THE EFFECTS OF ORALLY ADMINlSTRAT‘ED ENTEROCOCCI
ON CERTAIN PLASMA ENZYME LEVELS AND ANTEBODY
TITERS OF AXENIC, GNOTOBIOT‘C. AND
CONVENTIONAL CHICKENS

Thesis far the Dam of M. S.
MICHIGAN STATE UNIVERSITY
Lois Brenda Allen
1963

 

 

 

 

THESIS

   
     

  

LIBRARY
Michigan State

University

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:13 {:32 L”.’.".$ C“ C. 51.1.7.1 ‘ 3'73 1.1:.4“. D '17:.1332331'2'31 CH CST"; 3:]
II; c-- it‘s 11;..311 L U, IL; A 313-. 3‘1: C“ '22 T:1‘- . 1'! (IE:
3 9' I ‘ ['V' )'1f"‘ 1“ fl « '- ‘K? '32va .‘ Q r:- r5! 15‘4“? r‘f":

)0; -.-.'..::.s' G Loire ‘v‘ LIV fir.- '3 G 'niu.~.铧£.i - “ ‘hfl’am- biU

by 12:21:; 133mb slim

1:101:10 c.2123. commtfioafl (11.1.21! 2.9 raging from I, to 8 22:20:23

of age were orally 111212.213: 20:3 9:11-1:1151t‘“. .5. fat?“ :3 or g. £52335:

W21. :2 1.110 20.215. T2211 amouoo. oi’ti: s 52251.2W :2: to dcoact (22.132130 in
the level of 11152312.: :zrzmeu 1.115.022 121.112.: mozzazv 03‘ [wees-.10
(212212212115: zrcxhfiion. The 31:22:03? 1.113.: 21:13 3 (2? {33:3 01‘ £00235
were 02. scrvod at writ-.133 t". .519 mam-22222.21 afar 20.312.131.302}.

Host cmbohistm (1.1321021: :30... mod :v ...:2..:1:2 0:312 1221 hp: 2.122%.“6
cftrr 1:100: 11.52.213.011.” In {zmw 2m 3:3 :21162 imcmlatcd 001.111.131.191:-
(13212120313 t2 was was ar :1 .i-J. 1321326222.: 9 21:1 21'2”“: 01......02 2:2: 11: Lil 3
meta past 12“1oc112‘.22.t10:23. 222.22.21.25 the 31.212301 1:12.. 12:20:: 3 m 5 25002.10
after isms-1.1133921 thcro was :1 83.3.1.1. decrease: 33:2 9.2.1121 .233 tit-ms.
Fear-.1 311.111.013.121: 1mg (19110:. ...2:3 3.1 1122:: 21:: 21:. of motoiwisisic

dddtsm at. 3 to 4 11:20.5 {9:21. 3.10.1" '23:.021.

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5-21!

The 2'21: .221: (11‘ 012331131. 191' 3:19.13 01’ fez-.2220 ......:<W «.1 to .1 3 1313
at 3 1:002:23 after: 1:20:21 5.21.52. :1 and $1211 deemwascd to 1 x 1'37 beams
3 to 5 weeks post 11:91:19.1.1sn.

Tim's 2.22:1". a (1.22'12'20 .212 “213 lwsla 0.. es .232. 01‘- P2 0 I, 31.22;“ 0: 1:29.23

at 33-23: (3.19:1 t: 2:39 tow 1:22 to to 032:: “2.21:2: .1; 1:23-11 .1..-:c. .10 2.111203;

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The Moose .ntLe promo. ”food or Mohua but

maize effin- kzomlation is thought to rest... 1 z from

foo-.31 rati‘imftr 013.7143? cit-.47: ho 4.7.?) 4.3331" 43 not:

‘- t...” fmJLs-gfi. fig“? ‘41:} of; "from not only mt...

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produces mammnt ours-ma in the lava:

{3130-817}: eta; a, mi wwmw‘imsflts.

as r
1.3

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«.3? Lumouifl magi}. t1.

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lib-III»; -‘C.

TF3 Effigi‘g 0? 03‘.“th 51‘573'1145‘73337‘9 r'EWLQX’CvI 0:; (:1433
PEAS" £33.39; Lgbm mm sz'fi If! Tm‘fj‘: 0F

Mir-7:10, 01:13:71 7171c, . m as . «153.71. 1331.72:

Lois minim Allen

31123431“th to
”Jail 113m #1423“ Ufli‘mi‘flbhv "
‘3 partial Mil-1323621“ of “hi-13 F3111? cosmic!
{or we 110517043 of

H:- EYE“. (23.96517 $3273
1393;371:2033}. 0: )tiav-chaLOJ and mum Health

1963

I!!! author '
{4.0.1.8311}: ;
m. may.

all Hague;
55 lilo been a

01504/
J

H , 3

s ,7: a v.1 v- "um
and"... "Lumeium

Th. author wished to axyroal her aincaro approciation to
Dr. C. K. Smith for Lia patience and guidance during tho course
offlnlnudn

The frequm$ help of Else 5. Fittengm' and Mr. I. L. Dahljolm
his 81.0 m may appreciated.

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mom
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LIST 0? 1118135

Peg.

Th. formula for preparation of 100 g qnantitica of
thoohickudictIr-ZBQ........03....-

8m lntibody titer. and tho who: of 1m
good. 70. W in the feces of 0 chicken. in
mp mmmauon00..0.....0.00

Focal antibody production 1v gnotobiotio chick».
atoll! mm with Streptococcus faecflg . . . .

Anfibow titer. of gnotobiotic and conventional
chicken. rod 3;. W and of minocnlntd
conventionalchickcn. .0..000.00....0

 

Plum peptidae. actidtie. of gnotobicti. and
conventional chicken. administered g. M orally,
mdotuninoculatod conventional chicken. 0 0 . 0 0

Plasma acid phoaphataso vein... of gnotobzlotic and

17

25

28

30

conventional chicken. after inoculation with é. W

and of uninoculatod oonmtional chicken. . . . . 3 .

Plum. glutamic—oxalacotic tranaaninaa. activitiu of
uninccmlated conventional chickens, and of gnotobiotio
and conventional chicken. administered é. gaoogig

01'811’.0.0..00.0.0.00.00.000.0

Plum alumna-movie tranlaminas. activitie. of
uninoculatod conventional chickens, and of gnotobiotic
and conventional chicken. administered Q. faecggg

0311130...00....0...00.0...00.

32

34 .

36

5.

FIGURE
1.

2.

3o

1..

5.

LIST OF F 3.6 113335

Harm antibody titer and the umber of g. face. a

   

m. gum-51219 in the feces of group I chickens . . . . :16

Moan 1:131;in titers of gmtobiotic, inoculated and
911315031313th countrbicnel chicken. . Q Q t 3 Q g c g o 29

Mean Fluvial”. part-153.30: value. of mtobiotic and
cmV~vntioml chickez'm nénm—dstcrod g. M orally,
and of mfino-zmlntfid conventional cifzi-‘lsktixkfi . 0 . 0 . 0

2439.11 Values of pli'znna acid moagimtm... of gmbobiotio
and .cazwentia-n-gl chickens after ilwwlation with g. 8091 c
and of unimcul sat/ed cmrvantional (£116ch 3 . . . . 0 33

33cm) piazza glutrmio-mcalncatlo trmcmnnco vane.
01' usinomfiated cez'menticwsl chickens, md 01' gnatobiotic
and conventiomtl chickens administered fi. 33’s?! 3‘6”

mil-13.0.0.0...0...0......000 35

The 5:?“
had no ext-3" ~'
c ~ I
1.6..) d‘: 721%"
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00001031 021 ,

is 0. 11:11:10 regard-'1 tool 1000.10

1-,, ,, ,. ,_ .3 -
15.0 9.3-0.130 0....

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(1-031 0'... :3 $3.31.. -....a.10:;;.~.0.- 1,

In 1.2110 pain:

the term 11:31:13.0 13 used to (303-1010 22:11:10.16 11-1-21 0:0 1‘1311'1‘311 {1‘00 of

9.1;}; oil‘scrr £011: of Life,

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interdmmgcamy with 0:1: :15- 0
Tina G .v’im 1: mtul 00' .10.

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32:10:23 which 0011.11 10 100.. .1500 £3017. £0011

{'29 1.101”? $3.19 {1.10 r

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rate 11.6 Lit-2;?
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54:112.":03 313d coaveutisml rats we "first 121 “2.2143 tawny; of ccmfiee
mtg the 9.1242213!) freeman was 17 :3 granary tim- . $2-031- 00 31m $3432.:
mnmmrgx-J’te while '25, .4.) total $0244.13 :1 content £1430 04:1:1 0329-21331 tézrt.
of 1.1 o coavontiaxrl rat. 110:: an (1‘5512‘: 3303.333 2.121. 75 any 0141
(3:127er c214" 7149 have 17 93. more comm 2:1‘ 3'3“”: that. ‘2'. 4.132.103;qu
chick-23:21} ‘25. t t.‘ 0 tub]. serum {3 :.Jlmlz: n m: tent is 36" .‘J 143-75

flat .3 12 e ccrzvm. 4; ”my; €414. 3:241:53.

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11 your! Ila-x1 1/2- uhioh an. placed an tho gum... otool
qlindot m» mrmsauon;

m. Uotor was aux-mud 1n 2 11m- squuo-Puk‘ nun.

mm: A334 mm. A 2 x ooluuon or Poroootio ”145
no and no tho gomioido fixroughout this study. An anionic oynthotio

(Iota-cont, on mm 9.171 oultonato moreiolly known so laoconol6.
no Walton! into tho poroutio sold ooluuon.

mmw- mun-rm:- Mina-aw
MMtotISOOforoporiodofthmn-o. Wandongooooro

“unis“ Iv opening with 2 1 oquoouo pornoouo ootd elation
“madilooconolmohworo thonollowdtootondtorao
that“. u that “no tho do in tho ioolator mm m displaool
loin. o1: ohm-111M by filtration. m an wan otorfliood lg
plooinc 700 g mutt“ into uranium pm. Thou pm voro phood
nthoqlm.udthooponoudotthooynndorm1odvnhwlu
(unnumbu' 853 polyoom hpoandmmhort'llvinyl topo.
tho cylindoro m ploood in on autoclovo and oubjoctod to tho
following otormoauon proooduro: (a) mention of tho autoolovo
W to 28 inch” of um. (13) ochioaion of atom: to ohmbol'
an). o moan-o o! 10 poundo no roachod. (o) «amnion of tho

‘0 mm 8%!” Company, 321., Plo

5. Ioooo Meal Compaq. low Ion. How York

6. lotions}. mum. Division of Allied Chemical corporation.
low not, low not

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Brod ‘3 fl. (1957). Thou organ-o voro toolatod tron tho toooo
of conventional chickens. Lt tho beginning of o booting ported 10 I1
douhmmlmroot tho tootorganionwhiohhadbougromh
mum mum mm. minoonlatod diroctly into them-mun.
tutor of tho toot ohlokono.

W 2: M. Blood no obtainod by oordloo punotnro
ollovodtoolototflcrorlhmrtohamotmormnlquonuttoo
uoro phood in 50 n1 Erylormyor fluk- uhloh oontainod 20 I; of
pom-om onlnto ncoording to tho nothod of Stem 31;. g... (1950).
fho plum was ooparatod In: tho cello by centrifugation and on
W by loano of n capillary pipetto. Enmo analyooo voro
omiod out Mediatoly, but plasma for antibody analysi- was
otorod ot ~17 C until touted.

MWo Anus-n va- mwod by Nina: the wt
can. 13 brain hoax-t infusion booth for 24 hours at 35 G. Fonda

no ddod to tho broth to o oonoontrnuon of 0.5 i no suggested by
Moon: (1957) and gloao bond: uoro added to tho oulturo duel: no
ohohn tn 1 hour at 35 C to brook up olunpo of collo. rho culturoo
m thou incubatod 19 houro at 35 o and oontrifugod at ; rolotlvo
centrifugal foroo of 9750 x (Ix-adv. Tho oollo woro thou rompondod
in n 0.85 ‘ 88.01 solution o3 xfl 7.0 no suggested by Lino (1949).

Ibo oollo uoro washed throo tmoo in thin oalino solution bororo thaw
voro otoodu‘dizod to n donoity corresponding to Tubo 2 of McFarland
(1909). Tho standard ounponoion or bacterial collo van otorod ob l c.

0"."

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RESULTS

mmmmmmm

Inrabhiandfignrol tho amber of organism pox-grand
too» and the norm antibody titers of the group I snatchiotio
Mutant m given. Tho mum thorn and the MM of organim
1n the feces inoroaeod until 3 wk- after inoculation, after which
that was a deal-om in tho antibody than and a marked aux-mo
uthonmberofg.£§ggg}gm.w1nmo tacos. '

TABI! 2. 8m antibody titers and tho number of

m.w1nth¢feoenofth¢¢mtobioucm on. my
1 after inoculatiano

 

 

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that: nftc Antibody Moan Omani-o
Inoculation Titer: titer . per g fem
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o o
4
6 £
1 6A
a. 61. 1.9 x 109
2 256
128 192 2.7 I 1011
3 256
512 384 1.0 x 101’
4 320
320 320 1.0 x 1010
5 160
160 160 1.0 x 109

 

100 OF ANTIBODY TITER

  

A-A Antibody Titer

 

 

 

no"

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to

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F I

T1133 AFTER *~:ocummo:¢ (WEEKS

Figure 1. Mean serum antibody titer and the number of _S_. faec
zvzrepens in the feces of the chickens from group I.

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had We: are inmated twice that; tho pox-ind after mam.
Whoa plum antibody reached I peak at 3 min following 1300111an
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fiduhtbofmoat‘mkamincedatimntnuuroffl.
m de‘h an aim In mm. 3.

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M. thrice}. antibody motions Hart detected in the plum of
the group I? Wblotlu Mid Wilmal chicken. £ dam. fitter
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‘8
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C) A? 7 Ci 1’

TIRE AFioR IHOCULATION (DAYS)

Figure 2. Mean plasma antibody titers of gnotobiotic, inoculated and
uninoculated conventional chickens.

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Proceedings of. The Snow! symposiu- 0n Gnotobiotio toohnobg.
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Exp. Biol. and Mad. 93196499.

‘Owlm. H. B. 1953. Th. Origin Of FCC-3&1 Antibody. J. o: W.
£33590 _

Moo, 8. J. 1949. Th. Agglutimtian of Autoclavod Homolytio
Streptococci By Sam»: {You Patients with fihmtio Pm
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Hworland. J. 1909. Th. “331%”. ’0 A“ “Cd. Assoc.
£31176-1178. ‘

Won. 2.. no ’1: Ruth, M Ha R. Woli’o. 1954. m01p1t13
Production In Chickens, X Cellular Changes in Thai Spleen
During Antibody Formation. J. of Ismael. QM

Hakimdzm, 709 R. F. Ruth. mu! 3. R. "01!... 1954. m1p1m
Production In Chickmm. 1! 81a of Antibody Production.
J. O: W. £345.51.

Palmer, H. I. 1957. Agglutination. Mama). of Microbiological
f Methods. 3). 336-238. MoGrav—‘z’iill Book Company, Inc... lieu lurk,
' Toronto, Loam.

Wk. J. "a, R. V. Monte, :0 ‘0 Wm, and I. H. filddlo.
1953. Pcmtialitieo 01' tho Immocyu, with on kdditianal
Ramos to It: Dyafimction In Hodgkin's 018mm. Am.

I. I. load. 803.. 2138-38.

Reyniaro. J. A... 1949. Historical Revlon. Roaring om»
mum. Lobmd Reports No. 2. p. M.- Unimoity a:
flair. Du Prose. Rota-o %m, Ind.

Remain-I. I. A. 1959. Mata-y. Ann II. I. Aoad. Sci. fits-4:.

47

Rainier” Jo Ag. 3. ‘0 60’6“}. R. to Em. and H. “RM. 1%.
Reports No. 3. p. 3.33-5.33. Unimoity of Mr. Om Pnol.
Rot" Dame, Ind. .

Sign Chmiml Company. 1961 .. Technical Bulletin BO. 10‘.
3500 mm St... St. Louis 6, No.

Sign mammal 0071339 . 1961 b. Technical Mlotin No. 505.
3500 M311?! 5%. to ME 8, HO.

hula. P. 0.. and L. Reynolds. 1957. Pleadblo Pill! Apparatus
for the hearing and Use of Gomfroo Animals. 19:31. Microbiol.
j’ II)“ :‘1 2.

Trade. P. O. 19:99. The Use ctr-Plastics In The Design of 130an
«Systems. ‘33 R. I. head. 0: 531. £329" 1

magnet. 34., am! 3. Romania. 1953-1959. Studio!!! on Hamfimlnatod
Chicktml Scott ‘- ., . and tremtoc can nos: )
Pad Panic in. An‘ I: t. Ann. 9. m Medic
Encyclomdia. 1110. Rev York, I. I.

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”9188. 1!. Po. 811d D. U. FQaMtQ 1953. 03190638311631 013mm
02: Chicken Monocytoa Hamphagoa and Giant Call. In riom
01113.31". J. Iliamchm. Cybochm. I‘Q‘GSO

Whit... L. P. 1960. lawman in The magmatic Use of Eamon.
Enzyme: In Health and Discos... p. 331-351. C. 0. m...
Spoingfiold, Ill.

Hum, 69 8., and A. A. Mil-03. 1957. TOPIC! 3m “11“.. “mime.
of Bacteriology and Imumiw. p. m. the Killian.
m ”mirzfi cmflzm. BfiltWOQ Md.

ma. a. n. and a. pm... 1948. Prooipitin Production 1: 01102“.
a. or W. 23245-250.

um, B. 5. mgtu‘aflishoi. Personal memioation.

Putnam. B. 8. 1959. Sam Protons In Goran-o. Yamahmtoo.
Am. 3. I. head. of Sci. E!254P2(Jlu

“0mm, B. 5., K. “REIT-.94.“, (Kid 8. ‘. Gm. 1959.1960.
Effects of Frocaino Pencillin in cuckoo: Monooontaminatod
with glorttrfi... 11:3 mk‘fimo and {trogmmgw weer-.113.
Antihiot. Ann. p. SILL-f . 191-3511 Emyolcmm. Inc.
Hm I'm, N. I.

 

Wootmrm, I. 8.. and H. A. Gordon. 1960. Eloctrogahoro‘hio Stxfliel on
1h. 5m Protein Pattern of The comm-ea Rat and 117.3 momma
Upon Enema to A Commtioml Eactrarifl Flam. I. of Itmmol.

34:27-31.

Hroblouaki. I... and J. 8. Lanes. 1956. Scru- Glutomio-Pyrmrio
We. (SOB-1') in Hepatic Disease: A Prolininary
Rem. m. m. “84.. 52:801-811.

Wroblomski. L. G. Jarvis, and J. 8. Who. 1956. The magmatic,
Progawtio, Epidemiclogio Bigfii‘lazmoo of The Alteration of
8mm Glutnnio-Oxdacotio Transqmlnneo 1n Hepatitis. Ann
of Int. Med. 553783—8000

Zamecnik, P. 0.. E. L. Swtfizmx, and 0. Cape. 19:35. Papudaoo
Activity of Lymuh and Berna Arbor E'urna. J. Biochan.
£31391“...

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