SOME FACTORS INFLUENCING THE LEVEL OF REDUCING
SUGAR IN THE BLOOD OF BLACK-TAILED DEER
by
PHILIP EDWARD WHITEHEAD
B . S c , U n i v e r s i t y o f B r i t i s h Columbia, 1962
A Thesis Submitted i n P a r t i a l F u l f i l m e n t
of t h e Requirements f o r t h e Degree o f
MASTER OF SCIENCE
i n t h e Department
of
Zoology
We accept t h i s t h e s i s as conforming t o t h e
standard required from candidates f o r t h e
Degree o f Master o f Science.
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May, 1966
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ii
ABSTRACT
Some o f t h e f a c t o r s t h a t i n f l u e n c e the blood reducing sugar l e v e l i n
the b l a c k - t a i l e d deer Odeeoileus hemionus columbianus
(Vancouver I s l a n d genotype), have been i n v e s t i g a t e d .
(Richardson)
The d i s t r i b u t i o n
of reducing sugar i n the blood o f these animals was a l s o examined.
I t was found t h a t : feed i n t a k e during the hour preceeding blood
l e t t i n g , short periods o f f a s t , nature o f the feed, and sex o f t h e animal
apparently have no e f f e c t on t h e l e v e l o f blood reducing sugar i n deer.
Blood samples taken i n the evening generally had a higher reducing sugar
l e v e l than those taken e a r l i e r i n the day.
The means used t o r e s t r a i n
the animals during t h e blood l e t t i n g procedure was a l s o found t o have a
marked i n f l u e n c e on the l e v e l o f blood reducing sugar.
Deer r e s t r a i n e d
by p h y s i c a l f o r c e e x h i b i t e d s i g n i f i c a n t l y higher and more v a r i a b l e blood
sugar l e v e l s than those immobilized w i t h s u c c i n y l c h o l i n e .
The l e n g t h
of time required t o draw a blood sample from an animal a l s o i n f l u e n c e d
the blood sugar l e v e l .
The longer t h e time t o l e t a sample, t h e higher
the blood sugar l e v e l i n the sample.
The r e s u l t s i n d i c a t e t h a t the
degree o f excitement, f e a r , and p a i n experienced by the animals preceeding
and during t h e blood l e t t i n g procedure was the p r i n c i p a l cause o f
v a r i a b i l i t y found i n t h e l e v e l o f blood reducing sugar.
Wo reducing sugar could be detected i n t h e erythrocytes o f these deer.
iii
TABLE OF CONTENTS
ABSTRACT
i
TABLE OF CONTENTS
i
i i i
LIST OF TABLES
v
LIST OF FIGURES
vi
ACKNOWLEDGEMENTS
v i i
INTRODUCTION
1
EXPERIMENTAL
3
Animals
3
Nutrition
3
Housing
4
A n a l y t i c a l Techniques
4
Blood c o l l e c t i o n
4
Serum e x t r a c t i o n and storage
5
Hematocrits
5
Blood reducing sugar determination
5
EXPERIMENT 1
7
EXPERIMENT 2
32
EXPERIMENT 3
42
EXPERIMENT 4
47
RESUME
49
BIBLIOGRAPHY
53
APPENDICES
1
Glucose metabolism i n ruminants
59
2
Hormonal r e g u l a t i o n o f blood reducing sugar
61
XV
TABLE OF CONTENTS (cont.)
APPENDICES (cont.)
3
Succinylcholine chloride
63
4
Ingredients o f Peebles V l e r m i l k r e p l a c e r
64
5
Ingredients o f the weaning r a t i o n
65
6
Ingredients o f the adult r a t i o n
66
7
Absorption curve f o r t h e Nelson-Somogyi method o f
determining blood glucose
67
8
Standard curve f o r the Nelson-Somogyi method o f
69
f
determining blood glucose
9
10
Recovery o f glucose from serum
72
Blood sampling record form.
73
LIST OF TABLES
TABLE
PAGE
1
Treatment for experiment 1
2
Results of experiment 1
10
3
Student t test comparing serum reducing sugar
levels of fasted and nonfasted deer.
14
Student t test comparing serum reducing sugar
levels of deer immobilized with succinylcholine
and those restrained physically.
18
F test for determi ning the reproducibility of
serum reducing sugar levels by the two methods
of restraint.
20
Student t test comparing the serum reducing sugar
levels of male deer to those of female deer.
23
The influence of time of day on serum reducing
sugar levels i n deer.
26
Student t test comparing serum reducing sugar
levels of deer sampled at 0800, 1200, and 1600
hours to those sampled at 2000 hours.
28
Feed intake, serum reducing sugar levels, and
weight data of U34 throughout experiment 2.
35
Feed intake, serum reducing sugar levels, and
weight data of U43 throughout experiment 2.
36
Feed intake, serum reducing sugar levels, and
weight data of U16 throughout experiment 2.
37
4
5
6
7
8
8A
9
10
11
8
Feed intake, serum reducing sugar levels, and
weight data of U24 throughout experiment 2.
38
12
Reducing sugar levels i n sequential blood samples.
43
13
The distribution of reducing sugar i n the blood
of deer.
A comparison of blood sugar values reported for
deer.
14
48
50
vi
LIST OF FIGURES
FIGURE
1
2
3.
4
5
6
7
8
9
10
PAGE
The r e l a t i o n s h i p between serum reducing sugar
l e v e l s and feed i n t a k e t h e hour p r i o r t o blood
sampling.
12
A comparison between t h e serum reducing sugar
l e v e l s o f f a s t e d and nonfasted deer.
15
A comparison between t h e serum reducing sugar
l e v e l s o f deer r e s t r a i n e d p h y s i c a l l y and those
immobilized w i t h s u c c i n y l c h o l i n e .
19
A comparison between the serum reducing sugar
l e v e l s o f male and female b l a c k - t a i l e d deer.
24
Comparing serum reducing sugar l e v e l s found i n
deer a t d i f f e r e n t times during t h e day.
27
I l l u s t r a t i n g t h e r e l a t i o n s h i p t h a t can o c c a s i o n a l l y
occur between t h e serum reducing sugar l e v e l s o f
two animals sampled a t t h e same times on the same
day.
31
I l l u s t r a t i n g t h e changes i n serum reducing sugar
l e v e l s t h a t occurred i n deer on p e l l e t s and those
on hay.
33
The e f f e c t o f i m m o b i l i z a t i o n and blood sampling
on serum reducing sugar l e v e l s .
44
Absorption spectrum: Nelson-Somogyi method o f
determining blood sugar.
68
Standard curve: Nelson-Somogyi method o f determining
blood sugar.
71
vii
ACKNOWLEDGEMENTS
I wish t o thank Dr. I . McT. Cowan, Dean o f Graduate Studies, (formerly
Head o f the Department o f Zoology) f o r h i s permission t o undertake t h i s
p r o j e c t , f o r supporting i t , and f o r t h e use o f t h e f a c i l i t i e s o f the
Department o f Zoology.
My thanks a l s o t o Dr. A.J. Wood, Dean o f A r t s and Science at t h e
U n i v e r s i t y o f V i c t o r i a , (formerly Professor o f Animal Science at t h e
U n i v e r s i t y o f B r i t i s h Columbia) f o r the motivation he a p p l i e d during the
development o f t h i s work.
To Dr. H.C. Nordan, A s s i s t a n t Professor i n t h e Department o f Zoology,
I wish t o express s i n c e r e thanks f o r t h e i n t e r e s t , encouragement, guidance,
and philosophy that he provided throughout our a s s o c i a t i o n .
F i n a l l y I wish t o acknowledge t h e assistance o f Mr. R. Addison and
Mr. D. Leckenby, who made the load t h a t much e a s i e r t o c a r r y .
INTRODUCTION
1
INTRODUCTION
Subjective descriptions of the relationship of deer and habitat have
dominated game management literature for many years.
This material has
extended our knowledge of these animals and provided the understanding
essential for the development of needed management techniques.
However,
i n keeping with the contemporary movement of research to intensive investigation, deer, more than ever before, are being studied as unique physiological
systems manifesting profound cyclic phenomena.
Documentations of changes i n
antler growth, feed intake, body weight, hormonal levels, and other
physiological parameters, coincident with the seasonal sexual cycle are
becoming priminant i n the literature.
French et a l (1955), Cowan et a l
(1955), Kitts et a l (1956), Magruder et a l (1957) have a l l studied various
aspects of the growth and nutrition of deer.
metabolic rates.
Silver et a l (1959) reported
Rosen and Bischoff (1952), Kitts et a l (1956), Bandy
et a l (1956), Terri et a l (1958), and Touatt et a l (19#5) have carried out
studies on the haematology of deer under various circumstances. Investigations into bone and antler development have been reported by Bernhard et a l
(1953), Meister (1956) and Long (1959).
Robinson et a l (1965) studied the
reproductive cycle of male white-tailed deer.
Investigations into the
hormonal system of deer have been reported by Browman and Sears (1956),
Grieser and Browman (1956), Tyler (1961), and Dawson (1963).
French et a l
(i960) noted the response of white-tailed bucks to added a r t i f i c i a l light.
As one reviews literature of this nature i t i s apparent that i n some
cases the investigator has failed to appreciate the f u l l effect of his
experimental manipulation on the animal.
Techniques of handling,
2
r e s t r a i n i n g , and sampling, used w i t h success on domestic animals are o f t e n
misapplied t o w i l d species.
As a consequence r e s u l t s o f t e n r e f l e c t the
s t a t e o f a g i t a t i o n o f the animal r a t h e r than i t s normal, undisturbed,
physiological functioning.
I n i t i a l l y t h i s study was intended t o explore a l t e r a t i o n s i n the l e v e l
o f blood reducing sugar t h a t might occur i n deer during the profound
p h y s i o l o g i c a l changes t h a t are associated w i t h r u t .
The f i n d i n g s provide
information regarding the f a c t o r s responsible f o r v a r i a b i l i t y i n the blood
reducing sugar l e v e l s i n deer, and a l s o some i n d i c a t i o n o f the normal range
o f these l e v e l s .
They a l s o serve as a caution t o those engaged i n
measuring p h y s i o l o g i c a l parameters i n w i l d animals.
Goodwin (3956) pointed out t h a t the blood sugar o f adult ruminants i s
almost e x c l u s i v e l y glucose.
He suggested, however, t h a t the expression
*blood reducing sugar* i s more appropriate as i t acknowledges the other
reducing sugars (mostly f r u c t o s e ) thought t o be present i n small amounts.
This terminology was adopted and appears throughout the t h e s i s .
Expression o f reducing sugar l e v e l s as mgm.# i n serum r a t h e r than as
mgm.$ i n whole blood was done f o r two important reasons: 1. i t i s the
concentration o f sugar i n the serum and t i s s u e f l u i d t h a t the body c e l l s
respond t o , and 2. changes i n the d i s t r i b u t i o n o f t i s s u e f l u i d can occur
r a p i d l y , a l t e r i n g the packed c e l l volume, and subsequently the reducing
sugar l e v e l i f expressed i n terms o f whole blood.
EXPERIMENTAL
3
EXPERIMENTAL
1.
ANIMALS
Four deer, two male (U16 and U43) and two female (U24 and U34) of the
species Odecoileus hemionus columbianus (Vancouver Island genotype) were
used for these experiments.
The animals were captured as fawns i n the
spring of 1963 i n the vicinity of Courtenay on Vancouver Island, B.C.,
Canada.
Shortly after capture they were shipped, by air, to the Zoology
Vivarium on the campus of the University of British Columbia.
During the
period of experimentation from December 1964 to March 1965 the deer were
i n the late stages of rut.
Normally this i s a period when the animals,
particularly males, are d i f f i c u l t to handle.
However, the close care and
attention given them by the experimenter rendered them unusually tractable
throughout the experiments.
2.
NUTRITION
The experimental animals were fed a commercial milk replacer (Appendix
4) u n t i l they had reached a weight of approximately 15 pounds.
The weaning
process was then initiated and the animals were maintained on the weaning
ration (Appendix 5) u n t i l early September.
They were then placed on the
adult ration (Appendix 6) which was fed throughout the remainder of the
experimental period.
tions directed.
Some modifications were introduced as the investiga-
Animals U16 and U24 were given chopped a l f a l f a hay
exclusively from January 21 1965 to February 12 1965 and from January 21
1965 to March 23 1965, respectively.
4
3.
HOUSING
The animals used i n t h i s study were housed i n i n d i v i d u a l pens.
fawns they were held i n 4
f
by 2
heated room i n t h e Vivarium.
f
As
wooden pens l o c a t e d i n a ground l e v e l steam
At weaning the deer were moved t o l a r g e r
pens, 3* by 6* i n t h e same room.
By October 1963 t h e s i z e o f the deer and
the onset o f r u t n e c e s s i t a t e d a move t o the W i l d l i f e Unit (Wood et a l 1 9 6 l )
where they were housed i n a d u l t penes
These pens were l a t e r modified t o
include a slatted f l o o r .
4.
ANALYTICAL TECHNIQUES
( i ) Blood C o l l e c t i o n
Blood samples were taken from the j u g u l a r v e i n .
O c c a s i o n a l l y , when
t h i s proved d i f f i c u l t , samples were drawn from the r e c u r r e n t t a r s a l v e i n .
Blood was l e t i n t o untreated, evacuated, 10 cc. v a c u t a i n e r tubes*.
A
record was made o f t h e time r e q u i r e d f o r each aspect o f every blood
drawing o p e r a t i o n .
f
0 * time was considered t o be t h e i n i t i a l i n t r u s i o n
o f t h e t e c h n i c i a n on t h e animal i . e . as t h e pen door was opened.
Injection» time was then noted as t h e i m m o b i l i z i n g agent was administered.
t
The a c t i v i t y i n t h i s i n t e r v a l i n v o l v e d ushering t h e animal down t h e h a l l t o
the l a b o r a t o r y and t h e mln1ma,1 r e s t r a i n t necessary f o r i n j e c t i o n .
The
succeeding n o t a t i o n came as t h e animal c o l l a p s e d due t o the a c t i o n o f t h e
drug.
During t h e i n t e r v a l between ' i n j e c t i o n * and down time t h e deer
u s u a l l y stood q u i e t l y , apparently undisturbed.
,
T
Most a c t i v i t y i n t h i s
p e r i o d occurred as t h e animal fought t h e e f f e c t s o f t h e drug.
The time
t h a t blood was f i r s t released i n t o t h e vacutainer and t h e time t h e
* B.D. Vacutainer, Beeton, Dickinson and Co., Rutherford, N.J., U.S.A.
5
vacutainer was f i l l e d were a l s o noted.
See Appendix 10 f o r an example o f
the form used.
( i i ) Hematocrits
When circumstances allowed, t r i p l i c a t e blood samples were taken i n
heparinized c a p i l l a r y tubes f o r the purpose o f determining the packed
volume.
cell
These samples were a l l taken w i t h a s i n g l e i n s e r t i o n of the
vacutainer needle when a sample f o r glucose a n a l y s i s was drawn.
( i i i ) Serum E x t r a c t i o n and Storage
A f t e r c o l l e c t i o n , the blood samples were allowed t o stand undisturbed
f o r t h i r t y minutes.
The tubes were then Ringed* w i t h a wood dowel, f r e e i n g
the c l o t from the w a l l s of the v e s s e l .
They were then placed i n a c e n t r i -
fuge* and spun at h i g h speed f o r t e n minutes.
Following t h i s the serum was
c a r e f u l l y drawn o f f the c l o t w i t h a t h i n bore p i p e t t e and placed i n t o c l e a n ,
dry,
t e s t tubes.
The serum samples were immediately put i n a f r e e z e r and
stored u n t i l analyzed.
( i v ) Blood Glucose Determination
Of the methods f o r determining blood reducing sugar i n v e s t i g a t e d , a
Nelson-Somogyi technique modified t o s u i t our requirements was
chosen.
This method was found t o provide accurate reproducible r e s u l t s and i s ,
t h e r e f o r e , described i n d e t a i l .
0.2 cc. o f serum was p i p e t t e d i n t o 3.0 cc. o f d i s t i l l e d water.
D e p r o t e i n i z a t i o n w i t h 2.0 cc. o f 0.06 N ZnSO^ and 2.0 cc. o f 0.06_N Ba(0H)
followed.
I t should be noted t h a t the amount and concentration of these
s o l u t i o n s i s not c r i t i c a l but can be v a r i e d t o accomodate p a r t i c u l a r
s i t u a t i o n s (Somogyi 1945)•
As the amount o f glucose i n t h i s procedure
must not be too high, 2.0 cc. of the serum f i l t r a t e was added t o 2.0 cc.
* I n t e r n a t i o n a l Model HN, Needham Hts., Mass., U.S.A.
2
6
o f the combined a l k a l i n e t a r t r a t e and copper s o l u t i o n (Somogyi 1945).
The
t a r t r a t e and copper s o l u t i o n s were prepared separately and mixed as required
as the mixed reagents show some degree o f autoreduction.
(Somogyi 1952).
A f t e r heating i n a b o i l i n g water bath f o r e x a c t l y t e n minutes followed by
immediate c o o l i n g t o 20 C i n a stream o f cold water, the r e a c t i o n mixture
was combined w i t h 2.0 cc. o f Nelsons arsenomolybdate reagent (Nelson 1944).
The volume was made up t o 10 cc. ( a d d i t i o n o f 4.0 cc. o f d i s t i l l e d water) and
the c o l o r was allowed t o develop f o r at l e a s t 15 minutes.
The transmittance
was then measured w i t h a Beckman Model D.U. Spectrophotometer at 650 mu.
An absorption curve from 400 mu t o 1,000 mu o f the coloured end product
of the procedure was determined and appears i n Appendix 7«
Three standard glucose s o l u t i o n s at concentration o f 40, 70, and 100
mgm.# were prepared (Natelson 1961).
every s e r i e s o f unknowns.
One o f the standards was run w i t h
U l t i m a t e l y a number o f r e p e t i t i v e determinations
o f each standard s o l u t i o n was made and used t o prepare a standard curve.
(Appendix 8 ) .
(v) I n t e r n a l Standard
A standard glucose s o l u t i o n (70 mgm.$) was mixed w i t h a serum sample
at three d i f f e r e n t l e v e l s : 2:1, 1:1, and 1:2.
stored f o r a few days p r i o r t o a n a l y s i s .
The mixtures were frozen and
The r e s u l t s appear i n Appendix 9.
EXPERIMENT 1
7
EXPERIMENT 1
This experiment was designed t o i n v e s t i g a t e the e f f e c t s of feed i n t a k e ,
means of r e s t r a i n t , sex of the animal, and time o f day, on blood reducing
sugar l e v e l s i n deer.
Four animals were placed on a feeding regime such that each was
o f f e r e d 1/4 of 80% of i t s d a i l y i n t a k e ( c a l c u l a t e d from the average d a i l y
i n t a k e of the previous week), at each of O70O, 1100, 1500, and 1900
hours.
The d a i l y allowance was cut t o 80% of normal so that the deer would clean
up each feeding almost immediately a f t e r i t was o f f e r e d .
The animals were
allowed a week t o become accustomed t o t h i s program before the experiment
was begun.
By t h i s means some c o n t r o l was exercised over the amount and
time o f feed i n t a k e p r i o r t o blood l e t t i n g .
t a i n e d on t h i s regime.
Deer o f f f e e d
f
1
1900 hours the day before blood l e t t i n g .
Animals *on f e e d
1
were main-
received t h e i r usual allotment at
They were given no feed throughout
the day t h a t blood was being drawn, the next feeding o c c u r r i n g at 0700 hours
the succe<#ding day.
I n e f f e c t then, blood samples were drawn from these
animals a f t e r f a s t s of 12, 16, 20, and 24 hours.
The deer were r e s t r a i n e d during the blood l e t t i n g procedure by one of
two means; i m m o b i l i z a t i o n w i t h s u c c i n y l c h o l i n e or p h y s i c a l f o r c e .
When
the l a t t e r method was used, two or three t e c h n i c i a n s would s i e z e the animal
(recorded as time 'grabbed*), w r e s t l e i t down (time 'held*), and so hold i t
u n t i l the operation was completed.
Blood was drawn at 0800, 1200, 1600, and 2000 hours from the animals
as i n d i c a t e d i n t a b l e 1.
To assess the i n f l u e n c e of each v a r i a b l e , i . e . on feed* v s . ' o f f feed*
f
the data was grouped under these headings i r r e s p e c t i v e of the other
TABLE 1;
TREATMENT FOR EXPERIMENT 1
Date
Animals
Restraint
Feeding Regime
16/ 9/64
16.24
physical
on
22/10/64
34.43
physical
on
26/10/64
16.24
succinylcholine
on
27/10/64
34.43
sue c i n y l c h o l i n e
on
2/11/64
16.24
physical
off
3/11/64
34.43
physical
off
4/H/64
16.24
sue c i n y l c h o l i n e
off
25/11/64
34.43
sue c i n y l c h o l i n e
off
9
variables.
In the cases where i t was applicable a Student * t test was
f
applied to the data to determine the significance of any differences
noted.
(l)
Influence of Feed Intake
As can be seen from table 2 and figure 1 there i s no correlation
between feed intake the hour prior to blood letting and serum reducing
sugar level.
That monogastric animals exhibit a postprandial hyper-
glycemia associated with an increase i n glucose absorption through the
gut i s well documented.
Such a rise i s not found i n cattle (Hodgson
et a l 1932), or i n sheep (Allcroft and Strand 1933).
Moreover, i t has
been shown that the oral administration of glucose i n sheep (Schambye 1951)
and cattle (Bell and Jones 1945) does not cause an increase i n the blood
reducing sugar.
two facts.
The explanation for this relative constancy i s based on
Firstly, starch and cellulose, the principal carbohydrates
found i n the diet of ruminants, are rapidly degraded i n the rumen by a
variety of microorganisms to simple sugars; mostly glucose.
These sugars
are fermented to short chain volatile fatty acids which are then absorbed
into the energy metabolism of the animal.
The major source of ruminant
blood reducing sugar was once believed to result from acidic and enzymatic
digestion of the carbohydrate of ruminal bacteria and protozoa.
This
degradation occurs i n the abomasum and small intestine, and the liberated
glucose i s subsequently absorbed.
Heald (1952), however, investigated this
pathway and found, using sheep fed hay, that no more than 20 gms. of glucose
per day could be produced i n this manner, Later work by Annison et a l (1957)
supported this finding.
Although glucose can be absorbed directly from the
rumen as f i r s t shown by Rankin (1940), i t i s unlikely that any great amount
TABLE 2;
RESULTS OF EXPERIMENT 1:
PART 1
THE EFFECTS OF FEED INTAKE ON SERUM REDUCING SUGAR LEVELS IN BEER.
Feeding Program Animal
On
U16
Feed Intake (gms.)
426
426
426
426
U24
U34
U43
182
182
182
182
283
198
142
57
3 4 0
3 4 0
3 4 0
266
66
73
87
76
250
250
250
228
226
328
2 0 4
68
—
Serum Sugar (mgm.jS)
91.2
90.4
96.8
88.7
88.4
90.1
91.2
87.6
83.6
86.5
100.3
90.9
89.2
81.6
85.9
92.1
1 0 2 . 2
91-5
78.6
109.2
82.9
78.6
93.6
81.9
82.9
90.9
78.1
81.1
79.5
77.0
83.1
Average serum reducing sugar l e v e l *on feed': 2725.6 = 87.9 mgm./K.
31
TABLE 2:
(cont.)
Feeding Program
Off
Animal
U16
Hours of Fast
12
U24
U34
U43
U16
16
U24
U34
U43
U16
20
U24
U34
U43
U16
U24
U34
24
U43
Average serum reducing sugar level after:
(1) 12 hours fast
656.6 =82.1 mgm.#
8
(2) 16 hours fast
711.3 =88.9 mgm.#
8
(3) 20 hours fast
665.4 =83.2 mgm.#
8
(4) 24 hours fast
555.1 =92.5 mgm.#
6
Serum Sugar
(mgm.5)
85.2
85-4
92.1
75-4
81.6
79.0
81.6
76.3
83.0
83.1
90.4
80.7
103.0
83.6
102.5
84-9
87-6
78.6
92.1
75.2
82.4
80.9
79-7
88.9
86.2
97.1
98.4
87.3
88.7
97-4
FIGURE 1:
THE RELATIONSHIP BETWEEN SERUM
REDUCING SUGAR LEVEL AND FEED INTAKE DURING
THE HOUR PRIOR TO BLOOD SAMPLING.
160
150
140
130
120
110
E
J
100
<
O
90
o
z
80
Q
Z>
70
5
60
3
</>
o
Of
50
40
30
20
10
0
40
60
100
140
180
220
FEED INTAKE
260
(gm.)
300
340
380
420
460
13
follows t h i s route.
The glycemic l e v e l o f ruminants then i s not n e a r l y so
much the r e f l e c t i o n o f exogenous glucose absorption t h a t i t i s i n monogastric
animals but i s l a r g e l y a product o f gluconeogenesis.
Secondly, the l a r g e
s i z e o f t h e rumen and an associated slow r a t e o f passage r e s u l t s i n an even
flow o f i n g e s t a through the gut.
This flow i s r e l a t i v e l y unaffected by
temporary changes i n t h e f i l l o f t h e rumen.
This e x p l a i n s i n part why
there i s no s i g n i f i c a n t d i f f e r e n c e (0.200 =»P >0.400) between blood reducing
sugar l e v e l s when t h e animals were on feed as compared t o the l e v e l a f t e r
12, 16, 20, and 24 hours o f f a s t ( t a b l e 3).
Figure 2 i l l u s t r a t e s t h e
negligable e f f e c t o f these r e l a t i v e l y short periods o f f a s t on the blood
seducing sugar l e v e l o f deer.
The e f f e c t o f f a s t i n g on blood reducing sugar l e v e l s i n ruminants i s
variable.
A l l c r o f t and Strand (1933) noted t h a t a seven day f a s t had l i t t l e
e f f e c t on sheep.
Magee (1932), on t h e other hand, recorded a r i s e i n blood
reducing sugar i n goats during a seven day f a s t .
found a s u b s t a n t i a l increase i n the l a c t a t i n g cow.
Robertson (i960) a l s o
Hodgson et a l (1932),
however, noted a considerable decrease i n the l e v e l i n d a i r y c a t t l e during
a seven day f a s t .
Reid (1950) recorded t h a t 45 - 46 hour f a s t s have l i t t l e
e f f e c t on blood reducing sugar l e v e l s i n sheep but t h a t longer periods
produced s i g n i f i c a n t decreases.
Had t h e deer used i n t h i s experiment been
f a s t e d longer perhaps a s i m i l a r e f f e c t would have been noted.
TABLE 3: RESULTS OF EXPERIMENT 1: PART 1
STUDENT t TEST COMPARING THE SERUM REDUCING SUGAR LEVELS OF FASTED AND
NONFASTED DEER.
Animal
Treatment
Nonfasted
Fasted (12-24 hours)
X^
x-j-X-x
x?£
Xg
91.2
90.4
96.8
88.7
88.4
90.1
91.2
87-6
3.3
2.5
8.9
.8
.5
2.2
3.3
.4
10.89
6.25
79.21
.64
225
4.84
10.89
.16
85.2
85.4
83.0
83.1
87.6
78.6
86.2
1.0
.8
3.2
3.1
1.4
7.6
U24
83.6
86.5
100.3
90.9
89.2
81.6
85.9
92.1
4.3
1.4
12.4
3.0
1.7
6.3
2.0
4-2
18.49
1.96
153.76
9.00
2889
39.69
4.00
17-64
92.1
75.4
90.4
80.7
92.1
75.2
97-1
5-9
10.8
4-2
5.5
5.9
11.0
10.9
34.81
116.64
17.64
30.25
34.81
121.00
118.81
U34
102.2
91.5
78.6
109.2
82.9
78.6
93-6
14.3
3.6
9.3
21.3
5.0
9.3
5.7
204.49
12.96
86.49
453.69
25Q00
86.19
32.49
81.6
79.0
102.9
83.6
82.4
80.9
98.4
8723
4.6
7.2
16.8
2.6
3.8
5-3
12.2
1.1
21.16
51.84
282.24
6.76
14-44
28.09
148.84
1.21
U43
81.9
82.9
90.9
78.1
81.1
79.5
76.9
83.1
6.0
5.0
3.0
9.8
6.8
8.4
11.0
4.8
36.00
25-00
9.00
96.04
46.24
70.56
121.00
23.04
81.6
76.3
102.6
84-9
79.7
88.9
88.7
97-4
4.6
9.9
16.4
1.3
6.5
2.7
2.5
11.2
21.16
98.01
268.96
1.69
42.25
7-29
6.25
125.44
U16
Xx and X
n = 30,
x
2
n = 31, xx = 86.2,
2
2
x
<fx| = 1,688.75
- x = v 57-H(6l)/930 = 1.94
/
x
1.00
.64
10.24
9.61
1.96
57.76
—
= serum reducing sugar level (mgm./S)
= 1,680.80
Sx
X2-X-X
2
= 87-9,
d.f. = 59,
x
2
- x
x
= 1.1
s = 3,369-55/59 = 57-11
2
t = 1.7
1.94
= 0.876
,400<P< .200
FIGURE 2:
A COMPARISON BETWEEN THE SERUM
REDUCING SUGAR LEVELS OF FASTED AND NONFASTED DEER
SERUM REDUCING SUGAR
to
o
o
z
I
O
c
TO
CO
>
o
o
o
o
(mgrn. X)
oo
o
o
o
o
o
16
(2)
Means o f R e s t r a i n t
This study was designed t o i n v e s t i g a t e the e f f e c t o f the means o f
r e s t r a i n t used t o f a c i l i t a t e blood sampling on t h e l e v e l o f serum reducing
sugar i n deer.
Two methods o f r e s t r a i n t were i n v e s t i g a t e d : p h y s i c a l f o r c e ,
and immobilization w i t h s u c c i n y l c h o l i n e .
Deer p h y s i c a l l y r e s t r a i n e d were thrown and held down by the weight o f
two o r three t e c h n i c i a n s .
The process o f throwing t h e animals g e n e r a l l y
e n t a i l e d some f i g h t i n g , but there was l i t t l e struggle once they were down
and a good hold was secured.
I t u s u a l l y took l e s s time from the f i r s t
i n t r u s i o n t o drawing o f t h e sample using t h i s technique o f r e s t r a i n t compared
to succinylcholine.
A deer administered a s u i t a b l e dose o f s u c c i n y l c h o l i n e (Cowan 1962),
(Nordan, 1962) e x h i b i t s a reasonably c h a r a c t e r i s t i c response t o t h e onset
of p a r a l y s i s .
quiet.
The 2-3 minutes f o l l o w i n g i n j e c t i o n i t remains r e l a t i v e l y
Then, j u s t p r i o r t o c o l l a p s e , a b r i e f period o f s t i f f - l e g g e d
walking, chewing, back-humping and muscle spasms occurs.
An inexperienced
animal w i l l f i g h t t h e p a r a l y s i s and remain standing as long as p o s s i b l e .
I t s uncontrolled c o l l a p s e i s immediately followed by complete r e l a x a t i o n .
An experienced animal, on the other hand, does not f i g h t t o the same extent
and i t s c o l l a p s e i s generally more c o n t r o l l e d . I t i s worthy o f note t h a t
the animals used i n t h i s experiment were experienced.
Control o f t h e
musculature o f t h e jaw, neck, and e x t r e m i t i e s , u s u a l l y returned 15-20
minutes a f t e r c o l l a p s e , and t h e animal would regain i t s feet apparently
unaffected by t h e p a r a l y s i s .
The d i f f e r e n c e s i n behaviour e l l i c i t e d by t h e two methods o f r e s t r a i n t
are c l e a r l y r e f l e c t e d i n the response o f t h e blood sugar.
As can be seen
17
from t a b l e 4 s i g n i f i c a n t l y lower serum reducing sugar l e v e l s (0.001>P^>0.005)
r e s u l t when deer are r e s t r a i n e d using s u c c i n y l c h o l i n e as compared t o when
they are r e s t r a i n e d using p h y s i c a l f o r c e .
f i g u r e 3-
The d i f f e r e n c e i s i l l u s t r a t e d i n
Moreover, i t i s noteworthy that s i g n i f i c a n t l y l e s s v a r i a b l e
l e v e l s are obtained using s u c c i n y l c h o l i n e .
This c a l c u l a t i o n i s presented i n
t a b l e 5The d i f f e r e n c e i n blood reducing sugar l e v e l s obtained by the two
methods o f r e s t r a i n t might be accounted f o r i n two ways.
Firstly, the
degree o f muscular energy expended by the animals p h y s i c a l l y r e s t r a i n e d i s
considerably greater than when s u c c i n y l c h o l i n e i m m o b i l i z a t i o n i s used.
Solandt and Ferguson (1932) i n v e s t i g a t e d t h e e f f e c t s o f strenuous e x e r c i s e
of short d u r a t i o n on blood reducing sugar i n man.
They found t h a t 30-45
seconds o f standing running at t o p speed caused a n o t i c a b l e increase i n
blood reducing sugar f i v e minutes l a t e r .
Reichard et a l (1961) u s i n g -^C
l a b l e d glucose noted that there was an increased uptake o f blood sugar by
working muscle, compensated by increased hepatic glucose output.
The
struggle o f t h e p h y s i c a l l y r e s t r a i n e d deer, then, could account i n part f o r
the high blood sugar l e v e l s recorded.
Secondly, though i t i s impossible
t o assess w i t h c e r t a i n t y t h e degree o f f r i g h t and p a i n experienced by t h e
animals when r e s t r a i n e d by e i t h e r method, i t i s undoubtedly t r u e t h a t
p h y s i c a l r e s t r a i n t i s more traumatic than i m m o b i l i z a t i o n w i t h s u c c i n y l c h o l i n e .
Therefore, t h e d i f f e r e n c e i n blood reducing sugar l e v e l s obtained by t h e two
methods o f r e s t r a i n t , might r e f l e c t t h e degree o f s t i m u l a t i o n o f t h e
sympathicoadrenal complex.
The a c t i v a t i o n o f t h i s system has been shown
t o occur i n almost a l l types o f s t r e s s s i t u a t i o n s .
The f e a r and pain
associated w i t h both means o f r e s t r a i n t , then, t r i g g e r t h e complex r e s u l t i n g
18
TABLE 4:
RESULTS OF EXPERIMENT 1;
PART 2
STUDENT t TEST COMPARING SERUM REDUCING SUGAR LEVELS OF DEER IMMOBILIZED
WITH SUCCINYLCHOLINE AND THOSE RESTRAINED PHYSICALLY.
Animal
Treatment
Succinylcholine
X
U 16
l
Physical
X2=X-x
88.4
90.1
91.2
87.5
85-4
83.1
78.6
4-4
6.1
7.2
3.5
1.4
.9
5.4
19.36
37.21
51.84
12.25
1.96
.81
29.16
91.2
90.4
96.8
88.7
85.2
83.0
87-6
86.2
1.4
.6
7.0
1.1
4-6
6.8
2.2
3-6
1.96
•36
49.00
1.21
21.16
46.24
4-84
12.96
89.2
81.6
85.9
92.1
75-4
80.7
75-2
5.2
2.4
1.9
8.1
8.6
3.3
8.8
27-04
5.76
3.61
65.61
73-96
10.89
77-44
83.6
86.5
100.3
90.9
92.1
90.4
92.1
97.1
6.2
5-3
10.5
1.1
2.3
.6
2.3
7.3
38.44
10.89
110.25
1.21
5-29
.36
5.29
53.29
82.9
78.6
93.6
79-0
83.6
80.9
87-3
1.1
5.4
9.6
5.0
.4
3-1
3-3
1.21
29.16
92.16
25-00
.16
9.61
10.89
102.2
91.5
78.6
109.2
81.6
103.0
82.4
98.4
12.4
1.7
11.2
19-4
8.2
13.2
7.4
8.6
153.76
2.89
125.44
376.89
67.24
174.24
54-76
73.96
81.1
79.5
76.9
83.1
76.3
84-9
88.9
97-4
2.9
4-5
7.1
.9
7-7
.9
4.9
13.4
8.41
20.25
50.41
.81
59.29
-81
24.01
179-56
81.9
90.9
78.1
82.9
81.6
102.6
79.7
88.7
7.9
1.1
11.7
6.9
8.2
12.8
10.1
1.1
62.41
1. 21
136.89
47.61
67.24
163.84
102.01
1.21
and X2 = serum reducing sugar l e v e l (mgm.#).
^=29,
n2=32,
£x2 = 928.64,
s
2
x =84-0,
1
2
= 1,973-82,
= 2,902.46/59 = 49.19,
t = 5.8 = 3-222
1.8
x =89.8,
*2~\ =5-8
d.f. = 59
Sxj-xf, = \/49.19(6l)/928 = 1.80
. 005<P<.001
FIGURE 3:
A COMPARISON BETWEEN THE SERUM
REDUCING SUGAR LEVELS OF DEER RESTRAINED PHYSICALLY
AND THOSE IMMOBILIZED WITH SUCCINYLCHOLINE.
TABLE 5: RESULTS OF EXPERIMENT 1;
PART 2
F TEST* FOR DETERMINING THE REPRODUCIBILITY OF SERUM REDUCING SUGAR LEVELS
BY THE TWO METHODS OF RESTRAINT.
Animal
^
X
Xg
2
X
2
U16
88-4
90.1
91.2
87.5
85.4
83.1
78.6
7,814.56
8,118.01
8,317-44
7,656.25
7,293.16
6,905.61
6,177-96
91.2
90.4
96.8
88.7
85.2
83.0
87.6
86.2
8,317-44
8,172.16
9,370.24
7,867.69
7,259.04
6,889.00
7,673-76
7,430.44
U24
89.2
81.6
85-9
92.1
75-4
80.7
75-2
7,956.64
6,658.56
7,378.81
8,482.41
5,685.16
6,512.49
5,655-04
83.6
86.5
100.3
90.9
92.1
90.4
92.1
97.1
6,988.96
7,482.25
10,060.09
8,262.81
8,482.41
8,172.16
8,482.41
9,428.41
U34
82.9
78.6
93-6
79.0
83.6
80.9
87.3
6,872.41
6,177-96
8,760.96
6,241.00
6,988.96
6,544.81
7,621.29
102.2
91-5
78.6
109.2
81.6
103.0
82.4
98.4
10,444-84
8,372.25
6,177-96
11,924.64
6,658.56
10,609.00
6,789-76
9,682.56
U43
81.1
79.5
76.9
83.1
76.3
84.9
88.9
97.4
6,577-21
6,320.25
5,916.61
6,905-61
5,821.69
7,208.01
7,903.21
9,486.76
81.9
90.9
78.1
82.9
81.6
102.6
79-7
88.7
6,707.61
8,262.81
6,099.61
6,872.41
6,658.56
10,526.76
6,352.09
7,867.69
X-^ and X2 = serum reducing sugar l e v e l (mgm.#)
(x\ = 205,955.84,
f x | = 260,346.38
( ( \ ) = 5,945,794-56,
2
6? -
( ft)
n
( ^ X g ) = 8, 267,925.16,
2
d.f. =28
x
d.f. =31
2
2
n
F=$L
02
F = 63.66 = 1.919
33-16
* Natelson, S. Microtechniques o f C l i n i c a l Chemistry (1961) pp.493•
S p r i n g f i e l d I l l i n o i s , U.S.A.
C.C. Thomas
21
i n manifold physiological responses.
For example, heart rate, systolic
pressure, cardiac output, blood flow through the l i v e r , brain, kidney, and
musculature are undoubtedly increased.
On the other hand, the activity
of the intestine and genital system are slowed.
Of principal importance
the adrenal medulla i s stimulated to cause an increased release of
epinepherine.
Epinepherine augments the effects of stimulation of the
sympathetic system and e l l i c i t s an important hyperglycemic response.
It
also stimulates the adenohypothesis to release ACTH which increases the
release of adrenal cortical hormones, potent i n promoting gluconeogenesis
from protein (Turner I960).
Reid (1962) attempted to correlate blood
sugar and plasma C o r t i s o l levels i n sheep under stress but was unsuccessful.
He assumed that the hyperglycemia noted was an effect mediated by epinepherine
rather than by C o r t i s o l .
E l l i s (1956) stated that the metabolic effects of
epinepherine on carbohydrate metabolism are far too complex to allow any
precise conclusions as to the metabolic significance of the variations i n
blood sugar recorded.
The data presented i n this experiment indicate that the means used to
restrain deer during the blood letting procedure has a pronounced effect on
the level of serum reducing sugar.
Immobilization with succinylcholine
results i n lower and less variable serum reducing sugar levels.
22
(3)
The I n f l u e n c e o f Sex
The d a t a o f t h i s experiment ( t a b l e 6 ) , ( f i g u r e 4) i n d i c a t e t h a t t h e r e
i s no s i g n i f i c a n t d i f f e r e n c e ( . 2 0 0 > P > . 4 0 0 ) between t h e serum r e d u c i n g
s u g a r l e v e l s o f bucks as compared t o t h o s e o f d o e s .
This result
is
s u p p o r t e d by t h e f a c t t h a t no d i f f e r e n c e i n b l o o d r e d u c i n g s u g a r l e v e l s
between t h e s e x e s has been r e p o r t e d i n c a t t l e (Hodgson e t a l , 1 9 3 2 ) ,
calves (Voelker,
1 9 5 5 ) , o r between w e t h e r s and ewes ( R e i d , 1 9 5 0 ) .
i n d i c a t e d i n t a b l e 1 t h e m a j o r i t y o f t h e b l o o d samples f o r t h i s
were drawn i n l a t e O c t o b e r and e a r l y November.
in
As
experiment
A l t h o u g h t h e bucks were
e n t e r i n g t h e r u t t i n g p e r i o d a t t h i s t i m e t h e r e was l i t t l e e v i d e n c e o f
a g g r e s s i v e b e h a v i o u r , and no e f f e c t on serum r e d u c i n g s u g a r l e v e l s o c c u r r e d .
23
TABLE 6: RESULTS OF EXPERIMENT 1:
PART 3
STUDENT t TEST COMPARING THE SERUM REDUCING SUGAR LEVELS OF MALE DEER TO
THOSE OF FEMALE DEER.
Male
x X-x
91.2
90.4
96.8
88.7
88.4
90.1
91.2
87.6
85.2
83.0
87.6
86.2
85-4
83.1
78.6
81.9
82.9
90.9
78.1
81.1
79-5
77.0
83-1
102.6
79.7
88.6
76.3
84-9
88.9
97.4
5.1
4-3
10.7
2.6
2.3
4.0
5.1
1.5
.9
3.1
1.5
.1
•7
3.0
7.5
4.2
3.2
4.8
8.0
5.0
6.6
9.1
3.0
16.5
6.4
2.5
9.8
1.2
2.8
11.3
Female
£
1 =
Xg=X-x
26.01
18.49
114.49
6.76
5-39
16.00
26.01
2.25
.81
9.61
2.25
.01
.49
9.00
56.25
17-64
10.24
23.04
64-00
25.00
43-56
82.81
9.00
275.25
40.96
6.25
96.04
1.44
7.84
127-69
83.6
86.5
100.1
90.9
89-2
81.6
85.9
92.1
92.1
90.4
92.1
97.1
75.4
80.7
75.2
102.2
91.5
78.6
109.2
82.9
78.6
93.6
81.6
82.4
92.4
79.0
83.6
80.9
87.3
4.4
1.5
12.1
2.9
1.2
6.4
2.1
4.1
4.1
2.4
4.1
9.1
12.6
7.3
12.8
14.2
3.5
9.4
21.2
5.1
9.4
5.6
6.4
5.6
4.4
9.0
4-4
7.1
.7
19.36
2.25
146.41
8.41
1.44
40.96
4-41
16.81
16.81
5.76
16.81
82.81
158.76
53-29
163.84
201.64
12.25
88.36
449.44
26.01
88.36
31.36
40.96
31.36
19.36
81.00
19.36
50.41
.49
X^ and Xg = serum reducing sugar level (mgm.$).
*1 = 31,
nz - 30,
£xf = 1,141.73,
s
2
SQ_
= 2,103.49,
= 3,245.22/59 = 55-0,
t = 1^2 = 1.0
1.9
— 86.1, %2 —
88.0,
S x ^
.400<P<.200
x;,-*! = 1.9
d.f. = (31 + 30) - 2 = 59
V 55.0(6l)/928 = 1.9
/
FIGURE 4:
A COMPARISON BETWEEN THE SERUM REDUCING
SUGAR LEVELS OF MALE AND FEMALE BLACK-TAILED DEER.
SERUM REDUCING SUGAR
to
o
5
>
cn
m
X
>
s
(mgm.%)
00
o
8
O
O
25
(4)
The Influence of Time of Day
This experiment was designed to determine i f any change i n serum
reducing sugar level occurs during the day i n deer.
The animals were on
feed as outlined earlier, and blood samples were l e t at each of 0800,
1200, 1600, and 2000 hours.
As shown i n table 7 and figure 5 the serum reducing sugar levels i n
samples drawn at 0800, 1200, and 1600 hours are essentially the same.
The calculations presented i n table 8 show that the slightly higher levels
found i n samples drawn at 2000 hours are not significantly different from
the mean of the levels found at 0800, 1200, and 1600 hours.
This elevation
at 2000 hours, which was found i n the fasted (figure 2) and nonfasted deer
(figure 5) might be due to a mild stimulation of the sympathicoadrenal
complex.
Although the deer used i n this experiment were accustomed to
considerable activity i n the wildlife unit during the day, the disturbance
at night (light, talk, movement) associated with blood sampling was
unusual and could conceivably e l l i c i t e d such a response.
Reid (1950)
also noted i n sheep that the blood sugar level was higher i n the late
afternoon than i n the morning but could offer no explanationfcr this.
The
findings of this experiment are supported by the investigations of
Schuhecker (1925) and by Hitchcock and Phillipson (1946) who found no
diurnal variation i n the blood sugar of adult ruminants.
Teichman (1952)
as reported by Reid (1950), and Kennedy et a l (1939) reported a marked
diurnal variation i n the blood sugar levels of calves i n response to
feeding.
Preston and Ndumbe (l96l) studied the diurnal variations i n
the blood sugar of ruminating calves.
They noted a postprandial hyper-
glycemia similar to that found i n monogastric animals i n calves on a milk
TABLE 7:
RESULTS OF EXPERIMENT 1;
PART 4
THE INFLUENCE OF TIME OF DAY ON SERUM REDUCING SUGAR LEVELS IN DEER ON FEED.
Serum Reducing Sugar (mgm.#)
Time
0800
1200
1600
2000
91.2
88.4
83.6
89.2
102.2
82.9
81.9
81.1
90.4
90.1
86.5
81.6
91.5
82.9
79.5
96.8
91.2
100.1
85'.9
78.6
78.6
90.9
77.0
88.7
87.6
90.9
92.1
109.2
93.6
78.1
83.1
Average serum reducing sugar l e v e l a t 0800
1200
1600
2000
=87.6 mgm.$
= 86.1 mgm.#
= 87-4 mgm.$
= 90.4 mgm.$
FIGURE 5:
COMPARING SERUM REDUCING SUGAR LEVELS FOUND
IN DEER ON FEED AT DIFFERENT TIMES DURING THE DAT.
SERUM REDUCING SUGAR
o
o
o
o
(mgm.X)
oo
o
o
o
o
00
o
o
o
[
~1
O
>
-<
o
o
o
:
o
o
'-."fii rii^'Vl -
ri
••• -'-- - "-'*^»^^^' '^.^i
l
J
TABLE 8:
RESULTS Of EXPERIMENT It PART 4
STUDENT t TEST COMPARING SERUM REDUCING SUGAR LEVELS OF DEER SAMPLED AT
0800. 1200. and 1600 HOURS TO THOSE SAMPLED AT 2000 HOURS.
0800, 1200, 1600 Hours
x-^X-x
91.2
88.4
83.6
89.2
102.2
82.9
81.9
81.1
90.4
90.1
8655
91.5
82.9
79.5
96.8
91.2
100.1
85.9
78.6
78.6
90.9
77.0
81.6
2000 Hours
d
4.2
17-64
1.96
1.4
11.56
3-4
2.2
4.84
5.2
27.04
16.81
4.1
26.01
5-1
34.81
5.9
11.56
3-4
9.61
3.1
.5
.25
4-5
20.25
16.81
4.1
7-5
56.25
9.8
96.04
4-2
17.64
13.1 171.61
1.1
1.21
70.56
8.4
70.56
8.4
15.21
3-9
10.0 100.00
29.16
5-4
h
X2 X-x
88.7
87.6
90.9
92.1
109.2
93.6
78.1
83.1
1.7
2.8
.5
1.7
18.8
3.2
12.3
7.3
=
2.89
7.84
.25
2.89
353.44
10.24
151.29
53.29
X j and Xg = serum reducing sugar (mgm.$)
n
x
= 23, x
x
= 87.0,
^ x j = 827-39,
n| =
8, X2 = 90.4
s
1,409.52/29 = 48£6
2
=
Sxj-xf =
1
=
= 582.13,
/48.6(31)/184 =
lik = 1.188
2.86
^2~\
\ C l I = 2.86
.400<P<.200
=
3-4,
d.f. = 29
29
diet.
Calves on d r i e d grass, however, showed no response to feeding.
Another group on a concentrate r a t i o n exhibited a postprandial hypoglycemia,
presumably due t o a c t i v e fermentation i n the rumen immediately f o l l o w i n g
feeding.
The r e s u l t s of these studies on calves do not detract from the
f i n d i n g s o f t h i s experiment on deer.
I t i s u n l i k e l y that the energy
metabolism o f c a l v e s , even ruminating calves, as r e f l e c t e d by blood
reducing sugar l e v e l s can be compared t o any advantage t o that o f adult
deer.
I t i s p e r t i n e n t t o note t h a t blood sugar l e v e l s of animals sampled on
the same day at the same time o c c a s i o n a l l y v a r i e d i n the same d i r e c t i o n .
Figure 6 shows the blood reducing sugar l e v e l s o f U34 and U43 during the
f a s t i n g experiment.
Both deer were sampled at 0800, 1200, 1600, and 2000
hours, i n every case U34 was sampled f i r s t .
I n t h i s p a r t i c u l a r example
the reducing sugar l e v e l s o f both deer are almost i d e n t i c a l and t h e i r
coincident v a r i a t i o n i s s t r i k i n g .
ing
I n other instances, although the reduc-
sugar l e v e l s o f the deer are d i f f e r e n t the same coincidence o f v a r i a t i o n
occurs.
This phenomena i s l i k e l y due t o excitement o f the animals
preceeding blood sampling.
I f , f o r example, the deer were undisturbed the
hour or so p r i o r t o blood sampling the blood sugars o f both would be
r e l a t i v e l y low.
Had some unusual disturbance occurred, on the other hand,
then the blood sugars o f the deer would be correspondingly higher.
Under
the conditions of the experiment i t was impossible t o c o n t r o l t h i s 'extraneous
disturbance* f a c t o r .
I n t h i s regard i t i s noteworthy t h a t the second animal
sampled each day d i d not show c o n s i s t e n t l y higher serum reducing sugar l e v e l s
as compared t o the f i r s t deer sampled.
ways.
This can be accounted f o r i n two
F i r s t l y , and most important, the animals were accustomed t o some
30
disturbance during the day and blood sampling was carried out quietly in a
laboratory somewhat removed from the other animals i n the unit.
Secondly,
i t i s impossible to assess the starting or normal blood reducing sugar
level, therefore, i f the second animal had a lower normal level than the
first any increase due to excitement could be masked.
FIGURE 6:
ILLUSTRATING THE RELATIONSHIP THAT CAN
OCCASIONALLY OCCUR BETWEEN THE SERUM REDUCING SUGAR
LEVELS OF TWO ANIMALS SAMPLED AT THE SAME TIMES ON
THE SAME DAY.
5
U34
1
U43
"I
.5
i
• 'I
4?
I
••a
1
i-
I
I'
I
1
1
0800
1200
1600
TIME
(HOURS)
2000
EXPERIMENT 2
32
EXPERIMENT 2
This experiment was designed to determine i f a change i n diet from a
concentrate ration to chopped a l f a l f a hay would alter the blood reducing
sugar levels found i n deer.
As i n the f i r s t experiment the animals were placed on a feeding
program such that each was offered 1/4 of 80$ of i t s daily intake (calculated
from the daily intake of the previous week) at each of 0700, 1100, 1500, and
2000 hours.
As indicated earlier some control was thus exercised over the
amount and time of feed intake the hour prior to blood letting.
A week
was allowed for the deer to become accustomed to this regime before the
experiment was begun.
hour after feeding.
A l l blood samples were drawn about 0800 hours, one
The remaining feed of the
animal
being sampled was
weighed back and a record was kept of the feed intake the hour prior to
blood letting.
On the basis of the results of experiment 1 a l l animals were immobilized
with succinylcholine before blood samples were drawn.
10 cc. samples were l e t from a l l deer once they were accustomed to the
new feeding regime.
Following this U16 and U24 were taken off feed for
two days and then offered chopped a l f a l f a hay on the same schedule that the
concentrate ration had been given.
U34 and U43 were treated as controls,
remaining on concentrate throughout the experiment.
Blood samples were
drawn every second day, on alternate days, from both animals i n each group.
The deer were weighed regularly throughout the experiment.
Figure 7 illustrates the considerable variation found i n the serum
reducing sugar levels of the deer during the experimental period.
As can
FIGURE 7: ILLUSTRATING THE CHANGES IN SERUM
REDUCING SUGAR LEVELS THAT OCCURRED I N DEER
ON PELLETS AND THOSE ON HAY.
34
be seen from tables 8 and 9 no clearly defined trends are apparent i n the
levels of reducing sugar i n the control animals.
Both deer exhibit levels
ranging quite consistently between 80 mgm.# - 9 0 xogja.%.
U 4 3 showed the
greatest extremes i n levels with a low of 72 mgm.$ and a high of 133 mgjn.%.
The serum reducing sugar levels of the experimental deer (U16 and U24) as
shown i n tables 10 and 11, remained reasonably constant throughout the
experiment and were generally within the same range as those of the controls.
The data for U24 (table 11) might indicate a slight increase i n serum
reducing sugar level during the f i r s t week on hay.
Only three samples,
however, contribute to the higher level and considering the general
variability encountered i t i s unlikely that this i s significant.
The
relatively low levels obtained toward the end of the experiment (see figure
7) are no lower than those exhibited by this animal on the pelleted ration
i n earlier experiments (see table 3).
There was, therefore, no significant
change i n the levels of serum reducing sugar i n this deer after a period of
six weeks on hay.
The range i n the level of reducing sugar (85 mgm.# -
95 mgm.$) of Ul6 i s generally higher than that of the other animals.
As
indicated i n table 10, however, this deer showed no remarkable change i n
level of serum reducing sugar despite his serious loss of body weight on
hay.
The feed intake of the control animals (tables 8 and 9) was
relatively constant and apparently sufficient as they maintained their
weight with only minor fluctuations throughout the experiment.
The hay
intake of U16 and U24 was also reasonably constant (tables 10 and 11), both
however, suffered considerable weight loss.
In the case of U16 this was
serious enough to force his return to the pelleted ration after only three
weeks on hay.
During the last few days on this diet Ul6 was so weak and
35
TABLE 8:
Date
FEED INTAKE. SERUM REDUCING SUGAR. IND WEIGHT DATA OF U34
THROUGHOUT EXPERIMENT 2.
Feed Intake (gms.pellets)
19/1/65
20
21
22
23
24
25
26
27
28
29
30
31
1/2/65
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
1/3/65
834
902
963
753
753
743
755
858
865
825
946
427
747
699
811
815
806
881
806
515
629
847
761
805
906
714
578
772
946
860
914
868
669
829
728
859
1,004
1,004
890
562
516
875
Reducing Sugar (mgm./&)
Weight (lb.)
—
—
82.4
—
82.9
—
950
94-5
—
84-4
94
—
95
—
84-1
94-5
—
-
—
—
77-4
—
—
80.9
—
93
93
—
—
92.5
—
—
—
—
—
—
91.8
—
—
94.2
—
115-7
—
93
—
93
—
91
—
—
92
—
—
90.4
93
___
—
—
—
—
92
—
—
94.9
—
93
—
93.5
—
—
—
—
93
—
-
—
—
36
TABLE 9; FEED INTAKE. BLOOD REDUCING SUGAR. AND WEIGHT DATA OF U43
THROUGHOUT EXPERIMENT 2.
Date
19/1/65
20
21
22
23
24
25
26
27
28
29
30
31
1/2/65
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
1/3/65
Feed Intake (gms.pellets)
1,100
1,100
1,100
825
825
1,011
1,100
1,100
1,100
1,056
1,100
550
880
1,100
1,100
1,100
1,100
1,100
1,100
905
1,100
943
1,100
1,100
1,100
825
970
1,032
1,100
1,100
1,100
1,100
1,100
1,100
1,082
1,100
1,100
1,100
1,100
852
536
1,235
Reducing Sugar (mgm.%)
Weight ( l b . )
—
—
81.1
—
—
-
83.4
—
—
—
88.9
85.2
126.5
—
—
128
—
—
129
128
128
128
71.9
133-3
126
—
124.5
—
—
—
—
—
83.1
108.8
85.9
_ _ _
126
—
—
126.5
_ _ _
124
—
125
—
—
88.9
126.5
—
125.5
—
127
—
—
85.9
127.5
—
_
—
-
—
—
-
—
—
—
126.5
37
TABLE 10; FEED INTAKE. BLOOD REDUCING SUGAR. AM) WEIGHT DATA OF U16
THROUGHOUT EXPERIMENT 2.
Date
19/1/65
20
21
22
23
24
25
26
27
28
29
30
31
1/2/65
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
1/3/65
*
Feed Intake (gms. hay)
Reducing Sugar (mgm.%)
Weight (lb.)
92.7
j „
285
5336
349
350
472
482
631
580
810
403
756
648
500
333
246
231
420
274
602
702
622
590
162/549
819
1070
721
1243
1168
1268
1646
1865
1544
1805
1968
2268
2198
1837
2276
1969
2276
returned to pellets
79.2
109
79.7
108.5
—
100.8
—
100.8
92.7
94.6
. _
85.7
—
—
.I.,,, • -j-
—
105
105
103-5
102.5
99-5
95.8
96.4
97
107.6
97
115-7
92.1
—
—
96.4
97-5
96
91
94
88.9
96
96.5
88.9
_
.
—
82.6
100.5
99.5
38
TABLE 11:
P-3fee
19/1/65
20
21
22
23
24
25
26
27
28
29
30
31
1/2/65
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
1/3/65
FEED INTAKE. BLOOD REDUCING SUGAR. AND WEIGHT DATA OF U2A
THROUGHOUT EXPERIMENT""^
Feed Intake (gms.hay)
Reducing Sugar (mgnff^
75.2
_ _ _
234
235
389
464
414
372
737
632
699
344
667
499
416
570
452
630
669
478
676
564
559
610
774
602
773
947
507
901
831
91.2
107
80.9
105-5
—
89.2
92.7
106.8
92.7
—
88.9
106.5
104.5
103
101.5
—
87.0
97.8
101
83.1
100.5
_ _ _
—
,
N
,
M | |
80.4
—
_
82.4
_ _ _
79.7
958
77.9
_ _ _
98.5
100.5
98
—
962
891
105.5
_ _ _
939
845
1,059
756
726
756
479
1,112
Weight (lb.)
—
—
—
80.7
—
_
79.5
100
99.5
97-5
97
39
lethargic that he could not walk to the scale but had to be carried.
U24
lost weight consistently during the f i r s t three weeks of the experiment,
thereafter she maintained weight stasis.
There are three possible explanations for the inability of these
animals to maintain their weight during the early part of the experiment.
F i r s t l y , inadequate hay intake, secondly, the poor quality of the hay, and
thirdly, slow development of a rumen microflora capable of handling the hay.
Although no attempt was made to assess i n detail the nutritive quality of
the hay i t was apparent from the considerable amount of coarse material
l e f t by the deer that i t was poor.
This and the physical limitations of
the feeding arrangements undoubtedly resulted i n insufficient intake.
It
i s well estbalished that a considerable change i n rumen microflora i s
associated with any marked alteration i n diet.
The time required to effect
this change i s variable, being dependent on a multitude of factors.
Warner
(1962) noted that i n a sheep switched from a diet of hay alone to one of hay
plus concentrate a l l major changes i n rumen microflora were completed by ten
days.
He points out that this i s usually the period needed for adaptation
to a new diet i n ruminants.
Gouws (1965) studied the alteration i n c e l l -
ulytic bacterial species i n sheep associated with a change from lucerne to
t e f f hay.
He stated that the time lapse between a change of diet and the
attainment of a balance characteristic of the new diet varied from animal to
animal.
In one case he studied, the change was complete i n a week, i n
another there was no change after four weeks.
Although i t i s attractive to
account for the loss i n body weight of the deer during the f i r s t three weeks
as being due to the development of a rumen microflora capable of fermenting
hay, to do so i s complete conjecture.
40
The principal object of this experiment was to determine the effect of
a change i n ration on the serum reducing sugar level i n deer.
It i s
apparent from the data (tables 8 , 9 , 10, and 11) that there was no change
in this parameter associated with the alteration i n diet.
This finding i s
supported by that of Hibbs (1956) who noted that calves fed various ratios
of hay to grain had essentially the same level of blood reducing sugar.
Lambert (1955) reported similar results with calves fed various ratios of
calf starter and a l f a l f a .
There were two factors inherent i n the experiment that might have worked
to alter the serum reducing sugar levels of the deer.
the
diet.
One was the nature of
Annison et a l (1959) found that when sheep on a diet of hay plus
additives were turned out to lush spring grass their blood reducing sugar
levels rose from 45 mgm.# to 60 mgm.$.
He attributed this increase to
either or both the increased availability of proportionate or the abundant
supply of lactate.
Presumably the hay used i n this experiment did not
provide a sufficient alteration i n nutrients to cause a change i n the reducing
sugar level.
The second factor that might have led to a change i n the level
of serum reducing sugar was the stress that could have been imposed on the
deer by the substitute regime.
Reid (1950) and Wright (1962) showed that
underfeeding can produce a f a l l i n blood sugar levels i n pregnant ewes.
They also noted that blood reducing sugar levels f a l l during lactation.in
these animals.
the
Their explanation for the decrease i n levels found was that
demands of, i n the f i r s t case the feotus, and i n the second, milk
production, outstripped the ewes exogenous and endogenous sources of glucose.
During the experiment reported here, a stress, as reflected by the loss i n
body weight suffered by the deer, particularly U16, was undoubtedly imposed.
41
However, sufficient exogenous and endogenous material must have been present
to maintain the serum reducing sugar level as no decrease was found.
EXPERIMENT 3
42
EXPERIMENT 3
This experiment was designed to explore the possibility that the factor
of time during the actual drawing of the blood sample had an influence on
the blood reducing sugar levels determined.
On the basis of the results of experiment 1 no control of feed intake
was exercised.
The deer were immobilized with succinylcholine before blood samples
were drawn.
morning.
Each animal was immobilized on separate days generally i n the
As outlined earlier accurate accounts of the time required for
each aspect of every blood letting were made.
In this experiment
sequential rather than single samples were drawn from each animal.-
Once
the deer was immobilized and the vacutainer needle seated, a series of
samples were released at roughly 30 second intervals into separate, untreated vacutainer tubes.
It i s apparent from the data presented i n table 12 that the time lapse
between i n i t i a l intrusion on the animal and actual drawing of the blood
sample has a profound effect on the level of serum reducing sugar.
As
shown i n figure 8 all the deer exhibit the same general pattern although
the results are displaced one from the other both i n time and i n reducing
sugar level.
The data for U34 represents the trend and presents an almost
complete picture.
The f i r s t sample was drawn from this doe only three
minutes and forty seconds after the i n i t i a l intrusion of the technician.
This i s close to the minimal amount of time necessary to inject the animal,
have i t collapse, seat a vacutainer needle, and draw a blood sample.
The
second sample l e t slightly over a minute later showed essentially the same
43
TABLE 1 2 ;
REDUCING SUGAR LEVELS IN SEQUENTIAL BLOOD SAMPLES.
•Time* i s t h e i n t e r v a l ( i n m i n u t e s and seconds) f r o m i n i t i a l i n t r u s t i o n
o f t h e t e c h n i c i a n on t h e anima] t o t h e end o f t h a t p a r t i c u l a r s a m p l e .
Animal
Time
R e d u c i n g Sugar (mgm.%)
U16
4:02
4:40
8:41
9:34
12:45
13:30
14:21
15:14
97.1
102.6
118.1
117.4
113.0
113.5
115.3
114.8
U24
8:08
10:03
10:30
11:00
11:45
12:13
90.9
99.4
101.9
102.2
102.6
103-3
101.2
102.6
12:51
13:30
U34
3:40
4:52
5:42
6:35
9:01
9:50
10:07
11:10
U43
4:52
5:17
5:50
6:24
6:52
7:30
8:11
9:07
83.6
85.4
90.1
102.2
105.6
110.0
105.2
105.2
84/-4
83.6
86.2
91.5
91.8
98.1
97.1
99-7
45
l e v e l o f reducing sugar.
I t can be concluded that these l e v e l s are
reasonably c l o s e t o the normal f o r t h i s deer.
The f o l l o w i n g three
samples, drawn i n the i n t e r v a l from f i v e minutes t o nine minutes a f t e r
i n i t i a l i n t r u s i o n are t y p i c a l o f the r a p i d increase t h a t occurred i n a l l
the
deer w i t h the exception o f U24.
During t h i s i n t e r v a l U34 e x h i b i t e d a
25.8$ increase over the i n i t i a l serum reducing sugar l e v e l .
The l e v e l s
i n blood samples drawn a f t e r t e n minutes were g e n e r a l l y constant i n a l l the
deer.
The change i n the serum reducing sugar l e v e l s during the blood sampling
procedure found i n t h i s experiment might be due t o three f a c t o r s : f i r s t l y t o
the
excitement and a n t i c i p a t i o n experienced by the animals as they were
moved i n t o the l a b o r a t o r y , secondly t o the fear and p a i n associated w i t h
i n j e c t i o n , i m m o b i l i z a t i o n , and withdrawal o f the blood samples, and t h i r d l y
to the muscular energy expended by the deer throughout the procedure.
Unfortunately the r e l a t i v e c o n t r i b u t i o n s o f these f a c t o r s cannot be determined
from t h i s data.
adrenal complex.
The e f f e c t s o f a l l are mediated through the sympathicoAs pointed out e a r l i e r , the speed and extent o f the
m o b i l i z a t i o n o f glucose, as r e f l e c t e d i n the serum reducing sugar l e v e l ,
depends on the degree o f s t i m u l a t i o n o f t h i s complex.
That excitement might
have an e f f e c t on blood reducing sugar l e v e l s has been suggested by others;
Hodgson et a l (1932), i n c a t t l e Reid (1962) i n sheep, Wing (1955) i n c a l v e s ,
and Bandy (1957) and Touatt et a l (1965) i n deer.
No work has been done,
however, t o determine the r a t e and magnitude o f the change t h a t excitement
during blood sampling can e l l i c i t .
The response o f the deer i n t h i s
experiment was undoubtedly minimal as they had been handled e x t e n s i v e l y and
were r e l a t i v e l y accustomed t o i m m o b i l i z a t i o n .
The p a t t e r n f o r l e s s
46
experienced animals could 3how a curve attaining higher levels in a much
shorter time.
Moreover there might be considerable interspecies variation
in the rate and magnitude of change in the blood sugar.
Domestic species,
for example, might exhibit a slower and smaller increase under a particular
blood sampling.procedure than a wild species.
The findings here add a new
element to the factors that must be controlled i f useful blood sugar values
are to be obtained.
EXPERIMENT L
47
EXPERIMENT 4
This experiment was carried out to determine the distribution of
reducing sugar i n the blood of deer.
On the basis of the results of earlier experiments no control of
feed intake was exercised, a l l the deer were on pellets ad l i b .
The deer were immobilized with succinylcholine before blood was
drawn.
Each animal was immobilized on a different day, generally i n
the morning.
Two blood samples were l e t from each animal into separate
test tubes, the f i r s t being followed immediately by the second.
The
f i r s t tube was untreated, and serum was extracted as outlined earlier.
The second tube was heparinized.
Within minutes of being drawn, 0.2 cc.
of whole blood from this tube was pipetted into 3.0 cc. of d i s t i l l e d
water, thus laking the cells.
The level of blood reducing sugar i n this
sample was then determined i n the usual manner.
The results of the experiment appear i n table 13.
Although only a single determination per deer was done i t appears that
no reducing sugar occurs i n the erythrocytes of these animals.
Goodwin
(1956) using the same method of calculation, found that i n sheep the
corpuscular/plasma glucose concentration was 23-5$.
was roughly the same, being 24.2$.
This ratio i n cattle
In other words, he found that there
was a considerable amount of reducing material i n the erythrocytes of these
animals.
In vitro permeability studies, however, have shown that the
erythrocytes of most nonprimate adults are impermeable to glucose.
To
rationalize this inconsistancy Somogyi (1933) suggested that the corpuscular
glucose of sheep and cattle i s due to the plasma retained between the packed
cells.
Andreen-Svedberg (1933) on the other hand, considered the glucose
to be absorbed on the cells.
48
TABLE 13: THE DISTRIBUTION OF REDUCING SUGAR IN THE BLOOD OF DEER
Animal
Serum L e v e l
(mgm.%)
Hematocrit
U16
96.1
42.5#
55.3
56.5
U24
84.6
46.5#
45.3
49.9
U34
96.4
48.2^
49.9
47.6
U43
120.1
49.2#
61.0
59.6
Whole Blood
(mgm.$)
Calculated
Actual
Sample C a l c u l a t i o n :
serum reducing sugar l e v e l = 96.0 mgm.$
hematocrit o f 45$ (therefore 55$ i s plasma)
96 mgm.ff x 55% = t h e amount o f reducing sugar expected
100
i n whole blood sample assuming t h a t a l l the sugar occurs i n t h e
serum.
RESUME
49
RESUME
T h i s i n v e s t i g a t i o n has shown t h a t f e e d i n t a k e d u r i n g t h e hour p r e c e e d i n g
b l o o d s a m p l i n g , s h o r t p e r i o d s o f f a s t , n a t u r e o f t h e d i e t , and s e x o f t h e
a n i m a l , a p p a r e n t l y have no e f f e c t on b l o o d r e d u c i n g s u g a r l e v e l s i n d e e r .
I t was a l s o found t h a t b l o o d samples t a k e n i n t h e e v e n i n g had a s l i g h t l y
h i g h e r r e d u c i n g s u g a r l e v e l t h a n t h o s e drawn e a r l i e r i n t h e d a y .
r e d u c i n g s u g a r was f o u n d t o o c c u r i n t h e e r y t h r o c y t e s
No
of these deer.
Of t h e f a c t o r s s t u d i e d t h a t c o u l d p o s s i b l y i n f l u e n c e r e d u c i n g s u g a r
l e v e l s , o n l y t w o , t h e means o f r e s t r a i n t ,
samples were found t o have any e f f e c t .
and t h e t i m e r e q u i r e d t o draw
Deer p h y s i c a l l y r e s t r a i n e d e x h i b i t e d
h i g h e r and more v a r i a b l e r e d u c i n g s u g a r l e v e l s t h a n t h o s e i m m o b i l i z e d w i t h
succinylcholine.
The a n i m a l s a l s o showed a p r e c i p i t o u s i n c r e a s e i n
r e d u c i n g sugar l e v e l d u r i n g t h e b l o o d s a m p l i n g o p e r a t i o n .
It
i s interesting
t h a t t h e s e two f a c t o r s , means o f r e s t r a i n t , and t i m e r e q u i r e d t o draw a
sample, are r a r e l y noted by i n v e s t i g a t o r s r e p o r t i n g sugar l e v e l s i n a n i m a l s .
T a b l e 14 p r e s e n t s a c o m p a r i s o n o f t h e b l o o d sugar l e v e l s t h a t have been
r e p o r t e d f o r O d e c o i l e u s hemionus s p e c i e s .
I n e v e r y c a s e t h e d e e r were
r e s t r a i n e d by f o r c e , and b l o o d s u g a r v a l u e s were r e p o r t e d as mgm.% i n whole
blood.
Bandy (1957) found a s i g n i f i c a n t d i f f e r e n c e between t h e b l o o d
s u g a r l e v e l s o f y e a r l i n g d e e r on a h i g h p l a n e o f n u t r i t i o n and t h o s e o n a
low plane.
He a l s o n o t e d t h a t t h e b l o o d s u g a r l e v e l s o f fawns (20 - 100
days o l d ) and t h o s e o f a d u l t d e e r were s i g n i f i c a n t l y d i f f e r e n t .
Bandy
r e s t r a i n e d t h e d e e r w i t h p h y s i c a l f o r c e , but d i d n o t n o t e t h e t i m e r e q u i r e d
t o draw s a m p l e s .
The b l o o d sugar l e v e l s and v a r i a t i o n t h a t he found i n
adult animals are very c l o s e t o those reported i n t h i s
experiment.
50
TABLE 1 4 :
COMPARISON OF BLOOD SUGAR VALUES REPORTED FOR DEER.
Investigator
Bandy, P . J .
al
(1957)
Terri.
et
Animal
O d e c o i l e u s hemionus
columbianus
A. et a l
O d e c o i l e u s hemionus
virginianus
Y o u a t t , W.G. e t
al
(1965)
This report
(19585
This report
*
Restraint
# Det.
B l o o d Sugar
(mgm.%)
37-2 + 5-4
(whole b l o o d )
Physical
Physical
12
O d e c o i l e u s hemionus
virginianus
Physical
20
91.3 + 8 . 1
(whole b l o o d )
O d e c o i l e u s hemionus
columbianus
Physical
32
44-0 + 4.0
(whole""blood)
O d e c o i l e u s hemionus
columbianus
Succinylcholine
29
42.0 + 2.9
(whole b l o o d )
66.9 + 12.9
(whole b l o o d )
C a l c u l a t e d f r o m serum v a l u e s on t h e b a s i s o f a h e m a t o c r i t o f 50%.
51
T e r r i e t a l (1958) r e p o r t e d b l o o d s u g a r v a l u e s f o r d e e r i n
,
nutritionally
poor s t a t u s * .
The a n i m a l s were t h r o w n and b l o o d samples were l e t f r o m t h e
jugular vein.
T e r r i n o t e d t h a t t h e d e e r were e x t r e m e l y n e r v o u s ,
frothing
a t t h e mouth, and t h a t o c c a s i o n a l l y some were f a t a l l y i n j u r e d by t h i s
procedure.
The l e v e l s and v a r i a t i o n t h a t he f o u n d l i k e l y r e f l e c t t h e
and p a i n e x p e r i e n c e d by t h e d e e r d u r i n g b l o o d s a m p l i n g .
fear
The b l o o d s u g a r
l e v e l s r e p o r t e d by Y o u a t t e t a l (1965) a r e even h i g h e r and more v a r i a b l e
t h a n t h o s e r e p o r t e d by T e r r i e t a l ( 1 9 5 8 ) .
Youatt captured h i s animals
w i t h a n e t and s h a c k l e d them t o a r e s t r a i n i n g b o a r d d u r i n g b l o o d s a m p l i n g .
He n o t e d t h a t t h e d e e r were v e r y e x c i t e d b r e a t h e d h e a v i l y , and became
exhausted f i g h t i n g t h i s
procedure.
On t h e b a s i s o f t h e r e s u l t s r e p o r t e d i n t h i s t h e s i s t h e v a l u e o f t h e
b l o o d s u g a r l e v e l s o b t a i n e d by t h e s e methods o f r e s t r a i n t a r e open t o q u e s t i o n .
They most a s s u r e d l y a r e n o t n o r m a l l e v e l s , t h e r e f o r e t h e i r c o m p a r a t i v e v a l u e
i s d o u b t f u l , m o r e o v e r , any changes r e c o r d e d a s b e i n g due t o e x p e r i m e n t a l
manipulation are l i k e l y f o r t u i t o u s .
It
i s apparent t h e n , t h a t t h e blood
r e d u c i n g s u g a r l e v e l s r e p o r t e d f o r d e e r t o d a t e r e f l e c t t h e degree
of
e x c i t e m e n t , f e a r , and p a i n e x p e r i e n c e d by t h e a n i m a l s d u r i n g t h e b l o o d
s a m p l i n g p r o c e d u r e employed.
It
i s p e r t i n e n t once a g a i n t o p o i n t out t h e
r e l a t i v e l y l o w and c o n s i s t e n t b l o o d s u g a r l e v e l s t h a t r e s u l t when s u c c i n y l c h o l i n e i s used t o immobilize t h e deer during blood sampling ( t a b l e 1 3 ) .
The
u s e o f s u c c i n y l c h o l i n e , however, does n o t g u a r a n t e e c o n s i s t e n t , n o r m a l , b l o o d
sugar l e v e l s .
A number o f o t h e r f a c t o r s must be c o n s i d e r e d : t h e d e e r must
be c o m p l e t e l y f a m i l i a r w i t h t h o s e i n v o l v e d i n t h e b l o o d s a m p l i n g o p e r a t i o n ;
r e c o r d s s h o u l d be made o f t h e t i m e r e q u i r e d f o r each a s p e c t o f e v e r y b l o o d
s a m p l i n g , and o f t h e a c t i v i t y o f t h e d e e r t h r o u g h o u t t h e o p e r a t i o n ; and t h e
52
a n i m a l s s h o u l d be housed s u c h t h a t e x t r a n e o u s d i s t u r b a n c e i s n d n i m i z e d .
O n l y t h r o u g h t h e a d o p t i o n o f s u c h a n i n t e n s i v e program w i l l w o r t h w h i l e
d a t a r e l a t i n g b l o o d s u g a r t o e x p e r i m e n t a l m a n i p u l a t i o n be r e a l i z e d .
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53
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and plane o f n u t r i t i o n on t h e blood chemistry o f the Columbian
b l a c k t a i l e d deer.
Can. J . Zool. 34:477, 1956.
32.
Krebs, H.A.
GLuconeogenesis.
1964.
Proc. Royal Soc. S e r i e s B; 159-545,
-
33-
Kronfeld, D.S.
Growth hormone a d m i n i s t r a t i o n t o pregnant sheep.
C o r n e l l Vet. 47:255, 1957.
34-
Lambert, M.R., N.L. Jacobson, R.S. A l l e n , and M.R. B e l l .
The r e l a t i o n
of growth, f e e consumption and c e r t a i n blood constituents t o
changes i n the d i e t a r y o f young d a i r y calves.
J . Dairy S c i .
38:6, 1955.
35.
Lindsay, D.B.
The s i g n i f i c a n c e o f carbohydrate metabolism i n ruminant
metabolism.
Vet. Rev. 5:103, 1959.
36.
Long, T.A., C.E. French, and N.D. Magruder.
E f f e c t o f seasonal
r e s t r i c t i o n s on a n t l e r development o f w h i t e t a i l e d deer. Penn.
Agr. Exp. S t a t . Progr. Rept. #209, 1959.
37.
Magee, H.E.
Observations on d i g e s t i o n i n the ruminant.
J . Exp. B i o l .
9:409, 1932.
38.
Magruder, D., C.E. French, and T.A. Long.
Nutrient requirements o f
w h i t e t a i l e d deer f o r growth and a n t l e r development.
Pa. State
U. Agr. Exp. S t a . B u l l . 628, 1957.
39.
Mayrofer, D.K.
S e l f experiments w i t h s u c c i n y l c h o l i n e .
A new u l t r a s h o r t - a c t i n g muscle r e l a x a n t .
B r i t . Med. J . 1:1332, 1952.
40.
M e i s t e r , W.W. Changes i n h i s t o l o g i c a l s t r u c t u r e o f t h e long bones o f
w h i t e t a i l e d deer during t h e growth o f the a n t l e r s .
Anat. Rec.
124:709, 1956.
41.
N a t l e l s o n , S.
1961.
Microtechniques o f c l i n i c a l chemistry.
C C . Thomas,
56
42.
Nelson, N. A photometric adaptation o f the Somogyi method f o r t h e
determination o f glucose.
J . B i o l . Chem. 153:375, 1944.
43.
Nordan, H.C., A.J. Wood, and I . McT. Cowan.
Further studies on t h e
immobilization o f deer w i t h s u c c i n y l c h o l i n e .
Can. J . Comp. Med.
26:246, 1962.
44«
Ochs, S., B. Annis, and A.K. Mukherjee.
S u c c i n y l c h o l i n e and muscle
excitability.
S c i . 131:1679, I960.
45«
Pisty,,R.W., and J.F. Wright.
The i m m o b i l i z a t i o n o f c a p t i v e w i l d
animals w i t h s u c c i n y l c h o l i n e ; 2 .
Can. J . Comp. Med. 25:59,
1961.
46.
Potter, B.J., and I.G. J a r r e t .
convulsions i n sheep.
31:311, 1953.
47.
I n s u l i n t o l e r a n c e and hypoglycemia
Aust. J . E x p t l . B i o l . and Med. S c i .
Preston, T.R., and R.D. Ndumbe. D i u r n a l v a r i a t i o n s i n blood sugar
concentration i n ruminating calves.
B r i t . J . Nut. 15:281,
1961.
48.
Rankin, A.D.
A study o f absorption from the rumen o f the sheep.
Vet. B u l l , (abst.) 11:328, 1940.
49«
Reichard, G.A., B. I s s e k u t z , J r . , P. Kimbel, R.C. Putman, N.J. Hochella
and S. Weinhouse.
Blood glucose metabolism i n man during
muscular work.
J . Appl. P h y s i o l . 16:1001, 1961.
50.
Reid, R.L. Studies on the carbohydrate metabolism o f sheep. 1.
The range o f blood sugar values tinder s e v e r a l conditions.
Aust. J . Agr. Res. 1:182, 1950.
51.
Reid, R.L. Studies on the carbohydrate metabolism o f sheep. 3«
The blood glucose during i n s u l i n hypoglycemia.
Aust. J .
Agr. Res. 2:132, 1950.
52.
Reid, R.L. Studies on the carbohydrate metabolism o f sheep. 5.
The e f f e c t o f hyperglycemia and o f i n s u l i n on the r a t e o f
extrahepatic glucose a s s i m i l a t i o n .
Aust. J . Agr. Res.
3:160, 1952.
53.
Reid, R.L., and S.C. M i l l s .
Studies on the carbohydrate metabolsim
of sheep. 14.
The adrenal response t o p s y c h o l o g i c a l s t r e s s .
Aust. J . Agr. Res. 13:282, 1962.
54.
Robertson, A., H. Paver, P. Barden, and T.G. Marr.
F a s t i n g metabolism
of the l a c t a t i n g cow. Res. Vet. S c i . 1:117, I960.
55.
Robinson, R.M., R.G. Marburger, and J.W. Thomas.
The reproductive
c y c l e o f male w h i t e t a i l e d deer i n c e n t r a l Texas.
J. Wildl.
Mgt. 29:53, 1965.
57
56.
Rosen, M.N., and A.I. B i s c h o f f .
The r e l a t i o n o f hematology t o
c o n d i t i o n i n C a l i f o r n i a deer.
Trans. N.A. W i l d l . Conf.
17:482, 1952.
57*
Schambye, P. V o l a t i l e acids and glucose i n p o r t a l blood o f sheep.
Nord. Vet. Med. 3:1003, 1951.
58.
Schuhecker, K.
Beobachtungen uber den b l u t z u c k e r der ziege.
Biochem. Z. 156:353, 1925.
59.
S e t c h e l l , B.P. and G.L. McClymont.
Blood inorganic phosphorous
and serum potassium i n i n s u l i n hypoglycemia i n the sheep.
Aust. J . Agr. Res. 6:589, 1955.
60.
Shaw, J.C., A.C. Chung, and I . Bunding.
The effect o f p i t u i t a r y
growth hormone and adrenocorticotropic hormone on established
lactation.
Endocrinology 56:327, 1955.
61.
S i l v e r , H., N.F. Colovos, and H.H. Hayes.
w h i t e t a i l e d deer...a p i l o t study.
1959.
62.
Basal metabolism o f
J . W i l d l . Mgt. 23:434,
Soldandt, O.M., and G.C. Ferguson.
E f f e c t o f strenuous e x e r c i s e
o f short d u r a t i o n on the blood-sugar.
Trans. Royal Soc.
Can. 26; v:173, 1932.
63-
Somogyi, M. The d i s t r i b u t i o n o f sugar and r a t e o f g l y c o l y s i s i n
the blood o f some mammals. J . B i o l . Chem. 103:665, 1933.
64«
Somogyi, M., A new reagent f o r the determination o f sugars.
Chem. 160-61, 1945.
65.
Somogyi, M.
J . Biol.
Determination o f blood sugar.
J . B i o l . Chem. 160:69,
Notes on sugar determination.
J . B i o l . Chem. 195:19,
1945.
66.
Somogyi, M.
1952.
67.
T e r r i , A., W. Virchow, N.F. Colovos, and F. Greeley.
o f w h i t e t a i l e d deer.
J . Mamm. 39:269, 1958.
Blood composition
68.
T y l e r , C. The e f f e c t o f prolonged emotional disturbance on the vasopressor and o x y t c i c a c t i v i t i e s contained i n the p o r t e r i o r
p i t u i t a r y glands o f f a l l o w deer.
Arch. I n t e r n . Pharmacodyn.
131:301, 1961.
69
Voelker, H.H., N.L. Jacobson, and R,S. A l l e n .
Relationship of a n t i b i o t i c feeding and the r a t e o f growth t o blood reducing sugar
l e v e l s and glucose absorption i n d a i r y c a l v e s . A n t i b i o t i c s and
Chemotherapy 5:224, 1955.
70.
Warner, A.C.I.
Some f a c t o r s i n f l u e n c i n g the rumen m i c r o b i a l population.
J . Gen. M i c r o b i o l . 28:129, 1962.
58
71.
Whitlock, S.C.
Studies on the blood of w h i t e t a i l e d deer.
Mgt. 3:14, 1939.
72.
Wilber, C.G., and P.F. Robinson.
Aspects o f the blood chemistry of
w h i t e t a i l e d deer.
J . Mamm. 39:309, 1958.
73.
Wing, J.M., N.L. Jacobson, and R.S. A l l e n .
The e f f e c t o f various
r e s t r i c t e d d i e t s on the growth and c e r t a i n blood components
of young d a i r y c a l v e s .
J . Dairy S c i . 38:1006, 1955.
74.
Wright, P.L., A.L. Pope, and P.H. P h i l l i p s .
E f f e c t of p r o t e i n and
energy i n t a k e on lamb production and c e r t a i n blood constituents
of ewes.
J . An. S c i . 21:602, 1962.
75.
Youatt, W.G., L . J . Verme, and D.E. U l l r e y .
Composition of milk and
blood i n nursing does and blood composition o f t h e i r fawns.
J . W i l d l . Mgt. 29:79, 1965.
J. Wildl.
APPENDICES
APPENDIX I
Glucose Metabolism
G l u c o s e i s t h e m a j o r end p r o d u c t o f c a r b o h y d r a t e d i g e s t i o n i n t h e
m o n o g a s t r i c a n i m a l , and p l a y s a n i m p o r t a n t r o l e i n i t s m e t a b o l i c p r o c e s s e s .
It
i s e s s e n t i a l f o r t h e maintenance o f c e l l s b o t h as a p r e c u r s o r t o many
c e l l u l a r components and as a s o u r c e o f e n e r g y .
i s a precursor of l a c t o s e .
In the l a c t a t i n g animal i t
G l u c o s e a l s o p r o v i d e s 2 - c a r b o n fragments
for
f a t s y n t h e s i s and t h e n e c e s s a r y reduced coenzyme (NADPH) f o r t h e i n c o r p o r a t i o n o f t h e 2 - c a r b o n u n i t s i n t o l o n g c h a i n f a t t y a c i d s (Armstrong 1 9 6 5 ) .
It
has l o n g been c o n s i d e r e d t h a t g l u c o s e m e t a b o l i s m i s q u a n t i t a t i v e l y l e s s
i m p o r t a n t i n r u m i n a n t s as compared t o n o n - r u m i n a n t s .
(Baxter et a l 1955).
The f a c t t h a t l i t t l e g l u c o s e i s a b s o r b e d f r o m t h e d i g e s t i v e t r a c t o f t h e
r u m i n a n t , (Schambye 1951) and t h a t i t appears i n l o w c o n c e n t r a t i o n s i n t h e
b l o o d o f t h e s e a n i m a l s s u p p o r t s such a c o n t e n t i o n .
U t i l i z a t i o n rates
measured by
14
c
- g l u c o s e i n f u s i o n , and e x p r e s s e d p e r u n i t o f m e t a b o l i c body
s i z e , however, i n d i c a t e t h a t t h e r a t e o f u t i l i z a t i o n o f g l u c o s e i n r u m i n a n t s
i s e s s e n t i a l l y t h e same a s i t i s i n s i m p l e - s t o m a c h e d a n i m a l s .
1965) •
(Armstrong
G l u c o s e , t h e n , i s a a i m p o r t a n t i n t h e m e t a b o l i s m o f t h e ruminant as
i t i s the non-mmi nant.
(Lindsay 1959).
I n r u m i n a n t s d i e t a r y c a r b o h y d r a t e s a r e fermented r a t h e r t h a n d i g e s t e d
i n t h e rumen.
The u l t i m a t e p r o d u c t s o f t h i s f e r m e n t a t i o n a r e t h e v o l a t i l e
f a t t y a c i d s , a c e t i c , p r o p r i o n i c , and b u t y r i c .
These a r e absorbed i n t o t h e
p o r t a l b l o o d system t h r o u g h t h e rumen w a l l and p l a y a major r o l e i n t h e
energy m e t a b o l i s m o f t h e a n i m a l .
Some o f t h e a c e t a t e i s o x i d i z e d i n t h e
l i v e r , but t h e m a j o r i t y i s p a s s e d i n t o t h e p e r i p h e r a l c i r c u l a t i o n where i t
i s u t i l i z e d by t h e t i s s u e s a s a s o u r c e o f e n e r g y .
I t i s i n t e r e s t i n g t o note
60
t h a t many t i s s u e s can derive energy from o x i d i z i n g f a t t y acids and ketone
bodies, but nervous t i s s u e can u t i l i z e o n l y glucose as an energy source.
(Krebs 1964).
Proprionic a c i d i s e i t h e r o x i d i z e d o r converted t o glucose
i n the l i v e r .
L i t t l e butyric acid i s carried to the l i v e r , the majority i s
converted t o ketone bodies i n t h e rumen epithelium.
(Armstrong 1965).
Glucose i n the ruminant i s derived from non-carbohydrate sources by t h e
process o f gluconeogenesis.
precursors.
Proportionate and amino a c i d s are t h e p r i n c i p a l
The glucose formed i s metabolized i n e s s e n t i a l l y the same
manner as i t i s i n monogastric animals.
the elaboration o f l o n g chain f a t t y a c i d s .
L i t t l e , however, i s u t i l i z e d i n
The abundant supply o f r e a d i l y
a c t i v a t e d acetate i s the primary source o f these i n t h e ruminant.
I t has
a l s o been shown that glucose i s i n v o l v e d i n the synthesis o f l a c t o s e ,
g l y c e r o l , m i l k c i t r a t e , and some non-essential amino acids i n ruminants.
(Armstrong 1965).
61
APPENDIX 2
Hormonal R e g u l a t i o n o f B l o o d R e d u c i n g Sugar
The maintenance o f n o r m a l l e v e l s o f r e d u c i n g s u g a r i n t h e b l o o d o f
mammals i s a f i n e l y r e g u l a t e d h o m e o s t a t i c mechanism.
The l i v e r p l a y s an
e s s e n t i a l r o l e as i t f u n c t i o n s b o t h t o remove and t o add sugar t o t h e b l o o d .
The a c t i v i t y o f t h e l i v e r i n m a i n t a i n i n g n o r m a l l e v e l s o f s u g a r i n t h e b l o o d
i s i n f l u e n c e d by a number o f hormonal f a c t o r s .
1.
Insulin
I n s u l i n i s a s e c r e t i o n fo the B - c e l l s o f the i s l e t s o f Langerhans.
It
f u n c t i o n s t o l o w e r t h e b l o o d r e d u c i n g s u g a r l e v e l by p r o m o t i n g p e r i p h e r a l
u t i l i z a t i o n o f g l u c o s e , g l y c o g e n e s i s i n m u s c l e and l i v e r ,
and l i p o g e n e s i s .
The mechanism o f i n s u l i n a c t i o n remains open t o d i s c u s s i o n , however,
experimental
e v i d e n c e s u g g e s t s t h a t i t a c t s by a f f e c t i n g 1 . membrane t r a n s p o r t phenomena,
and 2 . o x i d a t i v e p h o s p h o r y l a t i o n r e a c t i o n s .
(Bard 1 9 6 l ) .
That i n j e c t i o n o f i n s u l i n l o w e r s t h e l e v e l o f b l o o d r e d u c i n g s u g a r i n
sheep was shown by R e i d (1951) and ( 1 9 5 2 ) .
The r a t e o f f a l l ,
however, was
c o n s i d e r a b l y s l o w e r t h a n t h a t r e p o r t e d f o r n o n - r u m i n a n t s , and t h e l e v e l d i d
n o t f a l l b e l o w 5 mgm.%.
Moreover, f u r t h e r i n j e c t i o n o f l a r g e doses o f
i n s u l i n (10 U/Kg.) d i d n o t cause a f u r t h e r r e d u c t i o n i n b l o o d s u g a r ,
merely prolonged the e x i s t i n g hypoglycemia.
It
i s noteworthy t h a t
but
spontaneous
d i a b e t e s i s uncommon i n r u m i n a n t s .
2.
Epinepherine
H y p e r g l y c e m i a has been r e p o r t e d i n sheep ( S a t c h e l l and McClymont
1955) and i n c a t t l e (Garner 1952) when i n j e c t e d w i t h e p i n e p h e r i n e .
importance o f t h i s
The
s e c r e t i o n : ! o f t h e a d r e n a l m e d u l l a becomes apparent when
62
a splanchnic s e c t i o n ( P o t t e r 1952) i s c a r r i e d out on a sheep - t h e blood
sugar l e v e l f a l l s t o nothing.
3.
Glucagon
Glucagon i s a s e c r e t i o n o f the o c c e l l s o f the pancreas t h a t operates
to r a i s e t h e l e v e l o f blood sugar by promoting glycogenolysis.
The s i t e o f
a c t i o n o f glucagon i s apparently the same as t h a t o f epinepherine. The
hyperglycemic r e a c t i o n o f glucagon a d m i n i s t r a t i o n has been demonstrated i n
sheep (Ho and Reber 1957)•
4.
Growth Hormone
In monogastric animals, growth hormone acts t o decrease p e r i p h e r a l
u t i l i z a t i o n o f blood sugar and t o increase glycogenolysis i n the l i v e r .
net e f f e c t , then, i s t o r a i s e t h e l e v e l o f blood reducing sugar.
The
This e f f e c t
was noted i n sheep (Kronfeld 1957), however, t h e elevated l e v e l d i d not
p e r s i s t d e s p i t e repeated i n j e c t i o n o f the hormone.
5.
ACTH and Cortisone
ACTH undoubtedly exerts i t s e f f e c t i n d i r e c t l y v i a increased r e l e a s e
of 11-oxycorticoids from t h e adrenal cortex.
The 11-oxycorticoids exert
t h e i r diabetogenic e f f e c t through increased gluconeogenesis from amino a c i d s
i n the l i v e r .
These hormones are a l s o i n s u l i n antagonists.
S a t c h e l l and
McClymont (1955) i n sheep and Shaw (1955) i n c a t t l e produced hyperglycemia by
i n j e c t i n g ACTH and C o r t i s o l .
APPENDIX 3
Succinylcholine
Chloride
Hunt (1906) was the f i r s t t o synthesis s u c c i n y l c h o l i n e .
The immobil-
i z a t i o n a c t i o n o f the drug remained undiscovered f o r f o r t y years.
Castillo
and deBeer (1950) f i r s t documented t h i s and since then numerous reports o f
i t s use i n v e t e r i n a r y and human medicine have appeared.
The structure o f
s u c c i n y l c h o l i n e , which appears below, i s v e r y s i m i l a r t o that o f a c e t y l choline.
HX
O
O
C H
H C-N CH CH OCCH CH COCH CH N -CK3
/
+
3
3
+
2
2
2
2
2
2
The mechanism o f a c t i o n o f s u c c i n y l c h o l i n e i s believed t o be enzymatic
i n h i b i t i o n at the myoneural j u n c t i o n ( P i s t e y and Wright 1961). The
s u c c i n y l c h o l i n e i n h i b i t s the a c t i o n o f a c e t y l c h o l i n e s t e r a s e r e s u l t i n g i n an
accumulation o f a c e t y l c h o l i n e .
This accumulation lowers the r e s t i n g
p o t e n t i a l o f the muscle, reducing i t s e x c i t a b i l i t y and the muscle i s
e f f e c t i v e l y paralyzed (Ochs I960).
Pseudocholinesterase, found i n the
plasma, hydrolyzes s u c c i n y l c h o l i n e i n t o succinylmonocholine and choline.
The succinylmonocholine i s then degraded i n t o s u c c i n i c a c i d and choline
c h l o r i d e (Bovet et a l 1949 as reported by P i s t e y and Wright 1961).
Acetylcholinesterase then acts t o degrade the accumulated
acetylcholine,
conduction i s resumed and the p a r a l y s i s i s terminated.
Mayrhofer (1952) induced s e l f p a r a l y s i s w i t h s u c c i n y l c h o l i n e .
Muscle
weakness, i n i t i a l l y o f the neck, jaw and diaphragm, p a i n f u l muscle t w i t c h i n g ,
and double v i s i o n , characterize the sensations that he a t t r i b u t e d t o the
a c t i o n o f the drug.
At no time d i d he become unconscious, and no change
i n pulse r a t e o r blood pressure was recorded.
e f f e c t s were experienced.
Moreover, no a f t e r o r side
APPENDIX A
Peebles V'LER M i l k Replacer
Ingredients
Guaranteed minimum a n a l y s i s
Dried skim milk
crude f a t
Dried butter m i l
crude p r o t e i n 24-0$
Dried whey-product
Vit A
Lecithin
Vit D
Sodiumbenzoat e
V i t B^
11 mgm.
Magnesium carbonate
V i t Bg
11 mgm.
1,500 U/lb
2
Dicalcium phosphate
I r o n sulphate
A n t i b i o t i c supplement ( o x y t e t r c y c l i n e and terramycin)
V i t A palmitate
Vit D
Vit
2
(thiamine)
Vit B (riboflavine)
Q
16. C#
3,000 U/lb
APPENDIX 5
Weaning Ration
Ingredient
lbs/ton
Ground barley
200
Ground wheat
585
Oat groats
390
Wheat bran
130
Herring meal
200
Soya meal
100
Lysine
Methionine
Skim
milk powder
Brewers yeast
Irradiated yeast
200
20
2
D i s t i l l e r s solubles
Dehydrated grass meal
Vit D
150
2
Dicalcium phosphate
10
Iodized s a l t
10
Manganous sulphate
Zinc sulphate
Chromic oxide
1
APPENDIX 6
Adult Ration (36-57)
Ingredients
lbs/ton
Com meal
600
Ground wheat
250
Bran
275
Beet pulp
200
Vitagrass
200
Soyabean meal
175
Herring meal
110
Bone meal
20
Iodized s a l t
20
Molasses
150
67
APPENDIX 7
Absorption Curve f o r t h e Nelson-Somogyi Method o f Determining Blood Glucose.
Beckman Model DU Spectrophotometer
Standard Glucose S o l u t i o n 100 mgms./S
Wavelength
Slitwidth
O.D.
400
410
420
430
440
450
460
470
480
490
500
510
520
530
540
550
560
570
580
590
600
605
610
615
620
625
630
635
640
645
650
655
660
665
670
1.95
1.15
0.68
0.44
0.30
0.22
0.158
0.150
0.150
0.140
0.140
0.140
0.065
0.065
0.065
0.065
0.070
0.080
0.090
0.110
0.14
0.17
0.22
0.28
0.32
0.40
0.075
0.075
0.065
0.065
0.065
0.065
0.065
0.065
0.065
.0752
.0809
.0894
.1007
.n?/,
.1278
.1421
.1586
.1733
.1898
.2083
.2291
.2557
.2865
.3242
.3675
.4179
.4660
.5186
.5686
.6126
.6308
.6478
.6676
.6799
.6946
.7305
•7471
.7696
.7878
.8069
.8327
.8539
.8827
.9136
Wavelength
675
680
685
690
695
700
710
720
730
740
750
760
770
780
790
800
810
820
830
840
850
860
870
880
890
900
910
920
930
940
950
960
970
980
990
1000
Slitwidth
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
0.065
O.D.
.9508
.9914
1.0269
1.0706
1.1079
1.1549
1.2218
1.2924
1.3468
1.3768
1.4089
1.4089
1.3979
1.3768
1.3279
1.3010
1.2596
I.2076
1.1675
1.1249
1.0809
1.0410
1.0000
.9626
.9318
.9031
.8665
.8356
.8097
.7852
.7595
.7375
• 7122
.6946
.6757
.6576
FIGURE 9:
ABSORPTION SPECTRUM:
NELSON-
SOMOGTI METHOD OF DETERMINING BLOOD GLUCOSE
1600
200 I -
560
600
640
I
I
700
750
.WAVE LENGTH
(mu)
800
850
900
<25C
69
APPENDIX 8
Standard Curve f o r the Nelson-Somogyi Method o f Determining Blood Glucose.
Beckman Model DU Spectrophotometer
S l i t w i d t h 0.065
Wavelength 650 mu
Cone ent r a t i o n
0. D.
Concentration
O.D.
Concentration
O.D.
100
.8539
• 8539
.8477
.8477
.8477
.8477
.8477
.8477
.8477
.8416
.8416
.8416
.8416
.8416
.8386
.8386
.8386
.8386
.8386
.8386
.8356
.8297
.8268
.8268
.8268
.8268
.8239
.8239
.8239
.8239
.8239
100
.8239
.8239
.8239
.8239
.8239
.8239
.8239
.8239
.8210
.8210
.8210
.8210
.8210
.8210
.8210
.8182
.8182
.8182
.8182
.8182
.8182
.8153
.8153
.8153
.8153
.8153
.8153
.8153
• 8153
.8125
.8125
100
.8125
.8125
.8097
.8041
.8041
.8041
.8041
.8041
.8041
.8041
.8013
.8013
.7986
.7986
•7986
.7986
.7986
• 7986
.7959
.7959
.7959
.7905
.7878
.7878
.7878
.7852
.7852
.7852
.7825
.7747
APPENDIX 8 (cont.)
Concentration
O.D.
Concentration
O.D.
Concentration
O.D.
70 mgm.$
.6055
.6055
.5918
.5918
• 5969
.5969
.5982
• 5901
.5884
.5850
.5850
• 5834
• 5834
• 5834
.5817
.5800
.5751
• 5735
.5735
.5735
• 5719
.5602
.5602
.5670
.5686
.5686
.5686
.5670
.5654
• 5654
.5654
.5654
• 5654
• 5654
.5654
70 mgm.#
.5654
.5654
.5638
>5638
.5622
.5607
.5607
.5591
.5591
.5560
.5560
• 5575
• 5544
.5513
.5513
•5513
.5513
.5498
.5492
.5452
.5376
.3468
.3382
53382
•3372
•3354
.3344
.3344
.3335
.3316
.3316
.3316
.3307
.3298
.3279
40 mgp\.%
.3270
.3270
.3261
.3261
.3261
.3251
•3251
.3251
.3251
•3251
.3233
.3233
.3233
•3224
• 3215
.3215
.3206
.3197
.3188
.3188
.3179
.3170
.3152
.3152
.3152
.3143
.3143
•3143
.3125
.3116
.3116
.3116
.3107
.3054
•3045
.3028
40 mgm.%
FIGURE 10:
STANDARD CURVE: NELSON-SOMGYI
METHOD OF DETERMINING BLOOD GLUCOSE.
OPTICAL
DENSITY
APPENDIX 9
Recovery of Glucose from Serum.
Sample
Recovery
Calculated
% Recovered
69-9
70.0
99.9
2:1
84.1
82.0
102.6
1:1
8753
88.0
99.2
1:2
89-5
94.0
95-2
70
mgm.%
serum
106.0
73
APPENDIX 10: BLOOD SAMPLING RECORD FORM.
BLOOD ANALYSIS DATA
DATE
ANIMAL
TIME
0800
RESTRAINT
WEIGHT
lapse
1200
lapse
Time approached
1600
ANECTINE DOSE
SEX
lapse
2000 lapse
.
Time grabbed/injected
Time held/down
Sampling begun
Sampling ended
Sample ringed
Sample centrifuged
Sample i n f r e e z e r
COMMENTS
GLUCOSE ANALYSIS
Time
0800 1
2
Date
% T.
O.D.
mgm. % Glucose
O.D.
mgm. % Calcium
3
1200 1
2
1600 1
2
2000 1
2
2_
CALCIUM ANALYSIS
Time
0800 1
2
3
1200 1
2
JL
1600 1
2
2_
2000 1
Date
% T.