avs of age. The cecui -ilzo showed 3n initially high

col.u'tt :.f eit'erococci ani a r3 robi;c bhctet i.a. Th,'ce wr,-r

replace-d by, ?rict-robic, typ-c at around 1i .i-ys of a,,; .

The normal bic t..rcial flora h a l3 crci rCepor-ed to

ini t[ of corni par-a.i tes I.ile:to'. n

et :-1.., 1'5'?). Anitr cr int. .rc.t-rin phenom,-non is the

,*jcr. .a-' a-i, or i'iC'r., :--, :n the p:i pul::i.'.n- of o;ie r t-L'i; s

of ihe flora during certain patihc':logic.l coriditioinc.

Balii-h ind Phillip
inf:-.-'tio.n i:. tHi. cr :.p., the couni t of er.tcr i c.::,'ci in ii-

ciaz.,d. iJ?qi --t al. l '1?7C reported z i-,ificant differ-

r ce:2- ini t!he i te ,it-inal mi ic.r fl' r. in turl:c,'s iil.T,',l1a-td

uit an in.t-1 ctiou r:fiLt':'ri .i z. "blue,:n ") n;i ir and t n-

inf:ci :Tled control tur::,y:.. Trie 'chlicii ,wr- -narl ac Iterized

bLy i- ris- int t ota.l microbial coiut of thi- intir.? intez'

a ci.-nificant if n'lC ease iin numiibai f ,: colifo.rnic, li'. cto;ze

,i-:,nf rnienters .and cl :.-stridia. L -":: ,-t,1 cilluc sp. ,Ic:cre'ased

with severe' infectious cni'--eriti: but in.:r asked when the

dii.c. a.r vaz t- ild. i-licroflora c anti,: similar to. these

findinries hi'.u,: been ol.,,ervd,.:. y riith and Jonrr-o 1i ''3,

1 6.67 and CO ata and foriCshiti i "-i.' ? in pi -.- in.oc-.ilated

e:,:.. riihi e-'. li ,ith an enLeri: t n .' .-.i.a-:.-izon aind

S-rle: IM.1 'i .i iot l 1 that in t- .1-: in f ct a on, a Ct nM -

lation of t- I. '.irthi of C. r. -., c.,. :2ic'rrT d iitii con-

cu rcnt lecr c-.c .I I ,tf .t,--ci ]i'r op. ai,. 3 -ricc.: i c- blood

giucoc- level during thi-- tih t- 7 h -ii y po, I.-iii cf.i tioA .

Thic ;'-:;.' b: relat,-d to -n i t.rf'i.rn e in b, 1- .::. aboli-si-i


and a role for the flors in tlhe pa.'thogferie'Ti of cecal

coccidioc.i:. Thus, in cecal coccidioi., tne problem to

be. tudliCd is iihcLher or not the bacterial flora present

in tohe intertine aid or hinder the initiation or the

disose and rubseqlucnt deelopt evelo ofr pathological cha:ngEs.

A i.articulai'r s=pcies or a colabination cf species nay (or

may not) help excystation of oocyscs, subsequent liberatic1,

and ix;'viv71l cf sp.-.r'o-.oites and inrvacioni or cocal epithle-

lium, development of shi-ontu.: and,'1r gam ctccytes and

thereby contribute to the tissue .*anaage.


-'not tELctic: tba :ter'ia- I in'i- :nid FLC-!'reC )

c*hiCi':,n- ',:i' i r i : in 1fl.?.< .b le plastic M fil m i.,lit.or

fr.-.l.ii in. tI ie: netlc. of 5Br dl .-,' -t i ( '- ). All

1i ..l1' .t'. a cc:: Jti : :... :u.ppli : we e obtainr rt om

Lit -". m? -cur:e.1 To, meith.' : CmpFi:.',,i in :t-rilzatio.n,

riin:tirt-ncf e, cmid : 'er P ict o of g-iitoc'iotic ,n jiro i' .iiient

L hAm.ri"' -ir d i 'I :p t t. r.: i ili t io' o)f fee:id up-

,pl ,i .andi-i tie ,: h rji.: : for ,- rte riirin th.. mic_ o-

biolo'v,:~1--l -zrtuv i ;re ihe same as thosee de:cribbd bt

E'r ,'le:, st ii. (I1c<.i .

D '.-- l.9. or l''-dav,-old embr :ona ted ihite Le iOrn

chliCon '-g u re"'- :ic a a ed ron a comr.ierci li har: hei '-

fre-i ofii -Irr.-A'ii ajnd [,l .l.-'ma infection and incuba teI

at t e i-Aborato:r'. f ior to ,i-trodu:ct:ionl into the isolator

C:limber.- all :S ,r' : cnr.dlo at1 ltea lc t t': ice to inri u.-

th.- ;i bilit;,- cf t .e n t. r.:,' 'The .:ur'fac e of th.e :Cr

sh,-ll "s :c- : t rl. li -.--d b, inn.r:in; tR..: a D (pack-l il ?,

t-ibul::' 'i' n t: i 'for min ut '. i czolut ion cf

1:. F,. Su.'. Li.1 i.:.on, '1 ort ti utentin Roa,i ,

Flora' ci ,t. ate HaI-ltc'ic.-,.r'i Cain .ec..ii -, F or1.i 3

mercuric chloride held at 37' C. The eCgg uier'. then drain

into the prciseri 1ized isol-tor b:y nre'onz of an eZ'e cllhte

and placed in a plastic tray .containinil a cotton todl.

After removal of the Ogg chute and ceaiini of the entry

port, the entire isolator u;a placed in i room held at

370 C .nid 80-S5'; relative ty for haLtching. After

h-tching, the eg- shells were removed anid the chicl-:ns

transferred to a plasti' urire-floored tasi':t 1 -ide the

isolator. Sterile feed and l -ter uere provided ad lib turn.

II. Froduc.tion of SPF ChiclIens

Chicl:ens hatching from fertile er _s obtainedl frcm

the same source as that from which eggs L;ere obiaiiid for

production of Enotobioti.' chCl.kc-Ls u;erE ini.d-iat.l;.' tran:-

ferred to noirficd Horsfall-BaueIr unit:. AltoC, thaer 10

siCch unit: urere kept in a room adijoining tios.: in which

the pla.l;ic film i:.olators were k:-pt for the prodlucti'

of g-notobiotic: chicl;ens. Air entering tlih Ho. ;fall-Bauer

units was st-.erilized by pa.sare throu;li sterilized fiber-

ilass filter media. 3tcril uat,:.r 'as csupplled in gallon-

cized bottles -tt-achedl to the inlet ti.iub of the.: unit and

the level controlled by r v-ity, flo;u. F,. : con -ist -td of

c.hickcn starter miash fre- of an' :u .ritibict Ics or .d-lcd

chemicals and iws of conpooii ior;j a; tin-. ltional Re:;'erch

Council (i;RC) standards. The fT.: u ... ateurized in a

hot air oven at 1.,':0 C for at l.: st 6j0 .. and -isT,

supplied to: tnh chicl:,-.: s in a m tal celf-feeder inside

:.-icf unit. TeLup'tr i.,ns cont rolled elc. t ric:a ll; and

ventilition ': as C['.z-'rcci nw:-E tive prc:.3Iur-. Be-fcre

.:,i .-ftc.r e:clih u:- the utnitc weere cleanedJ nd -I-c:r-uti .d

uith hot detergent solution rnd stem st-rili eid. A.s r.'

Spo'; ible, the uiit.s Wer-e ,*p-neid :nly ) time during ani

r 'Fe rirntri- t--to cntc-r rie'.;li-h- tc liedJ chliick for e:-:po ure

of the c hi.:: to E. tenill3- ooc: stz, andi to renov.: deid

Mird'd Fel ioodically,1, M l inia.l labor2tor, tect: tere

c.ondul.rcte to cihec! : tie pI tlthogen-frP -? nr.ture or' the birds:.

All experiment. l :!tiicil:c, i rai ed in there unit; rc .?

fnor itlr.i'.d bt,' t-:ndis rd lu tr:, at(rry methods for thrL. follou-

ing e r.; ifi.i p: thoens:

1. tlE '? 3pe. -ic .: E ir.:.pri3 c oausin COc ,ldii i
ih :'aC i2 -!cei ;

the c.:oi-ion incest : ii Iieiiainttn: cf c:nic: en

(A-cari1 q,. H- ;ter -i_:s, ., Bailli triri ?

Spec i es ) ;

3. 1an-i nen llsi an F rst,-.iireii cpec-:ies; and

4.. rnel- -.,-rIli .

III. F i'Ct 'i C r-." 1-ii -- C-iy'k-ir-

II.on l"-n!1.-,d chic : ns w; ere tr.--i C i' red fror, the

incub:t,.r -d r-e r':'-d in L i ctric. 11 1 ,'-l- 't, it t er:' b.rod-

erc. lincter- 11 ii ed c:hi,:-lrri :rter h wiitri rio noiti-

biotice oP ?iddid .l'i,:.,-iil. r .i f :','irn- i C requir-m.L tz,

and u:-:ter er ma,]e- ?v:il..ble i. t ~,tqn. Before and

iftcr c:'-ci u.-,; t-hir pens i:.'re clean.:.d, ccri.bbed wiit hot

d' terI. '- t or lutic., :. Liu. - .ii ii d.

IV. Sour-.c of Ei:.-r _n t7ie Ul

The cz.ic :train of '.. I;-n--1 11 icolatcd frou a

natural sasa- of cecsl coccidiosi.-; 'as u:cd tnrougIhout

this ierics of c.::periments. COc;.'stEs uere produced

accordini- to need in .:iis:eae-free 3-;eei:-old conventional

chicken. Fecal nmtirial from donor chicken uis col-

lected at 7-0 days after inoculaL.icn %:it;h a tsublethal

lose of oocy:ts and tlhe fe-:l debri: and other gross

particles removed b .- :eviin tlhr:ugh 30 and 80 nesh

ciev'es. The .:oc,:y t vrc their, sodimlented ,'v centrifiga-

tion allo'r d to srjortL1l Ite in 2"1 rotsium dichromate

solution at room te-iperatur'e and aftcr sporulation ;Cere

stored at lo C. Imc-dilately: before Lue in experimeTital

trials, all ooc-'sts ;ire curface-: terilized aith a '..5 .

solution of r-r-acet' i acid (Doll e l. l ?o13,. Steri it::

was tested ucinc standard bacteriolc-oicnl and mycological

proce'durec. Apprcr:.:i-n.itely 100,000 sporuiated oocy:,sts :ere

dmi iictered or ll for c:p r-.prinital infection of chici:ens,

usin t a asmill syrii .;:: : itli an at';tached c.:nrUlula to insure

depoziticon ito thli crop.

V. P-,t[-ln i, E- ,n--n- [ n .

Chici: ini c'iliatd 1.ltih oc.:';ts ujere I;cpt under

clo.c obj:ei".'v:tion. Blc i-'di or' ,ny their' clii.ical signs

vere nr-ted. E'lr,?l :-aiipl; s for i .l-c 1l cell volume' d.'ter-

Minltiot n u alid -.,rum n :0n.l"3Ic 1 e i:i:,i both before

Snc ul t -.tion .in t he 7tn ,:', after ,.-::r l.-'.re. LE a r,, rial

l.: ~ri'. mr,. on botl. th e contr. ol a d -:. :pr: r I ;ii

,;iic jl,1: .:-t n ,r pc '.-cc.rd.inii: to .he t.act.rii: ical3

rprc de crir-.e b':l : C hi lin.T .'.'i., aft-' :|

t. cc'.'zts and all th,:,; Zi. r ivir_,. cn th, 7,th dV:,- -ifter

.::p:.z.e L r.- nC 'rop' i:-il e:.:armined fi',r croF ?: lie ion of

ecal c :cidlio z i and thei , c. -::ni t it 7 e:,-ariiiid I

varI-: .i. dev. l p:. .T nir .?l ra3 ;, E.. t1 ci: 1 n: inf: ct in.'n

or':diL-:. in ea:'h in -oulated gronp war o.- pard .nd !vaded

for' macl i'cr :!:c : 1ic; :1- :. For hi -top:cath c'l:c l : :'-mlii --

ticOn, t iLsueI- oIin grr' o:' 1 1 :'i.n and c.Cial1

i ziz .- fr,:.m con ara' :le n ite. i'f :.'T cn'hicI:ir ,h''iii ino

'.:,." le ii:i' r'e fl;:.,'d in neutral ICi formna lin and

*:cti r,:.i it 5, or 10 .i- thicl:ness arid stain ed uith'

I i.-mlt'I'::'I -- '.:s in. Tne'r Y st emi i ar osc pic gra d1 in

i' l.ciis l d inro pi radini or rnr'.ic riir frasitism '*.uas

mcdelcd raft-.r H rt:. oni-S.,ith ,t al. '.1 9o ) az focllo .s;:

6 .-neie cf G-raling ofr Le'sione and Porarct it ism

E.-'t ei 'i' t1'. t re > pic 1 C"oo ,pic
L,:-. icn i, C ':dir, c, G riin. i'.
Frasi t ia i L.;.:icn3 Far i anti mI

Ilc d- t.-:' table fi. co--' clii'i t i :
leiE onS f:.Luid ,;7 1 : c.i r,:-
ful S,. rch

+ .3n-ll ui itr r .' Siill nir '.r of
Of le, ion --,i:. :: fl'
by c.'.r. -i l t:arc
V or- le-. i


['-i' oCr; c cr. ic
Gradinv: of
L,', sionr

lioder-ate numer' of01
1,-..ions uitn some
he-,or rn-'ti e

Hlum-rous lc. 1 ions
.;n. heor,-'orharge

l!ii.m' CIou lesions
i:ith severe ilinmor-
1h 'c rid C -3l1
enril rgerLut

-:t ; lcnt of
L- ion. an.-i
Par i i. ism


VI B-, ter c i lo -ic Proced riir.e:.

A. Drter -in tl Cion of" mcrc.,hil flora of tli- recum iL
c,-* -.-e i ti' CI -' 1 hIl -.' n

The development of cecal microbial floral in disease-

frc younct conveIenticna i ?hick.: and those ino.cuilated with

100,000 ?d o.cyst.- was studied as foliowus. iiely-

hatched '-liit. Leg.horn :chicks werie raised on standard

electric-ally-heated battery Erocder- and f,-d unsterilized

feed ani inter -1 lib li. i .n,. Four uninfect -.: control anid 4

infected.l chickens o're n-croncie-d a.t varicLo s a-we intervals:

d=-<-old (control on l., .-.-. 1.-ld, 1, 2, 3, 4 -ind 5 weeks

.of age. Irnoci ul. ationi 'i h jo.-:'-, r i I .1jistedJ so that the

day of necropsy, i!o l.1. bte d-.i'.; afterr exposure. At

nc.-rropsv, the v'iscera "'r. e:...o;c.d ao.l the cecal pouches

to'c:ther witli inche: ol interior '!o nll initeLtine and

G'radling of
I'Iri s i ism

Inall rnuni-br cof 2nd
unei''at ionr .-chlizi-nts
-,d, 'or e-ametocytes in
Lcatteireri pro'pi. p uith
acomi asSociaited tissue

flunerotls widely di s-
r i ruted -inetocytes
in localized fro.i uith
appreciable tirsuc
da ago

tlierli'Oi1 .schiszonts
and,' r Ranetocy ?es
iitl widespread d tiss3u?

p -.2t rior i.3r'e intent ine were tran Cirre to : terile

retr'i dish. Uhin:- aseptic pr'.:>e .irc., the p:',hes

i:re opened., the e- le'i le tip I 0.Oj:. m i o, f an i I I jocul-.r -

Vn : ne--41 u .- insertel into th.- cZ:' el tcon;i: tz .'t ic.drIrin,

n1.. t ie rnjteri'i l ocn the ioor. .-tre-a!:;-d c.i tli: surzi'3 of

eacr:! : i. ti,: v ]riou .li ne1di iue .ied in a t. -' m- ri petri

dic-h or i o.:ul-lat'ed ijnto,- a tut c.:- c 'f br,':, Z:, r sc' 1' ii z lid n '.-l

In ca-e of *.Iry ,'c-cal intentent: or 3 f *-..rmei *.c- l C.,'re, -

rc p of te-rile pH ".0, O.0'.' ri pr 0Fhzp1n te i.ffer i.; used

to .'ofrt-n tie material Pri.:'i Cto inos rtion c- the -sanprilini

loop. A\-:-r t.ez e -numer tt :i: c.n Di 1, 1 bra iin h.:.r-t

inrfutirn i.'a r; ent- .'cr ?c in Difco R a ide-id-;:-.trcze broth

..upleiriente-.i i.ith 1i p.p.m. neth,','lene bli. ani 1.' .;'-ir;

an.i cocli forms on r cCctn.. ey -,Cr. TI' e me-id.iiu' uc.--:d for

,lete ir ininr tio t aia:rtobic populationc: ii 1- Dii :oR brain

h-art inruc ion qla- ar up iplem'nt-.'l i.itl 0.1 .' ., e tein,:-

I,',.'ir-o .loriie and 0.1'. bovine ru n. ,. -- r i --:- L.

enuner- 1 on Dif':: u .ulphitit pol:'T :-.:in ?ulp~i.l' i ini:- -a. -r.

For f n -.:i.1 icol-tion, Dif *:,:, ;.3 b:cur,'au d ri- .:.-'troze .,,-r n, t

Difcc R F~-,a-sno-Le:.'in e ar c, ith r i ilt a iot. i-c i-re i; e.l; ,or

pro, .to:- can i :olat ion, Lifo .- 1E-:l ,n -h '*ir:d .iu a: ..ei. Tim.

ci. 1 ub-t utl ion Mil tc 1 hcour " 2 f t *:'ol i* ''f-7i.' .. ; -io r cI'or

lc'E tr-.ii a rnl ;:'. ,-our. f,:,r all .. -t : or *,i ,'f .:r

in'cubL-t ion, th e -' :c-lon ie c r.:.n ca of the unin fec tCd

.cO t.iC'. L O i n t. , 1 i fct-Cc '. C c "' '? c .iirt-.':i '0_i*1 n'-.n

nu mbr b *:al: ilit' rd.

1Dirco Lab-or.-t.ori s DE -tr.-,i. ;i.- .T n "''C...

B. D. t lrm'nr tin r of mi.'rob inl flor'T oi the cecum
in '.c -i i--,re'. -, SPF. -d c 'pI 'nt i c nIl chi ck- lns

At neertop;y, the primary isolation nedia used for

the recovery and ident:ii icat ion of bacteria, fungi and

PFLO from the ceca of the germ-fre.., SPF ind cronvention.-l

chicken.; are listed in Table I. The criteria listed by

Breed et A_1. (1957.1 erze cnplo'ed for characteri:zi-tion,

idenrtific-,tionL, and v.'erification of ba species.

Anacrol ic plates w.ere ilncutated in a G0sp-cch:RI A.iiaer!,lic

Jar r:6'0410 usin,7 Gaspack g1as generator envelopes for

generating:, h!dr'ogen and carbon dioxide sac. Mclthylene

blu' wia:s uI.ed as the indicator.

VII. Fac-:ed Cell Volume

H:1-,tocrit values Irer determined by drawing the

blood into a heparinized tube and centrii-

fuging the blood in an Irtrrn.t ional Ilicrocapillary

Cent-ifuie" for 5 minutes at 15,:''00 r.p.m. The packed

cell volume (PCV) was directly read usin? an Internati. ial

Capillr 1 :: Reader.-

VIII. Tr,,:-.1 -<-'r- PFr tein

Total serum protceiin ,as dcEtert'il-rel by tihe method of

./eich I1b.:-um (1''46) 1ith Lclii ht. mo-lific: ticn: A standard

1Baltimore Biolo-rical Ln'bor.tory, Bal timore,
[i-r,'l and.

-Inter.inati nal E.qipTi ent Compa-.ny, !eedmamn Heights,
rI; .-a i c r hu .- ts.

T .Yhbi I

Prim, ry IT.-.u,1 I. n ,.-.=i U.-e4i Cr tK Rec ler',
.nr, i ,' ntifi. t-. i i:' n ,o,. Ev .ter, i. -n F.i' F.,, .;i fl ro,,
idecr f -c 't i.Li t'ntori.:tI i i .' Fu' .i Crl: r
C ii C i' ?,i.i .' 1r-. ic -] l ii,-'I:c 1

Di fcc' .: id--.tie: 'o e broth lth 10 p. F.m 1.rnith:, .ne il e
,nd 1. ;, ?., -

D l cr.: lo zi':-' _-' ir Ait -. i ini. d btovi e bv i loc.od

Dire." b -in hi iart i','fu:"i--r' ._I'-- 1i t7l- 0,. C '.'. c.' :t ine--Hcl
aind ..1 tbo ir Ierurn

Di f'.'1 ?, i'ri 'i .van.'i: ; '-cuic -gr- r

DiCfcF, bl,.1 -.? id r.-- uii t 0. b Wooo Wnc.. a' r and na.m.-cin

Dif co -, lum bi. t.ra Ot r
D i f.c-R 1 t i 1 c 7 t 1 *!

Dif,.,-' pr'-,' .Irt i.'' bro '-n

Difcc."o ? Hic,.cn> ev.' _,car

D1f- h.:.W e Pa.-:n -Le'ine b:-.-e lir tr-iph,..' 1 t tr:. olium
chic'.oride id ntibiot il,-

Ditc..oR PLC: t:r t. ult antibiotl.:.-

D'Ifc: s'ai:..:iu de'.t' i 3 :C' i unti .iotic-:

Dirc,, s. iprn.t.-, 1eol "r.,;R::in 1 -].i l-. i: '. r

Dif....:, liioni-ir il' a-.^- hii l- 1-] '- ", -i :,r

D ,irc-," tI ,iii ;,lc ollat-: rnC.,iun Uiltilio t * --, :

Di -co : ,. 80, O .'i:le,: :" '-' : :.-: 1:J

DifC.r t ,; pt ic-: -; ,-- ,' c.y rcth

curve :as plotted uring v.-.rious dilutions :o a 10a bovine

serum albunrin solution and cor.respc.ndi r'L: optical densiity

in a Turrnr Spectrophotomnctcr at a nave length or 540 nnl.

In the procedure, 0.1 ml of url:nouwn serun and 8.0 ml of

stable biuret reazcenrt2 were mi:ed and incubated at 370 C

for 45 minutes. The optical dcrinity tias then read at

510 rnn. The total seCrum protein wa11 directly c:-lculated

from the -ta,'idari curve anJ expressed as grams per 100 ml

of seorum.

IX. S run El ect I'o I lre i

ElectrophoreEis of .;erum proteins uns done by

llicrcazoncr Electroph.horcii3 on cellulore-acetate menbrarnes

usin, pH .67, 0.075 ionic strength barbital buffer. Using

the applicator, 0.50 )il of the Zerum -ample was applied

onto the membrane -nd electrophorcsi:ed it 300 volts for

35 minutes. Tre membr.ric then was Stainerl in Ponceau-5

fixative dye solution for minutes and cleared in 33.

cvcloh-.:anone in absolute alcohol for 1 minute. After, thei mEembri'rnc was canried in a Ilicrozone R Denr it.-

meter and tie ch-:'-t tracing: evaluated to obtain the

co..ponent p 'rceintrtgs..

1G. K. Turi-ncr ;.T ai..itest Falo Alto, California.

2Hycel, Inc., Houston, Te::s.

35c--,i-ana I:n t run intc, Inc., Fu]llvrt on, C-~liforni3.

':. E-'.r',,i, re' 1.0 BE ,Ic',r .i ,I Fi, n 'I i

iniig.e ?p ic ie cr c.-tjiilat I o o bc a .::t riia or f'.rl-i

o b:' u ;i -lon iln t'cr l l i 'c f r r rr..: rI-t' : 1 ':<.po

l'ere -ele'?t-:'l *i th l:; l: of iicroi i il iC:i :ltt fromru 'L

o01 c:Clnl'. tio ,inal clici:ens. i:'.ii-i'. ehri 'V sev' C. :2Ci..iO10si2.

AIr.o.irmratel: I .'. ml 01. to broth' j cul t .I of

tie rErpective IrnliTin 1 A1 dmii st-rei oral -. u-ir.t-i a-

cL Ifili LI Iii *a:- 'f f. i c li: r th'. n i' r p. 0 5 mil

.:, h ..?.' :pr' :u pr.enIi,,-o un :" adniistOred. '. _.] i',ic,:!r'

W s Lr 't in? ife:. S t:iiuriudl de::tr --c: tI roth at .0 frc

4L hCLirs anT-1 l, 0.5 r of f-ne ro-li si a":r d ini ter-:.'i .ora / .

E' cI..; ill specie 's er.: dniii:t red lot.J.- before e E.

t,';- ,l ..3tc t ,te *i":i,; f'i.-1.:1: o c. n i'i 35 ;. rt'- iven

, Lr::''L r? prior to0 c'oc'st a:dini ic tr'ati':.n.


Dc'. loo-rint of 1 ie'o. j -il FlPr-r in CccI c r.mi .-,f C.n'.r t ri i oil
Chic:e. n Inocul.atcd ith E. tenrellr and Uniniculnted

The predominant aerobic. ans-EicObic bacteti'ial

specilec in the cccun -.t different ages or control chicl:s

and of tho 'se exposed to E. ten,'ll. oc:,,stz are listed in

Table II ani Table III.

In day-old ind 2-day-old conventional chicl::, cecal

organisms i. ,-re predor.mnantly enterococci (3. frc':i :1. 'ith

E. coli mal!:ing. up most of the remainder. In nioni'fect-d

chicl at 7 days of age, enterococci 'Jierc- not round to be

the preIdominlnt org-nisnm in the ccc:, these l.ein r..

by F. coli and Lactob-pm.i ll, -Zp. In i. t,-ne] -.--infected

chickens, the enterococci ,;irte c-restly outtnumlbcred Iy

other sp-cieZ, especially E. c,-l]i eaid E- tero:idJ -p., but

InIu bears of L-',',a ll'.-' 3p. erer greatly reduced as con-

parer- to noninected controls.

On the 14th day of -'e in nonnini-ctc:d ccnv'entic nal

clilc,:s, E. :a ] [-. :tb- l11 ,,. -r. trre p-'edorinnnt

'with f u w enterc, ccci, .-nd i ':-t -- il s p. 0r'e ri 'e.. In

the infected c hicl:-, F. ;oli 1l-.: the rist predcorninant

csp-cic. '-nd L-'ctt il 1 -. sp. .-re not detected.

A '1 l ?.e da ysz of -1e, E. n. 1- i :,ind Lm'tb.c % ill

sp. predor-cil ted, in the no-ninfected nr,,ir, jurile inr tl

infected prouip, 1. ,:,11 b ,:t.r.i.e: sp., enteroc occi aini

L'::t"h- ,ill .i e dt*etI ct-d,. A ctimulatition .:.

of E. e-c-i, FPcteroide s F. nid 1. fel:1lic !: s o.:berved

during. E. to.nil l- inf:-ct ion 1hile the ro:itl of L i: t,-

r-'., L F Lp. .s c-upprezzed. At 35 days of aie, the came

pattern rwas obb:,-rvcd e::-eFp t hat in chi.leen: infri.,ted ilth

E. re :l ll Ql pr': riri rT~:7r. and a!iIaer'obi': fere'l s:trrepr.oc ci

tere a. 3. detected.

Fui;-al isolate: 'were rare. In c.-d,-ol 1-cl non-

inf'c: ted criiCI:0:n C::.nlid-i 'Anrd Illh or specie: were detFe ted,

but in tl-i inrfec.ted -e.: cril;" iiao r pF Cies3 c-re s ien.

At 1, 3 a,-il 5 uwe -.: of Crndidj :peci-:'i wI re ,d tct te,

in nonin!,'ated g. rolp'iFs and itn '-, .:ek!-old infected c*hi.:lenr

m:,r c ,:ies wre detected. In 4-weekl-old .hi ?f:en?, no

funrl i ijr-e detected in eith-er roni nf-ecte.l -itd inl'ec'ted


EB-?t ri, l I :l t.--: in ;-ek-0 ,l SFF -m.l Cro.rvcnti.r-n-1
Chici ..i In'c.:u-l] t-. itih tt A !.- L'.- I ir-ic:i..:-,l:tc.

B,-Ecterial izol-ates in '-teak-c.ld SfF .ind "o.ivent .nAl

.lhi.Il:cnr iric.u:i Vted wlth E.. t-. j.1 ,^'r'e r,i,.t'.= or' less

sir il ar and are shon-r in Th L : VIII :nd I.-:. C. nor-

r i-, it:is ic..l lted 'ron:. infcc:t:.J -nd noninfect.ed SFF

, ich!i.lci 'I fl I'Cr infected conv.i nt ional. c hic-kenrsE m otrc fre-

quCnitly tlhan-- froma noanin r,. -te. conv eini: onnl chi c-ken:- A

stirIulation ..,f t grcir .th of C. r-e frin'cr,-i" Iwr both

in infected SPF and conv'irntional chicken; (Title X) In

noninr'ecte:l coinv'-ntio rnil chii clens, C. rerfi'rin~-- n t'.

icolatedi in only, 2 trialr while in infcctcd chickens thick wo i.solited in ill the trials. The number of

colonic of C. i.,i''rCi.n:--;nz Jvecloping from a standard

inoculum was hciicr for SPF chickens than for convenitinal

chickci:. :cc iu'rence of funi;al ;p--cies was Zlso fre'eiqucnt

in SPF chiclc-nn, e-sp'-cirilly in the noninfcicct.d ones.

Carid -i s pecie. I er. isolrate from nonii'cct-.d cont rol in

4 of the S trial, uhile Co-if'l:ida specie. ,orei- icolatdied

fron ,cnly 1 of the 8 trial: in conventional chnickr ken

ITables VIII and IX) ',' species r'-re in'frequent ind

i, re isolated only. in 1 fz'ron a rioninifectcd colivcin-

tioi-ial chlckr-i i-c,,OLilaited with E. coll C. n r''ri'i -.-ci,

and S. fecilic..

IFathclogTy due to f. toanell in Lacte.ria-, fiunii-

and PPLD-free chickens i; rt'ported in Tsble I7!. Clinical

s.mopt ora like bleerlding, aiiore:: uo l: ie, c l: i. drooping

e'or'e not nrot --d in i ny, of hL. ese chlc r.:-. i;o lior:it lity

occurred in 32 of t he:he chicl:er .- raiz.,.i -t teri :-, f'u,.i.i-

=-i FPLO-frer. The fimacru.opic : rdiin of :1- ions iat

ne.:'op y, in all theSe cai6e ti..-.L riCc t i'. ct 3incv thiCre '.J.a

no visible tthickc.' ninr i, he-iorril? cor'' fori-tion, Ci"

slouii-iin of ri'- f muce C-:.l -' n !rT.-- cnt n"r !L not

noted. Tlis appe.r rancc of ceca, li'vrr, I:idn:y-y small and

large inte tinll, burE-s of .'abri ciclc, ;plc:-n, i '-rt, ni.iuclic ,

ir.nd l1.uL ver'e n.orcial -i comTpvred to tie vi: '? Ir'

non iTi'.:'r;-,d controls r.-ii-e. 1 Iccteri- -, i-.r.r":i-, "FLC-Free

andi a31o tnm ce rai:1 i .-ec i i f pFr tho-, li-fr-';r and c on-

-nt i -,ll: -. Hi t.pathol_:' i l :,-, tic r.: u a; ni- ti2 l wu

d?'m-1~ ir licrTmo'rr'h '-X" loii,'kiri c- or tkhii',! i:e ir"if';;" of, th..:- ijt ,-.a3

e'.'i- nt .'-.i ni i r' o' o.pic e:r.- rniin'l'i' l v.e e -. I-1

sp:Fe.-c.r d -c -, r vr -ve ind .',iid r' :-.'', .a":- i v''lpFri_ rii since e

endinseinu T ~sl. ere :te.i in trh, ,'pi thr 1 i. cell11? or

tbr mE rj. ;Z a, :p. oec,.:--.11, C ..f the 3lani funii TIhe e e-nd.i 'e-

nous z t u tz ;-': ivere ident i _-l '. irrn-auri hi n.ts and

e arl v 1 .-ar:- tc ,t : Il1' lr '' e 2 n-i eC C r t i ri n l O lC CitZ

containing. mtiiure mero:imc: uwere son. C.l -occid)iosi

,- c .-cr-iibed t' T :T-cr 1-.' ;a.rid T,':::r. l. ', '.? 2 i i,:i-

ni:tc -: in r- e-.c ie : .C ,'-:en' a hcrrn :.p. ,-c. c: : **.-ment a]i t:.

: .jid r.fll 1 iiC.;0ilin cf C C I' l -] c:'oc'tc.

Ti-er.-e .LJ : no decr'e :a in ie tci' :ric t V'r ]u'-2 in

L.actez -, frungi- and r-PLC'- reer c, i. t:.: e:t..,. ed t. -. .

t vr.-- 19 L- 'Ta le .*:I) On the contr i', r hcrit crr i t val uc.

incrr e aza-* from a m,en prt:-infe.':.t i.: ol:.i-ne otf 27 to '. .n

th'- 7th d.ay pr st-infectioii.

It th 19. *e l1% C , ri _l i 1 .n r ," -" t ... -r'. n it C 'no,

pro''round bl.di.j n cri- :i u ';i T hie ) ii toIlo' i. al ,',

the t izs ue .:ar.age :as. n:rli i,-ibi- b,,; s itn _E -
erer e:cn, e p,,c-'. all;,' in.-".tii : j i ::.. :-. D i -.d ti n of

nuiC.o '" .s. rfit m ail. H-tmmt it vili. ." -0ho,.-.j -n ifnc"-rea3 e

of 1I'. in tiie-? chic cenk on tlh-: "tjh 'l po -infection.

In chliil:cns h1a-rborin f i' or cp?cier rioder-ate-

numbe.'rs of lezions I .30-ie heriorrha ;C Ind thu: ike in-

lias noted niacr'o.copiC"Ily Hit.'tlc ical..', t ere as

moderate tissue il'anage and l'ir- nunblrs of endjgenoiu

stages including oocyts u\-re seen.

In chicleen. mnecric'onta tinated.l ith anaerobi fec' l

stLreptc.occci 'nd :-pC::eDocd to t, '11 there were no

vi ible le.ion.? r hc .:r.!i hage, bu: hiitclo ically

noCu st i;.g s er,.I evi .ic-nt. Ti. ..z.: iiar.iare, -.lougI thing cf

rnmuco'sa, henlorrhalo, :arid lar'' 2nd .arne'iat ion 3,.CniCont

''ere also abi:.nt. He.-- tocrit ,values increased from a prc-

ce:poz-ur' volur-e cf 17".0 te a p':p't-c-:posuire volume of 71.5.

In m.;'lic Fne-is mi)jnoD nta-jnrt .ted .ith ana rerobic. fecal stli'r',n

cocci :nd a .3i.tpen.ion of killed C. oli 1 Cr-''rrinreur.

S. fec-alis, infection with tir. ti-n l1 did 1not produce any

clinical sy'mrptorm, dentl, vi1i1 ble le ions, Ihemorrhage or

thi cl:enin of c-c l rnuc,'z.a. Hirto logic illy, ino tisEsue

dr-nage-, large 2nd gcn rnatic-i cchlc cnts, or bleeding '. a

:limona. t rat ed, ibut i:Linatire sc hizonr- :'r- eoe .e in the

.plithlic li- cells of tc nucc~.'a. T'h1. nr, v. lues u 're

-7 at JnVy of 1 .I.c,:ulatioi '. e l "1 .n the 7I.h .: y post-infect i.n.

In chic':en n-1 :c.-:c n1. . i -1 i there' 'id'-.-

.:p., .-., r'**ri.c --? E,. l L ,ac o I:. c,. fo,.;'O '.". liklc A.

'* ro n-';- i .;, mod rate ni'IL'' 'l ofr ie ;i : '.'ii 'herorrhae werre

notice. '. 'Tl-1le VI). D.i' ,th .u- t.'. 1 c :.c i i..:i. occurred

oinly1 in c ut or 12' .1} i cl::n.' ; n. c io t.n.;ii ',td rl ith 2. .

c',: li s, and nIo ior i:.l it', c...iurred in L;heroc ion..-.cont i n tcd

j ilth >,thl-'r .p':ei' e. bi C a, I-cte' i In CiihiC. t.';,t, n, ;!i ccnt;i_ i-

nated ,'it-h ,"li'- th: r-: i'i. Ra t.-:td cic:' ':r tI 1- '.d

tP: cc. i'i. l te ii:s rn pidl F at r i all c c t l a t. ed bl,:o in

lh,3 c-:c,: i/a. chlrU .ct PriSt .c.llv pres.nit it Ph.ln;,- ,'ic,1c:eni .

H'-]:atc: cr'it v.l i -:. also ;:hc'~l 1 c.' .,jt.iir:.n iii *hickens-

io.ncc.t ru-tir atc i uith bacteri a -i.. iin cted. iIl E. ten'll :i t

,'-a"bj c :I ) Ci:Tm arat, lr a C ic. I, .e a C i ,oCnoc ,.il '~T i rna;te-

-it.ler .il'i F-rrtl r ie.-l l p., : lCri s C, ne, c r i .:

iev:lopei ro er.ti e t:o ?c r: p t h ; .l i hei n -lp.:,:, 1 t .
i-enrti- Chicket- wt b,,. : n'rtl.t 1n teL ilth F. *rl I ":r L-t, t,-

rcilluit. :r.. .iv:-lcrli cl.' tild tP rocd rata lvajcisr, n nid

tPi-,::- IC" rlc.:.,.'tami',i at*'_l: -i i. .tti a-rir'r ,.:bic f-c: Il CC: i

Pchot-..e littlJ i. .-,t ol,?':,' a.c:-.ert e: P C: :. I. t .: p ] 1 i-.

Chic::tc irfectiid .ilth E. t.:i-- 1 a pc.y:c it.Irri-

nated .t b :.r mc,: re Zpacie.. :,I biact..r'ia c:r funhi; zhc:.'..';,

le ;'i, -, int er d i 7.- atf- be lt ire.: -i t hi. e : mCrn,. c.,:,:.i ta tt i ted 'and

PFP r :, e:n,'enti.r-l chicl.e s i. Table 'II). H-a-;,y r rt:1 '

- a' and t;pi.Cal ;ricra c, plC nC icr>,c.; ic 1 l z iC h.-

ic'ted in chl ic:-etc prel 1;c i aii id th C. r. -rfri, :,.t' a-i, d

-. fec- r11i Deatl ,i, typical l ic:, i de'-v lo : ic c :-.:.n ii.;ro i

d-r_,-c.l.1 chii.ien. pol:.c,:! anint -a ed ith -,ti ',i C =jJ .o p.,

'-. r: 'fri '' :- r'.li and .. .: 1 r.'- .: :'r ed 1.0

.. ,- r'e 1l r -,ra r -t. i v'- ]:,', palh: ,l,:. c:,I natl iF' s t- iC-n-,

'..'ere mo er- ::v re' in chici,:,;.i p.r l;'o rt-t: n:.- ut.d it :' iletr il h

I. ---ID .... - a l -1 .. I I -1 ,. r _--" .' :,- i :L : -'id, t.

. -: 1. thin ini tAr.hocs p nta' ti r- r i ,ri- :

anrid C. ccl .i, L cti ci1 s c F' '1 -1 li .A : .ci*t.i.c

of b-ctcrin -:nd fnzmi fa'.'ored dev.: lopment of moderate

l C-ioi L-., but,'- lil not frvor development of Ereater p'thol-

o':,;. Ho'.cve.r, the p-ihology w3a morre s evCere than that

ec eit i-?r withi tnti bacterial :.pecie alone or fiunigu

*pocsie- alone.

Palholo -,ical miriife:totiions in S.. tcnclla-infe,?ted

SfF chic:'ens Wr-:-e typical of cecal oc.idiosis Table VIII).

Clinical .ynptoms like bleeding, anio'r::ia, droopiness and

mortality,' cri noted. Thirty-thrce of the total SJ

chick:en.- inocillsted lied, establishing a mortality rate

of 3 ilacrc.scopic as uiell as rici'roscopic legions were

otserv.d in all infected cnil:enc establishing an infec-

tion rinte of 100' The mean iross lesilo score was 2.6

an.3 there mean henatocrit value dropped frLon i pre-e-..p,:-.vre

'olume of 13.3 to 23,.7 on ''th day pos t-expo'ure.

Ini convent io nl c*hick.- kn yi:mptoms; were

identical '.ithi tho,- Fseen in 3FF .-hliclkenis (Table I:I.

Infe.: r't.c waS 100o1 -nd m-en gross lesion ccore ma 3.1.

T'etnty-tihree out of 104 inoculated chicl:en? died d i.le to

c -C I co..: ii ocis terinri ; a nor t lity rAt,. of 22.11.

Th. :n;c-.l hcn-tocrit v' '. l d'irpped from. a pre-,?xposure

volume of 30.3 to 20.1 on the ','th day.. post-:-.poutic. The

cl inicnil ;ymptconc, it o~-, le ioni in 1r1 t -OCLLii ,-']ii the

p' of large numbers of 21ii -eniertion sclizontt ,

c::'e : i"' -i.'- i'dat ion of the cecail mi.c hm:. hemorrhage

on his top ;a-i holon ic :-:imin.ntion confirmed th, preoZ. cc of

.cc-l cocc.idiolis s. dcscribcd 1...' TyL:zcr (1i29) nrd Ty-zzer

2 -C1). 11.T-2). I?' c-c:: im'.'in- a u;-c*. lHL :.n ci.:'Ce (f +r++

,o. +++, cn-..ic.-,.--'i. s .t:-c "c r :c- n *:.:: ildin. iccp ii he -

,i.t.Cl" l-,'ers f tr.h' 91 ill :i b ,."teri3 :c.iil often

'c c n aricr. t r3: a : aci r ..CO il 1 ThE! Coctal J ruiC

ro 'tei t *c ic' t iiitr i ;t,.C i arind t;he 3erii pr -otein rac' t i-:'n, F i n

in ', .t:d a. n.'. nfi. rf,' ,t d .. :' 7enti?,n.r l, ;FF an., b:tct F a: ri' -,

firi- Fn. FFLCO-fr, ? i:.n ar; rEp : rted in Ta tl.e II.

In *:,c.?. ,.:-t ,:'t.nial .i ii 2FF chicleni:: Inre.:1 tei ,i i E. tendl]

Chtre :, ,?. 'n Ltent r-educti.:. n o t cal .i.'ium pnr:. ini

?ouiir; it' :ticil -,l it,,uc r i';.iCti-'n .l a t :-',:-: in 3'ct,:-'ii-,

f .u'n i n-id FPLC-fre.,: .hi.ilcn" -.::p. i to E ten l. u,,,

: pared to io totiLal 2c.rLrI pfrte.-in ratic.n of .>-

ini'C cl. r. c'iitr-r:: .


The cec l. nicroflora vz': found to change in chickens

1 to 35 days of age. Also, changes in certain taxonomic

groups of b-:tteria occurred when rte chickens ,eire e.:por,-d

to standard doozes of sr'orulated occ,'czt of E. tr.-r.lla.

Earlier w.Jorkers reported chan-.:c ini the in-ligenouz micro-

flora dependini'g I'Ipoin the aze, feed, and cn'.,ironmr.nt of tihe

chiil:cns. Cli I~igs in cLrtariin grc'ups of c.bacteria rin the

ceca3 have teen reported in caces of bl,'

Joha.nzson and Snierler (198) and in cases cf "blu.ecomb" by

Taqi ct -.l1. (197l r ) Enterooccci (S. fec-l is. was the

dominant orrianisu of the, ceca of neuly-i-htched clii::'.,

E. coli :'is present in small nurnibers and bec csme dorinriii .

onilv in :uei:-old or oiler cliicken?. The~; results, as uell

as th1e occurrenc-e of larze numbers of L-pctchcCi.lliu Sp. and

.. Poli in chich-:.n aed, 14. to 31,5 d.-iys old, and the con-

stant isolation of r ic' rol.-'s sp. a ree with t'i,

obt..'incd yv earlier '.o:e:or. in cliic]:, ns (Huhtann :anil 196.5; Tirimms, 9 3) and in tur!+:' (liani ct -i].,


R-a.ll t al. ?17,.,) studied th, distrlLution of tarc-

teori in f,:-c.s of s-. ic and id. R' ifi-- l c,. l, I other

lactose ferriinte..r:;, lactose nonf'-ri'mentern 2, -r.b-r 1 or,-,f.cii

p iCl -:tridium tp., nt er o-:,ci a-l L: toi:-'?i.l : p.

T .-:; co'icl.ud thi3 dijl' b t-Uion ,:' I a.: teri l ,c:-i iS

zinir.- d:n iiin flce a ird c-r1f -n I: miit occcr i a:3 ii.:crzet

mi roc, oi, ':: ri t 2r t i t" a in 1 idi ual cell: i'i .5 iL-..'l- C .

J.loneS an i rift Fih: (1 ?'- r'ep:'rtei that. in ':,i -,i. -ct -: ri

,-' ij o, ie, ar-ni -.a. t .i. iu .' ,) azll., r'epvrtCa

Ize cc,,irr,.ce .t I ll, nic: of LJCtt Ia or th, Wall: i f tNJ

_,:tr'; t rin:t'sti'nal tract id 'cc.1:nts. .olac1 : et 9i. 2.9?,71i

u. i,'.r cc'.- eni.,:']ial ,dilution techn-+]'iqu.: r n:'' d more *.cacia-

tion o r-. ::rnp e an 0a I 1 1nim ? for "I.'-. r' al ,r'OiLpS

.'r :rg 'i Z.ii: T]i. the on c.:pt .[ ba le- ri occu':rri-. in

- ol.'ii in- fe' l :'-n 'les i'.-l i t r ,n- .":troirL :.tiri tract

:.aI'z. n'-?irii' fI.l : ci ic.--l iintrp'el a, i.n c.'f la d- riv, d

*r, c' cnT-'to'nal iilui ti,' fechriu41. ,ifficult ,e _a.s:

Cc i- ,r'matic, r:, '. 0 'C1'. Ui t.u t i .' ;l t1ut de CiC. ii. l.at3 -tnd

not i ri :: re the ,?..a i ual nuc'jat 'f ,' col i e' p, t C. -

t al. I'126 ) r-c p::rt'd 1 .?:.s fui i colat"'in La: ter.r id

*p. from''l SFF :iii after :. igot-ou. : LS t.bif' f rt e n. uc- to

eliIt the nltr'.app:, h .Ct-er'ia. a SirC. tIe diluit ior te-chniqu

h.: uch i:ri'it I l' i i.c th- Lec.lih ique of t :it g

c.lipl.'. usii. .i-. :taai.'d loop I c'..'::'t .pC l a .-t. u1;ei in

this: study. This >, qu-: h._.: b n u.:.e.7ssf1 i.. *:.mploye-d

api :,c','tll i'-eilt c"'V.'.',ti" ,il chic':".J r i l ir stud '

I'i' e' i .iti i 1 .. .in, ; of Hu-e f'- n,-rcr- -..f F :' :I I ,l 'i .

Tic a r'ar; ccurrCi: of p'i, .: I p rf'im .- hid

thouii,'t ubl. 1 t u.tonu in natrllre, has b- en "ariourly e::plaincd.

L,-v ot -1. C1i 75 ) ,reported tha.,t '. e. chii tC. "r_' in,~_n_')

liis p-reentt in day-old chicli in "inlfected" quarters lut

not in "unintif-ctc" quarters. They established a correln-

tion l.:trc- .', the -:feccts o:f i-antibiotic.s on the o,'ouv- biird

in the prscronce of this or'-.rnism. Hunt.'inirn and F-anLsack

(19%65.1 failed to substantiate thiir report. ;.qi et al.

I19' l b) foundii clo.-ti-rid ia :cCtoce nor nfermeni ter-s an.i.l -

trrai--E :p. present in turI'key. early in life, but: the

or-ganisns decreased in numbers or even irere absent iti older

turls:ey. They believed that such cl'rgaiimc on account of

thnir pticvcnic and in',v:i-;e prcrpertics, are eliminated

by tle drcfn-- nercl'lnisni. of the host, as reprortcd by Dubja.

et 1] 195,) Thi cc-ntenti ion L:eLuV- unlikely sinieiC TinmC

(196l r r-ported r--ular i"olitic.n o:f PE.cteroide. .p. and

,i. welchii ( r"'frill:e .o) from cec.a ,f chici:enz 1c days,

7 ueeks, and 5 months oli. There species there, hc..ever,

absent in the small intestine.

In the prr-enit study, C. nrrfr'in;"-ns uinn i.sol.ted

fron SPF cic::ens frequen:tl' Th e SFF ch irlckenr ii:-r

henlt;,, nn7,r-i no "infec t ion was noticed. They were r-iised

on pasteuriZ'ed fl A aid ,ntr. Al:.- t, "'i il 3 sp. rr'.re

isc.lated from tliese SEP chic,;:r.-,: 1 1 -.: fron conven-

tic nnl chick:ens. Ioroover, itin ,"'reemenr t :.'ith the report

by .nJohl in.-: en-id !u rle i l -d-. ) ini chi"ckcnt c*:.:t n-.-iedr to E.

tn-nallD%, a stilmulttion of ,roJt h of C. pr.f'rt'inr -:: occurred

b tth ,in 3'F and cnre'tit ion l ,l'c;,k, -- :ifter e:pc-'ipo r- L..o E.

t ~1'11i I'l c-onventic.rnal chic.en.: w it cc:- 9l c..:'c: id i : z,

C. fp rf i n' nri I.- re- ul arl,' 1: so. 1 a te in co.nt razt to. rinn-

e:po:ed c t. rol- iri whiL t Ch his o,'D ::nis 11 isZ l ted in

nl:,y 2 c, ,t o.f 3 t ij In 1 of' those t 'l.:il tne :.tIler

cnic,. of the e::periimental group utre J.delibrcite;i. inocu-

lated I itn thi: cr a-nil m. Tnle mechiani ms b. it hit:.h thie

initeztZinl bacteric l populi-p t ioni is caui-, to fi',ctunt-c

t,:cre :not se--,sert ine.d in tnis 1 tudy, but a['rctor-r su'c.i ac

int agon.-,Li m b-eti;:ren b cteria, conpet itin- fol- co:imon

n'utrtients, and *halinr'e i1 pH *-.rd o:.:ia tidcn-reduct io

potential ia :,' pisy an irp:ortpnt :irrt ,Hentg,,z, 19o7, 19or, ;

Lev e- t a 1'5"; iie.'ynell, I0o,,; Schaed.l r -t ., 19o:., .

Thfe pi'v- ent 3stud,' cotrfilrm the ob -.e-Pl';aion :of Jo nhasi'iZon

and Sa-les (19?.?) that :-jrowth of C. r,,tfrirnvrnt in the

cecal flor f citicK: lich. an incre .se durin r infect: ti1

with E. tenr ll a-i ;i.r:.iz,th oi f L ctob'' il, s p. s-hc t .% i

rEd.uctio. In 7-day-, 14-day-, 21-day-, '..-day.- and 35-

iray"-old crilck :en with cecal ?o.'cidi.:sis, a wcuppresDion

c: .:ro.itr- of L--toi : -,.- llu :p an a sti mul-iti:n of .:-roui th

o, r P t-': ides p. C. p rf ri I d .- i : c. noted.

Tlie fatc rl 3 i reccinsibl for i ':urtpr.:.' siont or an n croLic

ZpCr-ies during accal cocc.idio.i- "Jn.:lL .c I iL'L n -'itly

in no rh-,mal, h-ealthy c hici:. ns anrrd 2 t imri1i tion or a spc,:.ies

.*'*hi O _,:-CUrrs only Zpar el- in ni-. Li c-icen:. I1 not:'

;,'t l:t It. rialy t- that durin.l- thn prl' .r i t:t ani 'o

patent pt.riod .*f cec oal c.:occi io:ic, fa.ctotr :. f itorl'i,_-

-::rotth nuultiplir at ion or C. -r.frirr-rn ns ma-C tbe re-dlily

avoilibtl'e ;:ith a concc-iiitant reillcl; icn of tilhose fa'.'ol'ii

hjt'Couthl and multip i-i.:t ion of L-,.-n to. c ll sp. A dilff'er-

enc: in -ev.:-rity of tl;h ce Cc-al Ccc.cidio inis syd.lrome was

nrit itd betucen chicl;cni rioncocontaminaten d with C. p-'-rrrinjens

:i'.l chi-',: ns moi oci nrtamn i nateI d n .it ii L ctob. ci ] o llus p. In

Lh-; lforrner, tihe pith llo:i13c 1 r:iani fectations tiere more

sever.: Lhiirn tlh l tter. Al'c., m ie c'er. p'Atiology

du'.elocpedl in chlici.cnr mo'nocontaminsted either with 3.

f'-.'li.-, E. col or B-i-er oid 3 -.p. t ian in those m rino-

c-ntc-r.iir -ted with L- c-tc-o cil. 1lu sp. hen e:.:poser1 to ct -ind 'd

doses of tei-,11 .occCys- s. This inilicate7s that the in.-

Cluence of microbial fior't in the rpotholog-y of cecal

r. occidiocic ci1 epndc .'upon thle p..:ci-e in., o l-.lcd. This ir

illich more cl: -irly ,viid c t ,hen t h: pat lo-l : icnl man ifesta-

tion ini chic !hen: mronoc ont .rli;ri t,.d : iiti h -ir obic feral

.trerptoco.-,c i -ini also tho-.e m.i-noconi tminoted uttI C.

"-1 hi.n c i a it cnpor:d ulth that seen in hiicl:enr mono-

cornt.-ini-terd with otlhi.r specie. of microorcainisms. Little

p th iolo '1y 1 ': ;-cted' in c;hic,!:eiin harbolr-ijn cnly o an er- obic

.:cal s1 tru-pt c. ci nj, i .cul- iticor, o. h:at-kill,'d .. coll,

C. n- .-r 1 nd S. .- c.i di.- not inflr ence the de-

v' el parent of ." 'to ,i cor p-t rli.-r y.

l-thclo.;,: andI clinic i1 ,rptLon: vere neiotliiLible in

chic.:e;.- no:ic'..:.t in.t d i t.h C. I- i.-n "a- d. only mild

to ,n.odcr tc 1 ';ions d ":-lopd i,' chic'l-:-:ns m3nocontanmintedi

ith Pu.- 'pecie Uni -'r natural col lition ic., fungi seei

t': nii:ve little iinflrYuence in the developr ent of cecal

dicsis In r -i ;, o.. .ln-ri -inal n*?!hckcn funr:i occurre-d

on]. intfrequently. Th i mnar' be due to the '- nt a3oni nm

beLt ieen the f iunr arin b ac teri.i a iljici ic 1 iell :nomn in

rilem rbicl ,ic Balish cnd Piill ip ( '1 ) founl thiat ini

the preenc of b'act.ri, C. 3lbi'?ns ij: able to produce

n 'O : l rlo in the p:r. of cliic.'l: en but in the -bs2enact of

Ioti'r ic trf llora, Z. ?1 bic n pi odr .cd C crop I.- 1:i on:. In

noninc:c iulat".) S F chi-cl:~n C. -i, :'i ti us3 more flrequient

tliain in rtioninocul ated ci otvent ional clhi icken. Infet tion

.i hi E. t ri. ll -, .-.lid n':. influence t 0 e ',lroutIh of this fun1 uS

auind an inlibi tic. n cf i touth I L s'ent n in.?e iSoa 111i':c r o

f u'lii in chi -ien: lin.'iinr- ceal ioc i3s r:ac infrequent,

bo i h in S'F arid .:onv:e titon l e n"irnonr its.

A ,cripa'i s3.n of the pattiolorg of c '?al c,.cidiozis

in SPF, convent.ionil, oivn pro, 1c .: nt ar i in at d c-i hiceins r'"

in pla:1 ti filml isolator': cl-e.rl s. how:- i l' i-e influence of

tie enm i t--:riezlnt aid micr.flicors in -ecal cc?.:claioie i ,'n-

dr-o e. The- moot o al it rate rom' coccid i' r' i iC aI-2 liFier in

In fie.t-ed SPF c: icl:enii t Isn in i i fec ted. ct en'.:ltion -; i chlick.enc.

T! C-c'u?'rri n e and lthe n ur-.b r ,f :olo.: ie of y..i r.- t.,'

i' is l ; h irnr in .FF cii wienz t!.n in ? ,rn'.iLt ion I .1 clickc-:ri-.

Tfn, ir the onil'.' ubstantial dit ferenrcc in the ,icroif'r.

of tihe-e t',.. gre-oip.. In polyc :*:.'nt :i in *.-d chii l;en.- r-iised

in p .la tic.: i l icolatc ir ( t ri-. r.r t; n i de 1 .-n:i-it d .

"co''v. -r-nti C'nalizCd" ch,.cl::.n:' the- [r' l it,' i- tiLe ini Ilirh

nh-i.-n C r-., r '-i n '- :n ] .. r c -l.i ie.r- rin:-ml:,. of the

T ic roi lorni. A r po A ib.le role for' these orc:'lriiisriT ini cau ing

ae:ith ii chicken: cannot b: ruled out. t-ilichj aid Flililipt

'1966) reported 'va' cul r and r'n-'al invasion by cnterococci

in chicl;env and EDomernutnh and Ci'o:.-. ( 19I69) reported acute

septicemia ofr young and bacterial endocirditis of older

birds, due t.o C.. ,'11s. Dia'rhe:L, lecion- or h:monorrhagi

necrotic enteritis and dc-ath h:avc Lbeen reported in nruborrn

piglets die t, C. prj.rrri nrl n t'rype C infection IField and

iGib.on, 19.55; Barn es and [oon, 19&6 ; B.e.'sel'ind, 196 j).

In co nvention-il chicki-rni the lesions and reduction

in hlenLtocrit n.luez as a renrult of E. tL--n-lla infection

uere more noticeable than in SPF chickens, despite a lover

rate of nortality. Johnson nd Reid (1970) reported that,

in' certain cases, leFrions will be morc severe in ii.'e bird;

thr n ir dead birdc during. an outbrcrak of cecal coccidiosi.-

Trie ire of" hicks to recover 'rori the .-hock due t, the

1 .lr. number of developmental stages within the mucosal

players suffi-ic nl tly c:. l;,' i t.efoie Ecal1 bleeding occurs

may b-: influencinc- the dcath rttt i(E:rtl., 1963 The

i'd. of b -ti.erial species lil:c C. r='frini ecr and q.

fc,--lir in ths- initiation of this tu:pect,: d shock or in

the failure' of chicls to re-.'o'cr 2'!o thi. hock, if any,

1c not k no' i.

.Anothher f,.ature ,f cecal coccidiioni., un-rstricted

h:-,nor agn with ino coio-il :1 ti i".. :el.:urn ,eein in *-'i.ckeznz

harborin, o:inly, :. fe:-.1 i ..,ct r la Im -y hav.' ?, d iniite

P.:.c-l in' '**epin t lpe i 1C -i 'rc .n: f- .t1 rit,:- r''ly-

c.::., t:-.iic, ti: 'n '.lith ,. r..:rf.r_'i 11 arid _.. f,'e'?-.11- ,5r C.

ii ;:.:,-. r::n itch in cl- -i: l : Ecal coc, i.i :1c .- I

u li 'h Hi: ,.':.rt l' l .'.. i' i i'i L ',." *'.;I I t '.* ' r .,,.',: t-e: :p .: ur-,ll

*.:* thr.- 7th i. p, : t,-c-. :p : : ure ;:. -l '-, C i u rr,:i i r

rF Sur.:

TV i i ihiu ci th.-i.l.r io a.. F Irc-'i r b ny i -:'f I i

vari,:,u- t :.,:',cr':,,1 ic -r,-.,.ip: i: r,',i:,;e evi id I.i dhen I 1 *.-?:,-,:.]l

FFV.-Pr, -i- 'r e.- ,':hicr!:,:. i* n :;p.:."_ :,,e, -.i, t L" ,I '.,' ,.'. li':it- : '-

di'i r:'t .1 "c-- i ".,':, tr,. rqilhc..-,.i .m.c-i c l1i, .:.rf cc .:'e '' i':i .-" i r i

Tie to.:. E. t--.-.,e1 -. l.:. rr .rtali t,',. y.:, r.r d in thcl.: g .rcu .: -.

.il. :i,-. .-le ,:.r r.tul t i- 1 -?.i% C. r i.,c-teri i i zI.:,c; -le-

,l,:,: C llor t ali -.n I oo dy :.c0 1 co nt.en- r i- e n t.-d

,'l:/ in t .:- *?m,:. p? .l:,cOr.nt.iii nat t :1 tl h c .-:\er .a-,i:-j p.,

E. c l : C. r, r'-ir n. nd C. r-a1 .. ,riup m: Cn.-

: n i ir m jat.,. iitrh e. .- t l. ..-].p:.d ild ie .i:.n.-, but

did ct' cm r.oc.-t ii- or bic,.::i-fillej .cec 'Th. .c-.

c aii a' ,c :. c- Al ,,' rt' i tiil .' r1r- r iaii:. 1 ot e 1' .:.:- .

'"-.:: I.. chic :.n t .- .;-ri'," .'.ll- i : _..r. -d ~' i".

siceprt i.-: to E. t 1] tti iiic: cr . ., 2:r t .-

C1. Rc, ,c (1' ,.7'DI, C:. .1-' .t"- Lth- .:.,'-y' it-. iu ,: f F. ,'- i>,11

c, .cy -:' tb'-,rv d : :,.1 ,;r' ito,.r -.:--.:y r i. ,:,n in ,i-ii .k

at,:ed 4, 5 and ,'ecl.:. rind 's.? suc,:c s Lful e:':c.z-tat ion, in

1 d','- Or 1 ..: .--old c hic!:n 'h-, reaLcr susceptti-

bility of the 4;, ., and 6 iE:-l:-cold tchicl'cns w has thus

?scribed io the -icce..ful r.,-id -.c toit ion of ther

oc:,jcy:t and libcrction of 1aryg nunb-rs of sporociit es

in tlih c-,eal lui'en. Ti' ]o" l- .I.-t:pt i ility of da -,- ld

or w;ee!:-ol.l chickens ;'-- ]il:c,'iS.e ascribed to tniii less

Iucci iful :exY," t'ition t of ocyC'cts as a conz,'equeItn of the

less powerful 'indin~fiE action of thi-: immature gizzard and

,-iearth cf rufficiert tr'p.;in in t'rc new.jl:,'-haLtcl'ed or young

hick:cns. Tryps'in, bile, ind h'.-droa:cn,-ion concentrcration

r' c:orme of the fa.ctors cc.ritributLirin to the e:.:.'s station

(Levine, 1?42; Ikrdda, 1?5,, 10 '., 1960; Hibbert et l.,

1969). Hiib :rt _-t al. (196'?) fo.uid that oc :st s of man:,'

species of E -rin, incliiu lin. t e:--ll, i, f:iiled to -excy: .

if bile alone or t;'r.3in alcr- is present. 'The: also

found that ooc,.:ts of 2. Y.o'i .?nd C. ellir.,Oid .ls re-idily

e:x:c:'st in bo'i,.i: i il .: cc Ot-irni a hecv .' S.ip.iCn ion of

tb-ct'eri : an fun, -i. The cr.-cific action of thie bile :r

ti -e tLa te'ria :-.' f'L.--i in n.? e::c,'Et-tiio n is not :kncl:i n.

S..tnilr 'l -i 1 lt:r "' 'i1..ii of their :: .l l of the s-orn-

lated o:.t:y; ts : p'?ll. e in t'.- ferT.s of .x-3:.osed >cnic?!.ns

"3-s ob etIrv ty LOt -1 L, I1 :-'.. It is pO:; ibll

that enr::,ri' e i!:e Lryp:-in c -n ent ier renl il .: into the

ooc:.-t Ja ,:11 i : r11c ..:' 11,' c!-1 ,: The pre c:nt

cludy in.dic.itcd that n.:-velo .'. iat of coc.:l cocc idio;i3 Ais

r* p rul cf r4pid niji r: _t.r rate of e:-:c::tr: t ion scn tO

:' .r in .l :.'--. id ii '' .:r- 'rcl iT : O--C. ': .i L' t i al

. [.a..: -. i' n T'i .I reI' l ril' i :. i t :cd i'rom cec'- -f

: I', :' i r in. r t;,' l:-pi l li r ., : 1' :c '.::51 C, cI c c idi .-i;

01ii c. C
lic ic -: :'.:'ccc: fril in .:-i"'-cld *Nl'-: r r': T'ncL,:,Ocnt-i !'in ted

i t, t. 1'i In *c',ici:": uI .c: Cold, 1.- z: C'c : f-' .; ul

an -d : ... t t ti1 .-C IT. tc IC MVe C occurred I. d IMi ;; : .

C'e: 1 .- 11 .or both wa h'a rbo' ed in th: c ec: if

the 't '- [ fA' d c .: 1: tat. i 1 : C. rr[: i '. : t.d ith '. I

,d v lic-rie i:t f i- ,di -~: Ii'n th ne:.l- l?. t-:hed 1n-.'n-

t' i lr' : i': t e onl. ti r mi r ,; rga'i- ?i. Fi'ed'ii ri na l ,' '

c. .-li w nd _. *:11.i. It iu .s 1-' foun1 d that the p.-it"ij.-

li:.iLi.;ni r.anif:ct-tin: o f coccait.1i i wtere greater iY C.

:fr i t'!' cif'':' I::c i tc i 7li th either S. re-c:ills orC E.

col i Une her C. r-r 'ri 'e:.7 soids an :hi c:*C'; nation :.f

t'Ie .cUC": .," i t !.ki-n i[ d Ih.sI not be.-:E L t ,iid, ied.

r.! ,-I l t':'c.': r '.:iJLF 1.1 s.hubs qnuent i ber' t r ion of

Sp l me ? not be I he -ole r'o-..?.: riich will iAn-

in E. e i 1l.5 in-i' .:tioin H,,rtor' -. iit'i t l1 I I'[ l.

r p:,i't ed c i't'"n l t;: -.'.-- c t ion C '' 'r -i pc' .: trati :ri

of 1 ii. e .t- ?d 3sooc : it. : I ..u. 'C i,:ei: It i 17 :

!',o,-ln t t much o f th-: paI l h e: ,i In *:e.- 1 copii-.3 7i is

due to the m-rr: ...-I .-. o ..Ior "' i : of' t he la --. -nd

,?,.n tL -. :c. i-iort: r iii-:.,, d- tAr,: lie pi thel iA c 11 ,

cnusing slou-hing of the mucosa and hemorrhage. The role

of bacteria and or fungi in favoring the rapid development

of the enldogenouus Stages, if any, is not kno:.rn.

The study show-v that typical cecal cocci-

diosis originally described by l19291 and Tyz;er

Ct al. (19321 failed to develop in chickens raised

bact.eria-, fUmlii- and PPLO-free. ilo clinical symptoms,

mortality' or decrcase in hcuatocrit values fol.louin.- ex-

posure iuth standard doses of sporulated oocysts were

observe d. Lil;ewiise, no visible lesions were noted in

thcse chickens. Chickens raised simultaneoiuly i.nid: u r SPF

and conv:ntionral environments and inoculated urith the same

.tandnrd dose of sporulated ooc.sts o:h hibited the classi-

cnl ecal coccidiosi- syn.rome.

Thlrie was also a marlk:ed difference tbtwuen bacteria-

free and SPF or convent ional chickens based on hlistopatho-

logic findings. Ceca of the SFF and conventional chi]:K.

inoculated with E. te'c!ll' oocysts and necropsied on the

.th day post-e,:posure showed e::tensive denudation of the

nucosa, h.morrhai-rs arid large numbers of 2nd cn-neration

shiont.s. In severe cases, the hesmorrhLe:ic and tisCue

dfmnac even reached the muscularz laye-'ri, and the entire

mucozal layer ri-' seen to be? stri ppe do'.-n to the ba.c- of

the submucosa. Large 2nd generation scni-ontZ lying mostly

free and a feu within their host cells -ere the prc-ndmicant,

cniogenous stage seen in such c: es. But in the ceca of

b c, te'ri:-, f' i ..i id PPLO-f 'r e -!i,': c..' .: e t c E

t-.: 1 o.o:.'t .nt m l necr. ': .i.i on l 3th ;, o :t-c. t .cr. ; i -e,

o e::L:n 1 :.': di iNi-:t n n. ': i '.a, hi .:moirl'i c. '. i ri e

." !i.' i t -' _irt _n 'Ji:c rr j : onr seeri. 'ThE : ro; osai lay2r

iow.': r,-]. r c: le." i t ..;t and c.t fic.: r '. i -zl '. L ver'C Llni'ld iaSl a.-ed. ;

ni. -iIl -e :nd ci.-t:i---cn r: chi onit uere l o-:n. A -?ti en-

- 'lo:.-:rn, ti: t.: ,.L : ideni ified ?' iTim a tui LZe chi-. .nt : it re

::ti.:ti in ti l:. eitnelial c-llz ; f the r ucoz' : .zpeciall',

1io.. lliii-Inv t e giitd f[undi Tiii location cf th a.e;.,

.i'u--e:t- tin t ti--,' a n be t: l t 'c -m r ti i on :' hi:ont -

dev.eli. nr-, .cz; l7 ,l/y. liti- ,g t that in t.:icteri;.-,

'ifn i- -id P LOF' -fL*C Iic!':n the ic,:rree .rid rate of

lt .' r]n t if ,'.d.;e o. cta';-e. a .e m rarke-1.dl., inhi biLed.

T i' i nhil bit i':.n i d '.': 1 1:c ...i If cir-cd c T ta.;e ,

e cz,-..; 1 -iy t he1 p. t h- :7 .ni si ai. 1i cno ,'- t-]nic 2nld ne r -ati. ra n

' iizo.n m.;.' afM.i.3 pr 'otEctiion 1. tc host aind the pa- ti,-

1.'"-icil anifer t-t i:onc: ar.: no:.t .I-.-elc.pel. In r-act, the

ta:.ter i -, futltti- and FFLO-fre *:nhiic:enc e.;,i to:- L'e-have

-i. Sn b ariLicu 'line t Lhx a c : cat i I ? i '- 'c-l in the

c t:t inc; eicic:,e;' r;t .e: e ro n i the Pr:' c. lls

: t -'..-:'.:'- t i o Wi. I-i: t c h:1ii A i-t:e dii 'i elorp-

[ 'ent .Cin Lo i a t.- r ri i rd d. el :i ,1 F1. t 1C h ,

11 ti,; .li I0.. ;" .8Kl-0 bTo,.c: r,_-- c_ ;lilcl'...ri., O".at ': 'cd a

12-1 5 h utiL ay iii t': .".Fpe arnc'.- of 2nd ere r -t t i.:ii nero-

7:-, t i r tth C'e,- of 'Lt_ Cri .'- -f i C i_ C' : ' I H-:Iie er,

tie,' f i.i.d "o il' iti 1.i1: ,cOrP-e Cf i *n.'ction: in -ri.-free and c ioventinal chickens. Thic nact

experinenital pi'occdur'cs and re.-ullt are not conpl.ctly

I:noim to compC, arec their' rc.sults i n ai th,: fr tIe prPe ent

t udy.

!ait:enbach et -l. '19'66) lescrib'ing a method for

.terilizirin cocci.dian c.o,:;'sts employing Clorox: and uil-

i uric ,ci -dic}ror.ate solution, indicated that theY' coulll

prc.duce c, c.ccidiosis in "g iotobiotic chicl.ens."

Since the e:act nature cof their "gntol ,biotic c;:ici:eni"

ha:r not been indicated nor has the description of th

experimental pr.ce.iure amid results oft thc ji i o.rk co.en

given, it iZ nrot pozcible to compare tlecir r-esultz ritl

the present work. Thcir ,orl; u-. Lt o obtain

sterile o,.cyts for cultivation of the pI.ra itet il tircue

cult ure.

ilybcrg an-i :narpp (1 9",3, 1970b), e.:mining the

Docsts of E. tein.cll by reans of a scannruing, electron

microscope, reported certain structural alterationci- of

oocy,'s:ts. ioiLne an:l Honii l. 1'54' ) report .t that rhen

c:oc;,'st are tre-iteti wit.n sulfuric acid prini, to soliupr

'hy,'p', ilc'lori te, the outer layer lo the o-cy:Lst '-311 L .r..:-r

u'i'.i:ll'.d and apparcntl,' elev:,tet ai.- fi-cm 'he' inner

l.-,ver. Emploc.yni .,e t of C -uch .tricti.- 1] *.' 1 t.i-r'e oo.:z.' t

miiht influence pailtho.enesis 2i11 d.:'eve:lOFpncct c.f r.i'-aw-.

processe: of r.c c cidio i In tr...., t-h u.e of C.

per.-cetic acid for surf'.:ce stcriliczti.on of in.-.:rt itlA'teo

*:-.-e- "id ooce yct; his b en found t c. be- .'ry -,' rfect i. 'Doll

-. ,'" ,:; i ', .;1.,' -t aI. i':,,.. ; He- dc, e t l. 1? '1 ;

SprinT -r et :1., 1'1,0). lUlnd r t le lii hlt iicoi' zc,.op'e, no

diTi ', ,r chTi e t :, the oooy,'st 6.all I -I -:-. n I 'hen p':ra-

: -t '" aci d Ui2S LIed ri' ul.ri'I i e tCr' iii t i "ton an, t hlere f r'e

it is a': urmei that p r:icete:ic acid i i'ot it tinjuri'i t. the

co..:zt iIll e- n i.;. d accordinT- to the mTctlod of Doll

-'t i I i'?1 .1 and thi? met:lid i ui.: .i. duirin.-" tlo !..r-esent

: tLid:,. Th'- l'3t- bt irined in the pI e czci .tii .-l: can be

,ompIP-r'E-d t-: tho- *btairnci by Vic: anid burns (19 .6,

quoted b,- il-d ._t ,I., 19-.9) 6..ho b) zerv.e no m rc.t :li t;

in, bactcri C-fr'lee chi-ck: rthe-i e.po?:ez: tc E. te n.i11- c,.cys t

anid by Kem t l. !19'71 .'i h reportedly d l :'ini of the

.- '.-- .p r t of tlhe iendo;:'-n .iOu t '-e in rO ii-fr'e chi. 1 : .

In t pr trt otudy, th ere '.'a: also 'i delay in tie de-

'*:lcpmri:-t Of enl.c~ enou sta e.. b rv a r and tli: d.- lay I.i:'

be b ner .-f i ia I allc.'inri tai nocIt- to de.~'c ln 1 i't'ficilnt

r e.7- 1 sTr-ance and t h-reby inhibi t oti iie.'elo'mcin, of clinical

symptom: -ni O:th er itit'e tati:nt i : of th: dizense. It, is

=.1 o pcs.ibl that thie d.;.velopI nt [" the- Iir-: 2rnd

*.tenerati:rn s.cI..nt i" di l.ia:y : ,,,'r inibit. ed by the

ab.nce ,:'- mi L rcr r ill i- d to inin Al *.: .elor:nit

ofr, piti.holo; :, Inh .:.i t ion I -' de :c.'ll 1 -.: n t :i'id developme nt

,oi i.e r n e ii br oi-i' rln!-atc p- r" it : e bCein fo;.iti, in

Ci'n-iri'e h ,ct includin.- t ,i' C i: 'n (Il, 1i 1 '59

J.Tohnzon, Sl"'] l. Spt ii,: r' :.t 1. I1 i: rep r.ted .ur. ivinl

of H. rlm 12l'-: idi s r ithou1 t d1 Lca- e product ion in l.-rot abio-

tic hosts. Confirning the -synerzisftiic role of certain

bact ri'l specie- especiallyy C. crf r ein 1nr) and H.

melea7nri1- in the p'a.A-ct ion of infectious eiitroCiCpati-

tis in turkeys reported oy Br-dley' -t il. ('.19l4), Springer

et al. t:o0)) established thit tle role cnf bacteria is

more vital for the -urvival of H. rcl1 i.?nr'm-r l5arv\e Lhan

H. mpulea?'idiz in the pathogen-eis orf infectious entcro-


In E. tenell? infections, the e:::a'ct role of the

bacterial sppccies i, not no-,n, but the proec-n:. :tudy

indicates that for the dizesce ; to do. clop in

full, certain bacterial p e-cies ar-' e:7ccential cince no

mortality or gross lesions riere teen in hosts frec. of

any detect ble ,org'-nisjms Johncon et -l. (1l67) and

Roh)ovsl:y, andl Gricpe.ere (1t'67) found fc-line infectious

enteritis in the germ-free cat to be a mild, nonfatal

disease .:onsist.irg of leukopeni?:, thymic atrophy ani

lymphoid depletion but I ithour morphologic inte'stinal

lezionz. On the contrr-ry, CFF -cats developed clinical

si:; : f c typical inte'.tinal invc'lvenmnt a.: rcn in frani:

infe' entcritis. Sincea- L- ,e cecn of th. ':.hick:ns con-

tain the maior porl.t io of the lroii:.' ntin.l flcra

ITimrs, 19r,8), E. tpn.'ll- may have d.c,.lo;-pi n apparent

obligate rclatiorisiiin .'ith c r't-ain m-: i' rs of this micro-

flora for pat hoseresis to commencn.e a,.-. this z: lntion'lhip

would have influenced the over':ll host-parasite relationship.

T)r c? ,t i l i. n to f U:i i .r .u 1:. : c iO-7 r th

'- 1 ? :. -,ci'iii r',r i l n prod 1ucin c* 31? c. i1 sic s r.;'': i 'i i ir O I.r '

r I' r- i po:cl.-' d tv ,' t i 1 .3 '). ii- n f'". I p 'rt,:-id

IL., 1 TMrerCAc; f c'31 eoc;idiosis to be Aimi3r' toi

;io c' c : th ii .i core '' anid h: rrh-A e. .and

-am le"PH u1 .I ;Di: r'port1d a po.:i.ibe irrvolv'.M.nt Ofr

E ,ccli N the tii.._y c, 1e?? l ,co- i.4 iSi Th,?ir

it .dy 1 r -i,. th e p ,- .,-: tt ..Id :hi i',.d1 tIh?- .c. if r_'.. 3.:-

prre>-ut i 11->g- fi.WiriC.7P tIhrir.O Iirut tIm c* ur'- f cc-c0al

-".:*:idlio i- Th' ;y, m imed that thel r.:duced z v:-rit:.

in oile..1 :h c c di.P: to' 1 c: li f rm c-cnt ih thL-'

eum. The' rc uc id -'verit of ,e ai ca cidiA':'s. sr

'n da4 '-o,.i ohi,":en- ?, .lsc be Ikt tc t.hc- Sa-,T- reason,. '

Tvpical c -1..1 cccidi-,d L did not div-lop in

b.?ct-erii-, 'ni FPPLC'-rfr.c: -iiclkenc iinoc I'i ated uith

r .t:niidard doie of infective ooe.'zts : .f E. ten- la..

Hodi- v.r, rsnail nuntbert i- t.'.1 ll] I were able t c dev lop

at 2 slower rale in tir.-teri a-, funLgi- .anr I PFLCO-frIe-

chickeins as co mpriF:- '?e 'ii th~ r1 te i se r I in co inv'.nt i 'onal

c'ii .c!:c. In chicl.n'' n-once.jt- i .ated ulith C. perfrrin-

g;nc, . fc-alis, E. C,.li, B t- r,-,ii-sa sp., L[.T. toh- ,',11.r

:p., i,'nr ,p. or_' C. :--.1-i.-' n ..:pal r. ti,: E. 1 i- 1 1

oocy;,ts -res'ulted i nnii c .e l cocc-:idi 1i C.:c.:l c, ci-

di'.si i; Ch l'iii;crin c.l '- ntamin-ita d with bacteria ia.

conrtpani" blc :.ith th:it .- n .en in conv.-Ltiional or SPF chickens.

Hi lCher 1mortalityv bun lot.,-3r mein ir,..ross l,:ii.:'ns dLie toi

coccidiosi. seen in SPF chic!l.ens e::poscd to E. teni-11i,

compr -'i-,.--,d to t'.i in i ni i-'trio il chi.-l:ens.

Iridi-enou'i; i'l 'ra n cf tlihe Cc'~. i of, chic., ei- oLI various

ag im E nor nt i.:.l. Tn v'.'r V'.i..n: chi*,':en.; t(lu to 1

ti.el; of o.. ., rnt. ....;.'c i pr. '-. in tile coca while

in ol.-- r cl1hi.!::.n3 4 t- : -- L -- r oD, -.-.-. 1 s Sfp.

Dro',e o- inr ted .--t -, '; -- c. L-.-'t .. i llI' p.

and S. r I -- i : '-.r' p r.- l -C 1 .ir 'i fr.'ri co.:'.'e tiornle

chlic.-n:; a,:t 1, 2, ', 4 i "':c ks f a; I;oliation of


C. p'-rrrijL' ''-". L in'i.' ''iuentl in Ci',V.?in t i"n'al chi'?l'-ien ,

but freq.,ent in SFF chickEri. IIc- ul rticon o[" chiickens

with E. t --n-l 1a rezilted in ?t imulation c" i'.outh ,iof

C. per. ri' -?n-Z both in conventiouial aL-'Ld SPF birds. Ail C,

l.r-.:g nioLmmbci-- of E. coli and .ct='roridee.z p. but low

nurimber.: c' L" nctbcLa1C i l 4 p. i.cere ea-n in the ceca of

inrect'.:-d chicil:eLn-. Tie, indict ate that thel in.dii ennc.Z

bacteria s-id in r'apidi dicve lopnernt cf endoencu: : t age'r c:,

E. tenelli and prolducti on of *', p i C1 rec-dl rcciiiosi .


.FF-IiDI:' A

f* i. oC-- I .o 0 r-C o r'. 3 -f .- %D`. -D
'* N uT. t.r'. r r'a -4 L 30,

W- t =- '.. 0 71 LC V-- r rq 0
S .." -7 e -T .4 C".







-<. C..

i., I. ,


-i :0 0

L r' ..,_


C) U

C) .3 Ci

-a: W

(Ol C*.

U .A 0

., O *..

r- : ,

'2 'i'2





r-, c-


Ib ^

U. i,


*1 .-f L;


%D 0
0 0 3,

,O '. -

L r L'
,,,_ C ,'U


0 0

.-1 i -


: L'IJI I I II .,

1| ] |*. L- .|< .i
.. i- ii i r i i .I i'.'; -* i "

i- <~"t '*'I I'I 1 -i i- i" i Ti i,
..-_ -i .: I

-,H L ,--.-

C. '
1 I C.I
*- C. C,



f c.
1. C,
2Z'-* C *
.. *--T .0 -T Cci

0 C1 r'd f ...* .-- -

_-, .- CL,
.) *.* 0C. M

r- i
(* I r i .li

(n I C .,- .t U CI
*C.) .


4.) E I .-1 V -- 0 -

T-I 2 !H 'fl
rI .. .

'L [1 rcn C 4
I Il

r,.I ) C Ol

- ', 0 .r

C- -

f--- -;
l C r *- '.
S ( .1.
4-. c- *0 C -
C C. '
C j tj-- C C-

*1 1 r*
.^ k;.' '*: "''* . u 7*T 5
i-j & .i' * i ^

,',i '

1 .1.
--* c i

1 II

i ; '* r -4

. -i i.

1 i .- i




I -
a- -I
4- i

c rl
3 4.-

' '


l- 4.'l

L, C,]
1 .1 E. C i


Lj r':

*4 r i c'.'

--- c *-.I -. '1-

. -

rj tc'Z

C- -


r i .C
L .1
C. .

C -
l 1

c, .-i .z
S -I

S- i j -iL

c..144 S 4-.

l ~ F
i. ..

) (1' -i 4
r- i J '- *


.r L. t
C. C.
.- --1

t' l I. -._
0 z- C


C 1,
'-4 4-

i. U

,-1 i T

-_. *' .1.

C- -" ** C


C. C.

-1 0 c;
Q- C

-T 0., 'b

,I, tif

c *-i IAI

C C i
3 G,
", '1,

o ".
0 '.)

> > rj]

.* ... C
,. U 0I

_ .t (j

0 . ,
S' ..i

r) -
2. ri

-1 )
0. C

C. tl) Vi

.-1 C

, L *,.





1 r
[J 'j -
c'r i .

C..) -

-) 'r

-- '-.

'1) l -

r-4 0C


O. .-, -
.-- i ^i

U- *-
o - r4
f .0L

+ + +
+ -I +

ci 2 4j

.r j
-7 JI C C- f-l

I T'
E *D
iC, 4-t Li
11 L i
L) L 1 i

Q i L. i 0. r
C -. H
,n11 O
i' ,1* b, rn

4' r-i o
C 1 .-3 C-

:3I Ct

C-. IU 'K,

'C C)o

'3 '

;: 0c 1

:-, O 1.

'> 1.*'

0 O

H r-
rl .-

0 0 0

r -1 r 1
-,1 *- .,-1


i C
C. .

r-i i



.-I C', .
r1 (*J *3
C c a.

r C

., -. ti'

v-I C' "

2' C')..

+ +
-+- +

1. r T

Ca '- J I
C. .-. T -j
Ij C C

1Lr *C ,.- ..- r. .


s, *.) . -" .

,. : -,' -, .--*-.1 I.' --

I, .-- C-0

.- -- i
--j A C. "

+-' ; 2 C '" r,
"' "--. i L *' .

""- ,' I 1 -

,-, C. r. C ,
"!..- A ',

O, I..' .i- .. "

t-,1 ". L :1 ,

1-I *- .I *l A Ar-i

.-A r- j. I' l

':. I I r ,
,,-. L .
{_ '

-, __ 7 -1

l l . .- I r, .
Lr (L ':' 4-' ".T '; I"

S '.. .. ,' C

-'.- ., -' ** '- *, ,

.' I A -* .

-i -' '. 4

I 0 3
.,- L1
I i *

1* j v i 1

"ti .' ... -, -" ,
C-' -4 --I .. C' 0 'i
'- "- j I I
t'-. C -,-' '-*: :' -t rz -' ]t, L p LT, '1
'' *-. 'A -- t*, I 1 'i hi, ,'
- i i ' "

-1 i --r
I- ._-. ."-. -. *QI C- -

r- > .r. r, C '

- ., "
U, 4-1 44 *- ,ii-

C- j l-- []
O n 1. ,
C -t .- + ,C,

IJ *..* .

--, '.3

.-. r.C,
-1 . ,-
lr 1 -,

.L 0 *- .I-
C^ 'li ir u

.- '. I C4
'l i ,-

L.'J r-' 4-I (a

I > 12

O- 0-)s. + + i
Fj: & r 1 + n 0

4- 2l 4-i


University of Florida Home Page
© 2004 - 2010 University of Florida George A. Smathers Libraries.
All rights reserved.

Acceptable Use, Copyright, and Disclaimer Statement
Last updated October 10, 2010 - - mvs