Nutrition and breeding behavior of pocket mouse for space nutrition applicatio

Calloway, D. H.

Margen, S.

Ostwald, R.

English

NASA Technical Reports Server

INTEGRATED RESEARCH PROGRAM IN SPACE NUTRITEON 
Semi-Annual Report to the 
National Aeronautics and Space Administration 
NGR 05-003-351 
1 February 1970 to 31 July 1970 
Co-Principal Investigators: 
Doris Howes Calloway 
Professor of Nutrition 
Sheldon Margen 
Professor of Human Nutrition 
Rosemarie Ostwald 
Associate Professor of Nutrition 
Department of Nutritional Sciences 
University of California, Berkeley 
September 1970 
https://ntrs.nasa.gov/search.jsp?R=19710000940 2020-03-12T00:33:01+00:00Z
Abstract: Nutrition and Breeding Behavior of the Pocket Mouse 
1. Baseline data for the growth of - P. pe. fed a diet of mixed seeds 
were established. This was intended as a preliminary for the determination 
of the adequacy for growth of our semi-synthetic diet. 
2. The study of the effects of photoperiod on estrus behavior of E. pe. 
was terminated. The data of this 3-year study have been compiled and a 
manuscript for publication is in preparation. 
3 .  A study of the intestinal absorption and metabolism of calcium, mag- 
nesium, and phosphorus by - P. pe. has been undertaken. 
NOTE: Because of financial restraints during this project period a technician 
was available for five months only. 
Report - Work Completed 
Nutrition and Breeding Behavior of the Pocket Mouse 
I. Adequacy of the semi-synthetic diet for growth of P. penicillatus 
We have previously shown that the semi-synthetic diet developed by us 
is adequate for weight maintenance and general health of adult pocket mice 
(Spiller, Gene A, and Rosemarie Ostwald, J. Nutr. - 94:297, 1968). In order 
to test the adequacy of this diet for growth it was necessary to establish 
as a baseline growth of these animals on their usual diet. 
Ten litters of - P. E. (19 males, 13 females) born during summer and fall 
1969 were used in this study. 
diet routinely used by us (equal parts of sunflower seeds, millet, canary 
seeds and oats) and were supplemented with fresh carrots and lettuce 
5 times/week. By previous experience, these supplements have been found 
necessary for reasonable survival of the young, 
The dams and pups were fed the mixed seed 
In the absence of carrots, 
2. 
which we  b e l i e v e  t o  serve pr imar i ly  as source of water, t he re  is  l i t t l e  
growth and a h igh  m o r t a l i t y ,  probably due t o  f a i l u r e  of l a c t a t i o n .  I n  t h e  
absence of l e t t u c e  t h e r e  is  a high rate of m o r t a l i t y  a t  about 20-25 days 
of age preceeded by c h a r a c t e r i s t i c  o v e r a c t i v i t y  and convulsions.  We b e l i e v e  
t h a t  t h i s  may be  caused by a magnesium def ic iency ,  a l l e v i a t e d  by the  l e t t u c e .  
The pups were marked (1% gent ian  v i o l e t )  and weighed 3 times/week, s t a r t i n g  
a t  about 14 days of age. We d id  no t  a t tempt  earlier weighing f o r  f e a r  of 
d i s tu rb ing  the  dams unduly and s o  i n h i b i t  l a c t a t i o n .  The pups were separa ted  
i n t o  i n d i v i d u a l  cages when they reached 10-11 g. Ear l ier  sepa ra t ion  l e d  t o  
l ack  of growth and high mor t a l i t y .  
Figures  1 and 2 show the  growth curves.  There w a s  r ap id  growth t o  about 
Af t e r  20 days of age (0.6 and 0.5 g/day f o r  males and females r e spec t ive ly ) .  
t h i s  they grew a t  a s lower rate t o  45-55 days (0.16 and 0.13 g/day f o r  males 
and females r e spec t ive ly )  . 
There w e r e  14 deaths  during the  80 days of t h e  experiment. Table 1 shows 
the  age and s e x  d i s t r i b u t i o n  of t hese  l o s s e s .  
than females (10 males vs. 4 females).  The cause of t hese  deaths  are unknown. 
Visual i n spec t ion  a t  autopsy d i d  n o t  r e v e a l  any obvious abnormal i t ies .  Twelve 
of t he  1 4  deaths  occurred i n  only 5 of t h e  PO l i t ters .  I n  most cases t h e  
animals l o s t  weight dur ing  the  week preceding death and j u s t  d i d  n o t  seem t o  
t h r i v e .  
2 .  Mineral metabolism of P. Pe. 
Males seemed t o  be less s tu rdy  
We have previously shown ( S p i l l e r ,  Gene A. and Rosemarie Ostwald, 
J .  Nutr.  - 9 4 :  297,  1968) t h a t  a d i e t  conta in ing  calcium, magnesium, and 
phosphorus i n  the  ino rgan ic  forms usua l ly  employed is  t o l e r a b l e  f o r  pocket 
mice only i f  supplement wi th  c a r r o t s  as source of water. I f ,  howeverp 
calcium- and magnesium-glycero phosphate are used as source of minerals  
t h e  d i e t  is  adequate wi thout  such supplementation. I n  o r d e r  t o  understand 
3 .  
t h e  reasons f o r  t h i s  observa t ion ,  a s tudy w a s  undertaken t o  i n v e s t i g a t e  t h e  
i n t e s t i n a l  absorb t ion  of t h e  macro minera ls  i n  t h e s e  animals and t h e i r  
subsequent metabol ic  f a t e .  The p l a n  is  t o  measure t h e  i n t e s t i n a l  absorb t ion ,  
body r e t e n t i o n  and exc re t ion  of calcium, magnesium and phosphorus using 
r a d i o a c t i v e  i so topes  i n  E. e. f e d  four  d i f f e r e n t  d i e t s :  
1) Nixed seeds ,  on which they can be maintained i n d e f i n e t l y .  
2) The l e t h a l  d i e t  containing inorganic  salts .  
3) The same d i e t  wi th  a supplementation of c a r r o t s .  
4 )  Our semi-synthetic d i e t  conta in ing  calcium- and magnesium- 
g lycero  phosphates.  
As necessary p re l imina r i e s  we have s tud ied :  
a) Methods f o r  t h e  c o l l e c t i o n  of f e c e s  and u r ine .  
b) The amounts of calcium, magnesium and phosphorus excreted i n  
f eces  and u r i n e .  
c) The admin i s t r a t ion  of t he  tracers. 
d) Methods t o  determine the  amounts of r a d i o a c t i v i t y  excre ted  
and r e t a i n e d .  
Resul t s  
Feca l  p e l l e t s  can be co l l ec t ed  from mice kep t  i n  g l a s s  metabol ic  cages 
f o r  24 hours  wi thout  food. 
amounts of calcium and magnesium i n  the  exc re t a  has  been determined by atomic 
absorb t ion  measurement of d r i e d  and wet-ashed f eces  and i n  t h e  u r i n e  samples 
i n  1% LaC12 (Table 2 ) .  
Urine can b e  obtained by washing t h e  cages. The 
I n  o rde r  t o  adminis te r  t h e  r a d i o a c t i v e  tracer a s m a l l  p e l l e t  of t h e  
appropr i a t e  d i e t  w a s  t r e a t e d  wi th  an aequous s o l u t i o n  of t he  tracer, then 
d r i e d ,  counted and placed on the  back of t h e  tongue of t h e  mouse who swallowed 
i t  by reflex ac t ion .  
We have chosen t o  s tudy  i n i t i a l l y  Ca47 because of i t s  d e s i r a b l e  h a l f - l i f e  
and energy c h a r a c t e r i s i t i c s .  Figure 3 shows that body r e t e n t f o n  and decay 
4. 
of Ca47 can b e  measured by whole body counting i n  t h e  w e l l  of a Packard auto-  
g a m a  spectrometer ,  series 410A. 
We have a l s o  s tud ied  the  p o s s i b i l i t y  of measuring P32 by i ts  
bremstrahlung r a d i a t i o n  c h a r a c t e r i s t i c s .  
poss ib l e  i n  s p i t e  of t h e  f a c t  t h a t  the  peak of t h e  energy d i s t r i b u t i o n  occurs  
a t  t h e  lower end of t h e  scale where the  n o i s e  level of t h e  machine i s  high.  
With a wide mode of a n a l y s i s  (200-1000 o r  100-1000 Kev.) t h e  r e s u l t s  may 
be  even b e t t e r .  This  w i l l  make i t  poss ib l e  t o  use  P , a B - e m i t t e r ,  i n  
t he  same fash ion  as Ca47 f o r  t h e  s tudy of phosphate metabolism. 
Figure 4 shows t h a t  t h i s  is  indeed 
32 
Table 1 
Mor ta l i t y  of Young E. E. 
Age (days) 14-25 26-35 36-45 46-65 56-80 To t a l  
Male (19) 0 5 0 5 0 10 
Female (12) 0 0 1 2 1 4 
Male + Female - 5 1 7 1 14 
'I'able 2 
Excret ion of Calcium and Magnesium i n  P. Pe 
Animal Calcium IYagnes ium Calcium Magnesium 
mg/24 h r s  mg/24 hrs/mg dry  w t .  
Urine 
Feces 
1 293 94 
2 506 97 
3 501 - 
4 453 186 
2 687 1132 
3 407 1232 
4 455 832 
20 33 
9 27 
16 29 
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9. 
WORK I N  PROGRESS 
1. E f f i c i e n t  Subs t r a t e s  f o r  Energy. 
I n  an  at tempt  t o  determine t h e  minimal amounts of carbohydrate,  f a t  and 
p r o t e i n  necessary t o  prevent  t i s s u e  catabol ism,  s t u d i e s  were c a r r i e d  out  on 
s i x  obese sub jec t s .  
t h e  major f o o d s t u f f s  a t  low c a l o r i c  i n t a k e  i s  probably t h e  b e s t  model t o  
answer ques t ions  of i d e a l  energy s u b s t r a t e s .  
supply of energy source-- their  own adipose t i s s u e .  They are a l s o  a b l e  t o  
adapt  t o  f a t t y  a c i d s  as t h e  major energy source.  Experimental evidence has  
suggested t h a t  t h e  obese f a s t i n g  ind iv idua l  is  a b l e  t o  most e f f i c i e n t l y  burn 
f a t  and conserve p ro te in .  
u t i l i z a t i o n  of adipose t i s s u e  f o r  energy i s  always accompanied by a mandatory 
breakdown of c e l l u l a r  p r o t e i n .  The experimental  p l an  d i e t s  are ou t l ined  
below (Fig.  5). 
Obese s u b j e c t s  were s tud ied  because feeding such s u b j e c t s  
The obese s u b j e c t s  have a l a r g e  
We a l s o  attempted t o  asnwer t h e  ques t ion  whether 
The r e s u l t s  of t h e  experiment are s t i l l  being analyzed. However, t h e  
following r e s u l t s  can be commented upon: 
1) A l l  s u b j e c t s  l o s t  weight a t  about t he  same rate. 
2) The weight l o s s  appeared t o  be r e l a t e d  t o  c a l o r i c  d e f i c i t  and no t  t o  
t h e  s u b s t r a t e  f o r  energy being f ed .  
3) With weight l o s s  t h e r e  w a s  a gradual  f a l l  i n  metabol ic  rate. This  
decrease  w a s  due t o  ( a ) d e c r e a s e i n  active metabol ic  t i s s u e  and (b) 
apparent ly  an a l t e r a t i o n  i n  e f f i c i e n c y  of u t i l i z a t i o n  of s u b s t r a t e .  
The latter s ta tement  is  a d i f f i c u l t  t o  s u b s t a n t i a t e  and a more c a r e f u l  
a n a l y s i s  of t h e  d a t a  i s  underway before  w e  can be  c e r t a i n  of t h i s  
s ta tement .  
Analysis  of t h e  body composition changes is  no t  y e t  completed. Corre- 
l a t i o n s  between va r ious  techniques of measurement appear t o  be  low. 
Figure  5 
F i n a l  D i e t  P lan ,  Experiment B16 (Apr i l  - June, 1969) 
Metabolic Per iod  I I1 I11 I V  V V I  
No. DaysIPeriod 12  1 2  9 12  12 7 
Dates 4121-512 513-5114 5/15-5123 5124-614 615-6116 6/17-6123 
Subjec ts  D i e t s  
Mabel A. i/ 1 I/ 2 # 4 f 3 f 5  1000-cal e food 1) 
Ruth M. I/ 1 f 2 I/ 3 ii42) -- --- 
Ast r id  B. ill f 4 // 3 # 2 #5 1000-cal. food 
Meta D.  I/ 1 8 3 #43) j12~)  1/5 1000-cal food 
Malinda Lo // 1 a4 # 2 # 3 #5 1000-cal. food 
Mary F. a 1 #3 N ~ )  #44) #5 1000-cal e food 
Approx. gN/day 
D i e t  #l = 2000 K c a l ,  egg albumin + f a t  equal  t o  D i e t  113 12.4 
D i e t  #2 = 1000 Kcal, egg albumin and only CHO 12.4 
D i e t  #3 = 1000 Kcal, egg albumin and only f a t  12 .3  
D i e t  #4 = 1000 Kcal, a l l  p r o t e i n  (egg albumin, promine 
+ casal) 38.8 
D i e t  #5 = 377 Kcal, egg albumin 12 .3  
COMPOSITION OF DIETS AS SHOWN I N  FIGURE 6,  
1 )  1000 c a l o r i e  food d i e t  - balanced m e a l s  of low-calorie food i t e m s  t o  equal  about 
2) l e f t  5/24. 
1000 c a l o r i e s ,  100 gm p ro te in lday ,  and m i n i m a l  f a t .  
3) 5/15-5126. 
4) 5127-614. 
11. 
Figure  6 
Composition of Diets. 
D i e t  112: 1013 Kcal. 
1 2  gm N as egg albumin. 
R e s t  of Kcal as CHO. 
650 Kcal of t h e  1013 as CHO Kcal = 64.2%. 
Also 28 K e a 1  as f a t .  
Also 328 Kcal as p ro te in .  
D i e t  83: 1010 Kcal. 
12 gm N as egg albumin. 
R e s t  of Kcal as f a t  (50% b u t t e r f a t ,  50% saf f lower  o i l ) .  
662 K c a l  of t h e  1010 as f a t  K c a l  = 65.5%. 
Also 1 9  Kcal as CHO Kcal. 
Also 327 Kcal as p ro te in .  
D i e t  #4: 1015 Kcal - "all" p r o t e i n .  
946 Kcal of 1015 as p r o t e i n  Kcal 
(comes from 12.0 gm N egg albumin 
13.9 gm N promine D 
12.62 gm N c a s a l ) .  
Also 3 gm saf f lower  o i l  = 27 f a t  Kcal. 
+ 15 K c a l  from f a t  i n  3 p r o t e i n  sources .  
Also 28 Kcal as CHO Kcal i n  p r o t e i n  sources .  
2. Crea t ine  and Crea t in ine  Pool S i z e  and Turnover Rate. 
The experiment r e l a t i n g  t o  c r e a t i n e  and c r e a t i n i n e  pool s i z e  and turnover  
rate is  i n  progress .  
of heavy i s o t o p e  l a b e l l e d  compounds are under way. 
The human feeding  po r t ion  has  been completed and ana lyses  
3.  U t i l i z a t i o n  of Ketone Bodies i n  Brain During S ta rva t ion .  
The b r a i n  w a s  formerly considered t o  u t i l i z e  glucose only as energy f u e l .  
However, r e c e n t  evidence, e s p e c i a l l y  t h e  s tudy of prolonged s t a r v a t i o n  treat- 
ment of o b e s i t y  p a t i e n t s ,  have shown t h a t  ketone bodies  were t h e  major energy 
source a f t e r  a long s t a r v a t i o n .  Our pre l iminary  s t u d i e s  showed t h a t  t h e  b r a i n  
could u t i l i z e  a c e t o a c e t a t e  o r  B-hydroxybutyrate i n  v i t r o  and t h a t  t h e  amount 
of a c e t y l  Co-A formation w a s  increased  i n  t h e  b r a i n  p repa ra t ion  of s t a r v i n g  
animals.  
pathway i n  va r ious  condi t ions  i n  o rde r  t o  determine t h e  enzymes r e spons ib l e  
f o r  r egu la t ion .  
l u l a r  p a r t i c l e s  o r  his tochemical  prepara t ion .  
We measured t h e  amount of va r ious  me tabo l i t e s  of t h e  g l y c o l y t i c  
Enzyme l e v e l s  were a l s o  measured i n  e i t h e r  i s o l a t e d  subcel-  
No d e f i n i t e  and conclus ive  
mechanism could be  obta ined  a t  t h i s  moment because of i n c o n s i s t e n t  r e s u l t s  
and t h e  i n t e r f e r e n c e  of glucose and glycogen i n  t h e  prepara t ion .  
a t tempts  t o  i n h i b i t  t h e  u t i l i z a t i o n  of glucose f o r  t h i s  s tudy have not  been 
very  success fu l .  
The c u r r e n t  
Fur ther  s t u d i e s  t o  e l imina te  t h e  e f f e c t  of glucose and glycogen, using 
ev i sce ra t ed  animals and cytochemical s t u d i e s  f o r  va r ious  d i f f e r e n t  compartments 
of t he  b r a i n  should be performed i n  o rde r  t o  e l u c i d a t e  the  mechanism of u t i l i -  
z a t i o n  of ketone bodies  i n  t h e  b ra in .  
9 
FUTURE PLANS 
Work on i s o l a t i o n  of t h e  r a d i a t i o n  p r o t e c t i v e  f a c t o r  from a l f a l f a  has  
been temporar i ly  suspended pending sea rch  f o r  an  improved i s o l a t i o n  procedure 
I f  a d d i t i o n a l  funds are made a v a i l a b l e ,  work w i l l  be  continued on 
e f f i c i e n t  u se  of s u b s t r a t e s  f o r  energy. 
Current funds w i l l  be  s u f f i c i e n t  t o  complete t h e  s tudy of c r e a t i n e  
and c r e a t i n i n e  pool  s i z e  and turnover  rate. 


Integrated research program in space nutrition Semiannual report, 1 Feb. - 31 Jul. 1970

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