close

Вход

Забыли?

вход по аккаунту

?

Influence of certain dietary and hormonal factors on the development of resistance to neuro-invasiveness by vesicular stomatitis virus

код для вставкиСкачать
INFORMATION TO USERS
This manuscript has been reproduced from the microfilm master. UMI films
the text directly from the original or copy submitted. Thus, some thesis and
dissertation copies are in typewriter face, while others may be from any type of
computer printer.
T h e quality of this reproduction is d e p e n d e n t upon the quality o f the
co p y submitted. Broken or indistinct print, colored or poor quality illustrations
and photographs, print bleedthrough, substandard margins, and improper
alignment can adversely affect reproduction.
In the unlikely event that the author did not send UMI a complete manuscript
and there are missing pages, these will be noted.
Also, if unauthorized
copyright material had to be removed, a note will indicate the deletion.
Oversize
materials
(e.g.,
maps,
drawings,
charts)
are reproduced by
sectioning the original, beginning at the upper left-hand comer and continuing
from left to right in equal sections with small overlaps.
ProQuest Information and Learning
300 North Zeeb Road, Ann Arbor, Ml 481 06-1346 USA
800-521-0600
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
R eproduced with permission of the copyright owner. Further reproduction prohibited without permission.
UNIVERSITY OF CINCINNATI
May 20
/ 9
^°
I hereby recommend that the thesis prepared under m y
supervision h y
C arl Edward Du f f y _____________________________________
entitled _______I n flu e n c e
o f C e rta in D ie ta r y and Hormonal F a c to r s on th e
Development o f R e s is ta n c e t o N e u r o -In v a siv e n e ss by V e s ic u la r
S t o m a tit is V iru s
be accepted as fulfilling this part o f the requirements fo r the
degree o f _________ D octor
o f P h ilo so p h y ___________________________________
A p p ro ved by:
_________________
F o rm GGS— G. S. a n d E d.— 1M— 7-37
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
R eproduced with permission of the copyright owner. Further reproduction prohibited without permission.
INFLUENCE OE CERTAIN DIETART AND
\
HORMONAL FACTORS ON THE DEVELOPMENT OE
RESISTANCE TO NEURO-INVASIVENESS BY
VESICULAR STOMATITIS VIRUS
A d is s e r t a tio n subm itted t o th e
Graduate School
o f th e U n iv e r sity o f C in cin n a ti
i n p a r t ia l fu lf illm e n t o f th e
requirem ents fo r th e degree o f
DOCTOR OE PHILOSOPHY
1940
by
Carl Edward Duffy
11'
A. B. Ohio U n iv e r sity 1934
M. S . U n iv e r sity o f C in cin n a ti 1938
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
UMI Number: DP15734
IN F O R M A T IO N T O U S E R S
T h e q u a lity o f th is re p ro d u c tio n is d e p e n d e n t u p o n th e q u a lity o f th e co py
s u b m itte d .
B ro k e n o r in d istin ct print, c o lo re d o r p o o r q uality illu s tra tio n s a n d
p h o to g ra p h s , p rint b le e d -th ro u g h , s u b s ta n d a rd m a rg in s , and im p ro p e r
a lig n m e n t c a n a d v e rs e ly a ffe c t re p ro d u c tio n .
In th e unlikely e v e n t th a t th e a u th o r did n o t s e n d a c o m p le te m a n u s c rip t
a n d th e re a re m issin g p a g e s , th e s e w ill b e n o te d . A lso , if u n a u th o riz e d
c o p y rig h t m a te ria l h a d to b e re m o v e d , a n o te w ill in d ic a te th e d e le tio n .
U M I M ic ro fo rm D P 1 5 7 3 4
C o p y rig h t2 0 0 9 b y P ro Q u e s t L L C
A ll rights re s e rv e d . T h is m ic ro fo rm e d itio n is p ro te c te d a g a in s t
u n a u th o riz e d c o p y in g u n d e r T itle 1 7 , U n ite d S ta te s C o d e .
P ro Q u e s t L L C
7 8 9 E a s t E is e n h o w e r P a rk w a y
P .O . B o x 1 3 4 6
A n n A rb o r, M l 4 8 1 0 6 - 1 3 4 6
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
Acknowledgment
The s t u d ie s rep o rted in t h i s m anuscript were
c a r r ie d out in a s s o c ia t io n w ith Dr. A lb e r t B.
S a b in .
H is many v a lu a b le s u g g e s tio n s are d eep ly
a p p r e c ia te d .
C.E.D.
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
T ab le o f C ontents
I n tr o d u c tio n *
...........* 1
M ethods......................
.1 7
Experiment a l .............................18
D is c u s s io n .
Summary.
.............30
...............
£0
6 Photographs
5 T a b les
3 Charts
B ib lio g r a p h y
2 6 Ag ’4 0
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
Recent in v e s tig a tio n s (1 -7 ) w ith th e v ir u s e s o f v e s ic u la r
s to m a titis and equine en cep h alom yelitis have d is c lo s e d a type o f r e s i s t
ta n o e which cannot he a ttrib u ted t o p r io r exposure t o th e v ir u se s nor
t o t h e presence o f a n t iv ir a l sub stances in th e b lo o d .
I t was shown th a t
w h ile th e a b i l i t y or in a b i l it y o f t h e s e v ir u s e s t o invade the c e n tr a l
nervous system (C .N .S .) subsequent t o p erip h era l in o c u la tio n depended
on th e s p e c ie s o f th e h o st, other fa c to r s not h ere to fo r e observed were
a ls o in v o lv ed .
I t was noted th a t in c e r ta in an im als, th e young o f
which are not r e s is t a n t t o th e v ir u s e s when in je c te d p er ip h e r a lly ,
changes occur in c e r ta in c e l l s and t i s s u e s as th e anim als mature which
render them r e fr a c to r y t o th e v ir u s e s when in trod uced by c e r ta in p e r i­
p h eral r o u te s.
The changes are not g en era l in th e sen se that a com­
p l e t e r e s is ta n c e i s e sta b lish e d r e g a r d le s s o f th e ro u te o f in je c tio n
but ra th er occur in d e f in it e lo c a liz e d areas such a s in s p e c ia liz e d
n erve endings, c e r t a in reg io n s o f th e C .N .S ., in th e blood v e s s e ls or
in o th er t is s u e s such a s m uscle.
When t h e s e changes occur, s in c e th e
anim als remain s u s c e p tib le t o in tr a c r a n ia l and in tra n eu ra l in je c tio n o f
th e v ir u s e s , i t appears th a t th e above mentioned t i s s u e changes fu n c tio n
a s c o n s tit u tio n a l b a r r ie r s which prevent th e v ir u s e s from en terin g th e
O.R.S.
In v ie w o f th e f a c t th a t t h e s e c o n s t it u t io n a l b a r r ie r s appear
a s t h e anim al m atures i t would be o f i n t e r e s t t o know w hether, by vary­
in g c e r t a in d ie t a r y f a c t o r s w hich a re e s s e n t i a l f o r a growing a n im a l, th e
development o f t h e s e b a r r ie r s cou ld b e p rev en ted or d e la y e d .
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
2
S in c e a t l e a s t one o f t h e s e b a r r ie r s a r i s e s c o in c id e n t ly w ith
s e x u a l m atu ration i t was d ecid ed t o in v e s t ig a t e t h e in flu e n c e o f c e r ta in
hormonal f a c t o r s , which ind uce " p r e co c io u s” se x u a l m a tu ra tio n , on th e
development o f t h i s b a r r ie r .
T e s ic u la r s t o m a t it is v ir u s in j e c t e d in tra m u scu la rly in t o a l­
b in o m ice was s e le c t e d f o r t h e stu d y fo r th e f o llo w in g r ea so n s:
1) a l ­
b in o m ice a s th e y grow o ld e r e v e n tu a lly become 100 per cent r e s i s t a n t t o
t h e v ir u s when in j e c t e d .b y t h e above r o u te whereas th e number o f o ld m ice
w hich are r e s i s t a n t when in o c u la te d by o th er r o u te s i s v a r ia b le , 2) when
in j e c t e d by t h i s r o u te th e v ir u s produces r e a d ily d e te c ta b le c l i n i c a l
m a n ife s ta tio n s o f th e involvem ent o f th e C .N .S . o f n o n -r e s is ta n t m ice,
3) t h e mode o f spread and p a th o g e n e sis o f t h e v ir u s has been worked out
in c o n s id e r a b le d e t a i l , and l±) th e s u s c e p t i b i l i t y o f normal m ice a t d i f ­
fe r e n t ages i s known th u s p e r m ittin g c o r r e la t io n betw een th e d a ta ob­
ta in e d under t h e v a r io u s ex p erim en ta l c o n d itio n s w ith th a t a lr e a d y e s­
t a b lis h e d .
The i n f e c t i v e agent ( v e s ic u la r s t o m a t it is v ir u s ) u se d in th e
s tu d ie s t o b e r ep o rted in t h i s paper i s c la s s e d w ith th o s e o b lig a t o r y
in t r a c e l l u l a r p a r i s i t e s known a s f i l t e r a b l e v ir u s e s o r sim ply a s v ir u s e s .
All v ir u s e s have t h e 'f o llo w in g p r o p e r tie s :
1) th e y p a ss through f i l t e r s
which w i l l , r e t a in most ib a c t e r ia , !12) th e y cannot h e seen w ith t h e ord in ary
m icro sco p e, 3) th e y m u ltip ly o n ly in th e p resen ce o f l i v i n g c e l l s .
In n a tu r e v e s ic u la r s t o m a t i t is v ir u s c a u ses a m ild d is e a s e i n
h o r s e s and c a t t l e .
In t h e s e an im als th e le s i o n s produced by th e v ir u s
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
3
a re lim it e d t o t h e "buccal c a v ity 'a n d th e v ir u s does not invade t h e cen­
t r a l nervous system *
T e s ic u la r s t o m a t i t i s , caused by th e v i r u s , does not occu r in
n a tu re in t h e a lb in o mouse.
However, when e x p e r im e n ta lly in je c te d ; in t o
young w h ite m ice th e v ir u s has a ten d en cy t o in v a d e th e c e n tr a l nervous
system and produce a f a t a l a scen d in g f l a c c i d p a r a ly s is or a f a t a l en­
c e p h a l i t i s depending on th e r o u te o f in j e c t i o n .
For t h i s r ea so n th e
v ir u s has b een u sed in experim en ts d e a lin g w ith immunity t o v ir u s d i s ­
ea ses.
S in c e thei in tram u scu la r r o u te o f in j e c t i o n was used e x c lu s iv e ly
i n th e s tu d ie s r ep o rted in t h i s paper i t m ight b e w e ll t o c o n s id e r th e
manner by w hich v e s ic u la r s t o m a t i t is v ir u s i s in h ib it e d from e n te r in g
th e c e n tr a l n ervou s system o f r e s i s t a n t m ice when in tro d u ced by t h i s
r o u te .
I t has bebn p o in ted out e a r l i e r th a t t h e r e s is t a n c e t o in t r a ­
m uscular i n j e c t i o n o f th e v ir u s i s not n e c e s s a r i ly a s s o c ia te d w ith a
general r e s is ta n c e .
Old m ice w hich a re r e s i s t a n t t o in tra m u scu la r in ­
j e c t io n o f t h e v ir u s ar6 n e v e r t h e le s s s t i l l s u s c e p t ib le t o in t r a c r a n ia l
in o c u la t io n .
S ab in and O lit s k y ( 1 ,2 ) showed th a t a lth o u g h young and
o ld m ice dem onstrated g rea t d if f e r e n c e s in th e d eg ree o f s u s c e p t i b i l i t y
t o in tr a m u sc u la r --in je c tio n o f v e s ic u l a r s t o m a t i t i s v ir u s th e y were e q u a lly
s u s c e p t ib le t o in t r a c r a n ia l in o c u la t io n .
The same a u th o rs (2) showed
th a t th e s im ila r s u s c e p t i b i l i t y o f young and o ld m ice t o in t r a c r a n ia l
in o c u la tio n depended upon t h e manner i n w hich t h e v ir u s spread in th e
b r a in .
They were a b le t o show by m ic r o sc o p ic stu d y th a t th e apparent
l e s i o n s in t h e b r a in were co n tig u o u s w ith th e v e n t r ic u la r system* and w ith
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
4
t h e c e n tr a l carfal o f th e s p ih a l cord .
A lthough th e nerve c e l l s o f o ld
m ice were more r e s is t a n t t o in v a s io n o f th e v ir u s , a s in d ic a te d by few er
l e s i o n s in t h e b r a in s o f o ld m ic e , than were t h o s e o f young m ice never­
t h e l e s s th e v ir u s was a b le t o g a in a c c e s s t o t h e v e n t r ic le s and by u sin g
a v e n t r ic u la r mode o f spread was a b le t o a tta c k and m u ltip ly in a s u f­
f i c i e n t number o f s u s c e p t ib le c e l l s t o produce a f a t a l e n c e p h a lit is even
in o ld m ice.
The above a u th o rs concluded th a t t h e equal s u s c e p t i b i l i t y
o f young and o ld m ice to in t r a c r a n ia l in j e c t i o n o f th e v ir u s was due t o
th e v e n t r ic u la r or "open system " mode o f spread and not to th e eq u al s u s ­
c e p t i b i l i t y o f a l l neu ral t i s s u e .
S in c e th e l e s i o n s a re lim it e d t o a rea s
co n tig u o u s w ith t h e v e n tr ic u la r system and t h e c e n tr a l can al o f t h e s p in a l
cord i t i s n ecessary...to assume th a t th e v ir u s i s n ot p resen t i n t h e cereb ro­
s p in a l f l u i d f o r i f such w ere tr u e one would ex p ect to fin d l e s i o n s in
o th e r p a r ts o f th e b r a in w hich a re in c o n ta c t w ith c e r e b fo -s p in a l f l u i d .
The r e s is t a n c e o f o ld m ice t o in tra m u scu la r in j e c t io n o f v e s i ­
c u la r s t o m a t it is v ir u s bas b een shown ( 1 ,3 ) t °
dependent upon th e
mode o f spread o f t h e v ir u s from t h e m uscle t o th e c e n tr a l nervous s y s ­
tem .
The appearance o f p a r a ly s i s , fo llo w in g i n j e c t i o n o f th e v ir u s in t o
t h e c a l f m u scles o f th e hind l e g o f young m ice, f i r s t in th e in j e c t e d
l e g and th e n in t h e o p p o site member s u g g e sts th a t t h e v ir u s r ea c h e s th e
c e n tr a l, nervous system by sp rea d in g a lon g th e n erve supply t o t h e m uscle
and th u s in v a d in g t h e s p in a l c o r d . The c h i e f p a th o lo g ic a l l e s i o n s o f
were found
t h e c e n tr a l nervous systemAin th e a n te r io r horn c e l l s o f th e lumbar r e g io n
o f th e s p in a l cord fo llo w in g t h e in j e c t i o n o f th e v ir u s in t o th e m u scles
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
o f th e hind l e g .
T hese l e s i o n s c o n s is te d o f v a r io u s s ta g e s o f n e c r o s is
and e a r ly neuronophagia.
No le s i o n s were found in th e m eninges or
s p in a l and sym p ath etic g a n g lia .
From th e ahove fin d in g s i t would ap­
pear th a t th e v ir u s , when in j e c t e d in t o th e m u scle o f s u s c e p t ib le m ice,
e n te r s t h e p e r ip h e r a l n e r v es and by fo llo w in g th e e f f e r e n t f i b e r s g a in s
a c c e s s t o th e c e n t r a l nervous system .
S in c e th e v ir u s i s u n ab le t o in ­
vade th e c e n tr a l nervous sy stem o f o ld m ice when in j e c te d in to th e
m u scles o f t h e hind l e g t h e r e must e x i s t a b a r r ie r w hich h o ld s t h e v ir u s
back.
T h is b a r r ie r cou ld be s it u a t e d e it h e r in th e p e r ip h e r a l n e r v e s,
in th e s p e c ia l iz e d nerve en d in g s or in th e m u sc le .
The b a r r ie r t o
n e u r o in v a siv e n e ss may not b e i n th e nerve i t s e l f f o r a s has b een shown
M
'
'
(1 ) th e v ir u s , when in j e c t e d d i r e c t l y in t o t h e s c i a t i c nerve o f o ld ,
r e s is t a n t m ice in v a d es th e c e n t r a l nervous system and produces an ascend­
in g f l a c c i d p a r a ly s is such a s th a t seen in young m ice fo llo w in g in t r a ­
m uscular in o c u la t io n .
I t sh o u ld be n o ted , how ever, th a t in j e c t io n o f
t h e v ir u s d i r e c t l y in t o t h e n erv e may p la c e th e v ir u s in c o n ta c t w ith
broken o r damaged axones and th u s p rovid e a means o f e n te r in g t h e sp in a l
cord .
I t has been r e p o r te d (1 ,2 ) th a t v e s ic u l a r s t o m a t it is v ir u s i s
a b le t o m u ltip ly in t h e m u scle o f 15 day o ld m ice w ith t h e p ro d u ctio n
o f n e c r o s is o f t h e m uscle c e l l s to g e th e r w ith hypertrophy and p r o lif e r a ­
t i o n o f t h e sarcolem m al n u c l e i .
No ev id e n c e o f m u lt ip l ic a t io n was ob­
ta in e d when t h e v ir u s was in j e c t e d in to th e c a l f m u scles o f o ld , r e ­
s i s t a n t m ice and no l e s i o n s w ere produced.
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
6
The a b i l i t y ctf th e v ir u s t o m u ltip ly in th e m uscle o f young
m ice and i t s i n a b i l i t y t o do so in o ld m ice s u g g e s ts th a t t h e b a r r ie r
t o n e u r o in v a s iv e n e ss may be w ith in th e m u scle i t s e l f .
However i t has
been shown (4 ) th a t in th e g u in ea p ig , an anim al w hich i s a ls o s u s c e p t ib le
t o in t r a c r a n ia l in j e c t io n o f t h e v ir u s , a lth o u g h m u lt ip lic a t io n o f th e
v ir u s occu rred a f t e r in tram u scu lar in j e c t i o n th e r e was no e v id e n c e o f
in v a s io n o f t h e c e n tr a l nervous sy stem .
Here a g a in i f th e m u scle and
nerve en d in gs a re circum vented and in j e c t i o n made d ir e c t ly in t o th e
s c i a t i c n erve an ascending f l a c c i d p a r a ly s is d e v e lo p s as in m ic e .
P re­
lim in a r y l o c a l m u lt ip lic a t io n o f t h e v ir u s th e n i s not th e o n ly r e q u i s it e
f o r in v a s io n o f t h e c e n tr a l nervous sy stem .
I t sh ou ld be m entioned in
t h i s c o n n e c tio n th a t r a b ie s v ir u s i s a b le t o invad e a p e r ip h e r a l nerve
w ith ou t p r e lim in a ry lo c a l in c r e a s e ( 8 ) .
A summary o f th e p reced in g d is c u s s io n r e v e a ls th a t v e s ic u la r
s t o m a t it is v ir u s when in j e c t e d in t o th e c a l f m u scles o f young m ice under­
g o es l o c a l m u lt ip lic a t io n and th en in v a d es th e c e n tr a l nervous system
by way o f t h e p e r ip h e r a l n e r v es t o produce a f a t a l ascen d in g f l a c c i d
p a r a ly s is .
When t h e v ir u s i s in j e c t e d in t o th e c a l f m u scles o f o ld ,
r e s is t a n t m ice t h e v ir u s f a i l s t o p r o l i f e r a t e and does not in v a d e th e
c e n tr a l nervous system .
E v id e n tly a s m ice grow and develop a b a r r ie r
i s e s t a b lis h e d through w hich th e v ir u s cannot p a s s .
T h is b a r r ie r i s
a t p r e se n t b e lie v e d t o be lo c a te d e it h e r in th e m u sc le , in th e nerve
en d in gs o r in b o th .
The s u s c e p t i b i l i t y o f w h ite m ice o f v a r io u s ages t o in t r a ­
m uscular in j e c t io n o f v e s ic u la r s t o m a t i t is v ir u s has b een shown ( 1 ,2 ,7 )
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
t o b e a s fo llo w s :
100 p er cen t o f 2 week o ld m ice d ev elo p a f a t a l a s -
cen d in g f l a c c i d p a r a l y s i s .
At 21 days o f a g e '80 t o 90 per cen t are s u s ­
c e p t ib l e and a t t h e age o f 1 month 80 t o 90 per cen t a r e r e s i s t a n t in
th a t th e y show no s ig n s o f d is e a s e .
Between th e 5 th and 6t h weeks o f
l i f e about 100 per cen t a r e r e f r a c to r y t o th e v ir u s when in j e c t e d by
t h e above r o u te .
B ecause t h e change from 100 p er cen t s u s c e p t i b i l i t y t o a p p roxi­
m a tely 100 p er cent r e s is t a n c e o ccu rs betw een t h e 14t h and 35t h day o f
l i f e and b eca u se m ice s u c k le during t h e p e r io d when th e y a re d ev elo p in g
r e s i s t a n c e , experim en ts w ere s e t up t o determ ine:
1 ) th e e f f e c t o f th e
m atern al d ie t during l a c t a t i o n and 2 ) t h e e f f e c t at th e cliet o f m ice
weaned a t 14 days o f age on t h e developm ent o f r e s i s t a n t t o in tram u scu lar
in j e c t i o n o f t h e v i r u s .
B ecause se x u a l m a tu r ity o ccu rs during th e p e r io d
when m ice become a p p roxim ately 100 per cen t r e s is t a n t t o t h e v ir u s an
experim ent t o determ ine th e e f f e c t o f " p reco cio u s” s e x u a l m atu ration was
in clu d ed in t h e stu d y .
A rev iew o f t h e l i t e r a t u r e r e v e a ls th a t s u r p r is in g ly l i t t l e
work has been .done on th e r e la t io n s h ip betw een n u t r it io n and h o st su s­
c e p t i b i l i t y t o v ir u s d is e a s e s .
A lthough i t has been s u g g e ste d ( 9 »10)
th a t v ita m in s are im portant f a c t o r s in immunity t o v ir u s d is e a s e s th e r e
i s l i t t l e exp erim en tal e v id e n c e t o support such a b e l i e f .
R e c e n tly
Jungeblut and F e in e r (11) r ep o rted th a t v ita m in C i s an e f f e c t i v e agent
f o r th e p r e v e n tio n and cu re o f exp erim en tal p o lio m y e lit is i n monkeys.
However, th e f in d in g s o f Jungeblut and F e in e r were not s u b s ta n tia te d by
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
3
t h e r e s u l t s o b ta in ed by S ab in (12) who found th a t b o th n a tu r a l and syn­
t h e t i c p r e p a r a tio n s o f v ita m in C, a d m in istered im m ediately a f t e r th e
4
v ir u s was i n s t i l l e d in t o t h e n o s e , had no e f f e c t on th e co u r se o f ex­
p e r im e n ta lly induced p o li o m y e li t is in t h e monkey.
The l a t t e r author
showed th a t monkeys whose s t o r e o f v ita m in C was d e p le te d by fe e d in g a
d ie t d e f i c i e n t in t h e v ita m in r e a c te d in th e same way a s d id monkeys
r e c e iv in g an adequate d i e t .
Cowdry, Lucas and N e ff (13) s tu d ie d th e e f f e c t o f a d e fic ie n c y
o f v ita m in
and B2 on t h e r e s is t a n c e o f r a t s t o herpes v ir u s in j e c te d
in t r a c e r e b r a lly .
They concluded th a t r a t s d e f i c ie n t in v ita m in B^ and
Bg were s l i g h t l y more s u s c e p t i b l y to...the v ir u s than were normal r a t s .
Eowever t h e i r data are not c o n c lu s iv e s in c e t h e m inimal l e t h a l cer e b r a l
d o se o f th e v ir u s and t h e proper amount o f f l u i d w hich sh o u ld be in ­
je c t e d w ere a l l determ ined du rin g t h e co u rse o f e x p e rim en ta tio n .
If
t h e m inim al c e r b r a l l e t h a l d ose f o r t h e r a t had been determ ined b e fo r e ­
hand and k ep t co n sta n t du rin g t h e co u rse o f t h e experim ents th e r e s u lt s
m ight b e more im p r e ssiv e ,
K uczynski (.11) [q u o ted from P e r la ( 15 ) ] found th a t m ice fe d
d i e t s r i c h in v ita m in B^ were more r e s i s t a n t t o th e v ir u s o f y e llo w
f e v e r , in tro d u ced in t r a c e r e b r a lly , than w ere m ice fe d a stand ard d i e t .
Of 671 m ice f e d on a normal or standard d ie t (o a t s and m ilk ) 20 per cent
su r v iv e d when t h e v ir u s was in o c u la te d in t o th e b r a in .
When m ice were
f e d a d ie t r ic h in v ita m in B1 (wheat husks and m ilk) 75 per cen t o f 28
an im als test-ed, were r e s i s t a n t t o in tr a c e r e b r a l in o c u la tio n .
Of 137
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
anim als fe d a d ie t f r e e o f v ita m in B1 f o r a p erio d o f l^ .d a y s o n ly
X
t
1 6 .7 per cen t were r e s i s t a n t .
I t appears t o b e j u s t i f i a b l e t o conclude from Kuczynski* s
d a ta th a t d i e t s r ic h in v ita m in
d id enhance th e r e s is t a n c e o f m ice
t o in t r a c e r e b r a l in o c u la tio n o f y e llo w fe v e r v i r u s .
Whether h i s con­
c lu s io n th a t t h e r e s is t a n c e o f t h e nervous system, to a l l v ir u s in f e c t io n s
i s dependent upon th e p r e se n c e o f an adequate amount o f v ita m in B1 in
t h e d ie t i s j u s t i f i e d cannot be s a id a t th e p resen t tim e .
S in c e m ice d ev elo p r e s is t a n c e t o intram uscu lar i n j e c t i o n o f
v e s ic u la r s t o m a t it is v ir u s when th e y are growing r a p id ly exp erim en ts
were planned t o determ ine w hether th e developm ent o f t h i s r e s is t a n c e
would b e p rev en ted or r e ta r d e d b y e lim in a tin g c e r ta in f a c t o r s from th e
d ie t which are known to b e e s s e n t i a l f o r growth and developm ent.
In
t h e s e r i e s ' o f experim ents r ep o rted h ere th e r o l e o f n u t r it io n on th e
development o f r e s is t a n c e was s tu d ie d from tw o d if f e r e n t a s p e c t s :
l)
s in c e m ice s u c k lin g m others fe d a d ie t r ic h i n n a tu r a l f o o d s t u f f s b e­
come BO - 90 p er cent r e s i s t a n t a t 28 days o f age an experim ent was
planned t o determ ine w hether m ice su c k le d by m others fe d a d ie t d e f ic ­
ie n t in c e r t a in vitaffiin s w ould a ls o become r e s i s t a n t , 2) a second s e t
o f experim en ts was planned t o determ ine th e in flu e n c e o f an i n s u f f i c i e n t
amount o f a d i e t c o n s is t in g o f n a tu r a l f o o d s t u f f s and o f d i e t s d e f ic ie n t
in c e r ta in v ita m in s on t h e development o f r e s is t a n c e in prem atu rely
weaned m ice.
D ie t s d e f ic ie n t in v ita m in B^ and r ib o f la v in (v ita m in B^)
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
10
w ere s e l e c t e d f o r th e s t u d ie s p r im a r ily b eca u se t h e s e v ita m in s are
known t o b e e s s e n t i a l fo r growth and developm ent ( 1 6 ,1 7 ) .
However a
d e f ic ie n c y o f e it h e r o f t h e above v ita m in s g iv e s r i s e t o o th e r p h y si­
o l o g i c a l and p a t h o lo g ic a l changes w hich may p la y a part in th e d ev el§
opment o f r e s is t a n c e in t h e grow ing a n im a l.
B efo re ta k in g up a d i s ­
c u s s io n o f t h e above m entioned p h y s io lo g ic a l and p a t h o lo g ic a l changes
i t might b e w e ll t o d is c u s s a t t h i s p o in t t h e n a tu re o f th e above
v ita m in s .
V itam in
and r ib o f l a v in (v ita m in B^) are tw o members o f
what i s now term ed ’th e B com plex.
They are b o th p r e se n t in y e a s t .
V itam in B^ i s o f t e n term ed th e a n t in e u r it i c v ita m in .
b e r i i n man and p o ly n e u r it is i n a n im a ls.
I t p r e v e n ts b e r i­
The v ita m in i s h e a t - l a b i l e
b e in g d e str o y ed at 120°C i n a n e u tr a l o r a lk a lin e medium.
A common
method f o r t h e p r ep a ra tio n o f a d ie t d e f i c i e n t in v ita m in B^. i s t o add
y e a s t w hich has been a u to c la v e d under 15 - 16 pounds p r e ssu r e t o a mix­
tu r e o f c a s e in , la r d , c o r n sta r c h and s a l t s .
However a d ie t made up in
t h i s manner i s not on ly d e f i c i e n t i n v ita m in B^ but a ls o in o th e r heat
l a b i l e components o f th e B com plex, such a s v ita m in B^, w hich p r e v en ts
p a r a ly s is in r a t s and c h ic k s .
S in c e th e B^ d e f i c ie n t d ie t u se d in th e
p r e se n t s tu d ie s was made up a s in d ic a te d above i t t o o was d e f i c i e n t in
more th an v ita m in B^.
Throughout t h e rem ainder o f th e paper t h e d ie t
w hich was u sed t o stu d y t h e in f lu e n c e o f a d ie t d e f ic ie n t in v ita m in B^
w i l l b e term ed "a d ie t d e f i c i e n t in th e h e a t - l a b i l e E-complex" i n order
n ot t o im ply th a t t h e on}.y d e f ic ie n c y was in v ita m in B^.
i
R ib o fla v in or v ita m in B0 i s a w a te r - s o lu b le y e llo w -g r e e n
^
\
f lo r e s c e n t p igm en t. I t i s found i n y e a s t and i s not d estro y ed by
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
11
a u to c la v in g a t 120°C.
I t i s n e c e ssa r y f o r growth in c h ic k s and r a t s .
I t i s a component o f an o x id a tio n -r e d u c tio n system (Warburg*s y e llo w
enzym e)(18) in l i v i n g c e l l s .
I n t h e p r e se n t s tu d ie s a d ie t d e f ic ie n t
in r ib o f l a v i n was made up by adding r i c e p o lis h in g s c o n c e n tr a te ( r ic h
i n v ita m in B^) t o a m ixtu re o f c a s e in , l a r d , c o rn sta rch and s a l t s .
I t i s e v id e n t from t h e co m p o sitio n o f t h i s d ie t th a t i t i s d e f ic ie n t
in o th e r f a c t o r s o f t h e B com plex b e s id e s r ib o f l a v in .
I n order not
t o g iv e th e im p ression th a t th e d ie t was d e f ic ie n t o n ly in r ib o f l a v in
t h i s d ie t
has been term ed na / d i e t d e f i c i e n t in th e B-eom plex except B ^ .w
No attem pt w i l l be made a t t h i s tim e t o p r e se n t a rev iew o f
a l l t h e r ep o rted p h y s io lo g ic a l and p a t h o lo g ic a l changes w hich have been
a t t r ib u t e d t o a d e f ic ie n c y o f v ita m in B^.
seem t o be r e la t e d t o th e
p r e se n t problem w i l l be d is c u s s e d .
lo n g been known th a t a d e f ic ie n c y
m uscular symptoms.
Only th o s e changes which
I t has
o f v ita m in B le a d s t o d e f i n i t e neuro­
In r a t s and b ir d s t h e s e symptoms a re m a n ife sted a s
s p a s t ic p a r a ly s is , a t a x ia , o p isth o to n u s and c o n v u ls io n s.
I n a d d itio n
t o th e above a " lo ss o f a p p e t it e o ccu rs i n anim als fe d a B - d e f ic ie n t
d i e t w ith a consequent, l o s s o f w e ig h t.
I t i s d i f f i c u l t in r e t r o s p e c t
t o a t t r i b u t e th e changes d e s c r ib e d by t h e e a r ly w orkers s o l e l y t o th e
la c k o f v ita m in B s in c e th e r a t io n s u sed in t h e experim en ts were de­
f i c i e n t in more th an one e s s e n t i a l n u t r ie n t , nam ely, p r o te in ( r i c e d i e t s )
m in era ls' and o th er v ita m in s .
The ex p erim en ta l anim als were t h e r e fo r e
s u f f e r in g from a m u ltip le d ie ta r y d e f i c ie n c y .
L a ter in v e s t ig a t o r s
have e lim in a te d from t h e i r d i e t s , a s
f a r as p o s s ib le , on ly th e B complex or c e r t a in o f i t s c o n s t it u e n t s .
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
12
S t e m and F in d la y (19) s tu d ie d th e changes i n th e n ervou s system o f
r a t s fe d a d i e t d e f ic ie n t in v ita m in B]_.
They found th a t d e g en era tio n
o f th e m y e lin sh e a th s o f p e r ip h e r a l n erv es occu rs when r a t s a r e fe d
th is d ie t.
Zimmerman and Burack (20) found th a t th e r e was e x te n s iv e
d e g e n e r a tio n o f th e m y elin sh e a th o f th e s c i a t i c n erve in dogs de­
p r iv e d o f w a te r -s o lu b le B v ita m in s .
D avison and S ton e (21) a ls o found
e x t e n s iv e d e g e n e ra tio n o f th e m y elin sh ea th o f th e s c i a t i c n erve in r a t s
f e d a d ie t d e f i c i e n t i n v ita m in B^«
The f a c t th a t d e g e n e ra tio n o f th e m y e lin sh e a th o f p e r ip h e r a l
n e r v e s o ccu rs i n an im als f e d a d i e t d e f ic i e n t in v ita m in B^ assumes
g r e a t im portance from th e v iew p o in t o f th e p r e se n t s tu d y .
I t w i l l be
r e c a lle d th a t when v e s ic u la r s t o m a t i t is v ir u s i s in j e c t e d in t o th e
m u scle o f r e s i s t a n t m ice i t does not invade th e p e r ip h e r a l n e r v es and
th u s does n o t invade th e c e n t r a l nervous sy stem .
I f s im ila r degenera­
t i o n o f th e m y e lin sh e a th o f th e p e r ip h e r a l n e r v es occu rred i n m ice fe d
a d i e t d e f i c i e n t i n B^_ i t m ight be p ex m issa b le t o assume th a v tn e v ir u s
c o u ld g a in a c c e s s t o th e axones o f p e r ip h e r a l n e r v es and th u s invade
th e c e n t r a l nervous sy ste m .
On th e o th e r hand i f th e b a r r ie r t o neuro-.
in v a s iv e n e s s i s d is s o c ia t e d e n t i r e l y from t h e p e r ip h e r a l n e r v e s and
i s lo c a t e d w ith in th e m u scle o th er e f f e c t s o f a d e f ic ie n c y o f v ita m in
m ust be c o n s id e r e d .
I t has r e c e n t ly b een shown th a t c o ca r b o x y la se i s e s s e n t i a l l y
a d ip h o sp h o rie e s t e r o f v ita m in B]_ ( 2 2 ) .
w hich a c t i v a t e s c a r b o x y la se .
C ocarboxylase i s a c o -fe im e n t
The l a t t e r enzyme i s im portant i n th e
carbohydrate m etab olism o f p la n t c e l l s in t h a t i t c o n v e r ts p y ru v ic a c id
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
13
t o a c eta ld eh y d e and carbon d io x id e .
As f a r a s t h e w r ite r i s aware no
i
s im ila r fu n c tio n o f vitam in
has been dem onstrated in anim al c e l l s .
However i t i s not beyond th e realm o f p o s s i b i l i t y th a t B^ may fu n c tio n
i n a s im ila r manner in animal c e l l s .
A f a i l u r e o f th e c e l l s o f th e
m u scle t o become r e s is t a n t t o t h e v ir u s might w e ll accompany any d i s ­
tu rb an ce in t h e carbohydrate m etab olism .
Zimmerman, C o w g ill and Fox (23) found th a t marked degenera­
t i o n o f th e m edu llary sh eath s £ihd a x is c y lin d e r s occu rred when dogs
w ere fe d a d ie t d e f ic ie n t in r ib o f l a v in .
However Zimmerman and Burack
(2 0 ) d id not o b serv e changes in th e p e r ip h e r a l n e r v e s o f r a t s d ep rived
o f t h e above v ita m in .
E v id e n tly dogs and r a t s do not respond s im ila r ly
t o a d e f ic ie n c y o f r ib o f la v in .
From th e ev id e n c e found in th e l i t e r a ­
tu r e i t i s d i f f i c u l t t o p o s t u la t e how a d e f ic ie n c y o f r ib o f l a v in might
a f f e c t t h e p e r ip h e r a l n erv es o f m ice and th u s ren d er them s u s c e p t ib le
t o in tram u scu lar in j e c t io n o f v e s ic u la r s t o m a t it is v ir u s .
However r e ­
cen t in v e s t ig a t i o n s have r e v e a le d th a t r ib o f l a v in i s a component o f an
o x id a tio n -r e d u c tio h system in l i v i n g c e l l s .
R ib o f la v in , a tta ch ed t o a
p r o te in m o le c u le through a phosphoric a c id nu cleu s^ ,is now known to be
a part o f Warburg’ s y e llo w enzyme.
c a r r ie r i n c e l l r e s p ir a t io n .
T h is enzyme s e r v e s a s an oxygen
I t i s c o n c e iv a b le th a t a d e fic ie n c y in
th e d ie t o f r ib o f l a v in would c o n seq u en tly le a d to d istu r b a n c e s in c e l l
m etab olism and in t h i s manner in t e r f e r e w ith th e normal development o f
r e s is t a n c e in th e grow ing an im al.
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
14
I n one s e t o f experim en ts m ice w hich had su c k le d m others fe d
a d ie t d e f ic ie n t e it h e r in v ita m in
s is t a n c e t o th e v ir u s .
o r r ib o f l a v in were t e s t e d fo r r e ­
I t i s e v id en t th a t w h ile th e anim als were suck­
li n g th e y were dependent upon th e mothers* m ilk a s a so u rce o f th e
above v ita m in s .
The r a te at w hich th e m other was d e p le te d o f r ib o ­
f l a v i n and v ita m in
ta k e .
would o f cou rse b e determ ined by t h e vitam in in ­
There appear t o b e no r e p o r ts i n th e l i t e r a t u r e on th e r a te o f
d e p le t io n o f v ita m in s in th e mouse.
Su re (24) has r e p o r te d th a t l a c t -
a tin g r a t s when put on d i e t s d e f ic ie n t in t h e above v ita m in s on th e
day o f p a r tu r it io n a r e d e p le te d o f t h e i r s t o r e o f v ita m in s in about 2
w eek s.
In th e p r e se n t s t u d ie s th e l a c t a t i n g m ice a llo w e d to fe e d on
a d ie t o f n a tu r a l f o o d s t u f f s f o r 2 days a f t e r d e liv e r y and w ere th en
put on t h e d e f ic ie n t d i e t s .
The e f f e c t o f a d ie t d e f ic ie n t i n v ita m in E on th e d ev elo p ­
ment o f r e s is t a n c e t o in tram u scu la r i n j e c t i o n o f v e s ic u l a r s t o m a t it is
v ir u s was s tu d ie d b eca u se i t has been shown ( 25) th a t when la c t a t in g
r a t s a r e f e d such a d ie t about 75 per cen t o f th e s u c k lin g young de­
v e lo p p a r a ly s is betw een th e 2 1 st and 2 5 th day o f l i f e .
O lc o tt (26) and
Pappenheimer ( 2 7 ) have s tu d ie d th e p a t h o lo g ic a l changes in young r a t s
due t o a d e f ic ie n c y o f v ita m in E.
They found th a t th e a lt e r a t io n s are
s t r i c t l y lim it e d t o v o lu n ta r y m u scle.
The l e s i o n s in t h e m u scles con­
s i s t e d o f e x te n s iv e d e g e n e r a tio n o f t h e m uscle c e l l s .
A lthough m uscular
d ystroph y due t o a d e f ic ie n c y o f v ita m in E has not b een r e p o r te d in th e
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
15
mouse t h e stu d y was n e v e r t h e le s s undertaken s in c e a la c k o f v ita m in E
seems t o p la y some r o le in th e w e ll-b e in g o f m uscle c e l l s and th e
m u scle may p la y a part in t h e developm ent o f r e s is t a n c e t o v e s ic u l a r
s t o m a t i t is v ir u s .
I t has been shown (2 8 -2 9 ) th a t young r a t s kept a t co n sta n t
w eigh t by u n d erfeed in g c o n tin u e to grow in th a t c e r t a in organs or p a r ts
in c r e a s e in w eight at th e expense o f o t h e r s .
The t e s t e s , su p r a r e n a ls,
e y e b a lls , c e n tr a l nervous sy stem , and s k e le t o n co n tin u ed t o grow w h ile
o th er organs e it h e r l o s e w eig h t or m a in ta in t h e i r o r ig in a l w e ig h t.
It
w i l l b e n oted th a t m u sculature i s not in c lu d e d among t h o s e organs or
p a r ts w hich in c r e a s e in s i z e when r a t s a re kept a t a c o n sta n t w e ig h t.
I t has b een n o ted above th a t t h e i n a b i l i t y o f v e s ic u l a r
s t o m a t it is v ir u s t o invad e th e c e n tr a l nervous system o f o ld m ice f o l ­
low in g in tram u scu lar in j e c t i o n may be a s s o c ia te ^ w ith th e f a i l u r e o f
t h e v ir u s t o m u ltip ly in t h e m u scle.
I f th e m u sculature o f young m ice
k ept on a p a r t ia l in a n it io n d ie t was found t o r e a c t in t h e same manner
a s th a t o f r a t s i t might b e th a t a r e t a r d a tio n in th e developm ent o f
r e s is t a n c e would r e s u lt i f m ice were u n d erfed . : An experim ent d e sig n e d
t o determ ine th e e f f e c t o f u n d erfeed in g on th e development o f r e s is t a n c e
t o in tra m u scu la r i n j e c t i o n o f t h e v ir u s was c a r r ie d out in th e p r e se n t
s e r i e s o f s tu d ie d .
The r e s u l t s appear l a t e r in t h e pap er.
I t has b een p o in te d out e a r l i e r th a t m ice become about 100
p e r c en t r e s is t a n t t o in tram u scu la r in j e c t i o n o f v e s ic u l a r s t o m a t i t i s
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
16
v ir u s during t h e p eriod when th ey reach sexu al m aturation ( 2 8 -3 6 days
o f a g e ).
The w r it e r i s not f a m ilia r w ith any ex p erim en ta l s t u d ie s in
w hich an attem pt was made t o c o r r e la t e se x u a l m a tu ra tio n and immunity
1
t o v ir u s d is e a s e s . I t should be-'noted however th a t th e in c id e n c e o f
p o li o m y e lit is d e c r e a s e s a f t e r th e age o f p u b erty .
T h is d ecrea sed i n c i ­
dence m igh t, how ever, ^be due t o immunity t o th e d is e a s e acquired by
h avin g come in c o n ta c t w ith s u b in f e c t iv e d o se s o f t h e v ir u s .r a t h e r th a n
t o s e x u a l m a tu r ity ..
I n ord er t o stud y t h e r e la t io n s h ip betw een se x u a l m atu ration
and development o f r e s is t a n c e t o in tra m u scu la r i n j e c t i o n o f v e s ic u l a r
s t o m a t i t is v ir u s an experim ent was c a r r ie d out t o determ ine w hether
m ice which had been caused t o become " p r e c o c io u sly ” s e x u a lly mature would
be more r e s i s t a n t th a n normal m ic e .
duce e a r ly se x u a l m a tu ra tio n .
Pregnant mare serum was used t o in ­
The r e s u l t s o b ta in ed in t h i s experim ent
appear l a t e r i n t h e pap er.
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
17
i
t
Methods
V ir u s, an im als, d ie t s , and hormonal f a c t o r : The New J ersey s t r a in o f
v e s ic u la r s to m a titis v ir u s was used in th e present s e r ie s o f s tu d ie s .
In every in sta n ce fr e s h ly passaged v ir u s was employed.
The b ra in s o f
m ice succumbing t o in tr a c e r e b r a l in je c tio n o f th e v ir u s were ground w ith
sand and s u f f ic ie n t b roth to make a 10 per cent suspension.
The suspen­
s io n was ce n trifu g ed at about 1000 R.P.M . fo r 5 m inutes in .a h o rizo n ta l
c e n t r ifu g e - The supernatant f lu id was d esign ated as th e 1 :1 0 d ilu t io n .
Tw o-tenths c . c . o f t h i s susp ension was in je c te d in to th e c a lf m uscles
o f th e hind l e g o f th e m ice t o be t e s t e d .
The minimal cereb ra l le t h a l
dose (M .C.L.D.) was determined by in je c tin g 1 0 -6 and 10~7 d ilu tio n s o f
th e susp en sion in tr a c e r e b r a lly in to groups o f 3 mice ( 0 .0 3 c . c . per
m ouse).
The t i t e r o f th e v ir u s suspension was u su a lly 10 “ 7 but occas­
io n a lly o n ly 10"6.
Each 0 .2 c . c . in je c te d intram uscularly th e r e fo r e
contained from 1 t o 10 m illio n M.C.L.D.
The alb in o m ice were o f inbred sto ck supp lied by a lo c a l d ea ler
who r a is e d them on th e "Mn d ie t l i s t e d below .
Female mice were obtained
during th e 3rd week o f pregnancy and d e liv e r e d in th e lab oratory so th a t
an accu rate record o f th e age o f th e o ffs p r in g was a v a ila b le .
B efore undertaking any experim ents i t was n ecessary t o d eter­
mine th e s u s c e p t ib ilit y o f th e above s t r a in o f mice at variou s a g e s.
Pre­
lim in ary t e s t s showed th a t t h e s e m ice rea cted t o intram uscular in je c tio n
o f th e v ir u s in th e same manner as th e m ice p rev io u sly stu d ied ( 1 ,2 ) .
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
18
"M" d ie t
Y ellow C om Meal
Ground Wheat
"
O ats
A lf a lf a M eal
Bone Meal
D ried M ilk (skimmed)
Tankage
NaCl
CaCo^
45$
20$
12$
2$
2$
8$
4$
2$
1$
Meat Scrap
4$
100$
To stu d y th e in f lu e n c e o f a d e f ic ie n c y o f c e r t a in d ie t a r y f a c ­
t o r s on th e developm ent o f r e s i s t a n c e a p u r if i e d , a r t i f i c i a l v ita m in -fr e e
d ie t was prepared from t h e f o llo w in g in g r e d ie n ts :
C asein (V ita m in -fr e e ) 1
C om S ta r c h
Lard
McCollum* s S a lt
M ixture
22$
54$
20$
4$
100$
1.
C a sein Company o f America
Depending upon t h e p a r tic u la r d e f ic ie n c y t o h e s tu d ie d e ss e n ­
tia l
a c c e s s o r y fo o d f a c t o r s w ere added t o o r exclu d ed from t h e above d i e t .
The co m p osition o f th e a r t i f i c i a l d i e t s used w i l l b e found
i n T a b le £ ,
A com m ercial p r e p a r a tio n o f pregnant mare serum (Gonadogen—
The Upjohn Company) was u se d t o ind uce "precociou s" s e x u a l m a tu ra tio n .
E xperim ental
I n flu e n c e o f u n d erfeed in g on t h e development o f r e s is t a n c e t o v e s ic u la r
s t o m a t i t is v ir u s in p rem atu rely weaned m ice:
B e fo r e a tte m p tin g t o
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
T able I
P u r if ie d , A r t i f i c i a l D ie t s
D iet
F itam infr e e
sto ck
d ie t
Dehy­
drated
brewer*s
y e a s t1
(a u to elaved)
100 gms.
1 . Sup­
plemented
a r tific ­
ia l
100
2 . De­
f ic ie n t
in h e a tla b ile
B-complex
"
3 . De­
fic ie n t
in B complex
except
100
"
4 . De­
f ic ie n t
in v i t a ­
min E
100
«
C on stitu en ts
Dehy­
P ic e
Wheat
p o lis h ­
drated
germ
brewer’s in g s con­ o i l 4
y ea st
cent ra t e^
(tintr e a te d )
8 gms.
0 .$ gms.
8 gms.
6 gms.
8
Cod l i v e r
o il5
«
4 gms.
0 .3
"
4
"
0 .5
«
4
"
4
•
1 & 5: Mead Johnson and Company
2 : 10 hours a t 16 pounds
3: The Borden Company
4: F u rn ish ed th rou gh c o u r te sy o f G eneral M ills
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
determ ine t h e e f f e c t o f a d e f ic ie n c y o f any s i n g l e d ie ta r y f a c t o r i t
was d ecid ed t o le a r n f i r s t t h e in flu e n c e o f u n d erfeed in g w ith a d ie t
r ic h in n a tu r a l f o o d - s t u f f s and v ita m in s on t h e development o f r e s i s 1 ■* .
•
1*1 •
ta n e e t o in tra m u scu la r i n j e c t i o n o f t r h e ; •;*•':&in’c"gj14 day o ld m ice
• 5
• * '** '*•
••• ■ •
a re 100 p er c e n t s u s c e p t ib le t o in o ciH ^ ‘4 bn.'l3y jtT^s ronfce but become
r e s is t a n t d u rin g subsequent w eeks th e quest le d arbjie**whet h e r o r not
m ice weaned a t 14 days o f a ge and fe d j u s t enough fo o d t o m a in ta in
t h e i r weaning w eig h t would become r e s i s t a n t l a t e r .
To answer t h i s ques­
t i o n m ice w ere ta k e n from t h e i r m others when 2 w eeks o ld and f e d a mix­
t u r e c o n s is t in g o f 1500 gms. o f th e "M" d i e t , 1000 c . c . o f w hole m ilk ,
1
2
150 gms. o f dehyd rated brew ers y e a s t , and 60 gms. o f Cod l i v e r o i l .
At th e tim e o f w eaning 6 t o 8 m ice were p la c e d in a m etal box c o n ta in ­
in g e x c e l s i o r .
T h e ir in d iv id u a l w eig h ts w ere record ed d a i l y .
The
amount o f fo o d g iv e n a group o f m ice was w eighed and record ed each day
so th a t i f on an en su in g day a g e n e r a l g a in in w eight was n o ted t h e
amount o f fo o d g iv e n th a t day was d im in ish ed .
I f an o .c ca sio n a l mouse
began t o g a in w e ig h t i n e x c e s s o f 7 gms. i t was removed from t h e group
and g iv e n l e s s fo o d f o r a day o r tw o .
i t was retu r n e d t o t h e grou p .
When i t had l o s t th e e x c e s s w eight
I n th e e a r ly p art o f t h e experim ent m ice
w hich w ere removed b ecau se o f e x c e s s w eig h t were g iv e n no fo o d on t h e
day o f rem o v a l.
T h is 'p r a c tic e was d is c o n tin u e d b ecau se i t was found
th a t 24 hou rs o f s t a r v a t io n was u s u a lly f a t a l t o t h e s e young m ice.
S ib ­
l i n g s o f t h e u n d erfed m ice w ere weaned sim u lta n e o u sly and g iv e n th e same
d ie t ad lib it u m .
1 . Mead Johnson and- Company
2 . Mead Johnson and Company
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
Reproduced
with permission
of the copyright owner.
INFLUENCE OF U N D ERFEED IN G O N THE
DEVELOPMENT OF RESISTANCE TO INTRA­
M U SC U LA R IN JECTIO N O F V E S I C U 1 A R
ST O M A T IT IS VIRUS
WELL-NOURISHED
MICE
Further reproduction
C M *diet + w hole milk. + i j e a s t
+ c o d - l i v e r o i l .) a d lib.
•,wt.
.
5 .9 gm s,:
ZoVo
pcoroA yzcdl
prohibited without permission.
2 8 DAYS_
UNDERFED
LITTER
z'
:
MATES
(*M*d i e t +whole milk. + i|e a st + cod-liver
o i l ,) q.s. t o main tain weigKt
Wt'
87% I
« t. :
5'99">s. nC>cay,£J 5.99ms.
i
3 6 - 3 9 DAYS
Reproduced
with permission
of the copyright owner.
CHART
E F F E C T OF UNDERFEEDIN& O N
fig u re
I
DEVELOPMENT OF
W
S u s c e p tib ility curves o f under-feot a n d welln o u r is b e d m ic e w e a n e d a t 14 d a q s
100
DIET + WHOLE MILK + YEAST + COD LIVER OIL
w
9o
>i
15
Further reproduction
weaned a t 14 d ay s
d ie t a d libitum
« 7o-fl
c£ 60-f
W
5: .
o 4 o-H
a? 1
//
7
Ui
5
prohibited without permission.
weaned a t i4 d aq s - diet q.s. maintain wt.
o 20
ui
S
111
/
A
IO-
a
AGE IN DAYS
m/ ad. lib. (9 mice) 28 days old
"3
/
)r
T .
/
/* q.s. maintain wt.
/
(1 3 m ic e )
w/
lU
N
>- 3otil
/
q.s. maintain w eight
(13 m ice)
".
^ so
10
2
fig u re
G row th cu rv es of u n d e r f e d a n d w ell-nourished mice
w e a n e d a t 14-d a g s
20
RESISTANCE
I
1
2
3
4
5
6
ad-lib. (19 mice) 36*39 dags old
7
8
9
10 11
a
13
DAYS AFTER INOCULATION WITH VIRUS
20
The accompanying photographs show th e appearance o f t h e under­
f e d and w e ll-n o u r is h e d m ice a t th e age o f 1 4 , 2 8 , and 39 d a y s.
I t w ill
b e n o te d th a t grow th has occu rred in t h e m ice h e ld a t weaning w eig h t in
th a t t h e y a re la r g e r and appear more m ature th a n th e y d id a t 14 days o f
age.
The 28 and 39 day o ld m ice have la r g e r h ead s, t h e i r h a ir has grown,
t h e i r e y e s a re l a r g e r , and th e s k e l e t a l s tr u c tu r e i s more d ev elo p ed .
When t h e u n d erfed m ice a r e compared w it h t h e w e ll-n o u r is h e d ones i t i s
e v id e n t th a t t h e r e i s o n ly a s l i g h t d if f e r e n c e i n le n g t h .
i s a c o n s id e r a b le d if f e r e n c e i n w e ig h t.
However, th e r e
M ice weaned a t 14 days o f age
u s u a ll y l o s e w eigh t f o r about 3 days and th e n b e g in t o g a in (Chart I ,
F ig u r e 1 ) .
In t h e group w hich was g iv e n fo o d ad lib it u m th e w eaning
w e ig h t was a t t a in e d on th e 2 0 th day o f l i f e .
From th a t tim e on th e y
g a in e d r a p id ly and a t 28 days o f age w ere alm ost a s heavy a s unweaned
m ice o f th e same a g e .
The d a ta o b ta in e d i n t h e s e exp erim en ts a re p r e se n ted g r a p h ic a lly
i n Chart I .
The cu rv es (F ig u r e 2) in d ic a t e th e p ercen ta g e, o f m ice w hich
w ere p a r a ly z e d on d if f e r e n t days a f t e r in o c u la tio n o f t h e v ir u s .
The r e ­
s u l t s r e v e a l s i g n i f i c a n t d if f e r e n c e s i n t h e s u s c e p t i b i l i t y o f th e 2
g r o u p s.
At t h e a g e o f 28 days^,87 p er cen t o f th e u n d erfed m ice d ev elo p ed
p a r a ly s is a s c o n tr a s te d w ith o n ly 11 p e r cen t o f th e w e ll-n o u r is h e d a n i­
m a ls .
The d if f e r e n c e i s even more s t r i k i n g in th e 36 t o 39 day o ld m ic e .
N in e ty -tw o p er c en t o f t h e un derfed d ev elo p ed p a r a ly s is w hereas 100 p er
c en t o f th o s e g iv e n fo o d ad lib itu m w ere r e s i s t a n t .
"When th e two mal­
n o u r ish e d groups a r e compared i t i s s e e n t h a t a p p ro x im a tely t h e same p e r c e n t­
age
o f t h e 28 and 36 t o 39 day o ld m ic e d eveloped p a r a ly s is .
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
However,
21
th e o n s e t m s e a r l i e r In th e younger group.
P a r a ly s is appeared
a s e a r ly a s th e th ir d day a f t e r in o c u la tio n in th e 28 day o ld
an im als and by t h e 6 th day 60 p er cen t were p a r a ly z e d .
In th e
36 t o 39 day o ld group s ig n s o f C .N .S . involvem ent d id n o t appear
u n t i l th e 6 th day and by th e 8 th day o n ly about 50 p er c e n t were
p a r a ly z e d .
I n flu e n c e o f m atern al d i e t on developm ent o f r e s i s t a n c e :
S in c e
m ice s u c k lin g m others on a d ie t r ic h i n n a tu r a l f o o d s t u f f s ("M"
d ie t ) become 80 t o 90 p e r c e n t r e s i s t a n t t o in tra m u scu la r i n j e c t i o n
o f th e v ir u s a t th e end o f th e su c k lin g p e r io d , i t was d e c id e d to
determ ine w hether m ice su c k le d by m others fe d d i e t s d e f i c i e n t i n
th e above v itam in s would a ls o develop t h i s r e s i s t a n c e .
In t h is s e t
o f exp erim en ts th e m others were m a in ta in ed on th e "M” d i e t throughout
pregnancy and f o r 2 days a f t e r d e liv e r y , when th e y w ere g iv e n th e
d i e t s l i s t e d i n T ab le I .
The o f f s p r in g were p e r m itted t o rem ain
w ith t h e i r m others f o r 2 8 days and th en co n tin u ed on th e r e s p e c t iv e
d i e t s u n t i l th e end o f th e t e s t p e r io d .
A l l m ice w ere o b serv ed f o r
a t l e a s t 21 days a f t e r I n o c u la tio n ox t h e v ir u s .
D if f e r e n t groups
o f m ice were t e s t e d a t 4 , 5 , and 6 weeks o f a g e .
The d ata o b ta in ed
i n t h e s e s tu d ie s a re p r e se n te d g r a p h ic a lly in Chart I I .
Growth and
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
22
s u s c e p t i b i l i t y cu rv es o f m ice su c k le d by m others fe d d i e t s r ic h in
n a tu r a l f o o d s t u f f s a re in c lu d e d f o r comparison*
P u r if ie d , a r t i f i c i a l v ita m in -fr e e d ie t supplem ented by a d d itio n o f
y e a s t , cod l i v e r o i l , and wheat germ o i l :
A t o t a l o f 41 m ice,
which had been su c k le d by 8 m others fe e d in g on t h e above d i e t , were
t e s t e d (Chart I I , F igu re 3 ) .
S e v e n ty -th r e e p er c e n t o f 15 m ice
t e s t e d a t 4 weeks o f age proved to be r e s i s t a n t .
o f t h e s e 15 m ice was 9 .7 grams.
The average w eig h t
At 5 weeks o f a g e 93 p er cen t o f
14 m ice (average w eig h t 1 2 .9 grams) d id n o t show e v id e n c e o f C .N .S .
in volvem en t and a t 6 weeks o f age 100 p er c en t o f th e 12 mice t e s t e d
(average w eigh t 12 grama) were r e f r a c t o r y .
A lthough t h e m ice i n
t h i s group were n o t q u ite as r e s is t a n t a t 4 w eeks o f age a s m ice
which had su c k le d m others fe d a d i e t o f v a r io u s n a tu r a l f o o d s t u f f s
(Chart I I , F ig u r e 2) th e y d id , however, d e v e lo p r e s is t a n c e norm ally
i n subsequent w eeks.
P u r if ie d , a r t i f i c i a l d ie t d e f i c i e n t in t h e h e a t - l a b i l e components o f
th e B -com plex:
N ine m others s u c k lin g 54 m ice w ere put on t h i s d ie t
2 days p o st partum .
However, due t o c a n n ib a lism f i r s t by th e m others
and l a t e r by th e more a g g r e s s iv e o ff s p r in g o n ly 8 o f th e o r ig in a l 54
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
23
m ice w ere a v a ila b le f o r t e s t i n g .
A ll o f t h e s e S m ice proved t o be s u s ­
c e p t ib l e a t tb e age o f 4 weeks (Chart I I , F ig u re 4 ) .
The naturd o f th e
i n f e c t io n in' th e s e m ice was d if f e r e n t from th a t o ccu rrin g in o th e r m ice
in th a t o n ly 2 developed t h e c h a r a c t e r is t ic p a r a ly s is .
That' d ea th o f
th e o th e r 6 was due t o in v a s io n o f th e C .N .S. by th e v ir u s was ev id en t
from th e f a c t th a t v ir u s was reco v ered from t h e s p in a l cord and .brain or
from t h e s p in a l cord a lo n e o f t h e s e m ice a t th e tim e o f d e a th .
A pp arently
th e v ir u s , once i t had g a in ed en tran ce t o t h e C .N .S ., spread r a p id ly t o
in v o lv e v i t a l c e n te r s o f th e cord and m edulla b e fo r e p a r a ly s is , co u ld
d e v e lo p .
Growth was markedly reta rd ed in t h e s e 8 m ic e .
At 28 days o f
age t h e i r average w eight was o n ly 4*3 grams a s compared w ith an average
• w eigh t o f 9*7 grams fo r 4 week o ld m ice w hich had su ck led m others fe d
t h e .p u r ifie d , a r t i f i c i a l d ie t supplem ented w ith y e a s t , cod l i v e r o i l , and
wheat germ o i l (Chart I I , F ig u re 1 ) .
B ecause p a r a ly s is 'w a s not a co n sta n t o ccu rren ce in t h i s group
o f m ice, Chart I I , F ig u r e 4 in d ic a t e s th e day o f d eath r a th e r th a n th e
day o f onset, o f p a r a ly s is .
P u r i f ie d , a r t i f i c i a l d ie t d e f i c i e n t in th e v ita m in B-com plex excep t B^:
Two days p ost partum 9 m others su c k lin g 62 m ice were p la c e d on t h i s d i e t .
C annibalism occu rred in t h i s group a s in th e p r e v io u s o n e.
However, th e
m o r t a lit y was not so h ig h s in c e 33 o f th e o r ig in a l 62 m ice su r v iv ed
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
t o b e weaned a t 28 d a y s.
At t h e age o f 4 w eeks 13 o f t h e m ice were
t e s t e d , and a l l w ere s u s c e p t ib le t o in tra m u scu la r i n j e c t i o n o f th e v ir u s
(Chart I I , F ig u r e 5)*
At 5 w eeks 90 p er c en t o f 10 an im als w ere r e s i s ­
ta n t and a t 6 w eeks 6 7 per c en t o f 10 m ice showed no s ig n s o f C .N .S . in ­
v o lv em en t.
In t h i s group, a s i n t h e group o f m ice w hich su ck led m others
fe d a d ie t d e f i c i e n t in t h e h e a t - l a b i l e components o f t h e B -com plex,
grow th was r e ta r d e d .
The average w eight o f t h e 1 3 m ice t e s t e d a t 4 weeks
o f age was 5*3 gram s, at 5 weeks o f age th e a v era g e w eigh t was 7 .5 grams
and a t 6 weeks t h e average w eig h t o f 10 m ice was o n ly 8 .1 grams (Chart I I ,
F ig u r e 1 ) .
P u r if ie d , a r t i f i c i a l d ie t d e f i c i e n t i n v ita m in E (Wheat germ o i l ) :
Two
days a f t e r d e liv e r y 8 m others s u c k lin g 56 young w ere p la c e d on t h i s d i e t .
F o r ty o f t h e young su r v iv ed t o b e t e s t e d a t 4» 5» end. 6 w eek s.
was n ot r e ta r d e d i n t h i s group o f m ice (C hart I I , F ig u r e 1 ) .
Growth
At t h e age
o f 4 w eeks 54 p e r c en t o f 15 m ic e , whose a v era g e w eight was 1 3 .6 grams,
were r e s i s t a n t .
However, a t 6 weeks o f age 100 p e r cen t o f 10 m ice (a v er­
age w eigh t 1 5 •4 grams) w ere r e f r a c t o r y (C hart I I , F ig u r e 6 ) .
A summary o f t h e d a ta in Chart I I r e v e a ls th a t when l a c t a t in g
m ice a r e fe d a p u r if ie d , a r t i f i c i a l d i e t w hich h a s b een supplem ented by
t h e a d d itio n o f y e a s t , cod l i v e r o i l , and wheat germ o i l th e s u c k lin g
young grow w e l l and d evelop r e s i s t a n c e t o t h e in tra m u scu la r i n j e c t i o n o f
v e s ic u l a r s t o m a t i t i s v ir u s .
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
Reproduced
with permission
CHART H
of the copyright owner.
INFLUENCE OF MATERNAL DIET DURING LACTATION ON DEVELOPMENT OF RESISTANCE
TO INTRAMUSCULAR INJECTION OF VESICULAR STOMATITIS VIRUS IN THE O F F S P R I N G .
FIGUR.E
FIGURE
I
Growth, curves o f mice sucKhnq mothers
FIGURE 3
2
C S LS + Y + C LO + W&O
"M" d i e t o r ' M ' d i e t * w h o l e m i l h t c l o * y e a s t
on v a rio u s d ie ts
2 week.* o l d ( 3 3 m i c e )
3 w eek s o ld ( s s m i c e )
’ M" d i e t
Further reproduction
s4 '5
0£
O
Zio
F
0
-CSLS+Y^CLO
<n10011
v» 90
5K 6o
2
io
|
60
(def. in vitamin 6 j
9iu 4500
3 4«
CSLS-*YtCL0+W&O
4 weets old (ismicel
' \C S L S*R PC tC L 0+ W & 0
(del. in B-comple* e x c e p t B , )
ui 10
> csis*autoclaved yeast
,
* CLO ■* W & O
.
4 w e e k s old ( i s m i c e )
5 w e e k s o l d (i& m i c e )
(def.in heat-labile 0 complex j
h
z
I 0
O
DAYS A F T E R I N O C U L A T I O N
IN D A Y S
prohibited without permission.
FIGURE
4
FIGURE
c s L S + a u to c ia v e d y e a s t + c l o + w g o
( d e f .i n h e a t - l a b i l e B - c o m p ie < )
i— i- i
s w e e t s o ld ( itw io e )
| X *6w ee>:s o ld d a m icel
i— i— i— r— i— i 1— i 1— r ~ i
I
4
01 io
IO 13 Ife iq 12 25 2B 3i 34 31 40 43
AGE
|
io
2.
3
S
6
*7
8
9
io
II
II
13
DAY5 AFTER INOCULATION W IT H VIRUS
WI TH V I R U S
S
FIGURE
C S L S + R P C + C L O +-W GO
( d e f . i n B* c o m p l e x e x c e p t B | )
6
CSLS + Y * CLO
(d e f - in Y i t a m i n £ )
4 weeks old (i3m ice)
> : 6o
a to
4 w e e k s o ld
4w
eeks old(8m
ice)
O3o
>■ 3o
s w e e k s oid
OAYS A F T E R
IN O CU LA TIO N
W IT H Y l R U S
DAYS A F T E R I N O C U L A T I O N
CSLS
RPC
CLO
WG O
Y
•
■
•
=
•
S w eek s o ld ( is m ic e )
(io m ice)
WITH V I R U S
(is m ice)
6 w eeks o ld ( lo rm c e )
DAYS
CASEI N, CORNS TARCH, LARD S S A L T S ( NUCOLLUM' S)
RICE POLISHIN&S C O N CE N T RA T E
COD LIVER OIL
WHEAT & E R W OIL
DEHYDRATED BREWERS YEAST
A FTER
IN O C U LA TIO N
W ITH
V IR U S
14
25
When l a c t a t i n g m ice a re put on a p u r if ie d , a r t i f i c i a l d ie t de­
f i c i e n t e it h e r i n t h e h e a t - l a b i l e B-com plex or in th e B-com plex except
B^ t h e r e i s a marked r e ta r d a tio n o f growth i n th e s u c k lin g young and a
d e la y i n th e development o f r e s is t a n c e t o in tra m u scu la r i n j e c t i o n o f th e
v ir u s .
Young m ice which su ck led m others put on a d ie t d e f i c i e n t i n
v ita m in E tw o days a f t e r d e liv e r y , a lth o u g h t h e y co n tin u ed t o g a in w e ig h t,
d id n o t d ev elo p r e s is t a n c e t o v e s ic u la r s t o m a t i t i s v ir u s in a normal
manner.
I n flu e n c e o f v a r io u s d i e t s on t h e developm ent o f r e s is t a n c e in -prematurely
weaned m ice:
In t h i s s e t o f experim ents m ice w hich had su c k le d m others
m ain tain ed on th e "M" d ie t w ere weaned a t 14 days o f a g e .
Four groups
o f ap p roxim ately equal numbers o f s i b l i n g s w ere g iv e n th e d i e t s l i s t e d
in T ab le I .
o f age.
D iff e r e n t groups o f m ice w ere t e s t e d at 4 , 5> and 6 weeks
The m ice were c o n tin u ed on th e d ie t u n t i l th e end o f th e t e s t
p e r io d .
P u r i f ie d , a r t i f i c i a l d iet-su p p lem en ted by a d d itio n o f y e a s t , cod l i v e r
o i l , end wheat germ o i l ;
I t has been p o in te d out e a r l i e r th a t m ice
weaned a t 14 days o f age l o s e w eig h t f o r about 3 days and th e n b e g in t o
g a in .
A fte r th e i n i t i a l l o s s in w eight t h e prem atu rely weaned m ice fe d
th e above d i e t grew r a p id ly {Chart I I I , F ig u r e 1 ) .
The a v erage w eight
o f th e 16 m ice t e s t e d a t 4 weeks o f age was 1 2 .3 ©rams*
S ix t y - t h r e e per
cen t o f t h e s e 16 m ice proved t o be r e s i s t a n t (Chart I I I , F ig u re 3 ) t o
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
in tra m u scu la r in j e c t io n o f t lie v ir u s .
At 5 weeks o f age a l l o f t h e 12
I
an im als t e s t e d (average w eigh t 1 3 .2 grams) were r e f r a c to r y and a t 6
weeks 91 p er cen t o f 11 m ice,w h ose average w eig h t was 1 4 .8 grams d id not
d ev elo p s ig n s o f C .N .S. in v o lv em en t.
P u r i f i e d , a r t i f i c i a l d ie t d e f ic ie n t i n th e h e a t - l a b i l e components o f th e
B -com plex:
d ie t.
A t o t a l o f 39 m ice were weaned a t 14 days and p la c e d on t h i s
I t was found th a t b o th growth and developm ent o f r e s is t a n c e w ere
r eta rd ed in t h e s e anim als (Chart I I I , F ig u r e s 1 and 4 ) .
The average
w eigh t o f t h e 15 m ice t e s t e d at 4 weeks was 8 .6 grams, and o n ly 20 p er
cen t o f th e s e anim als were found t o be r e s i s t a n t .
F i f t e e n m ice whose
average w eigh t was 6 .7 grams were t e s t e d a t 5 weeks o f a g e .
cen t w ere r e f r a c t o r y .
Only 33 p er
At 6 weeks o f age 9 m ice were t e s t e d .
age w eig h t o f t h i s group was 1 0 .7 grams.
The aver­
S ix t y -s e v e n p er cen t showed
no e v id e n c e o f C .N .S . in v o lv em en t.
P u r if ie d , a r t i f i c i a l d ie t d e f ic ie n t i n t h e v ita m in B -com plex excep t B^:
The weaned m ice w hich w ere fe d t h i s d i e t , l i k e th o s e in t h e p r e v io u s
group, f a i l e d t o g a in w eig h t in a normal manner (Chart I I I , F ig u r e 1 ) .
The a v era g e w eigh t o f t h e m ice t e s t e d a t 4 w eeks was 8 .2 gram s, at 5 weeks
th e average w eight was 8 .8 grams and at 6 w eeks 10 . 7 gram s.
At 4 weeks
o f age 67 p er c en t o f 15 m ice t e s t e d d id not d evelop p a r a l y s i s .
Of t h e
1 8 m ice t e s t e d a t 5 weeks 6 7 p er cen t w ere r e s i s t a n t and a t 6 weeks 89
p er c en t o f 9 m ice were r e f r a c t o r y (Chart I I I , F ig u re 5)*
The r e l a t i v e l y
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
27
h igh degree o f r e s is ta n c e (6 7 per cent) o f th e 4 week o ld m ice i s sur­
p r is in g when i t i s r e c a lle d th a t 100 per cent o f th e m ice which suckled
mothers fe d t h i s d ie t were s u s c e p tib le a t 4 weeks o f age (Chart I I ,
F igu re 5) •
I t would appear th a t during th e 1 4 days o f su c k lin g some
f a c to r was obtained in th e m ilk which enabled th e s e anim als t o develop
r e s is t a n c e .
I t should be n o ted , however, th a t at 5 weeks o f age on ly
6 7 per cent o f th e weaned m ice fe d th e above d ie t were r e s is t a n t as com­
pared w ith 100 per cent o f th o s e weaned and fe d th e supplemented d i e t .
P u r i f ie d , a r t i f i c i a l d ie t d e f i c i e n t i n v ita m in E (wheat germ o i l ) :
When m ice w ere weaned a t 14 d a y s o f age and put on t h e above d ie* i t was
found th a t th e y g a in e d w eig h t i n t h e same manner a s m ice w h ich were weaned
and g iv e n t h e supplem ented d ie t c o n ta in in g wheat germ o i l (Chart I I I ,
F ig u r e 1 ) .
The average w eig h t o f 15 m ice t e s t e d a t 4 w eeks o f age was
9 . 7 grams.
Only 20 p er cen t o f t h e s e an im als proved t o b e r e s i s t a n t t o
t h e v ir u s .
At 5 weeks o f age 100 per cen t o f 10 m ic e , whose average
w eig h t was I 4 . 3 gram s, w ere r e f r a c t o r y .
t e s t e d a t 6 weeks was 13*8 gram s.
not d ev elo p p a r a l y s i s .
The a verage w eig h t o f 6 m ice
Only 33 Pe r cen t o f t h e s e an im als d id
B ecau se o n ly 6 s ix -w e e k o ld m ice w ere a v a ila b le
f o r t e s t i n g t h e apparent h ig h degree o f s u s c e p t i b i l i t y o f t h e s e anim als
cannot b e c o n sid e r e d s i g n i f i c a n t u n t i l la r g e r numbers a r e t e s t e d .
I t appears from t h e s e d a ta th a t p rem atu rely weaned m ice when
f e d a p u r if ie d , a r t i f i c i a l d i e t , w hich has b een supplem ented by th e addi­
t i o n o f v ita m in s , become r e s i s t a n t t o in tra m u scu la r i n j e c t i o n o f v e s ic u la r
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
Reproduced
with permission
CHART m
of the copyright owner.
INFLUENCE OF DIET OF PREMATURELY WEANED MICE (WEANED AT 14 DAYS OF AGE) ON THE
DEVELOPMENT OF RESISTANCE TO INTRAMUSCULAR INJECTION OF VESICULAR STOMATITIS VIRUS
FIG U R E
FIGURE
I
FIGURE 3
CSLS TY T clo tw e o
2
*M
diet +whole m
ilk,* c
G r o w t h curves o f p r e m a t u r e l y w e a n e d
m ice F e d v a r i o u s d i e t s
l o
+ y
IOO
— 9o
W
2
cs
ls
80
A 70
u
♦ y-» c t o ( e *
°*60
♦ CLO+WOO
Further reproduction
4 w eeks o ld ( Ibm icel
4o
CSLS ♦ RPC
♦CLO«WGO
(def. in B-compie*
except 80
c s L S ta u to d c u e d
yeast+ cio+w G o
(def. in A e a tlabile B-complex}
20
5 w e ek s old (iq m ice}
17 20 23 26 29 32 35 3B 41
AGE
IN
0
DAYS
prohibited
FIGURE
1 2
DAYS
3 4
AFTER
5
6
7
8
F IG U R E
4-
C S L S T a u t o c l a v e d y e a s t t c l o ■» w & o
( d e f . in h e a t l a b i l e B -c o m p le x )
C SLS
9 10 H H 8 14
INO CU LA TIO N
t
R P C
t
W ITH
V IR U S
0 1 2 3 4
DA Y S A F T E R
5
6
7
6w e ek s old
(it m ice)
s weeks o ld
( i i m ice]
9
10 II
12 13 14
IN O C U L A T IO N
6
W ITH
V IR U S
FIGURE
5
CLO
* W OO
(def. in B- c o mp l e x e x c e p t B i )
6
C S J.S + Y + C L O
( d e f . in v i t a m i n E )
without permission.
in
4 w e e k s o l d ( i s mi ce}
^/6w ksoid_(6 m ice£
5 weeks o ld
(is m ic e ) '
a,
O4 0
5a 3o-
4w ks. old ( i s m i c e }
6 w e e k s o ld
( q m ice)
V-
z
5 wks. old (ismice)
few eeks o ld (Q m i c e )
u
Of
sw e e k s old
G
om i c e )
Ui
a
0
I
2 3
4
DAYS A F T E R
5
& 7
8
9 10 II I I 13 14
I N O C U L A T I O N WI TH V I R U S
0
1 2 3 4 5 6 7 0 9 IO II I I 13 14
D A Y S A F T E R IN O C U L A T IO N WITH V I R U S
0
1 2 3 4 5 6 7 8 9 IO H 12 13 14D A Y S A F T E R . IN O C U L A T I O N W IT H V I R U S
CSLS
RPC
CASEIN, C
O
R
NSTAR
C
H
, LA
R
D S. SALTS (M
ECOLLUM's)
CLO
W& O
Y
COD L IV E R O IL
WHEAT G E R M O I L
DEHYDRATED B R E W E R ' S Y E A S T
RI CE P OL I SH I NG S
CONCENTRATE
s t o m a t it is v ir u s in th e same manner a s do prem atu rely weaned m ice fe d a
m ixtu re o f n a tu r a l f o o d - s t u f f s and v ita m in s .
When mice are weaned at 14 days o f age and put on a p u r ifie d
a r t i f i c i a l d ie t d e fic ie n t in th e h e a t - la b ile components o f th e B-complex
both growth and development o f r e s is ta n c e are retard ed .
Although re­
ta r d a tio n in t h e development o f r e s is ta n c e was not marked in th e mice
weaned and fe d a p u r ifie d , a r t i f i c i a l d ie t d e f ic ie n t in th e B-compler
except Bj^ s t i l l on ly 6 7 per cent were r e s is t a n t at 5 weeks as compared
w ith 100 per cent o f th e weaned mice which r e c eiv ed th e supplemented
d ie t .
In t h e group o f weaned m ice w h ich were fe d a p u r if ie d , a r t i ­
f i c i a l d ie t d e f ic ie n t in v ita m in E t h e r e was a d e la y i n th e development
o f r e s is t a n c e alth o u g h growth was not r e ta r d e d .
I n flu e n c e o f v a r io u s d e f i c i e n t d i e t s on t h e r e s is t a n c e o f fu ll-g r o w n m ice;
The e f f e c t o f d ie ta r y d e f i c i e n c i e s on th e r e s is t a n c e o f fu ll-g r o w n m ice
was not in v e s t ig a t e d e x t e n s i v e ly in t h e s e s t u d ie s .
However, d a ta was ob­
ta in e d from s e v e r a l groups o f m ice which had b een fe d d e f i c i e n t d i e t s ,
as in d ic a te d i n T ab le I I ,
fo r
a p erio d o f 6 t o 8 w eek s.
The d a ta (T able I I ) r e v e a l th a t none o f t h e an i ma l s fe d th e
d e f ic ie n t d i e t s were s u s c e p t ib le t o v e s ic u l a r s t o m a t i t is v ir u s fo llo w in g
in tram u scu lar in j e c t i o n .
S e v e r a l o f t h e m ice fe d th e a u to c la v e d d ie t
develop ed s ig n s o f B^ d e f ic ie n c y e it h e r b e fo r e o r a f t e r i n j e c t i o n o f th e
v ir u s and s e v e r a l d ie d d u rin g t h e t e s t p e r io d .
However, t h e v ir u s was
not reco v ered from e it h e r
th e
b r a in o r th e s p in a l cord o f t h e s e m ic e ; and
i t must b e concluded th a t
d e a th was due t o t h e v ita m in d e f ic ie n c y and
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
T able I I
E ffe c t o f D e f ic ie n t D ie t s
on R e s is ta n c e o f Full-G-rovm M ice
D ie t
D e f ic ie n t in B complex excep t B^
Time
on D iet
6 - 8 wks.
No. M ice
$
R e s is ta n t
8
100
It
D e f ic ie n t in
v ita m in E
n
7
* ( 1IMW d ie t + w bole
m ilk + y e a s t + cod
l i v e r o i l ) In g red ­
i e n t s mixed and
a n to c la v e d 6 h r s .
a t 16 l b s .
n
10
V
V
* D e f ic ie n t in b e a t - l a b i l e B-com plex
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
n ot t o t h e v i r u s .
I t sh o u ld be p o in te d out th a t th e a u to c la v e d d ie t in
a d d itio n t o b e in g d e f i c i e n t i n th e h e a t - l a b i l e B -com plex was a ls o d e f i c i e n t
i n v ita m in A.
S in c e cod l i v e r o i l was added t o th e d i e t b e fo r e a u to -
c la v in g t h e d ie t was p rob ab ly a ls o d e f i c i e n t in v ita m in E.
I t would appear th e n th a t once m ice have d ev elo p ed r e s is t a n c e
t o in tram u scu lar in j e c t i o n o f th e v ir u s th e y do not a g a in become su sce p ­
t i b l e when fe d th e d e f i c i e n t d i e t s u sed in t h i s s tu d y .
I n flu e n c e o f " p recociou s" s e x u a l m a tu ra tio n on t h e developm ent o f r e s is t a n c e ;
The purpose o f t h i s experim ent was t o determ ine w hether s u c k lin g m ice in ­
je c t e d w it h a gon ad otrop ic hormone w hich in d u c es •’p r eco cio u s" s e x u a l mat­
u r a tio n w ould become r e s i s t a n t at an age when most m ice a r e s t i l l su s­
c e p tib le .
B egin n in g on th e 5 th day o f l i f e 20 m ice w ere in j e c t e d su b c u t-
a n e o u sly w ith 1 u n it ( C a rtlan d -N elso n ) o f a com m ercial p r e p a r a tio n o f
pregnant mare serum (G onadogen).
m ice w ere 21 days o ld .
The in j e c t i o n s w ere c o n tin u e d u n t i l th e
On t h e t w e n t y - f ir s t day o f l i f e t h e anim als were
weaned and t e s t e d f o r r e s is t a n c e t o t h e v i r u s .
serv ed a s c o n t r o ls .
T w elve normal l i t term inates
D escent o f th e t e s t e s o r opening o f th e v a g in a were
th e c r i t e r i a used t o d eterm in e sex u a l m a tu r a tio n .
The m others o f t h e s e
m ice w ere f e d a m ixtu re c o n s is t in g o f 1500 gms. "M " d ie t + 1000 c . c . w hole
m ilk + 150 grams y e a s t + 60 gms. cod l i v e r o i l .
The d a ta o b ta in ed i n t h i s
stu d y a r e found i n T a b le I I I .
I t w i l l b e s e e n t h a t opening o f t h e v a g in a o r d e sc e n t o f th e
t e s t e s occu rred by t h e 2 1 s t day i n 100 p er cen t o f t h e t r e a t e d m ice w h ile
none o f t h e u n tr e a te d lit t e r m a t e c o n tr o ls showed e v id e n c e o f sex u a l m a tu r ity .
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
T able I I I
In flu en ce o f "Precocious” Sexual M aturation on
tb e Development o f R esista n ce in Su cklin g Mice
shov/ing
opening o f
vagina or
descent o f
t e s t e s at
21 days o f age
i
Treatment
%
No. o f
Mice
Av. w t. at
tim e o f t e s t
20
9 .3
100
25
12
3 .9
0
25
r e s is t a n t
1 u n it o f
3-onadogen
d a ily
(1 6 u n its
per mouse)
None
(l i t t e r mat es o f
above
animals)
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
30
D escent o f t h e t e s t e s or openin g o f t h e v a g in a occu rred a s e a r ly a s th e
1 7 th day o f l i f e in some o f th e t r e a t e d a n im a ls.
M ice n o rm a lly become
s e x u a lly mature a t 28 t o 36 days o f a g e .
A lth ough "precociou s" s e x u a l m atu ration was induced i n 100 per
cen t o f th e t r e a t e d m ice t h e s e anim als were n ot more r e s i s t a n t than th e
lit te r r a a te c o n t r o ls .
S e v e n t y - f iv e p er cen t o f 21 day o ld m ice i n each
group were found t o be s u s c e p t ib le t o th e v ir u s f o llo w in g in j e c t i o n in to
th e m u scle.
I t appears from t h e d a ta o b ta in ed in t h i s experim ent th a t m ice
t r e a t e d w ith pregnant mare serum r e a c t t o t h e v ir u s i n th e same manner
a s do normal m ice o f th e same a g e , i . e . premature development o f r e s is t a n c e
i s not a s s o c ia t e d w ith " p recociou s" s e x u a l m a tu ra tio n .
D is c u s s io n
B efo re b eg in n in g a d is c u s s io n o f th e d a ta o b ta in e d in th e p r esen t
exp erim en ts i t w i l l b e w e ll t o r e s t a t e th e purpose o f t h e s t u d ie s .
V e s i­
c u la r s t o m a t it is v ir u s when in j e c t e d in t o th e c a l f m u sc le s o f 14- dgy o ld
a lb in o m ice in v a d es t h e c e n tr a l n ervou s system by way o f th e p e r ip h e r a l
n e r v e s and g iv e s r i s e t o a f a t a l a sce n d in g f l a c c i d p a r a ly s is i n 100 per
cen t o f th e anim als ( 1 , 2 ) .
However a s m ice grow and d ev elo p th e y become
in c r e a s in g ly r e s i s t a n t t o th e v ir u s when in j e c te d by t h i s r o u te .
At 1
month o f age o n ly 10 t o 20 p e r cen t a r e s u s c e p t ib le and betw een t h e 5 th
and 6 th week o f l i f e th e y a r e about 100 per cen t r e s i s t a n t ( 1 , 2 , 7 ) •
The
p r e se n t in v e s t ig a t io n s were undertaken t o determ ine w h eth er th e development
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
31
o f t h i s r e s is ta n c e could he in flu en ced by ce r ta in d ie ta r y d e f ic ie n c ie s
or by inducing e a r ly sexu al m aturation.
I t was observed th a t mice which were weaned at 14 days o f
age and kept at t h e i r weaning weight by underfeeding w ith a d iet r ic h
in n a tu ra l f o o d s tu ffs and v itam in s were s t i l l h ig h ly s u sc e p tib le to th e
v ir u s at 28 and 36 t o 39 days o f age.
S ib lin g s on th e other hand* weaned
at th e same tim e and given th e same d ie t ad lib itu m developed r e s is ta n c e
in a normal manner, i .e .^ a t 36 t o 39 days o f age th e y were 100 per cent
r e s is ta n t.
I t i s th u s evident th at when normal growth and development
are suppressed by in a n itio n r e s is ta n c e t o intram uscular in je c tio n o f
th e v ir u s does not develop in a normal manner.
become r e s is t a n t t o be explained?
How i s t h i s f a ilu r e t o
I t may be r e c a lle d th at p h y s io lo g ic a l
changes which ta k e p la ce in th e m uscle c e l l s o f m ice as th ey grow and
d evelop may be r e sp o n sib le fo r p reventing th e v ir u s from entering th e
p erip h era l nerves and thus gain a c c e ss t o th e c e n tr a l nervous system .
I t was noted th a t th e c a l f m uscles o f th e underfed m ice used in t h i s
p a r tic u la r study were much sm aller than th o se o f w ell-n ou rish ed m ice o f
th e same age.
I t might be th a t th e supp ression o f m etabolic a c t i v it y
o f th e muscle c e l l s , due t o a la ck o f n u trien t m a te r ia l, may have in t e r ­
fe r e d w ith th e development o f r e s is t a n c e in th e m u scle.
However at th e
p resen t tim e i t cannot be s ta te d d e f in it e ly th at th e f a ilu r e o f th e c a l f
m uscles t o develop was th e s o le co n trib u tin g cause o f th e f a ilu r e o f
th e underfed mice t o become r e s is t a n t .
I t has been pointed out e a r lie r
th a t th e b a rr ie r t o n eu roin vasiven ess fo llo w in g intram uscular in j e c t io n
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
32
o f t h e v ir u s may be s itu a te d in th e p erip h eral nerve or nerve end ings.
T his b ein g th e case th en p o s s ib le changes in th e p erip h era l nerves or
in th e nerve endings, due t o u nd erfeeding, must a lso be co n sid ered .
At p resen t i t i s not known whether such changes do occur in m ice which
are h eld a t a more or l e s s constant w eig h t.
However i t has been r e ­
ported ( 2 1 ) th a t d is in t e g r a tio n o f th e m yelin sheaths o f p erip h era l
nerves occurs in r a ts on a t o t a l in a n itio n regimen.
I t should be noted
th a t because o f th e nature o f th e d ie t fe d th e s e m alnourished mice
(h ig h in v itam in s A, B-com plex, D) th e anim als may have r eceiv ed s u f­
f i c i e n t vitam in s even in th e sm all amount o f food g iv en them.
A lthough i t has been shown th a t underfeeding d e f i n i t e l y r e ­
ta r d s th e development o f r e s is t a n c e in m ice t o intram uscular in je c tio n
o f v e s ic u la r s to m a titis v ir u s th e present s t a t e of our knowledge does
not permit one t o s t a t e by what mechanism t h i s f a ilu r e t o d evelop r e ­
s is ta n c e i s brought ab ou t.
7/hen m ice were perm itted t o su c k le mothers which had been
put on a d ie t d e fic ie n t in th e h e a t - la b ile components o f th e vitam in Bcomplex 2 days a fte r d e liv e r y th ey did not develop r e s is t a n c e t o in tr a ­
muscular in j e c t io n o f t h e v ir u s .
At 4 weeks o f age a l l o f th e 8 mice
t e s t e d proved t o be s u s c e p t ib le .
Only 2 o f th e se m ice developed th e
c h a r a c t e r is t ic ascending f la c c id p a r a ly s is but th e v ir u s was recovered
from th e b ra in and cord or from th e cord a lo n e o f th e other 6 at th e
tim e o f d ea th .
I t would appear th a t once th e v ir u s gained entrance t o
th e c e n tr a l nervous system i t spread r a p id ly t o in v o lv e h igh er cen ters
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
33
o f th e s p in a l cord and m ed u lla b e fo r e p a r a ly s is co u ld d e v e lo p .
I t w i l l be n o ted (Cbarfc I I , Bigur® 1) th a t grow th was markedly
r eta rd ed in th e m ice w hich su c k le d m others fe d th e d ie t d e f i c i e n t in
th e h e a t - l a b i l e B -com plex.
of lif e
The anim als d id not grow a f t e r th e 1 0 th day
(w e ig h ts were ta k e n from th e 1 0 th t o 2Sth d a y ).
A pparently
th e la c t a t i n g mouse i s d e p le te d o f th e h e a t - l a b i l e B -com plex more
r a p id ly th an th e r a t s in c e d e p le tio n a p p a re n tly occurred a f t e r th e
m ice had b een on th e d ie t f o r & d ays.
The apparent r a p id spread o f th e v ir u s in t h e c e n t r a l nervous
system o f t h e m ice which had b een su ck led by m others fe d a d ie t d e f ic ie n t
in t h e h e a t - l a b i l e B-com plex s u g g e s ts th a t th e f a i l u r e o f t h e s e anim als
t o d e v e lo p r e s is t a n c e may have been a s s o c ia t e d w ith changes in th e ner­
vous sy stem .
As a lrea d y in d ic a te d e a r l i e r in th e paper v ita m in B^ seems
t o b e n e c e s s a r y f o r th e m aintenance o f h e a lth y n e u r a l t i s s u e .
I t may
w e ll be th e n th a t v ita m in B^ i s a ls o n e c e s s a r y f o r th e normal d ev elo p ­
ment o f t h e n ervous sy ste m .
I f t h i s i s th e c a se th e n i t may be l o g i c a l
t o assume th a t th e f a i l u r e o f t h e s e m ice t o d ev elo p r e s is t a n c e was a s ­
s o c ia t e d w ith a f a il u r e o f t h e p e r ip h e r a l n e r v es t o d e v e lo p in a normal
manner.
I t must be remembered however th a t v ita m in B^ may p la y some part
in t h e carb ohydrate m etab olism o f th e c e l l and as p o in te d out e a r l i e r
a d istu r b a n c e o f m etabolism m ight in t e r f e r e w ith th e developm ent o f r e ­
s is ta n c e .
A marked r e t a r d a tio n o f grow th and a d e la y in t h e development
o f r e s is t a n c e t o in tram u scu la r in j e c t io n o f v e s ic u l a r s t o m a t i t i s v ir u s
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
34
occurred in mice which suckled mothers fe d a d iet, d e fic ie n t in th e
B-complex except B^.
At 28 days o f age 100 per cent o f th e mice
t e s t e d were s u s c e p tib le .
A ll o f th e se m ice developed th e character­
i s t i c ascending f la c c id p a r a ly s is .
I t i s in te r e s tin g t o n ote th at 5
week o ld m ice which had suckled mothers fed th e above d e f ic ie n t d ie t
were 90 per cent r e s is t a n t .
T h is might suggest th a t mice feed in g on;
a d ie t d e f ic ie n t in th e B-complex except B^ are ab le t o develop r e s i s ­
ta n ce t o th e v ir u s .
In view o f th e fa c t th a t th e 5 week old mice
proved t o be h ig h ly r e s is ta n t i t i s d i f f i c u l t t o form ulate an explana­
t io n fo r th e high degree o f s u s c e p t ib i lit y o f th e 28 day o ld m ice.
Re­
ta r d a tio n o f growth may have been a fa c to r but th e d iffe r e n c e in th e
average weight o f th e two groups ( 2 .2 grams) would not in d ic a te t h i s
t o be th e s o le cau se.
only 7 .5 grams.
The average weight o f th e 35 day old m ice was
In other experim ents normal 14 day old mice w eighing
7 .5 grams or more have been found t o be s u s c e p tib le t o th e v ir u s when
in je c te d in tram u scu larly.
The on ly apparent exp lanation fo r th e f a ilu r e
o f th e 28 day old mice t o become r e s is t a n t i s th at a la ck o f th e Bcomplex except B^, caused a distu rban ce in t i s s u e m etabolism which in
turn retard ed th e normal development o f r e s is ta n c e in th e m uscle c e l l s .
The grovith curves (Chart I I , F igure l ) o f th e m ice suckled
by mothers fed a d ie t d e f ic ie n t in th e h e a t - la b ile B-complex show th a t
growth was retarded from th e 10th day o f l i f e .
I t would appear th a t in
th e la c t a t in g mouse th e h e a t - la b ile B-complex and th e B-complex except
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
35
B-j_ a r e d e p le te d a t about th e same r a t e .
I t h as been shown (1 6 , 25) th a t both v ita m in Bj_, and
r ib o f la v in , a r e n e c e ssa r y f o r s u c c e s s f u l l a c t a t i o n i n th e r a t .
The r e s u l t s o f t h i s phase o f th e stu d y su g g e st t h a t th e same i s
tr u e f o r th e m ouse.
Of an o r i g i n a l group o f 54 m ice su ck led by
m others fe d a d i e t d e f i c i e n t i n th e h e a t - l a b i l e components o f th e
B-com plex o n ly 8 were a l i v e a t th e end o f th e s u c k lin g p e r io d , and
o n ly 33 o f an o r ig in a l 62 m ice su c k le d by m others f e d a d ie t d e f i c i e n t
in th e B-com plex e x ce p t B^ su r v iv ed t o be weaned.
I t i s in t e r e s t in g
t o n o te in t h i s c o n n e c tio n t h a t com plete f a i l u r e o f la c t a t i o n o ccu rred
(33) when a lb in o r a t s , f e d a d i e t d e f ic ie n t i n y e a s t , w ere fe d
supplem ents o f v ita m in B^ and r i b o f l a v i n .
However, i f , i n a d d itio n
t o th e above v ita m in s , t h e f i l t r a t e f a c t o r (34) (th e f i l t r a t e f a c t o r
p r e v e n ts n u t r it io n a l d erm a to sis i n c h ic k s ) was added to t h e d i e t , s u c c e s s ­
f u l la c t a t i o n o c cu rred .
I n th e p r e se n t s e r i e s o f s t u d ie s n u t r it i o n a l m uscular dystrop h y
was n o t o b served in any o f th e m ice which w ere s u c k le d by m others fe d a
d i e t d e f ic ie n t in v ita m in S .
E v id e n tly s u c k lin g m ice and su c k lin g r a t s
behave d i f f e r e n t l y to a d e f ic ie n c y o f v ita m in E s in c e r a t s s u c k lin g m others
on a d i e t d e f i c i e n t i n E d ev elo p p a r a ly s i s .
A lthough t h e m ice su c k le d
by m others fe d th e above d i e t d id n o t show any c l i n i c a l symptoms o f a
d e f ic ie n c y o f v ita m in E, n e v e r t h e le s s th ey were h ig h ly s u s c e p t ib le (54
p e r c e n t) t o th e v ir u s a t 2 8 days o f a g e .
However, a t 5 and 6 weeks o f
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
-
36
age th e m ice had d evelop ed normal r e s is t a n c e t o t h e v ir u s*
Growth
was n ot r e ta r d e d in t h e s e m ice and th e y were normal in appearance*
I t would appear th a t f a c t o r s o th e r th a n grow th a r e in v o lv e d i n th e
developm ent o f r e s is t a n c e .
At p r e se n t no e x p la n a tio n can be o ffe r e d
f o r th e r e ta r d a tio n i n t h e developm ent o f r e s i s t a n c e n o ted i n m ice
w hich had su c k le d m others fe d a d i e t d e f i c ie n t in v ita m in E*
It
m ight be th a t d e g e n e r a tiv e changes may have occu rred i n th e m nsole
c e l l s which were n o t m a n ife sted c l i n i c a l l y but th e r e i s no e v id e n c e
a t hand t o support t h i s s u g g e s tio n .
A s t r ik in g d if fe r e n c e i n th e d eg ree o f r e s i s t a n c e was n o ted
b etw een th e m ice weaned a t 14 days o f age and f e d a d ie t d e f i c i e n t i n
th e h e a t - l a b i l e B complex and th e an im als weaned a t th e same age and
f e d a d i e t d e f i c i e n t in t h e B com plex ex cep t B^.
The l a t t e r group
d ev elo p ed r e s is t a n c e in an alm ost normal manner w h ile i n th e former
group th e r e was a marked r e ta r d a tio n i n t h e developm ent o f r e s is ta n c e *
I t must be remembered th a t b o th groups o f m ice w ere su ck led fo r 14 days
b y m others fe e d in g on a d i e t r ic h in n a tu r a l f o o d s t u f f s .
I t would
appear th a t when m ice a re a llo w e d to s u c k le fo r 14 days and are th en
w eaned, r e s is t a n c e d e v e lo p s even when th e B-com plex ex cep t
e lim in a te d from th e d i e t .
is
However, when h e a t - l a b i l e components o f th e
B -com plex are e lim in a te d from th e d i e t o f 14 day o ld m ic e , th e d ev elo p ­
ment o f r e s is t a n c e i s markedly r e ta r d e d .
I t i s a p p a ren t, th en , th a t
th e h e a t - l a b i l e components o f t h e B-oom plex must be in c lu d e d in th e
m atern al d ie t and in th e d ie t o f prem atu rely weaned m ice i f r e s is t a n c e
i s t o d ev elo p norm ally*
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without perm ission.
37
The h ig h degree o f r e s is t a n c e a t S8 days o f age o f th e
prem aturely weaned m ice w hich had been fe d th e d i e t d e f i c i e n t in
th e B-com plex e x ce p t
i s s u r p r is in g when i t i s r e c a l le d th a t
100 p er c e n t o f t h e m ice, w hich had su ck led m others fe d th e same
d i e t , were s u s c e p t ib le a t 1 month o f a g e .
I t would appear th a t when
v ery young m ice (2 days o ld ) are d ep rived o f th e B-com plex e x ce p t B^,
th ey f a i l t o become r e s i s t a n t t o th e v ir u s a t a tim e when normal m ice
a re h ig h ly r e s i s t a n t .
However, i f m ice, p e r m itted t o s u c k le m others
fe e d in g on a d i e t o f n a tu r a l f o o d s t u f f s , a r e weaned a t 14 days o f age
and g iv e n a d ie t d e f i c i e n t in th e B-com plex, th e y become r e s i s t a n t in
an alm ost normal manner.
The m ice which w ere weaned a t 14 days o f age and g iv e n a
d ie t d e f i c i e n t in v ita m in E, a lth o u g h th ey showed no symptoms o f E
d e f ic ie n c y , were more s u s c e p t ib le a t 28 days o f a g e th a n w ere m ice
which had su c k le d m others f e d a d ie t d e f ic ie n t in th e same v ita m in .
I t m ight b e concluded th a t th e unweaned m ice r e c e iv e d seme f a c t o r in
th e
m ilk (perhaps v ita m in E) which en ab led them t o become more
r e s is t a n t than th e prem atu rely weaned m ice.
_
S in c e th e m ioe weanedat
14 days o f age and fe d t h e d ie t d e f ic ie n t in v ita m in £ d id n o t show
any symptoms o f £ d e f ic ie n c y , no e x p la n a tio n can b e o f f e r e d f o r th e
d e la y i n th e developm ent o f r e s is t a n c e .
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
38
F u ll-grow n m ice, which are r e s i s t a n t t o in tram u scu lar
i n j e c t i o n o f v e s ic u la r s t o m a t it is v ir u s , d id not lo s e t h i s r e s i s t ­
ance when f e d a d ie t d e f i c i e n t in c e r t a in components o f th e
or in vita m in £ f o r a p e r io d o f from 6 t o 8 weeks*
B-com plex
T hese fin d in g s
su g g e st th a t once m ice become r e s i s t a n t t o th e v ir u s , th e r e s is t a n c e
i s n o t a f f e c t e d by a d e f ic ie n c y o f th e above vitam ins*
I t may w e ll
be t h a t c e l l u l a r changes w hich occu r during growth and development
are n o t r e v e r s ib le .
The f a c t t h a t s e v e r a l o f th e m ice w hich were
fe d a d ie t d e f ic ie n t i n th e h e a t - l a b i l e components o f th e B-com plex
d evelop ed s ig n s o f B^ d e f ic ie n c y (weakness o f e x tr e m itie s and trem ors)
b e fo r e or s h o r t ly a f t e r i n j e c t i o n o f t h e v ir u s m ight su g g e st th a t
even when th e p e r ip h e r a l n e r v es a r e damaged, th e v ir u s n e v e r t h e le s s
does n o t invade th e c e n t r a l nervous system *
In order th a t m isle a d in g g e n e r a liz a t io n may not be made
from th e above o b s e r v a tio n s , i t sh o u ld be p o in te d ou t th a t w h ile
c e r t a in d ie ta r y d e f i c i e n c i e s appear to in f lu e n c e th e developm ent o f
r e s is t a n c e t o v e s ic u la r s t o m a t it is v ir u s when in j e c t e d by th e in t r a ­
m uscular r o u te , i t cannot b e s a id th a t th e r e s is t a n c e t o a l l v ir u s e s
th a t
would be a f f e c t e d in t h e same manner. I t may a ls o b e /th e r e s is t a n c e
t o v e s ic u la r s t o m a t i t is v ir u s when in j e c t e d by o th er r o u te s m ight
be a ffe c te d d iffe r e n tly .
S tark ey and Leathern (35) have r e c e n t ly shown t h a t pregnant
mare serum i s cap ab le o f producing a s tim u la tin g e f f e c t on th e gonads
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
29
o f IE day o ld m ic e .
immature fem ale m ice,
They found an o v a ria n w eig h t in c r e a s e i n th e
f o l l i c u l a r m a tu ra tio n was induced but th e
form ation o f l u t e i n t i s s u e predom inated.
c r e a se in i n t e r s t i t i a l t i s s u e and
They a ls o found an in ­
a r e s u lt in g in c r e a s e i n a c c e s s o r y
s e x u a l organs in th e immature m a le s.
In th e p r e se n t stu d y no h i s t o l o g i c a l s t u d ie s were made o f
th e s e x organs o f m ice t r e a t e d w ith pregnant mare serum.
However,
d e sc e n t o f th e t e s t e s o r opening o f th e v a g in a occu rred a s e a r ly
a s th e 17th day when 1 u n it (C artlan d -N elson ) o f pregnant mare serum
was in j e c t e d d a ily b e g in n in g w ith t h e 5 th day o f l i f e .
By t h e 2 1 s t
day o f l i f e d escen t o f th e t e s t e s or opening o f th e v a g in a was observed
i n 100 p er c e n t o f t h e in j e c t e d m ic e .
A lthough ev id en ce o f "precocious**
s e x u a l m atu ration was o b ta in e d in a l l t h e t r e a t e d m ice, th e y w ere j u s t
a s s u s c e p t ib le to in tra m u scu la r in j e c t io n o f th e v ir u s a s were t h e ir
u n tr e a te d lit te x m a te c o n t r o ls .
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
40
SUMMABY
The p r e se n t s e r i e s o f in v e s t ig a t i o n s was undertaken t o
stu d y th e in f lu e n c e o f c e r t a in d ie t a r y and hormonal f a c t o r s on th e
developm ent o f r e s is t a n c e , in t h e a lb in o m ouse, to Intram uscular
in j e c t i o n o f v e s ic u la r s t o m a t i t is v ir u s*
E xperim ents were conducted
t o determ in e th e e f f e c t on th e developm ent o f t h i s r e s is t a n c e o f
(1 ) th e m atern al d i e t during l a c t a t i o n ,
(2 ) th e d i e t o f m ice weaned
a t 14 days o f a g e , and (5) a g o n a d a tro p ic hormone which in d u ces
" p recociou s" s e x u a l m aturation*
The d a ta o b ta in e d in th e d ie ta r y s t u d ie s are suusnarized i n
T a b le s IT and V*
The r e s u l t s (Table IT) r e v e a l t h a t when la c t a t in g m ice were
f e d a p u r if ie d a r t i f i c i a l d i e t w hich had b een supplem ented by th e
a d d itio n o f y e a s t , cod l i v e r o i l and wheat germ o i l , th e o ffs p r in g
grew and d evelop ed r e s is t a n c e t o in tra m u scu la r i n j e c t i o n o f th e v ir u s
in th e same manner a s d id m ice which w ere s u c k le d by m others f e d &
d i e t r ic h i n n a tu r a l fo o d s tu ffs *
HJhen l a c t a t in g m ice were fe d a p u r if ie d a r t i f i c i a l d ie t
d e fic ie n t
e it h e r i n the h e a t - l a b i l e components o f th e B-oomplex or
th e B-cam plex ex cep t v ita m in B^, th e r e was a r e ta r d a tio n o f growth
and a marked d ela y in the developm ent o f r e s is t a n c e i n th e o ffs p r in g *
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
Reproduced
with permission
Table IV
In flu en ce o f Maternal D iet during Nursing P eriod on Development o f R esista n ce in O ffspring
(V. S . V irus Intram uscularly)
of the copyright owner.
Age o f Mice
4 weeks
5 week 3
6 weeks
No.
No.
No.
%
*
*
r esista n t t e s ­ Av. resista n t t e s ­ Av. ■resista n t t e s ­ Av.
ted wt.
ted wt.
ted wt.
D iet
Further reproduction
1 . "M" d iet or "M" d iet + whole milk
+ y ea st + CLO
2. CSLS + 0L0 + TOO + yeast
3.
4.
"
+
”
+ yeast (d e fic ie n t in E)
" + « + WGO + RPC
(d e fic ie n t in B. complex except B^)
prohibited without permission.
5. CSLS + CLO + TOO + autoclaved yeast
(d e fic ie n t in h e a t-la b ile B)
'
82
75
Gm.
13.4
2 1
15
9 .7
JA
15
9 .5
0
13
5.3
0
8
4 .3
3&
18
Gm.
15.0
2 1
14
12.9
100
12
12.0
15
1 3 .6
100
10
15.4
10
7.5
64
11
8.1
80
2 2
Gm.
No survivor '3 on d iet
CSLS « c a se in (v ita m in -fr ee ) + sta r ch + la r d + s a lt m ixture (McCollum’ s)
CLO =» cod l i v e r o i l ; TOO = wheat germ o i l ; RPG = r i c e p o lish in g s concen trate
Av. w t. = average weight
Reproduced
with permission
Table V
Development o f R esista n ce in Prematurely Weaned Mice R eceiving
Q u a n tita tiv e ly or Q u a lita tiv e ly Inadequate D ie ts
(V. S . V irus Intram uscularly)
of the copyright owner.
Age o f Mice
D iet
Further reproduction
1 . "M" d ie t + whole m ilk + y e a st +
CLO ad l i b .
+
2 . "M" d ie t + « . «
CLO q .s . m aintain wt.
«
6 wfiftks
4 weeks
5 weeks
No.
No.
No.
%
*
*
r e s is t a n t t e s ­ Av. r e s is ta n t t e s - Av. r e s is ta n t t e s - Av.
te d w t.
jsdu.
A9<L LffiL._____
Gm. \
Gm.
Gm.
\
100
12
J
1
6
.8
•
•
a
•
•
•
• «»
9
19
JZS
+
prohibited
without permission.
23
6 .6
8
1?
6 .9
*• »
•••
•••
£2
16
1 2 .3
100
12
1 3 .2
3k
11
140
n
+ m + « + epc ( d e f ic ie n t
in B complex except B^)
£z
15
8. 2'
£2
18
8 .8
£2
9
1 0 .7
5. CSLS + CLO + y e a st (d e f ic ie n t in E)'
20
15
9 .7
100
10
140
21
6
13.8
n
+ « + WGO + autoclaved y ea st
(d e fic ie n t in h e a t - la b ile B)
20
15
8 .6
12
15
6 .7
62
9
1 0 .7
3 . CSLS + CLO + WOO +‘ y ea st
4*
6.
.
Mice suckling mothers on "M" d ie t , weaned at 14 clays o f age
"M" d ie t « mixture o f natural foods described in t e x t; other abbreviations— same as in Table IV
41
_
Young m ice w hich su c k le d m others fe d a p u r if ie d a r t i f i c i a l
d ie t d e f i c i e n t in v ita m in E (wheat germ o i l ) , a lth ou gh th e y con tin u ed
t o g a in in w e ig h t, d id not d e v e lo p r e s is t a n c e to th e v ir u s in t h e same
manner as m ice w hich had su c k le d m others fe d a supplemented d i e t con­
t a in in g wheat germ o i l .
M ice w hich had su c k le d m others fe d a d ie t r ic h in n a tu r a l
f o o d s t u f f s w ere weaned at 14 days o f age and fe d t h e d i e t s l i s t e d in
T ab le V.
The d ata o b ta in ed in t h i s ph ase o f t h e stu d y (T ab le V) r e v e a l
th a t prem aturely weaned m ice fe d a d ie t r i c h in n a tu r a l f o o d s t u f f s and
v ita m in s g a in ed in w eight and d evelop ed r e s is t a n c e t o th e v ir u s in a
normal manner.
However, m ice weaned a t 14 days g iv e n th e same diert,
but o n ly in amounts s u f f i c i e n t t o m a in ta in t h e i r weaning w e ig h t, d id
n o t become r e s i s t a n t .
At 36 t o 39 days o f a g e t h e s e underfed m ice were
s t i l l 92 per cen t s u s c e p t ib le a s c o n tr a s te d w ith 100 per cen t r e s is t a n c e
i n t h e i r s i b l i n g s w hich r e c e iv e d t h e same d i e t ad lib itu m .
P rem aturely weaned m ice g iv e n t h e p u r if ie d a r t i f i c i a l d ie t
supplem ented by t h e a d d itio n o f y e a s t , cod l i v e r o i l and wheat germ
o i l were not q.uite as r e s i s t a n t a t 4 w eeks o f age a s th e weaned m ice fe d
t h e d ie t r i c h in n a tu r a l f o o d s t u f f s and v ita m in s .
However, a t 5 snd 6
w eeks o f age th e y had d ev elo p ed normal r e s i s t a n c e .
When m ice were weaned a t 14 days and put on a d i e t d e f ic ie n t
in t h e h e a t - l a b i l e components o f th e B -com plex b o th growth and d e v e lo p ­
ment o f r e s is t a n c e were markedly r e ta r d e d .
A lthough r e ta r d a tio n i n t h e development o f r e s is t a n c e was not
marked in t h e weaned m ice fe d a d ie t d e f i c i e n t i n t h e B—complex ex cep t
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
B^ s t i l l o n ly 67 p er cen t were r e s i s t a n t a t 5 weeks o f age a s compared
with. 100 p e r c e n t o f th e weaned m ice w hich r e c e iv e d th e supplem ented
d ie t.
I n th e group o f weaned m ice w hich w ere fe d a p u r if ie d a r t i ­
f i c i a l d i e t d e f ic ie n t in v ita m in E th e r e was a d e la y i n th e development
o f r e s is t a n c e a lth o u g h growth was not r e ta r d e d .
F u ll-g ro w n m ice, w hich a re r e s i s t a n t t o in tra m u scu la r in j e c ­
t i o n o f v e s i c u l a r s t o m a t it is v i r u s , d id not l o s e t h i s r e s is t a n c e when
m ain tained on a d ie t d e f i c i e n t e it h e r in c e r t a in components o f th e B com plex or in v ita m in E f o r a p e r io d o f 6 t o 8 w eeks.
I t i s apparent t h a t a d e f ic ie n c y o f c e r t a in d ie t a r y f a c t o r s ,
e it h e r in th e m atern al d ie t du rin g th e n u r sin g p erio d o r in t h e d i e t o f
prem atu rely weaned m ic e , can r e ta r d or i n h i b i t t h e developm ent o f at
l e a s t one ty p e o f c o n s t it u t io n a l b a r r ie r t o involvem ent o f th e c e n tr a l
nervous sy stem b y a n e u r o tr o p ic v ir u s .
However, i t has not been p o s s ib le
t o break down t h i s r e s is t a n c e by th e same means once i t has been acq u ired
by fu ll-g r o w n m ic e .
E vid en ce o f " p recociou s" s e x u a l m a tu ra tio n (d e s c e n t o f t e s t e s
or opening o f v a g in a ) was ob served on th e 21s t day o f l i f e i n 100 per
cen t o f m ice in j e c t e d w ith pregnant mare serum (m ice u s u a lly become
s e x u a lly m ature betw een th e 28th and 36t h day o f l i f e ) .
However, pre­
m ature developm ent o f r e s is t a n c e t o in tra m u scu la r in j e c t io n o f v e s ic u la r
s t o m a t it is v ir u s was not a s s o c ia t e d w ith " p recociou s" s e x u a l m atu ration
s in c e 21 day o ld tr e a t e d and u n tr e a te d l it t e r m a t e s were e q u a lly s u s c e p t ib le .
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
BIBLIO GRAFH T
1.
S a b in , A .B . and O lit s k y , P .K ., J . E xper. M ed., 6 6 :1 5 , 1937
2.
'__________ , i b i d . , 66 :3 5 , 1937
3 * ______________ ;_________________ , i b i d . , 6 7 : 2 0 1 , 1938
4*
_______________ j_______________ , i b i d . , 67:229, 1938
5« _______________________________ , An. J . P a t h ., 1 3 : 6 1 5 , 1 93 7
6 .__________ ___________________
, P r o c . S o c . Exper. B i o l , and M ed.,
38:595, 1938
7.
~
____________
i b i d . , 38:597, 1938
8.
H u rst, E.W ., B rain , 59 :1 , 1936
9.
G ersten b erg er, H .J ., Am. J . D is . C h ild ., 2 6 :3 0 9 , 1923
10.
11.
K u ttn er, A.G. and Wang, S .H ., J . E xper. M ed., 6 0 :7 7 3 , 1934
T u n geb lu t, C.W. and E e in e r , R'.R., J . Exper. M ed., 6 6 :4 7 9 , 1937
A
'
12.
S a b in , A .B ., J . Exper. M ed., 69:5Q 7, I 939
13.
Cowdry, E .V ., L u cas, A.M. and i l e f f . C .E ., J . I n f e c t . D i s . , 5 7 :1 7 4 ,
14•
K u czyn sk i, M ., The A lim entary E a cto r in D is e a s e , The Hague, G.N a e ff,
1937
15.
P erl,a, D ., Arch. P a t h ., 2 5 :6 9 4 , 1938
16.
Osborne, T .B . and Mendel j L .B ., J . B i o l . Chem.:,
t,
1
Gyorgy, P . , Biochem, J . , 2 9 :7 4 1 , 1935
17.
18.
6 3 : 2 3 3 , 1925
19.
Warburg, 0 . and C h r is tia n , W., Biochem. Z t s c h r ., 2 5 4 :4 3 8 , 1932
I
S te r n , P.O . and F in d la y , M.,v J . P a th , end B a c t ., 3 2 :6 3 , 1929
20.
Zimmerman, H.M. and Burack, E . , A rch. P a t h ., 1 3 :2 0 7 , 1932
2 1.
D avison , C. and S to n e , L ., A rch. P a t h ., 2 3 :2 0 6 , 1937
22.
Tauber, H ., S c ie n c e , 86:180, 1937
Reproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
1934
23.
Zimmerman, H.M ., C o w g ill, G .E ., and Fox, J . C . J r . , A rch. N eurol.'jand
P s y c h ia t ., 3 7 :2 8 6 , 1937
24.
Su re, B ., J . B i o l . Chem., S 0 :2 9 7 , 1928
25.
Evans, H.M. and B urr, G .O ., J . B i o l . Chem., 76:2.73» 1928
26.
O le o t t , E .S ., J . N u t r it io n , 1 5 :2 2 1 , I 938
27.
Pappenheimer, A.M ., Am. J . P a t h ., 1 5 :1 7 9 , 1939
28.
Jaclcson, C.M ., E f f e c t s o f I n a n it io n and M a ln u tr itio n upon Growth and
S tr u c tu r e , P . B la k is to n t s Son and Co. 1925
29-
__________
30.
______________, i b i d . , S:B 1, 1929
31.
, Arch. P a t h ., 7 :1 0 4 2 , 1929
, i b i d . , 8 :2 7 3 , 1929
32.
Rudin, D ., A nat. E e c ., 6 3 :1 7 5 , 1935
33.
S u re, B ., J . N u t r it io n , 1 9 :5 7 , 1940
3 4.
Elvehjemj-
C-.A., Koehm, C .J . and O leso n , J . J . ,
J.B io l.
Chem., 1 1 5 :7 0 7 ,
1936
35*
S ta rk ey , W.F. and Leathern, J . H . , Am. J . P h y s i o l ., 1 2 7 :7 5 1 , 1939
R eproduced with perm ission of the copyright owner. Further reproduction prohibited without permission.
Документ
Категория
Без категории
Просмотров
0
Размер файла
2 527 Кб
Теги
sdewsdweddes
1/--страниц
Пожаловаться на содержимое документа