PROJECT REPORT 
COMMITTEE OR FOOD RESEARCH 
QUARTERMASTER FOOD AND CONTAINER INSTITUTE 
FOR THE ARMEQ FORCES 
CHICAGO ILLINOIS 
298W&M& AH® BXTSLOPNKRT BRANCR 
MILITARY FLAJHflHO PIVSSIOM 
OPTICS or TNS 
0OASTRRMA8TSS OBMSBAL 
T53FEiATrirr"i gyrmiTrw—~ 
Rutgers University 
LOCAL I Tfr' ' '' ' 
Hew Brunswick* New Jersey 
DIVISION 
Arts & Sciences 
NIP artNent ; 
Bureau of Biological Research 
OFFICIAL INVESTIGATOR, 
James Allison 
COLLABORATORS 
John A© Anderson. 
REPORT NO, 
1 
PI LE NO. 
.P-1017 
CONTRACT NO. 
Wll=483«qm~195 
For PERIOO eOVERIN6 
1 July 1947 - 1 Septc 1947 
1 July 1947 
T»TL|t 1 x] PR0§RE*S REPORT 1 j PNA»E NlfONT [ J ANNUAL REPORT [ JteWHNATION REPORT 
Utilization of Proteins Correlated with Appetite and Intestinal Flora© 
SUMMARY 
The following experiment involving wheat gluten is a part of a 
study to determine the'nutritive value of a variety of proteins in 
normal and protein depleted dogs.® The first column in Table 1 records 
the nitrogen balance indexes of wheat gluten in five normal dogs 0 
The index varied between Qc3i and Qn54<> averaging 0o44© Following- 
the determination of the. indexes' in normal dogs- the animals were 
depleted in proteins by feeding a protem--free diet© Dogs 36 39* 
66;. and 52 were fed the protein*free diet until the albumin/gtobulin 
ration wa3 reduced to approximately 0o5o This low ratio* determined 
by salt fractionation is found in dogs in a severely depleted stateo * 
The nitrogen balance indexes in these depleted dogs varied from 0o7 
to Oc76s much higher than control values© The albumin globulin ratio 
for dog 60 was Q07 indirating that the depletion was not as severe 
no in the other four animals.o The index was increased to 0o57 when 
dog 60 was depleted but the increase was not marksdo 
After the nitrogen balance index of wheat gluten had been determined 
In the depleted state* the dogs were fed 0o62 grans of wheat gluten 
nitrogen/day/kulo of body weight for thirty days© The value 0o62 was 
selected because it represented the amount of wheat gluten nitrogen* 
based on relative nitrogen balance indexes*.that would be equivalent 
to 0o35 grams of casein nitrogen© An attempt was made,, in other words*; 
to feed sufficient wheat gluten nitrogen so that the same degree of 
positive nitrogen balance woi:id produced as was obtained by feeding 
0-35 grams of of body weight© 
(Continued) 
TO?".* 12-121 It v;as hoped, in this v*ay, to compare various proteins 
on a constant nitrogen balance rather than a constant 
nitro-sn intake level. The nitrogen invested over a per- 
iod of 30 recorded in column 3 of Table I, represents 
the actual intake oi the s expressed on a sq. Th body 
surface area basis. The intake was similar in all dow s 
except oO which rel sed to eat as much as the others. 
The b >dv nitrogen gained, listed in column 3, is the diff- 
erence between the nitrogen invested and that excreted. 
The jer cent oi nitrot an invested which s represented 
in vody nitrogen gained is recorded in churn 6. nhe 
per cent is small b t essentially he sai e (20 0 
in the four doss which were seriously depleted. mhe per 
cent (.am f*7 0 was mch less in the dog 76o, with the 
low nitrogen balance index, in the depleted state. Thus, 
the nitrogen balance index is one measure of the efficiency 
of the protein in repletion. The total circulating 
plasma protein nitrogen gained is recorded in column 7 
and the per cent body nitrogen gained which is represented 
in plasma protein nitrogen, averaged 3.If. The fraction 
of nitrogen that entered into tie formation of plasma pro- 
tein was, therefore, quite small. 
Data, summarized in previous reports, demonstrated 
the red ction in the excretion of urinary nitrogen which 
accompanies depletion m proteins. The excretion of 
urinary nitrogen r. fleets in par', kt least, the magnitude 
of the protein stores of the animal. The average urinary 
nitrogen excretion of dogs $6, 39# 96 control 
periods feeding a protoin-free diet was 2.3 m 
of body surface area. 
After depletion the average excretion ,n.-s reduced to 
1*33 gn/day/ sq. m.. Thirty days of re Action on the wheat 
gluten did not increase, rather ecro.sed, this excretion. 
The average excretion on a pro''.c in-free diet f or example 
of these three doc a f ollowing repletion was A 25 um/clay/ 
sq. m. Repletion with wheat gluten, therefore, did not 
increase the excretion ox ody nitrogen above that *0'nnn 
in the depleted state. nhese data ca be interpreted 
to mean that wheat gluten did not re A. nish those protein 
stores associated with the excretion of catabolic nitrogen. 
P-1017 4k1 
Cotninued  
Repletion over period of 30 day s 
i Balance 
i'll Ui C? X 
Dog 
Index 
* 
Plasma 
# 
Nitrogen 
Nitrogen 
Body 
B.K.G.xlOO 
Protein 
P.N.G.xlOO 
Control 
Depleted 
Ingested 
Excreted 
Nitrogen 
N.I. 
Nitrogen 
pro 
N.I* 
.ga ined 
gained 
.D* 14 • D. 
(B.N.G. ) 
fp.II.G. ) 
(1) 
(2) 
(3) 
(4) 
(5) 
(6) 
(70 
to 
gra/sq.ri. 
gxn/ sq.N* 
gm/sq.N. 
gm/sq. I.:* 
gm/ sq. M. 
Pt 
Jo 
36 
o.47 
0.76 
332.6 
291.5 
62.3 
19 
1.5 
2.2 • 
CC 
o .51}. 
0.70 
351.5 
290.0 
78.3 
22 
f 
O 
3*4 
oo 
Co 
0.49 
0.76 
293.2 
239.3 
70*9 
24 
2.4 
3-3 
Oc. 
°*?1 
0.70* 
136.3 
103.7 
32.7 
24 
1-7 
5-3 
DU 
0.42 
0.57 
344.0 
313.4 
25.6 
7 
1.1 
2*8 
1 
* Estime 
ted* 
— 
— 
Table I 
Nitrogen balance indexes of wheat gluten before and after depletion 
in proteins and wheat gluten nitrogen ingested (Hi), nitrogen 
excreted, body nitrogen gained (B*N*G*), per cent of nitrogen Ingested 
represented by (B*H.G.), plasma protein nitrogen gained, and per 
cent of body nitrogen gained represented by P.H.G. after 30 days of 
reple tion on wheat gluten. 
P-1017 #1 PROJECT REPORT 
COMMITTEE ON FOOD RESEARCH 
QUARTERMASTER FOOD AND CONTAINER INSTITUTE 
FOR THE ARMED FORCES 
CHICAGO ILLINOIS 
RI8XARCK AMD BX'ftLOPIlSrr BRANCH 
MILITARY PLARMING DIVISION 
OPPICX or THE 
QUARTERMASTER GENERAL 
T?R5PimtlN6 INSTITUTION 
Rutgers University 
LOCALITY 
N5W Brunswick, New Jersey 
DIVISION 
College of Arts & Sciences 
DEPARTMENT ” 
Bureiu of Biological Research 
OPFI Cl AL INVEST! GATOR 
James B. Allison 
COLLABORATORS 
John A. Anderson 
REPORT NO. 
c. 
PILE NO. 
. 1. r«i ni 7 
contract" no. 
188-3^105 
(*0R P fRIOD COVERING “ 
1 Sept, 194-7-1 Dec* 191+7 
— — 
INTTIATI ON 0ATE 
1 July 19il-7 
T,TLE! r PR0§RESS Report [ j phase report [ Iannual report [ Jtermimation report 
Utilization of Proteins Correlated with Appetite and 
— T-nf.n nfl 1 FI * 
SUMMAiY 
This investigation is an outgrowth of observations 
noted from time to time while gathering data on the 
nutritive value of proteins in normal and depleted dogs. 
An observation commonly noted by ourselves and others 
was that minor changes in the composition of a ration 
may change the texture of feces for one or more days 
following ingestion of the altered ration. In our exper- 
ience this change in fecal structure may be induced by a 
modification in chemical composition of the ration, causing 
no alteration of its physical texture, he also noted at 
times, especially in depleted dogs, feces having odors 
which suggested that the abundant and varied microbial 
flora of the intestinal tract might be undergoing con- 
siderable modification. 
This progress report deals with the relative numbers 
of different bacteria present in the feces of dogs employed 
in nitrogen balance index tests of egg albumin and of 
casoin, and in feces from these same dogs after transfer 
to our basic protein-free diet. The protein-free ration 
which was fed at a level of 70 cal. per kilogram of body 
weight had the following composition: 
Grams 
Calories 
Sucrose 
3-50 
ill. Op 
Dextrose 
5.go 
23.6p 
Dextrin 
2.05 
ll.l+Ot 
Lard 
2.3’) 
21.00" 
Salt mixture 
0.30 
-- 
Agar 
0.47 
- - 
Vi a ter 
21.1+9 
— 
M>. 
- i » 
70.00 
JSslS17, #2 
C(JMO FORM 19.191 t 
5 Apr I! 46 1 1*1 (**vi%»4) The necessary vitamin supplements were added at the' time 
of -feeding. In these experiments Swift*# egg albumin 
was added to the protein-free diet at a level 
of 0.99 girt per kg* of body weight and Labco casein 
at a level of 1.10 gms. per kg. of body weight#1 
Observations were made on the feces of. thirteen dogs 
(five on egg albumin and eight on casein) through the 
latter portion of the protein test period and for a 
longer portion of the protein-free period than previous 
investigators found necessary for a change in bacterial 
flora- 
In none of these dogs did the intestinal flora under- 
go any noteworthy change in passing from the protein 
period to the protein-free period. 
A typical microscope slide stained by the Gram method 
would show approximately the following distribution of 
orgamisms? Gram positive forms, of the total 
nurnbe‘r; Spore formers, aorobic and anaerobic,- the latter 
most abundant 2-2cK; micrococci and stophylococci,- 
diplococci (Diplococcus ovalis, most common) 
Streptococci, non spore forming anaerobic' (acidophilic) 
forms, gram negative forms: Escherichia and aerobacter 
most abundant by far (cultures show- mall numbers of 
Proteins and Alcaligenes .forms)., spirals,* 
yeasts, occassionally present* Many cells staining Gram 
negative are doubtless old and dead Gram positive organisms* 
The reasons for lack of significant change in bacterial 
flora under the conditions studied are very likely,» (1) the 
small amounts of protein required for the nitrogen balance 
index and (2) an inadequate quantity of suitable carbohy- 
drate (dextrin) to support development of an-acidophilic 
flora,. We /intend to check these two points-* Although 
the numbers of different kinds of microorganisms varied 
from dog to dog, in general there was marked similarity- 
While the number of animals studied is comparatively small 
there are good reasons for believing these results to be 
typical of the conditions studied, namely,- low protein 
intake, just sufficient for nitrogen equilibrium,- followed 
by a proteihrfjgfp, diet- 
P-1017 #2 
Continued The outstanding conclusion from the work done thus 
i*ar is that no changes in intestinal flora occurred during 
our previous nitrogen balance index studies of proteins 
Undertaken for the formulation of rations. Although 
comparatively few investigators have’ studied the intestinal 
flora of the dogs, there is little doubt that findings 
in the dog are applicable to man. 
P-P-1017 #2