VARIATIONS IN
T~
AND DOSAGE
OF ED«ERIA TENELLA FOR
D&UNITY PRODUCTION IN CHICKENS
by
WILLIAM EDWARD BABCOCK
A 'l'HESIS
subnitted to
OREGON STATE COLI.J!X}E
in partial .tu11'11lment o1'
the requirements tor the
degree o1'
MASTER OF SCIER::E
June 1951
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AOKNOWLEDOIIEN!
I 'Wish to e:xpresa 'Ill' sincere appreciation to Dr. E. 11. ]):told.naon,
Professor of Ve,ennary lledicin•, for his maa;,y helptul suggeetions aDd
generous aaeiatance in th• organisation and ocapletion ot· thia imeati•
gationJ and to Dr. J. N. Sbaw, Head, Deparlment of Veterirlax7 Medicine,
tor his interest in this project.
TABLE OF CONTENTS
ACKNOWI..EDGMEHT
INTRODUCTION
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PART I - SIZE OF DOSE AND 'tnfE ~UIRm FOR DamNITl
'1'0 DEVEL{)P •
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IIATERIALS AND UTHCDS
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TRIAL I-A
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TRIAL I-B
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DISCUSSIOJ
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TABLE I • SOWARY PART I
PART II - HUIIBER OF DOSES AND DOruNITI PROOUCED
'l1UAL II-A
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TRIAL II-B
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nliAL II-c
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'lRIAL II-D
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TRIAL II-E
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DISCUSSION
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TABLE II - Sl.BOIARI PAR'! II
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PAR! III- Unmtmf 'tDIE R DIRED POR n.DruNITI TO DEVELOP
CCNPARING A RELAmELY OLD AND tOOt«; CULTURE
!RIAL III-A
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TRIAL III-B
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'lRIAL nr-c
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'!'RIAL III-D
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'!'RIAL UI-E
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TABLE OF CONTENTS (CONTINUm)
TRIAL III-r
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'1tUAL III-Q
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TRIAL III-H
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DISCUSSION
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FIGURE I - TillE REQUIREl) FOR DamRITt PRODUCTION FRCil
OLD AND YOUI«i CULTURES OF El¥ERIA TJNELLA
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TABLE III- SmawtY PART' In
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BIBLIOORAPHY
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SUIGlARY
VARIATIONS IN TIME AND DOSAGE OF EWER!A TENELLA FOR
IWUNITI PRODUCTION IN CHICKENS
INTRODUCTION
the prcducUon of imunity to coccid.ioeia in chickens wa1
obaerYed by Johnaon (3, p.)O) in 1927.
Man7 ~rkers have re~t.ed
studies irwol't'ing the production of coccidial SII!Wlnity since thia
first report ot Johnson's, but the vast majority of these atudiea baa
been directed toward the developaent. of a practical prograa ot arti­
ficial immunization in poultry of coJIIDleroial and tarm flocksJ cone...
quentlJ' they are ot a rather general nature.
Very little information
ia arulable concerning the baaic principles ot immun1 t7 production
and the relationship of the varioa.a factors which •1 influence this
reaction.
The size of the doaage neoesaary to produce immunity and the re-­
la t.i.onahip of the dosage to the time required tor immunity production
are two impcrtant factor• in coccidial immunity.
'l'hia il'lTestigation
-.s designed. to gbe data which would provide further umerstanding
of these two factors.
To study this probl• Eimeria tenella was
choaen since the available literature on the aubject of U.unity ia
largelJ' concerned w1 th this species.
A :rertew of the literature revealed onl,y t11o r eparts which
applied directly to this investigation.
Jankiewicz (2, p.2)) pobll­
abed an abstract of hie work with Eimeria tenella in
~ioh
he claaa1­
tied on a percentage baais the i""'Pnity produced frOIIl single do••• ot
2
varling eize.
In this 110rk the time allowed for 1mmun1t,J production
was held constant~
·ickwsre (8, pp.l25-l30) has reported a atud,r on
Kiaeria tenella in which the time neces&ar.J for irm:nuni ty to be pro­
duced from a. slngle constant dose was determined.
Other reports
touch upon the subject but their very general nature make them of
little value in an accurate quantitative evaluation of the question.
'l'bia investigation was conducted a.a a aeries of fifteen experi­
aerrt.al trials 1 w1 th results of one determining the procedure for the
next.
For convenience ood clarit.Y the series was dirlded into three
basic parts, eaoh part designed to give data on one or mere inf'lu..
encing factors
or
isaN.nity production.
Part I conceru the compari­
son af large and aaa.ll immuniaing doses to determine the effects of
dosage size upon the time required for immwlit.Y to be produced.
Part II oonoerns the effects frOIIl varJing the nuaber of individual
daily dosea (the total dosage alwats constant) upon the clinical
s1J11Ptoma and resulting 'JrrUDunity.
1111lunizing procedure which
part.
'WU
'lbe results prorlded a
sa~isfactor7
used as a standard method in the third
Part III concerns the time r.quired to produce i.Dinunit7 from
the standard illlaunizing procedure using a rela.tivel¥ old culture in
oomparieon to a relat.ivelJ' roung culture.
The general aim was to keep the intluencing factors as constant
as possible eo that the results could be olearlr interpreted for an
accurate evaluation
or the
dosage and time factors.
The criterion
used to determine the degree of suaceptibillt,y after attempted imllu.•
niz.at1on was the mortality and morbid! ty resulting from a large
challenge dose of Eiaeria tenell;a OOOJllta.
ItATlmiALS AND JIK1'flODS
A total of
968 chioka
were uaed in thia imeatigation.
1he7
wre aeoured aa daT-Old obicka ti'OII the Department of Poultr7
Huabandr71 Oregon State College,, wlt.b the exception or thoae 1ft tour
trial• ot Part III, which •re purcbaaed 1'Jtola a ca.ercial pwl.t17
batcher7.
(
Both Siql...Coab White Leghorn (SC tt.) and Hew Haapshire
) chicle• were ued.
!be chickBns were reared in aU.,..tal cagea equipped with one­
halt inch hardw&N cloth tloora
ment waa thoroughlr waahed and
am
electricaU,y heated.
~•infected
All equip­
before tbe start ot each
trial.
'lb.e chicks wre ted an aU-.uh ration aad receiYecl tap water.
Feeder• and drinking rountaiu wre ateaa aterillsed before each trial
and c1ur1ng the trials they wer-e aaaigned to indiYidual lota; thua each
lot of chicka waa maintained as an isolated unit.
Fecal uaminationa wre ma.de periodicalJ.r to detect the occur­
rence of accidental coccidial intecUon.
All of these ena5 natiou
wre negatiw tor ooc7sta ao:l the absence of coccidial infection waa
further emphasised b7 the constant auscept1b1Ut.r ot the control birds
to Eimeria tenella.
The cultures ot Eiaeria tenella used were 1'1-ca atook culture•
propagated and aaintained bt the Depart.lllen.t of Veterlnar7 Jledicine for
inYeatigational purposes.
Two =lturea were uaeda l-l6SO and. 9-20SO.
4
!b& onl.¥ c.U.t.terence between the culwree •• age al.nce 9-20.50 wu
proPa&ated with sporulated oooyate troa 1•1650.
In these trials cul­
turt age was tbe ntlllber o.t days bet'Aen the date a auaoeptible chick­
en wae inoculated tor propagation of the culture and the date thie
oul ture was used for hwun:tzation.
For the 1JIIIIUnizing doses the stock cultures were diluted with
phfaiological eallne solution and placed 1n l2S ml. pyrex glass drop- ·
piag bottlee equipped with a grourut glaaa pipette stopper.
tion was adjusted until each drop fr• the
pi~tt.
'lbe 4Uu­
contained the
appraxiJBate desired nuaber of eporulated ooc,.ta. · 'l'he drQpa wn
controlled b.r means o! a rubber bulb and allowed to drop troa the end
of the pipette when held in a .f trt.i.cal position.
To determine the
nUIIber- ot OOO)'IIt.e cont..,.ned in one clrop trom a giYen pipette, the en­
Ure drop was scanned in a counting oeU.
Since sporulation was the
onl.7 meane of determining YiabiU.t1 of the oocJSta, onl.Y •porulated
ooc.rata were counted.
He:rea.tter, the uae ot the word oocyata in
reference to counta wl.ll refer to aporula ted ooc,veta only.
All cou.nta were made with a Sedgeld.ck-Ra.tter counting ·cell.
To
· aalce the count 0.45 .ml. ot potaaaiwa dichr011ate solution with a spe­
cific
~aYity
ot 1.25 was placed in the counting cell.. One drop of
the dilUted ettlture waa added. .troa the vertically held pipette and the
coY8r
gla~a
placed upon the cell.
.After allowing one or two Jlli.mltee
tor the .o ooysta to rise to the t.op, tbe entire area • • scanned wi\b
low-powr aicroacopy and the ooo,ysta were counted.
':be final count
waa. the average of eeveral individual oounta, therefore the nuaber
of oocrats per drop waa al1ra1• an approxi..IGation.
The adjusted dilu­
tion • • then held at roara temperature.
'!he inmmn1zing inoculations were given between 8t00 A. lt. and
9c00 A. M.
Since the ceca normall.T emptt their content• in the earlt
morning, it waa thought that administering the immunizing doaea at
this hour would facilitate the maximum number of aporozoitea in
reaching the ceca.
IJDmuni zation
twlve to sixteen da1s old.
a atarted when the chicks were
Birds were considered one day old
tnnt1-four hours after the.Y were remot"ed fr011 the hatching tra1••
The !aaunizing &:>sea
ere inoculated per oa.
The bird''s head
waa held back with the beak forced opon and the desired number of
drops was dl"opped into the mouth from. the verticall.Y held pipette.
'lbe culture dilution was agitated before inoculating each bird to
keep the oocrats in a unifol"'l suepenaion.
The undiluted etock culture wa1 uaed tor the challenge.
The de­
aired doaage was administered direotl¥ into the crop w1 th a one 1111.
glass pipette.
'a letzq and Hughes {71 p.493) have indicated that earl.J' re•ia­
tance following a primary inoculation with Eimeria tenella ooctsta
11a1 not be a true acquired 1amnity but aerel.¥ a phenomenon due to
the inactivation of the ceca by the pathology resulting from the
primary inoculation.
The degree and dur tion of the cecal pathologr
was reported to be roughlJ proportional to the size of the primary
inoculation.
The primary inoculation used by aletzky and Hughes
s considerably larger than the ialllunizing doses used in this stud¥.
6
BowYer, to deteraine that the ceca were not inaotiYatecl the presence
or aDsence of norJU.l aeoal droppings was recorded at the tt• ot
challenge.
Normal cecal dropp1nga wre obaened at this tiM in all
trials, and ae tar as could be determined, phrsiological activitr of
the
ce~a
was not altered bt the immunizing inoculations.
'!be trials were all conducted on the saae general pattern ld.th
an attempt
to standardise as Jl&l1l tactara as possible.
'l'be birda of
each trial were diYided at random into lots and placed in the Mtal
oapa,
When tbe.r reached. the age at whioh illaun1zation wu started,
all ot the lota in each trial with the exception ot the non-t..unised
control lot were g1 nn a1a1lar imnamislng inoculationa.
'ftle challenge was alao ei.Uar tor each lot but the tille of chal­
lenge ..-.ried.
!he control lot waa challenged at the t.i11e the last
inoculated lot • • chal.lenged.
The challenge inoculation was adain­
ieterecl bet•en 8a00 A.M. and. 9a00 A.M. ao that the Ume interYal
betwen t.aunlsation a.nd challenge could be accuratel7 deterrained.
Tbia tiae intarYal waa upreaaed as days.
The aain critel"ion for eYaluation of the 11111uni t.t produced ••
aurrl.Yal to the challenge.
All birde tla t died were autopaied and
the cause of death determined.
Death waa considered to be due to
E1aer1a tenelJ.t when the gross and. alcroscopic poat-.ort.em leaiona
wre tJPiCal tor cecal coccidioaia and leaiona suggestift of other
diaeaae conditione were absent.
All
J~Crtalitiea
reported in the re­
aulta of the Yarioua trials are fatalities !1'011 lieeria ten!lla
7
infection. Jlo:rtalltiee from other cauaee are epeoitioallT indicated.
ln. thie paper the use ot the teru imrawdtr or imraune doee not
uan reeiet.ance in the •nse that no intect.ton waa producec.t, bQt re­
tere to aufficS.ent ree1etanoe to wit.hatand a challenge wi t.b a l&l"ge
IWIUer ot oooyata wl.tbout the occurrence ot mortal.it.7 or severe
morbidity.
8
PART I
SIZB OF DOSE AND 'l'ID REQUIRED
OR DOruNITY TO DEVELOP
TG eecure data pertaining tl> the ettecte ot the dosage aise, two
1eparate trials were designed.
Trial 1-A emplo.J8d a aeries ot 8llall
da1.l,y doaea tor 1aunizat1on, each doee the same size.
Trial I-B
&leo eaploJ8d a aeries ot dail,y doses but with a progresaiYe increase
in the else ot the dose g1 Yen.
The total <bsage of '!'rial I-A waa
a11a.Uer than the total dosage of Trial I•B but the number of doses
and the t1• involnd in gi'dng the dosage were the same in both
trials.
Culture 1-1650 was used tor both t.lunisat.ion and challenge.
It •• 91 da.ya old '~Gtn the iawdsatton was started in Trial I-A and
l40 da.ra old when started in !rial I-B.
S.ventl""'tive fll auad ale chicks wre uMd.
'l'h•T Wl"e dhi.dacl
into tive lota ot titteen chicka each and at twelve dqa ot age the
tirat ilau.nising inoculation was gi-nn to all birds in Lots 1, 2, 3,
and. 4•
Lot S was ll&int.&ined aa the non-inoculated susceptible con­
trol.
'!he dose uaed •• one drop ot culture dilution containing 175
ooo.rate &iven d.ail¥ for
tive dail_r doaea.
aa da.rs,
exoept to Lot l which reoeivecl onl,r
'l'be resulting total 1araunizing dosage • • 1050
oootata tor Lots 2, .3, and 4 and 87S oocyata for Lot l.
!he challenge dose • • 0.1 111. ot the stock culture or appro:xi­
aatel.T 100,000 OOCJata.
Lot l waa challenged twnt.r-tour hours after
9
the fifth 1aaunizing inoculation at Wiich time Lots 21 31 and 4 re-o:
ceiYed their sixth da1l7 :lwuni ling inoculation.
Lots 2 1 31 and 4
wre challenged on the aecom, fourth, and sixth dar following the
'lbe control~ Lot ' ' wu chal­
dxth and. ·last iJDamnllina iliOOulatioa,
lenged at the salle time aa Lot 4 -.b-.a the birds were 23 days old,
A al1ght amount of blood waa preaalt in the cecal dropping• of
Lota l, 2, 31 and
4; awen d.qs after the iaunization was started.
Blood waa obaeM'ed for three da..rs but other than for 1 ta appearance,
tbe bil"da were not Tiaibl.T a.t fected.
Bl.ood waa aeen in the cecal
dropping• in all lots ti'Ye dar• after the challenge.
Deaths occurred 1n Lots l, 31 and S on the seventh poet-challenge
da7 and in Lot 2 on the eighth poaWhallenge day.
1n Lot
Lot
No deatha occurred
4. '!he per cent aortality waa ae follows Lot 1, 26.67J
2, 6.67J Lot ), 13.33J Lot 4, o.OJ and Lot
s,
46.67.
'tRIAL I-B
Orae hundred S<mL chicks were used in thial trial.
di'Yided into fin lots of 20 ch'i cka each.
Lot
They were
Two cull birds,
o~~tt'rca
.3 aDd one tr• Lot ' ' were destroJed before the iaaunisation • •
atart..d.
At twl'Ye dqa
ot age the tint S..u.niz1ng inoculation waa
giYen to all bil"da in Lota 1, 2, 31 and. 4.
the non-inoculated auaceptible control.
Lot S •• uintained aa
The culture dilution uaed
contained 125 ooctat• per drop and differed troa the pre'Yioua dU.u­
tion 1n that it contained 2.5, per cent potaeaiua dichrou.t..
10
The schedule tor 1Juwn1ut1ob waa eimilar to Trial I-A except
that \be dOle lin was inereaHCi p.rogressivel.T each dq. . 'lhe first
doae • • one drop of the dilution 1'ollo111ed b1 two dail.T doee1 of two
drops each.
!be fourth and fifth
dailJr d.oeee were tour cb"opa each.
Hownr, due to the potaaeium diohl"cu.te t.be birds were reluctant to
nallow this larger Tolue and eoae
a portion of the dilution.
Lota 2, 3,
~d
ot tbe birds choked and
inhaled
The aixth dail.r dose, llhl.ch was given to
4, waa to haTe been eight dropa but due to the un­
auccesatul att..apta with the fourth and firth dail,y doees the drop
llethod waa not ueed.
Iutead, the equivalent or eight dropa
(0.5 llll.) was placed dtrectl¥ 1ntQ the crop with the pipette.
'fbe
inhalation of thia foreign aaterial proYed irritating to the rea­
pirator7 tract and aome tracheitia and pneaonia reaulted.
! total
of 14 .death• occvred in the tov lo.ta b'om thia caue but all of the
deaths occurred before a.JIIPtou of BiMria t.enella infection were
Yiaible.
It wae iapoasible to determine accurately the nUIIber of oocyata
the birda reoeiTed froa the fourth and fifth inoculations .but it waa
estimated that 50 per cent of the ooc,yat1 were lost.
'l'be total
~
nizing dose waa therefore eatimated to be 112' oocysta per bird tor
Lot
l and 2125 ooc1at• per bird tor Loti 2, ), and 4•
'lhe challenge dose waa the eaae ae that given in 'l'l"ial I-A.
Lot 1 waa challenged 24 houra after the fifth 1MUnis1ng inoeula.tion.
At t.hia aaae tiae Lota 2, 31 and. 4 received the a1xth
ftising inoculation.
Lota 2,
dail7
iDim­
3, and 4 wre challenged on the aeoond,
u
tourth, and ninth day respectively .t ollo'idng the sixth and laat
'lhe control, Lot ·S1 was challenged at the same
nizing inoculation.
t1rae as
1ma~
•r• 26 dqs old.
Lot 4 when tbe birds
Some lUood was present in the cecal droppings nine claf• after
the fir,t i.Dauniziq inocUlation.
ahd waa alightl.y more
cop1o~s
·Blaod was obsernd for three d.aya
than that seen in Trial I-A.
Except
fOr th!e 1 however 1 the bit'd.& W$!"e not vJ.sibly aftected b,Y the ~
niaing inoculations.
Bl.ood was pl"eeent in the Qecal, droppings in Lota l and ;, tift
daya after the challenge.
mood was ni'.>t obeernd in Lots 2 and 3
unt.il &ffven d.a3a atter the chall.age and ita preaence was never de•
tected 1n Lot 4.
Deathe ocou.rred. 1n tote l. and: S on the fifth to seventh poet­
challenge da,ys.
No death~ OCCUi"Md in Lots 2,
cent aortaU ty tigur"ed
on tta
b&ti.a
·Ot
3, and 4• '!he per
the nwaber of bir<le reaaini.tlg
in each lot alter the lose from pnewaold.a and tracheitis had. occur­
red was as follows:
Lot
S;
Lot 1, 22.22J Lot 2, 3, and 4, o.OJ and
J2.63.
DISCUSSION
Proa the results in T.riala 1-A and I-B, !t was obTioua that the
alze ot the 1mmunising dose would materiall,y affect the tiaa requiMKl
tor -.wtT to be produc•d·
The larger doae of 2l2S OOCJ'Sts pro­
duced auttiO-ient 1mmun1t7 by the second dSI' following the u.muni.zing
inoculationa to preTent mortality fJ'cm a challenge which b.a4 caused a
12
aortallt.r or
~2.63
per cent in the tataoeptible control
bird.a~
ualler doae ot 1050 OOOJeta did. not produce sutt1c1ent
'l'be
~t7
to
prnent. aort&Ut7 until the sinh da7 following the 1aaunid.ng inocu­
lation•.
Upon coaparing the 110rt&lit.iea ot tbt Yarioue inoculated
lota to the control lot., it waa erldent that
duced resardl.eaa ot the doeage aise.
ceiYed. the aaal.lest iJEunising doae
ao• protection wae pro­
Lot 1 of '1'1-ial I-A, lllhich re­
(875 ooc,.ta) auftered a •r­
talit,y ot 26.67 per cent, 11hereu tlw control lot auttered a
mortalit,y ot 46.67 per cent.
It waa untartwate that the deaths tr011
ocourNd in Trial
I~BJ
affected the reaul.ta
pneUD~enia
and traoheitia
hownr it was not telt that the,y aaterlall,y
or thle
trial.
theae death• occurred before the
As preYioualt stated all of
•.YJ~Ptou
ot cocoidioaia wre •••·
It is of interest to note that one chick affected with tracbeitia wu
preaent in Lot 4.
W&S
'Ibis b1rc:l •s cont.inuall.,y gaaping and ita growth
aenrel,y retarded but nen in it.e walcened condition, autficient
u.mitT as dneloped to wi thataad the ohalhnge.
tion of the trial the chick was killed and ex••ined.
At. the temina­
No leaiona of
cecal ooco1dioa1a were obaerY&d.
A ooaparatlft .-&rJ of the trial• in Put I ia &1 Yell in Table I
oD
the toUowlng paae.
TABU: I
SUDA.Rt PART I
I-B
TRIAL
OHIOKENS
•
Breed
Wwaber
4ge - Fir•" hmmising dou
CULTOltB
Age .... Firail
:tm.U!lUmg
1S
12dqa
1•1650
dose
:OOWNIZATION
Bamber or da1l1 dosea
Lot l
Lots 21 3., and .4
llwBber r4 . qocyst• per dose
1
I
:1'
9lda1'8
1·1650
140 days
s
s
l?S
17J
17S
250
6
175
4
'
6
Total. i.mmwl111ng dose
U:»t 1 • .
. ~t· 2, 3, e.nd 4
S01fL
98·
l2 dq$
6
~s
250
2S<>*
175
175
250*
1000
875
1125*
1.050
2125*
CHALLENGE DOSE
Days from last immuniaing
dose to challenge
lt,t 1
.
2
3
4
1
1
4
2
4
9
a
6
Jl(llTALITJ FR<JI COCCIDIOSIS
(total obaUenged,/llortality.Per cent)
IDi l
,
a
4
O.ontrol
IICBTA.L M FROJI O'.rHER CAUSES
('rotal birda/llorial1V.hr cent)
*
**
Eatw~$4,
See ten
see text
7S/o- o.o
PAR! II
BUMB»l C'l DOOES AND DDlUNITY PRODUCID
!ftle reaul.ta ot Trial• I-A and I-B indicate t.ha t tht aise ot t.be
Senn1s1A& doae intlueacea the t i • required tor 1awnn1tt to be pro­
duoed.
In t.hia part a group of tr1ala wen conduo ted to detera1.n.e
the ettecta ot Yarting the nUIIber ot iQdiyidual doeee CGIIprilillg a
ataadarclhaun1s1Dg
do••·
A etaad.ar4 u.mist.ng doae ot 1600
ooc78ta waa aeleoted ainoe thie tiSU:re was the approximate average
or the closagee uaed in Part I and appa.NAU, would be autticient to
produce
~ty.
'!be beat aethod thu deterain.ed bec&M the atan­
darcl l•urdsing procedure tor Part III.
miAL II-A
ODe hundred tiYe SCWL chioke wre u1ed tor this trial.
!bet
were clidded into tift lots ot 21 birda each and at 1'1tteen dqs ot
age the tirat i•nnizi.ng inoculation wae giYen to all blrda in Loti
1, 21 3, ancl 4.
Uble control.
Lot 5 waa maintained aa the non-inoculated auacep­
A dilution ot culture 1-1650
'RS
made with phpio­
logical aallne solution and •• adjusted to contain 200 ooc.rat• per
ctrop.
'l'he age of the culture at the
waa 182 dq8.
beg~
or the 1-wd.sation
!be ilawdzation wa• d1rlcled into tour dail¥ doaeaJ
the tirat and aecond doses being 200 ooc,..ta each, the tJdrd dose
400, and the fourth due 800.
•
15
'!be challenge wu 0.1 AU.. (approxillateq 100,000 ooc.rsta) ot the
Lots 1, 2, 3, and 4 were challenged on the
atook culture 1-1650.
first, aecond, third, and toUI"th c1a,r respectinly tollowins the lut.
1he control, Lot 5, wae challenged at the
s..nmis1ng inoculation.
..... tiae ·aa Lot 4 when tl'a birde wve 22 4aya old.
A ...U a110unt of blood
waa present in the cecal droppings tJ"'OIl
all ot the inoculated lote on the ••••nth day following till tiret ia­
allDisiag inoculation.
Blood waa present in the cecal droppings tor
three dqe but. otber 'ri.aible •JDlPtoU ot tha infection were absent.
Blood was preaent in the cecal droppings of Lots 1 and S on the
tilth
d.a.r followln& the oballenge. It appeared in
Lot a 2 and
3 on
the aixth d.a.,y and Lot 4 o.n the •••nth da,y following the challenge.
Deatbe ocouned in
Lo~a
dar• No dea.tba occurred. in
l and 5 on the filth poet-challenge
Lot 2, but Lota
3 and 4 lost one bird
eaoh on the aixth and teftntb poat-challenge dqa nspect1vel7.
per cent acrtalitr waa aa follo•a Lot 1, 28.57; Lot 2,
'!be
o.o;
Leta 3 and 4, 4.?6; and Lot 5, 57.~
'!RIAL II-B
'.three 1ota of 20 SCWL ohioka each were used tor thie trial.
'1'bree additional lota
ot 20· birds each f!o011 the s8Dle hatch wre uaed
in Trial II-c, 11b.iob wu coa&lcted aiault&Mousl.T.
OM ·lot ot 16
ohioka was uaed aa the control tor both tr1al1.
'ftle bird.a were giYen the tirat isawd.zing doae at fifteen da.ra of
a&e.
1be iDimuaising dilut.ton -.. the eama ae that uaed in the
16
prftioua trial and t.be age ot thia Olllture waa 2)2 dqa when imaun1­
u.tton waa at.arteti in both TPiala II-B and II....C.
The tmuniu.tion
waa dirld.ed into three dail,y doaea ot 200, 400, and 1,000, tot.aunc
the standard 1600 ooc,.ta.
'l'be challenge waa the aame as that uaed in the prenou trial
and waa adad.niat.ered to Lote 1, 2, and 3 ott the second, third, and
tourt.h da7 reapectiYel.T tol.l.ow1ng the lut i.wdsing 1Aooulation.
'l'be control lot was challenged one da7 atter Lot 3, at llbioh t1ae the
birds wre 22
A
-u
daT•
old.
8IICWlt o.f blood was present in the cecal droppings tl"Oia
Lota l, 2, and 3 on the aeYenth de¥ following beginning o.f the illau­
n1sat.ion.
Jfo
~her
81Jlptou were obseM'ed and the blood was present
tor onl.r three dqa.
sal.'l amounts of ·blood were obsened in the cecal dropping• of
Lot l on
the: titth
dar after the challenge.
Moderate 8Jil9Wlta were
present in the cecal droppinga ot Lots l, 2, and
following the challenge.
3 on the aixth
da,y
!he control lot paaeed large amount• ot
blood in the cecal dropping• 9n the aixtb post-challenge daT.
Deaths occurred in Lot l on the tittb and sixth post-challenge
daT•· One bird died ita Lot 2 aix dare followina the challenge, while
Lot 3 au.ftend. a.o mortalit7.
Deatha occurred in the control lot on
the sixth and ••••nth poat,-challenge dqe.
was aa tollo•a
43.7S.
!he per cent mortallt.T
Lot 1, lS.OJ Lot 2, ,.OJ Lot
3, o.OJ and Control,
17
m.IAL II...C
!bia trial varied froa the preceeding one in that the standard
1600 OOCfata ware g1ven 1n t1 Ye dailf doses, tb e first two doaee be­
ing 200 OOCJBt• each, tollo•d by three daily dosea o.t 400 ooc.rsta
each.
'ftle challenge inoculation of 100,000 oocyata wae g1Yen 1n
tote 1, 2, and 3, one, two, and three dqe reapectivel,y follow1Dg the
fifth and last immunizing inoculation.
'l'he control lot waa ehallen­
ged with Lot 3 at which tiM the 'b1rda were 22 d.aya old.
Aa in the precMding trials a aall. amount of hemorrhage reeul•
ted troa the munizing inoculationa.
'fh1e appeared on the enent.h
cla.r following the tirat inoculation and occurred in amall amounts in
Lota 1 1 2, and 3 tor three ooneecutiYe c:lqa.
obaerYed.
No other •111ptoae nre
Huorrhage occurred in moderate amounts in all lote on the
eixth dq following the challenge.
Death occurred onl1 in Lot 2, one bird d11ng on the sixth poat­
challeDge daf·
1be per cent mortallt1•• as follower
Lot 2, 5.0J and Lot 3, 0.0.
Lot 1, O.OJ
1be control lot 'lilich also aenecl aa
the control for '!'rial U-B suffered a mortal.i t1 ot 4:3.7' per cent.
TRIAL II•D
'ftda trial was conducted aiaulataneou:aly with Trial. II-E.
total of
147
1ft ohicb
A
nre uaed in the two trials. 1be birds wn
di'f'ided into aenn lots ot 21 chioke each.
'l'l'ial II-D and three lota tor Trial II-E.
'lhree lots " " used for
One lot waa used aa the
18
nora-:tm•rdsed control for both trials.
t.o dilutions of culture l•l6SO were made, one containing 600
OOC78ta per drop and the other 1,000 ooeyat.a per drop.
waa 276 da.rs old when the
~sation
'lhe culture
waa started in these t.riala.
'l'he tirat u.wd.Bing doae ot 600 OOCJIIta was g1Yen "When the birda
wre 16 d&18 old.
'lbia waa followed one day later by the aeool'ld
doae of 1,000 oooyata.
'lhe challenge dosage waa ittcreaeecl to
1;o,ooo oocyst•
since the
reduced mortality in the control lot of the predoua trial incU.cated.
that cultUl"e l-1650 waa loaing ita pathogenicity.
Lota 1, 2 1 ar¥1 .3
were challenged on tlB third, fourth and fifth day nYapectiftl.Jr fol­
lowing the laat htmun1sing inoculation.
'lhe control lot waa chal•
lenged one d&7 follold.ng the chal.l.ange of Lot 3 at lfhich time the
bir4e were 2.3 d&7a old.
A ...U.
ount. of blood waa present in the cecal droppings ot
the inocula ted lots tift da,ya following the first im.unising inocu­
lation and
118.8
seen tor tlree consecutiw d&7S but other aympt.oae
were not observed.
Blood was seen in moderate amount• on the aixth
poet-challenge da,y in the cecal droppings of the three inoculated
lota an<1 on the fifth poet-challenge da11n the control lot.
No loaaea occurred in tote 1, 2, and 31 whereas \he control lot
a\lttered a 1110rt.al1t7 ot 42.86 per cent.
The deaths ocoUl"Ncl in the
eont.rol lot on the aixth and ••·• •nth d8,1 following the challeage.
19
1RIAL II-I
Tbia trial waa identical with '!'rial II-D go•pt that tM birda
rM•1••d both the tannising do••• on. the a&M day.
'!he tiret. date
ot 600 OOOJ"t• • • g1wn at 9t00 A.M. and waa followed. b7 the aeoond
doae
ot
1,000 OOOJ8ta at 10& 30 A.M.
'!be birda were challenged with the aame doeage used in the pr..
YiO\la trial but the challe03e inoculation was delated one da,r longer.
Lota 1, 2, and 3 were challenged on the fifth, aixth, and anentb day
respectiYel¥ following tbl 1mal&nldng inooulationa.
and Lot
'lbe control lot
..
3 wre challenged aimul.ta.Moual.T when the birda were 23
c1qa
of age.
A ...U amount of blood •s obaerved in the cecal droppings on
the titth dq following tht inocula tiona but other a.vaptou of coo•
o1d.1os1s •re absent.
Blood waa present in. the cecal droppings ot
all four lots tift dqa following the chall.enge.
lo deatha occurred in Lota l and 2.
One bird (4. 76 per cent)
died in Lot 3 au dqa following tht challenge.
Aa prmoual7'
atat.ed the mortaliV tar 'the control
lot waa 42.86 per cent.
:.
DISCUSSIOJ ·
Verr Uttle difference could. be e..n among the five different
lJ!IDW\ising procedure• in coaparing the reaul.ta ot thia group of
triala.
All ot the procedure• produced 'daible a,_ptou ot cocci­
dioaia, but in all oaaea the I.JDIPtou were liJaited to a uall UOUDt
of blood paaaed 1r!th the cecal clroppinga tor a period not exoeediaa
three daya.
The preaenc• of thia hemorrhage did not redule the ngor
of the chicks and no detrillental efteota could 'be attributed to it.
Soae deane of iau.nit7 resulted from allot the prooedurea uecl
and 1tith t.be exception of
Lou 1 in Tr1ala U-A and II-B nrr good
protection waa p.rodu.oed.
Upoa
chaUe~e
the aYerage JDOJ"t.ality in the
inocula ted lots •• 4.12 per ~t u ooapared to
the control lots.
Oth.r tban 1Jl lAte l
46.06 per cent ln
•
et !r1ala II•A and II-B 11111ch
had a mortalttr of 28.6 and lS.O per cent reapectlwl.T, the clitfc­
ence in aor'talltr percentages of the inocula ted lot• waa not great
eoough to be
ot aignificance.
!here waa ·an 1ndicati.on 1a Trial• II-B aGd. II...C that culture
l-l.6SO wae loaing pathogen1cit.Y•
•• 236 data·
The age of the culture at that t t .
1bia reduced patb.ogen1c1tJ waa d•on.atrated again in
Triala II-D and II-E
eftft
t.bough the challenge doae had been in­
creued by SO per cent.
A comparative euaary ot the Trials 1n Part II 1• gi'Yea 1n
Table II on the following page.
TABLE II
SWMARY
PART II
n-B
II-A
TRIAL
GHlGilmS
Bne4
U•E
II~
Il.o.C
BH
Age • la\ ~ dose
SCWL
· SCIL
l5 dqe•
lS days
15 days
.&.ga· • lat :ftmmnizing doatt
nn.roBIZl'tiON
lS2 daya
da1fJ
232~
3
s
2
1000
- 1.000
em:.~
• l ...l6So
)lumber ~
cto.ea
Bamber oocysta per doae
1
'Z16
da)"8
600
l6
&
c1qa
Z76 day8
2*
600
200
200
200
200
200
3
4
5
400
800
400
l.OOO
400
1600
1600
400
400
1.600
1600
"1600
100#000
ltlO.ooo
100.000
150_,000
150,000
2
l
2
3
,
4
K<afALM f'B.CU
0
'
Lnl
OOOCmiOSI~
'
4
Control
l
2
3
4
­
:
3
4
21/ 648~57
20/;J•lS~O
21/1...;. 4~76
21/ 1.- 4~76
20/o- o.o
211 o- 0~0
2
a
4
Udap
2
.·. Total~ do:ae
CBliJ.BIIGB
'lumbar of oocyst8
~- .fJ"'OIl lan batmi trtng
dose to cllallenge
I.ot ].
2
*·
232
'SC\R ·
2'i/l2-57.l4
Both doses given #8M• day'
Total. challengacJ/Ilortal.ty-Pe cent· Jlorlality'
20/1- 5.,('}
16/7-4}.75
.
3
4
s
21/_o.. o.o
21/o- o.o
20/o-o.o _ 21/o- o.o
s.. n..a ~.86
20/o-o~o
20/l~s.o
5
6
~~:0·
2l/o...<t•o
21/1-4-76
. . II-»
-~
22
PAR'l' III
MIND4tJK TDLE ~UIRJtD FOR DOOJNITI TO DEVELOP
CCilPARitG A RELATIVELY OLD AND IOORG CULTURI
'l'be trial• oompriai.ng thia port.iDn ot the im'est.igat.ion •re de­
aigMd to det.el'lline the ainimua tf.• required tor 1aawn1t.1 to be
produoed when using e old and a ,young culture in the U.Wd.ag
procedure.
Tbe general pattern wa• similar to that. uaed. in the pre­
Tioue trials.
to etandard1se all ot the 1nt1u­
An effort. wae made
enci.as factors except the t.iae intenal troa the last. ilawd.nng
inoculat.ion to the challenge.
A at.andard iBD.unld.ng procedure waa used tor all t.he trials in
thie part or the investigation.
Th1• procedure was the same ... that
used. in '!'rial II-D which waa one dose of 600 0007ste tollowd in one
da,y b7 a second doae of 1,000 OOCJ'8ta.
New Hampshire chicks wre UMd in all the trials, but. it was nat.
poesible to secure all ot the bircle troa the Poultr7 DepariaentJ
therefore, chicks pvohaaecl 1'roal a
o~eroial
used in four of the eight tr1&la.
Ia each trial the 1aawds1ng
poultrr hatche17 wre
procedure •s begun when tbe chick• wre fourteen daT• ot age.
Two trials wre alwqs OOJ¥haot.ed aiaultaneoua}¥, one using the
old culture l.-1650 and the other, the ,young culture 9-2050 tor the
1Jaun1ming proced\11"8.
Two dil.utiou were aade troa cultUN 9-2050,
one containing 600 OOC78t• per drop, the other containing 1,000.
'!'he
dilutions of culture l-1650 u.aed were the two prepared prnioual7 tor
23
Trials II-D and II-&.
culture.
The birds were challenged with the )'Owager
'ftle challenge dose was 0.2 ml. ot the stock culture or ap­
proxia&tel¥ 120,000 oocrsts.
en the dar old ohicka were reoeiYecl, the.r wre diYided into
aeYen lots, tbre'e lots to be inoculated with the old culture
trial
am
tor one
three lots to be inoculated with the TOUD& culture tor the
other trial.
'l'he seftnth lot • • not inOculated and serYed as a
suoepti ble control tor both trials.
The control lot waa al•18 challenged at the NID.e t1ae that t.he
last lots (Lota 3) were c ballenged except in the last two trials 11ben
the control lot was challe..ecl with Lot 3 ot Trial III-G.
'!RIAL Ul-A
1hree lots ot lS chicks each •re u•ed•
With culture 1-1650 -.hich was .309 dqa olcl.
'l'beT were inoculated
Lots l, 2, and l wre
challenged on the third, fifth; and lixth dar reapectiYely tollowS.ng
the s.-unizin& procecbare.
Lot a l and 2 lost one chick each ti'Oill the
challenge, a JIIIC)rtallt7 of 6.67 per oeot reapectiftlj.
no 110rtallt7•
Lot 3 8Ufferecl
'lbe control lot (14 chicks) had a mort.al.1t7 ot 64.29
per cent.
1RIAL III-B
'Ibis trial was coDiucted. aiDn&ltaneouaJ..y nth
lota of snenteen chicks each were aaed.
Trial
III-A.
Three
1her •re inooulatecl wi 'U1
.
~
culture 9-20SO mich •• 62 dqa old.
Lot.• 1, 2, and 3 were challen­
ged on the third, titt.h, and aixt.h da7 reapect1Yel¥ tollowi.ng the 1a­
JIIlUD1a1ng procedure blt no aol"t.ality reaal.t.ecl
trca the challenge. Aa
prerloull' atat.ed, the oont.Jool lot auttered a aorta11t7 ot 64.29 per
cent.
'l'RIAL III-o
'!he chick• ued io thia and the tollow1ng trial were pu'Ohaaecl
troa a oa.erolal poultey hat.oher7.
'ftlree lota of titteen chicka
each wre inoculated with culture l-1.6SO llhich waa 344 da7a old.
Lot. 1, 2, and
3 were oballengecl on the tirat, eecoDd, and third da7
reapect1Yel.¥ following the tawun1sJ.n& prooeclure.
The per cent ..or­
t.allt7 •• aa tollowt Lot 11 66~67J X.Ot 2, 40.0J aad Lot 3, 26.67.
'lbe control lot (1.8 oh1cka) had a aortalit7 ot 44.44 per cent.
miAL III-D .
'1hrM lota ot fifteen ohicke were inocU& ted with cultun
9-20SO.
At that tiae the culture •• W dqa old.
Lote 1, 2, and 3
were ch.U.nged on the tiret, eecoDd, and third da7 reapect.1Ye)J
tollowiDg the
~sing
procedure • . '!be per cent .art.allt.7 •• aa
tollowaa Lot 1, 60.0; Lot 2, 6.67J Lot 31
atat.ed, the oontrol, 44.44.
o.o
andt aa prnioul.T
\Ma tz'iial and trial IU•F were a dupllcation .o f the t1110 pre­
noaa trials ex.cept. that chie'ka !Jrca the Pollltry Depart.nt wer-e qaed.
Three lota ot twel'V'e cbicke each nre inoculated with ·c ulture 1...16.50
which waa ~' <lap old. tote 1, 2, and J were challenged
tirat, aeconcl 1 and third
proc!tdure.
~
reepecti"f'ely fbUowlng the iawd.&it'l
'!he per cent m.. t.al.ity • • ae tollow1a
Lot 2, .50.0J Lot 3, 2,.0.
on the
Lot l..; 91.67;
the cont;ol 'l ot (12 ch!ol(t) _had a •rtali•
ty of 66.67 per cent.
1bree lots of t-..l:nt cblcka wre inoculated w1 tb fulture 9-20SO
-
wh1cb wae lJ.S dqs
olct~
.
~
'
Lots 1, 2 1 and 3 were challenged on the
first, eecond, and t.h1r4 daT reap.ot11"ely following the i.-wd.s1ng
proo•dure.
,-o.o;
Lot 2,
'!be per ce• 110rtal.ittYas &I tollowaJ Lot 1, .58.)3J
Lot 3,
o.o;
and, aa prrw1oual.T etated, the contl"'l,
66.67.
'l'l\e chioks uaed tn thu and the toUold..~ trial were purobaaecl
~
'
from the eam.e
Ill-D~
cc:~~ael"Cial
•
I
1.
hatchery ae the chick• in 'l'l'!ale III...O and.
'lbree lot• of fit\Mn ch1cke each were inoculat8d with oul•
ture l-1,.50' which was '9) c:ia71 old. Lota 1, 21 uct 3 ._,.. che.Ueqe<l
on the fourth, titt.h, and aixth daT reap.ctiYelf following \he
26
t..unistng procedure,
'l'be per cent •rtallt7 •• a a followat
40.0; Lot 2, 6.67J and Lot 3, 20.0.
Lot 1,
1he control lot (lS chicks) had
a 111Dl"tal1t,T of ' ' · ' ' per oeat..
!RIAL Ili-11
'lhrM lota of fitte~n obiea each"" inoculated with cultur.
9-20~
wtdoh waa 146 dqa old., Lot. 1, 2, and 3 •re challenged oa
the thir4, fourth and tilth daT t-eapeot.iftl,r following
141
procecbare.
1be per cent aortalit7 •• •• tollowa
the
t-nt s- ·
Lot 1., 6.67J
tot 2, o.OJ Lot 3, o.OJ and, •• prniouaq atated, the control,
,,.,,.
DISCUSSION
'.ftte reaction to the 1mr•u:ds1ng procec:luHa •• l1aited to tbe
appearance of bloocl in the o.oal clropplnga.
In the lote wbe!'e the
chal.lenp •• dela7ecl loag ·•nouch (two ott aore da71 toUolllnc the
iMUnising prooed.un) the blood reaul\ing froa the s.-ud.si.ng pro­
cedure appeared on the •••entb dq tollold.ng the first 1aooulation.
!be h..,mage proG.cecl troa culture 9-20SO • • mere pronounoed. thall
that troa n.lture l-1.650, but in all i!lltaftcea the reaction wu 11­
llited to the appearance ot blood, and the general l'igor of the cblcka
waa
not affected.
In trials III-I and III-a, .no heaonhage wu ob­
..ned troa the iamunis1- procedure, this further indicated that
cultur-e l-l.6SO • •
losina
pathogenicity with ita 1norealing
•a••
27
1he hemorrhage resulting trom the challenge waa regular)¥ obaer­
Yed on
~
fifth poet-challenge da.r.
'l'he
~~&jority
of the aortaUty
neul. ting troa the challenge occurrecl on the sixth poet-challenge
Deaths occurred u earl,r aa the Atth and aa late aa the
day.
aeYenth poat-challe.np day.
1he
miniDJJt
tiM reqdred for the production of .s.-unity 11b.en
culture l-l6SO ._. eaployed waa el.x dqa following the S..Unizing
pl"'Oedure.
dqf
'lhia occurred in 'trial In-A lfhen the culture waa 309
ot age. 'lbia reMtlt • • not 4upllcated 1n Trial III-G llhen the
culture • • 393 c:lqa of age aince tbPee birda (20.0 per cent) auo­
ouabecl to the challeoge.
In Trial III-A there waa an indication that
couiderable protection had de'Yeloped in three
d&J•,
bird (6.67 per cent) auocuabed to the challenge.
ainoe
onlT one
'!hie protection waa
aot experiencec.t in Trial.l IXI...C and. In•E when the culture waa 344
and
365
da:r-
ot
age reapect1Yel7, the liDrtality being ~proxiru.tel.y
2S per cent in each trial.
GoOd protection • • deYelopeclin tift
d&7a 'llben the culture waa botb 309 and 393 d&TS of age.
In both ift­
etancea one bird (6.67 per cent) died troa the challenge.
'Jhe ad.niiNJil tlae required tor culture 9-2050 to de'ntlop 5mann1­
t7 wu three daTa following the 1Biaun1d.ng
procedure~
!hia result
wa• repeated in three of tour different viala w1th the age ot the
c\llt.ve ranging fJ"'Ol 62 to 1.46 d.afa.
'l'here waa no indication ot a
decrease 1n activitr aa waa seen 111. th culture 1-1650 but it llboald be
pointed out that culture l-1650 did not. begin loai.ng pathogenicity
28
and ant.t&enioity until atter it had reached an age of more \han 2SO
and.
340 daya reapectinly.
to coapare the aotiTit.t of the two culture• with respect to the
tiM allowcl between the u.ntsing procedure and tbe challenge, the
lote ot the Yarioua triala were arranged according to the t.i• inter­
Yala..
'!he n\lllhel" ot ohioka aunirtAS the oballen&e at each t1.M
·intern.l •• uaed aa an indication ot the s.-anity produced.
intora t.ion 1• preaentecl graphioall.T in Figure 1, page 29.
1h1•
Since the
low challenged at orw U.. interYal wul.d. be trca Yarious trials,
the age of the oul.turea
daya.
liU
not conatant and could Y&rf aa auch u 84
1be difference in the age
ot the two cultures (247 ciqa) ••
Conatant and tor each group of birda :rece1Ting the 70Uft8el' Culture
and challenged at a g1Ten inter'tal there was a co:r:reeponding group of
birds which had receiTed the old culture.
Four lota, a total ot
ei&ht \rials.
'9 chioka,
HI'T8d ae the control tor the
!be nuber ot au:rrlTing bi:rda waa 42.83 per cent.
1bia percentage waa conaidered the e tandard. :rate ot euni"f'al. tor
auaceptlble chicks to the ch..U.nge g1Yen.
1he anrage percentage ot
auniYOrs in the lots receiTing the 1mmwl1sing procedures ard chal­
l~eci
at Yari<*a interTala waa COJ:Iparecl to thia atandarcl 42.83 per
cent, the -d ifference in.dicating the J..,n1 t7 present.
Pour lots were challenged one da7 following the t.au.oising pro­
cedure.
Two lota (27 ohiob) had reoe1Yed culture l-1650 and the
aVYinl • • 20.83 per cent.
'l'wo lot• (27 ohickll) had receiftd
29
TIME REQUIRED FOR IMMUNITY PRODUCTION FROM OLD AND
YOUNG CULTURES OF EIMERIA TENELLA
100
_j
~
>
0:::
::J
(/)
1-­
z
w
u
0:::
w
Cl..
3
I
2
DAYS FROM IMMUNIZING PROCEDURE
2
CHALLENGE
1
TO
A- CULTURE 1-1650 ( 309 TO 393 DAYS OLD)
8-CULTURE 9-2050 ( 62 TO 146 DAYS OLD)
C- CONTROL
I -600 OOCYSTS FIRST DAY AND 1000 OOCYSTS SECOND DAY
2-0.2 ML . STOCK CULTURE 9-2050 ( 120,000 OOCYSTS)
FIGURE I
culture 9--20SO and the surdval wae 40.85 per cent.
the 11\&rfiTal percentages were
group.
In both groupe
1••• than the atan4ard tor the control
1.'h1a _. interpreted to indicate that no protection had de­
'ftloped trca tru. '••anJziQ8 procedure 8114 the ooc1eta giYenin thia
procedure aotuall,y increaeed the aeveritr or the challenge.
Four lots -.zoe challeqecl two da,. following the S.,.,ni siq
procedure.
Two lot• (27 chicka) reoe1Yed culture 1·1.650
n.niftl wae
ss.o per cent.
am
the
Two lota (27 chick•) received culture
9-2050 and the aurrinl wu 71.67 per cent..
In both groupe the
percentage wae greater than the control et&D:lard but not enough to
indl.oat.e a significant production of t.awd.ty.
Snen lot• were challenged three
procedure.
4&3• following the t..unising
Three lots (42 chicks) rece1nd C\.llture 1-16~ and the
eurTiYal • • 80.5S per cent.
Four 1otll (59 chicks) receiTed culture
9-2050 and the 1uniYal waa 98.33 per cent.
Onl.T two lots were challenged tour da.ra tollo1d.ng the 1•nnizi.ng
procedure.
One lot of fifteen chlcka received culture l-1650 while
the other lot or fifteen chicks received c\.llture
9-20~.
'lhe nrrl­
Yorl were 60.0 and 100 per cent reapect1'ftl7•
Pour lots were
procedure.
cbaUe~~gecl
tiTe dqa tollowi.ng the hwun1zing
Two lots (30 chicks) receiYed culture 1-1650 and the •ur­
Tiftl waa 93.33 per cent.
Two lots (32 chicka) receiTed culture
9-20SO and 100 per cent aurrlTed b
chall.enge.
lbree lot.s •re challenged: six ctqa following the SJWUnJsing
procedure.
fifO lota (30 ch1cka) received . culture 1-1650 and the ev­
Tival .a 90.0 per cent.
c.te lot (17 chica) receiYed culture 9-20'0
and 100 per cent IW'vi:nd the chal1qe.
the reeulta obtained boJa theae trial.a demonstrated. that the
,ounpr culture could atiaulate the production of autticien\ 1...­
Ditt in \hree daTa following the bllaun1z1ng procedure to preftnt
aortall\7 froa the challenge.
'l'be older culture required aix da1•
to atiwalate an oqual degree of i11111Ul1t7. When the challenge _,
given before 1mmnlt1 had deTeloped, the 30\&nger culture routinel,y
cleaonatrated a higher degNe of protection than the older culture.
x.amitr to coccidioaia deTe1opa onl.1' after \he in.geat.lon ot
'liable ooc7ata reaulting in active infection.
prodUcing
~t7
All o\her Mthoda ot
have • t with failure tbua tar.
Ita product.ion
ia aaaociation 1d tb the irrt aion of auaceptible boat cella
(epithelial cella of the cecal m.ucoaa) b7 the growing paraaitio
toru occurring in the ditferen\ d..,..lopaental atagea ot the para-
a! te 'a ille c,.cl•.
lation ot the
~
'lbe antigenic tact.or reaponaible trr the atiau­
reaponae, 1a therefore, the presence t either
these growlng forma or their utabolltea in the. boat cella.
Siace
the presence of hw.oral ant1bod1•• cannot be deiiOnatratecl 1n cocci..
dial iaunity (6, pp.)66-369) the ~ naponee 11 aaawaed to be ot
a local cellular nature 1nYo1rlng the boat cel.ll.
11Dm1tt
'!be degree ot
or antibody developed, uawdng that all ot the chick• haft
a shlilar abilltr to reapond to a given antigenic atillul&tion, would
32
4ttpend. -opoA the aDIO'IInt
be~
ot ant.! eft pne:ent. in the Uaauea or t.he
nwfl:o!o
ot host 4ell• idYolved.
'l'b• llf• C.JCl• ot a oocoidl-. 1e 41:dded into aeYeral <htvelop­
unta.l et.agee ,
AI 'the CJCl• .PJ'OS"•••e through these a\agea there 1a
a PI'OI)I'e•t1'f'e aultJ.pUcatJ.on ot puaal.t!a tonu and a progreea1ve in­
crenae 1n the DQIIIMI" of
euaceptlb~
boat. o:eUe 1nYOlvect. Sln.9• the
die.ue 1a 1elt• l.1m1UAa (J; p. )26), a Jiwn l'UJr4l)er ot viable ooe,ate
tbeoretiO:allT 110Uld noclUO t. &lvetl number of paraef.\10 tortaa 11bJ.ob
wul4 lnYolve a gi'Yen nwaber ot •u•eptible host oeU. in a g1ftn
.. •'
period of t:Sae.
1h• <U.Itereace obeel"Md 1n tb • U... t>equiled .tor- t.be production
ot s.utaltt when ulna the
ol<l OAci ~he fOUft&
"-of hoet uU• 1uvolw4 at. • glven
•r ~aion
'three po•e1b1Ut1ea that
tibla tbeorr a.re OOtUJ14eJ'M.
Ftnt tbe
tvlt
ti••
culture could be attn•
ft
-.v haft be•
't'
ot
epot~tN 00071\fl
proent in tbe old.ft' cul­
reduced •1t.h age and th number ol paraaitle fol'ftll
ol" h t. oeUe 1nVolft4 durina t.he 4ewlopPnt.a1 etagea would not be
qtttoien.t, 1.0- procblce U.Wlit1 WltJ.l. l&te ln the Ute o,.cle.
~·~'
oult.ure eontaiftiag
ghate~
The
awlbera of eporulat.ed ooc,.ata
woulA produce t.ho l'tJQ.\Llrred tluaber of
pllt'u1~1o torae 1nvol1'1ng the
•1llt1cient tal.lll.ber of hMt cella e&J""lter 1n \he lite cycle.
To deterlltiM if tbia waa the caae t.he culture dilu ione wre r .....
ooW'lted betore atartlng !riale XII•ll and Itt-r.
Cicn~
Mile at that t1mD were not
aianifl<~oaotJ.t
Th• a.veraa•• of the
dlttereot, troa the
33
average of the counts made when the dilutions
Y~&re
prepared.
The ad­
ditional time required for i.anunit1 product.i.on from the older culture
could not, ther.fore, be attributed to a decrease in oocyst numbers.
The second poaeibiUt1 would be a decrease in the number of
viable aportU.ated OQOJ•ts aaaociated with 1mreanng age.
1he number
of OOCTSta produced troa a given incculation would be an indication
of the number of viable ooctets present.
A comparison o! direct teeal smears trom. Trials III-G and III-H
ahowed a marked difference in OOC18t production.
Numerous oocyata
were present in !rial III-H seven days following the first immunizing
inoculation.
No oocyata were observed at this time in Trial III-G.
Eight dqs .t'ollolld.ng the first immunization a few OOCJata were
obsened in Trial III-G whereas, large numbers were obsened in
Trial III-H.
On the ninth d.a7 OOCJeta were quite numerous in Trial
III-G but the number preaent was considerabl1 smaller than the num­
ber present in Trial II.I-H.
Dickinson (l• p • .39l) observed that the number of oocysta pro­
duced from inoculations with old cultures of Eimelj.a tenella 1tas
conaiderabl¥ 8Dialler than the number produced from inoculations with
Yery young cultures.
A third poe aibiUty 110uld be that the older OOCJ&ta had a re­
duced actiut.r and consequently reproduction was slower, mare t i •
being recpired for the coapletion of the various stages in the life
CJCle.
34
Blood resulting from the i.aaunizing procedure was obserYed on
the seventh dar following the first iDIIlunizing inoculation with the
exception of Trials III-E and III..O.
In Trial III-E bloOd resulting
from the U.unizing procedure was not obserYed but its presence would
have been ma.sked by the hemorrhage from. the earl,y challenge if it
were delqed longer than the eighth d.aT·
Blood from the immunizing
procedure could have been detected as late aa the eleventh da,y fol­
lowing the first immunizing dose in Trial III-G blt ite presence wu
not obeerved.
Ooc7st prcduction was also dela,.ed in this trial.
A
tew ooc7sts were s ..n on the eighth dS¥ while in Trial III-H numerous
OOC1St8
were present on the seventh da.Y•
Dickinson (1, p.)9l) has
damonatrated a dela¥ed reproductive c7cle from inoculations with old
cultures of iippri! tenella.
Ot these three poesibilitiee a combination of the latter two
wuld present the moe t logical explanation.
In either case fewr
parasitic toru would be produced invol'ri.ng fner host cella at a
given tilDe thus explaining the elower developnent of 1neun1ty when
the older culture was used in the imDmrd.zing procedure.
Considerable variation lias obsel"'Yed in the suaceptibillty of the
control chicks from the two ditterent sources.
The control chicks ·
obtained from the Poultry Department were more susceptible than those
obtained fl'Olll the CO.DD.e!'Cial poultry hatchery, the aortallty being
65.48 and 48.89 per cent respectively.
The chicks from the Poultry
Department exhibited rapid and -..n growth with earl1 physical de­
velopllent.
!he feathering was SIIOOth and the general activit.r of the
35
ch1cka wu sood.
VerT tew oull birda appeared in these groups.
'ftle
ohlcka troa the ooamercial source exhib1 ted slow an1 uneven growth.
'ftle ph7aic al deoveloJ:a nt • • retarded when compared to the other
chiob"
Feathering waa rough and a oo neiderable number ot birds de­
veloped wings which were longer than their llodiea and would be
olaaeitied aa culls.
The general aot.irit7 ot the chicke waa not
•1gn1ttcantl.T ditterent from the other groups.
'lbe variation in
aueceptibillty did not substantiate the coaaon conCept. that slow
de-veloping, retarded chicka are mere ausoeptible to ooccidioaia.
Roaenberg, (4, p.472) deaonatrated a aimilar variation in the
auacepti'bilit.Y to Eimeria tenella intecUon in the progen,y from two
different
group~
of · bite Leghor.na .
hom his obaervationa he con­
cluded that reaia.t aftce and ausoeptibillt1 to thia infection ia
hereditary in the dlicken.
Although the tindinga ot this atudf are
not &fticient to warrant a concl.ueion that reaiatance or auacepti­
bilit1 ia inherited. they indicate the poaaibllit1.
A comparative
aWIIIIal"(
ot tbe trial• in Part
Table III on the following page.
In
1a given in
TABI&ni
StJl8WtY PART
TRIAL
CHICKENS
Soorc8*
DOIUHDA'fiON
CUlture
.A.g• of culture :l.n days
Days tram tmmuniaing procedure
to challenge
Lotl
2
3
ID-A
ni-B
PD
PD
m
In-o
III-D
ec. ',
Cca
In-B
nt-F
ttt-G
III-R
PD
PD
C<a
ea.
J.-1650 9-2050 l.-l6SO 9-2050 1-1:650 9-2050 l.-1650 9-2050
309
62
344
97
365
llS
393
146
3
5
6
3
5
6
1
2
3
1
2
l
2
3
3
llORTALITI FR£11 COCCIDIOSIS
1
2
'
4
5
6
3
4
5
'total. cballenged/llortality
Per cent llortali'Y
lot l
2
15/1
6.67
17/0
15/1
17/0
15/0
17/0
6.67
3
ContrOl
*
**
o.o
o.o
o.o
o.o
14/9**
64.29
12/11
!2./7
58.33
lS/6
40.0
15/1
50.0
l2/6
12/6
50.0
15/l
6.67
15/0
12/3
32/0
15/3
15/0
15/lo
66.67
15/9
60.0
91.67
40.0
15/6
15/1
6.67
15/4
15/0
26.61
o.o
25.0
18/8**
44·1.4
PD- Poultry Department, Com- Commercial Poultry Hatchery
Served as control for previous trial which was conducted aimul.taneous]¥
o.o
12/8**
.66.67
20.0
-6.67
o.o
o..o
15/8-H
53.33
~
37
1.
1be ai&e ot the amunizin.g doe• attected the tiM required
tor the produotion ot iwunitT to B1ae£1a tenella.
An 1wuni sing
doae of 10'0 aporula ted oocyata requirec:l tour dqa lonaer to produce
1aun1t1 than did a dose ot 2l2' aporula ted ooc.rata.
2.
fheae tz.iw <*onatrated. that 1600 aporulated ooc.rete 111\en
properl.1' adllin1itered pro4uced a practical U.WUt.r that withatood
aevere ohall•n&••
3. !he act.irlt7 ot a culture waa found to deoreaae u the age
of the cul\u:re inoreaaed.
a culture that • •
Reduaed pat.hogenioit.r wae deaout.ratecl ill
236 da.r• ot
age.
Reduoed antigenicitr waa erldent
in the aaae culture 'tlben it waa 344 clqa ot age.
4. The Jiiniaua time required tor the proc:btction ot t..unit.Y
trOll a atandard 1-unhing procedure (600 ooc,.ta tiret da7, 1,000
000,18ta aeoond da.r) • • .S..x da¥• with a culture more than 300 dqa ot
age and three dqa 111. th a culture leaa than lSO dap ot age.
s.
Chicka troa different eourcea nried in waceptibillt7 1ft..
dicating a poaaible iaherent re1iatanc e to coccidioaia.
BIBLIOGRAPHI
1~
:Dicldnaon; E.. M. A factor in delayed production ot E1m.eria tenella
oocysts.'. Poultry Science 25 J)9l.. 1946.
2..
Janldewics, H, A~ Symptome and immunity: toUorlng graduated doses
· of EiJaeria tenella. Journal of Parasitologr 28 ( Su.pplaJlent) t
23. 1942~
3.
Johnson,. w. T, Immu.nity or resistance of the chicken to coccidial
infection. Corvallis, Oregon. Oregon Agricultural Experiment
Station, 192?. .)lp. (Station bulletin 230)
4.
Rosenberg, M. M. A !Jtudy of the inheritance of resistance to
Eimeria tenella in the domestic fowl. Pou.ltey Science 20 t472.
1941.
5.
Tys11w, Erneet Edward. Criteria and methods in the investigation
ot avian coccidiosis. Science 75 tJ24-328. 1932.
6.
.
. , Bans 'lheUer, and E. Elisabeth Jones.
CoecidloS!s in gallinaceous birds. II. A comparative atuttr
of specttea of Eimeria ·of the chicken. 'lbe American Journal
of HYgiene
15 .t319-393. 1932.
.
7.
Waletlk;y, E. and c. o. Hughes. Factora involved in tests for ac­
quired inmtll'lity in E;blleria tenella infections of the· chicken.
Annals of the New York: A<*iCt~q ol · Science 52 t478-495. 1949.
S,
Wickware, A. B. Studies on reaistance to Eimeria tenella infecticm.
Canadian Journal of COmparative Medicine 11=125-jjd. 1947.