THE EPP2CT OB EHantB-SUFPLEKXNTATlCN
OF MILK KSPLACERS QK THE G-ROV.'TH 0? CALV.

By
George F %

Fries

AN ABSTRACT

Submitted to the School of Graduate Studies of Michigan
State U ni ve rs it y of Agriculture and Applied S cience
in partial fulfillment of the requirement
for the degree of

SOOTCR OP PHILOSOPHY

;oparteent of dairy

195c

M f*r*v"s

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+ V . A st

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-Ov

(T/9-

ABSTRACT
Two experiments were carried out to determine the ef­
fect of supplementing milk replacers with enzymes.

In the

first experiment, a vegetable milk replacer was predigested
with malt diastase, papain, and a combination of these en­
zymes.

Fifty-five pounds of milk were fed through the 20th

day and 30.5 lb. of the treated milk replacer were fed
through the 40th day.
days.

The experiment was terminated at 60

Decreased starter consumption and growth from 20 to

40 days and an increased mortality rate indicated that
papain produced deleterious effects.

Malt diastase produced

no improvement in gain during the periods it was used.
In the second experiment, soybean flour-corn, soybean
flour-cerelose, and skimmilk-cerelose milk replacers were
supplemented with pepsin but were not predigested.

All

other aspects of the experiment were similar to the first
esqperiment.

Pepsin produced no improvement in growth with

any of the replacers.

The skimmilk-cerelose replacer pro­

duced significantly greater weight gains than the other
replacers during the first 20-day period.

In the second

20-day period the soybean flour-cerelose replacer produced
significantly smaller gains than the other replacers.
Starter consumption by this group was also significantly
depressed.

THE EFFECT OF ENZYME-SUPPLEMENTATION
OF MILK REPLACERS ON THE GROWTH OF CALVES

By
George F. Fries

A THESIS

Submitted to the School of Graduate Studies of Michigan
State University of Agriculture and Applied Science
in partial fulfillment of the requirements
for the degree of

DOCTOR OF PHILOSOPHY

Department of Dairy
1958

ProQuest Number: 10008537

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ACKNOWLEDGMENTS

The author wishes to express his appreciation to Dr.
Earl Weaver, Professor of Dairying, and Dr. N. I. Ralston,
Professor of Dairying, for award of the Graduate Assistantship and provision of the facilities for conducting the
study; to Dr. C. E. Huffman, Professor of Dairying, for
suggesting the problem and for his continued interest
throughout the study; to Dr. C. A. Lassiter, Associate Pro­
fessor of Dairying, for his guidance and counsel; to Dr. D.
E. Madden, Assistant Professor of Dairying, for advice on
statistical analysis; to Dr. E. P. Reineke, Professor of
Physiology and Pharmacology, Dr. R. U. Byerrum, Professor
of Chemistry, Dr. R. S. Emery, Assistant Professor of
Dairying, and Mr. J. P. Everett, Graduate Assistant in
Dairy, for critically reading the manuscript; to Mr. R. E.
Reid and Mr. L. A. Reid for assistance in caring for the
animals; to the other staff members and graduate students
who provided advice and assistance at various times during
the study.

TABLE OF CONTENTS

Page
INTRODUCTION .......................................

1

REVIEW OF LITERATURE ...............................

3

Corn-Soybean Flour Milk Replacers ............

4

The Effect of Age on the Utilization of Nu­
trients .................................

5

Development of Enzyme Systems in the Young

8

. .

Pepsin ...................................
Gastric acidity
Trypsin

........................

..........

. . . . .

Other proteolytic enzymes

9
10
12

. . . . . . . .

Pancreatic amylase

13
13

L a c t a s e ................................... 14
M a l t a s e ................................... 15
S u c r a s e ................................... 16
Pancreatic lipase

......................

16

Supplementation of Diets with Enzymes and
Hydrolysates .
........................... 16
Studies of enzyme deficiencies ..........

17

Use of enzymes and hydrolysates in feeding
young a n i m a l s ......................... 18
Summary of the Literature Review

............

EXPERIMENTAL PROCEDURE .............................

20
21

TABLE OF CONTEXTS

(Continued)
Page

R E S U L T S ...........................................

27

Milk and Milk Replacer Consumption..........

27

Starter Consumption

31

........................

Hay C o n s u m p t i o n .............................

33

v/eight G a i n .................................

33

M o r t a l i t y ...................................

35

D I S C U S S I O N .......................................

36

S U M M A R Y ...........................................

45

LITERATURE CITED

47

APPENDIX

.................................

54

LIST OF TABLES

Table
1

Page
Composition of the experimental milk re­
placers ...................................

22

Chemical composition of the feeds used in
the two experiments......................

23

3

Daily feeding schedule

24

4

The results of the first experiment . . . .

28

5

The results of the second experiment

29

2

...................

...

LIST OF APPENDIX TABLES
Table
1
2
3
4

Page
Summary of analysis of variance:
The first
.
e x p e r i m e n t ............................

55

Summary of analysis of variance:
ond e x p e r i m e n t

56

The sec­

Data for the individual calves:
The first
e x p e r i m e n t ...............................

37

Data for the individual calves:
ond e x p e r i m e n t

39

The sec­

INTRODUCTION
Milk is one of the most expensive items in the diet
of the young calf.

For this reason a large number of at­

tempts have been made to develop feeding regimes which
would reduce the amount of milk required for raising calves.
Generally, these attempts have been successful with calves
over 30 days of age.

With calves under this age it has not

been possible to develop a milk replacer which would sup­
port normal growth.

The so-called milk replacers currently

in use are composed to a large extent of milk by-products
and, therefore, cannot be considered true milk replacers.
It has been shown that calves are unable to digest
vegetable milk replacers at birth and that the ability to
digest these replacers increases with age until it reaches
the adult level at 30 to 35 days of age.

Since growth was

subnormal initially and improved as digestibility improved,
it is reasonable to assume that the primary fault of these
milk replacers is their indigestibility.

The young of

several species have been found to be deficient in one or
more of the digestive enzymes.

These enzyme deficiencies

could explain the inability of the young calf to utilize
vegetable milk replacers.
The following experiments were carried out to study
the use of amyloytic and proteolytic enzymes and the use
-

1

-

2
of a simple carbohydrate instead of starch in vegetable
milk replacer formulation.

REVIEW OF LITERATURE

The nutrient requirements of the young calf are in­
fluenced to a large extent by the degree of rumen develop­
ment.

At birth the rumen is nonfunctional and the nutrient

requirements of the calf resemble the requirements of
simple-stomached animals.
during this period.

Milk meets these requirements

Replacement of milk with feeds of

vegetable origin has not supported a normal growth rate.
As the rumen becomes functional the requirements become
less specific.

At this time replacement of milk with vari­

ous feeds of vegetable origin has generally been success­
ful.
The current studies are concerned primarily with the
period preceding the development of a functional rumen.
The literature concerning the rumen development of the calf
has been thoroughly reviewed by Roller (1955)*

This review

includes discussions on the use of corn-soybean flour milk
replacers, the effect of age on the utilization of various
nutrients, the development of the enzyme systems in the
young, and the effect of supplementing diets with various
enzymes and hydrolysates.

- 3 -

4

Corn-Soybean Flour Milk Replacers
The milk replacer with which the present study con­
cerns itself has been evolved from the formulas first de­
scribed by Roller and Huffman (1953)*

All replacers con­

tained finely-ground corn and rolled oats as the primary
sources of carbohydrate and soybean flour as the primary
source of protein.

In addition all replacers contained

10% distillers solubles and 10% whey which was the only
animal product used.

A number of minor ingredients were

used to supply the various vitamins and minerals.

The in­

gredients used were considered to be the ones which had
shown the most promise in previous studies (Roller* 1935)*
These milk replacers did not support satisfactory growth
until the calves were approximately 30 days of age.

After

this time the growth rate was similar to the rate of the
controls fed whole milk.

The term "critical period" was

applied to the period of low growth rate by these workers.
In later studies the milk replacer formulas were sim­
plified and were similar to the basal replacer used in the
current study.

In comparison with a basal replacer that

did not contain distillers solubles and whey, Huffman et al.
(1934) found that inclusion of 5% brewers dried yeast or
10% distillers solubles did not improve the growth rate.
The use of 3% whey improved growth rate and physical

5
appearance of the calves but the difference was not statis­
tically significant.

In a following study Noller et al.

(1956a) confirmed the results concerning whey.

Use of

lactose in amounts equivalent to that contained in the
whey did not produce these results.

Fries et al. (1956)

confirmed the results concerning distillers solubles and
also found that supplemental vitamin

had no beneficial

effect.
The results of digestion trials involving these types
of milk replacers will be discussed in the following sec­
tion.
The Effect of Age on the Utilization of Nutrients
While it appears to be general knowledge that very
young animals (i.e., calves under 30 days of age or equi­
valent ages in other species) cannot utilize certain nu­
trients as well as older animals, there have been very few
studies in which this point was specifically examined.
Several studies have indicated that a number of carbo­
hydrates which are utilized by older pigs are not utilized
by the very young pig.

Johnson (194-9) found that glucose

or lactose in purified diets fed beginning the second day
after birth pi^oduced satisfactory growth in pigs.

Sucrose

resulted in acute diarrhea, kidney hemorrhage and death
within a short time.

Fructose was also found to be

6
unsatisfactory.

The inability of the 2-day old pig to

utilize sucrose has been confirmed by Becker et al. (1954-a).
In another study with pigs during the period from 0 to 9
days of age, Becker et al. (1954-b) found that glucose or
invert sugar produced satisfactory growth and survival
rates.

Pigs that received fructose or sucrose lost weight

and had a very low survival rate.

Diarrhea was severe with

the use of fructose and sucrose, less severe with invert
sugar, and there was none with glucose.
Two studies have been reported in which various carbo­
hydrates were compared for the pig in the period from 7 to
35 days of age.

Becker et al. (1954-a) found that glucose,

lactose, sucrose, dextrin or starch produced about equal
gains during this period.

However, less diarrhea was noted

in the groups receiving lactose and starch.

In contrast,

Hudman et al. (1955) reported that lactose was superior to
glucose, sucrose, corn syrup solids and corn.

Corn starch,

oat groats, corn flakes and gelatinized starch produced
gains considerably lower than lactose.
Using 9-week old pigs Becker and Terrill (1954-) found
that glucose, sucrose, dextrin and starch produced equal
gains when fed at the rate of 50% of the diet.

A slight

depression in growth occured when lactose was fed at this
rate but the depression did not occur when the diet was
composed of 25% lactose.

7
Shaw et al* (1917) have studied the effect of age on
starch utilization hy calves.

Calves from 4 to 7 days of

age were able to digest only 20% of the starch they were
fed.

At 3 to 4 weeks of age over 90% of the starch was

digested.
Flipse and coworkers have published a number of stu­
dies on the ability of the young calf to utilize various
carbohydrates.

In 28-day trials calves which had received

glucose as the carbohydrate increased 3.3% in body weight
while calves receiving dextrin and starch lost 8.8% and
6.9% of their initial weight (Flipse et al., 1948).

The

calves which were fed glucose were in better condition than
the other calves but showed a paralysis which was curable
by either potassium or biotin.
In another study Flipse £t al. (1930b) compared diets
in which the carbohydrate used was glucose at the 60% level
and in which part of the glucose was replaced by corn syrup
or lactose.

Lactose at the 5* 10 and 30% levels were about

equal and were superior to glucose alone and glucose with
various levels of corn syrup.

Corn syrup at levels of 10

and 30% produced gains superior to glucose, but corn syrup
at the 43% level resulted in a loss of weight during the
experimental period.
The presence of 10% lactose in diets utilizing starch
as the principle source of carbohydrate increased the rate

8
of gain to approximately double the rate for starch alone
(Flipse et al*, 1950a)*

After feeding test meals contain­

ing a single carbohydrate, it was found that glucose and
lactose caused significant increases in blood sugar while
starch did not.

This would indicate that starch is not

utilized to the same degree as the other carbohydrates
studied*
Noller et al. (1950b) have studied the influence of
age on the digestibility of vegetable milk replacers simi­
lar to those described in the preceding section.

Digesti­

bility of dry matter, crude protein and nitrogen-free ex­
tract was low at two weeks of age but increased with age.
By five weeks of age the coefficients of digestibility of
the various fractions had increased to near the levels that
one would expect for adult animals.

In the same experiment

the digestibility of the various components of whole milk
was high at all ages.
Development of Enzyme Systems in the Young
Most of the studies concerning the enzymes present in
the contents and tissues of the alimentary tract have been
of a qualitative nature.

In quantitative studies a variety

of methods for determining enzyme activity and for express­
ing units of activity have been employed.

In addition,

most methods of determining enzyme activity are not

9
specific for a given enzyme.

For example, the term "tryp­

sin" usually refers to all proteolytic activity of the
pancreas and would include activity of proteolytic enzymes
other than trypsin.

Therefore, no attempt will be made to

discuss these studies in other than the most general terms.
Pepsin.

The appearance of pepsin in the development

of the fetus appears to vary greatly depending upon the
species.

Pepsin has been found as early as the fourth

month in the development of the human fetus (Keene and Hewer,
1929; Langendorf, 1879)*

It has been reported in cattle as

early as the end of the third month (Moriggia, 1879) and in
the stomachs of sheep embryos as small as 6 inches in length
(Sewall, 1878).

Langendorf (1879) has reported that pepsin

is present in the stomachs of the rat and the rabbit fe­
tuses.

However, Wolffhugel (1878) was unable to find it in

the rabbit fetus.

Pepsin has not been detected or was

found only in small traces in the fetus of the dog (Grutzner,
1875)* the c&t (Sewall, 1878), and the pig (Mendel, 1906;
Grutzner, 1875; Langendorf, 1879; Sewall, 1878).
Zweifel (1874) and Schmidt (1914) reported that pepsin
was always present at birth in the human.

Hammarsten (1374)

has reported that the amount present at birth may vary
greatly.

With a premature 7-months child which had been

maintained 14- days, the gastric mucosa exhibited only traces
of pepsin.

Werner (1948) found that the pepsin activity

10
of full term infants was only 10% of the activity of the
adult.
Hammarsten (1874) found no pepsin in the stomachs of
dogs under one week of age.

During the second week it be­

gan to appear in small quantities, but the amount was not
significant until after 3 bo 4- weeks of age.

Results

similar to this have been reported by Gmelin (1902).

He

found no pepsin at birth and significant quantities did
not begin to appear until after 18 days of age.

In pigs,

Kvasnitskii and Bakeeva (194-0) were able to detect the pre­
sence of pepsin in stomach contents at birth.

However, it

did not reach its highest activity until 49 days of age.
Pepsin has been reported to be present at birth in the
rat (Langendorf, 1879)» absent in the cat (Hammarsten,
1874), and present (Langendorf, 1879) and absent (tfolffhugel,
1876) in the rabbit.
Gastric acidity.

tfhile hydrochloric acid is not a

digestive enzyme, it will be discussed because its produc­
tion in sufficient quantities is required for optimum pep­
sin activity.
Grutzner (1879) was unable to find acid in the fetal
stomachs of cattle, sheep, pigs and dogs.
fetal development was not reported.

The stage of

In contrast, Moriggia

(1879) reported that the stomach contents of the bovine
fetus were always slightly acid after the third month.

11
Hydrochloric acid has been found to be present in the hu­
man fetal stomach as early as the nineteenth week of de­
velopment (Keene and Hewer, 1929)*
Gmelin (1902, 1904) studied the effect of increasing
age on the hydrochloric acid production of the dog.

In

both studies it was found that the level of acid was low
at birth and increased rapidly after 18 days of age.

In

the second study Pavlov fistulas were installed in order
to eliminate any direct effect of the food on the stomach
acidity.
Stomach contents of premature infants have been re­
ported to be lower in acid content than the contents of
full term infants (Miller, 194-1)*

In general, the acid

content was relatively high at birth.

It dropped to a

low at 10 days of age and then increased.
Kvasnitskii and Bakeeva (194-0) found that hydrochloric
acid did not appear in significant quantities in the pig
until 20 to 50 days of age.

Lewis et ad. (1955a) recorded

pH values of 4-.2, 5*5> 3*9* 3*6, 4.0 and 4.2 on 6-hour
fasted baby pigs stomachs at 1, 7, 14, 21, 28 and 55 days
of age•
A pH of 4.4 for the combined contents of the omasum
and abomasum of calves at birth has been reported by
Parrish and Fountaine (1952).
a pH of 6.4 was recorded.

For the rumen and reticulum

Since the omasum does not secrete

12
hydrochloric acid, a somewhat lower pH would be expected
if the abomasum had been considered alone.
Trypsin.

Langendorf (1879) was able to demonstrate

the presence of trypsin in the human fetus as early as the
fifth month of development.

These findings have been con­

firmed by Ibrahim (1909)» Keene and Hewer (1929) and
Schmidt (1914-).

Zweifel (1874-) and Schmidt (1914-) re­

ported that trypsin was present in the newborn infant but
the level was not high.

In quantitative studies of the

duodenal juice of infants, Vazquez (1991) found that tryp­
sin was present at low levels at birth.

The levels in­

creased with age and at 3 weeks the level was approximately
the same as the level at 3 months of age.

Berner (194-8)

compared the tryptic activity of premature infants (2.5
kg.) with that of full term infants (3*0 kg.).

The pre­

mature infants showed little or no activity while the full
term infants showed about 85% of the activity of the adult
human.
The only data for species other than the human was
reported by Langendorf (1879)*

Trypsin was usually found

in the pig fetus after the 100 mm. stage of development
and in the bovine fetus after the 250 mm. stage.

In rats,

both the fetus and the newborn exhibited trypsin but the
activity was low.
rabbit.

Trypsin was present in the new-born

13
Other proteolytic enzymes.

Ibrahim (1909) found that

enterokinase appears in the human fetus at about the same
time as trypsin.

The presence of erepsin (a mixture of

exopeptidases in the intestinal juice) has been demonstra­
ted in the newborn human (Cohnhiem, 1909), premature in­
fants (Jaiggy, 1907), and as early as the fifth month of
fetal development (Jaiggy, 1907; Keene and Hewer, 1929;
Langstein and Soldin, 1908).
Pancreatic Amylase.

Much of the information concern­

ing pancreatic amylase is conflicting.

This conflict is

probably due to the variety of techniques and to the small
numbers used in many of the studies.

Langendorf (1879)

was not able to detect pancreatic amylase at the 6-months
stage in the human fetus.

However, Keene and Hewer (1929)

found it as early as the twenty-second week of fetal de­
velopment, but it did not appear in all individuals as
late as full term.

Zweifel (1874) has reported that pan­

creatic amylase was absent in the newborn and did not ap­
pear in significant quantities until the end of the second
month of age.

In contrast, considerable diastatic activity

of the intestinal contents of the newborn has been reported
by Schmidt (1914).

In quantitative studies of the duo­

denal juice from 1 to 21 days of age, Vazquez (1931) found
the amylolytic activity was either low or absent during
this period.

14

There is a paucity of data concerning species other
than the human.

Langendorf (1879) found pancreatic amylase

to be present in the 100 mm. pig embryo and the 230 mm.
bovine embryo.

Both the rat fetus and the newborn rat ex­

hibited amylolytic activity and the level was high at 3 to
4 days of age.

Pancreatic amylase was not detected in the

newborn rabbit.
A quantitative study using pigs has been conducted by
Kitts et al. (1936)*

The pancreatic amylase activity was

approximately 100 units per kilogram body weight at birth,
increased to 1500 units at the sixth day of age and re­
mained at this level through the twenty-second day of age.
It had increased to 4300 units at 37 days of age.
Lactase.

Lactase is one of the last of the digestive

enzymes to appear in the development of the fetus.

Ibra­

him (1910) and Ibrahim and Kaumheimer (1910) reported that
it was not present until the seventh or eighth month in the
fetal development of the human.

In some premature infants

it was absent in the stool on the first day after birth
but it appeared soon thereafter.

Keene and Hewer (1929)

were not able to detect lactase in the human fetus except
at full term.

Plimmer (1906) did not find lactase in the

rat fetus 2 days prepartum but did find it 12 hours prepartum.

Lactase was reported to be present in the pig em­

bryo by Mendel (1906) but the stage of development v/as not
given.

13
Prom his studies of a large number of species,
Plimmer (1906) came to the conclusion that lactase is pre­
sent in high concentrations at birth in all mammalian spe­
cies and then decreases with increasing age.

He also

found it to be present in adult omnivores and carnivores
but not in adult herbivores.

However, Mendel (1906) was

not able to demonstrate lactase in the adult pig.

In re­

cent studies with the pig, Bailey et al. (1936) found lac­
tase to be present at a high level at birth and it remained
at this level until about 20 days of age.

The level then

began to decrease rapidly and by 30 days of age the level
of lactase was negligable.

Similar results have been

found with rabbits and cattle (Heilskov, 1931)*

lo the

bovine the lactase activity decreased about 30% during the
period from 3 to 30 days of age.
Maltase.

In the human fetus, maltase has been found

as early as the fifth month by Ibrahim (1910) and the
sixth month by Keene and Hewer (1929).

However, it may

not be present in all individuals at full term (Keene and
Hewer, 1929)*
Mendel (1906) has qualitatively shown the presence
of maltase in the suckling pig.

In quantitative studies

with pigs, Bailey et al. (1956) found that the level of
maltase was negligible at birth.

It increased to a maximum

16
at 25 days of age and remained at this level until the ex­
periment was terminated at 50 days.
Sucrase.

The behavior of sucrase is very similar to

that of maltase with the exception that it appears at gin
earlier stage of development in the fetus (Bailey et al*,
1956; Ibrahim, 1910; Keene and Hewer, 1929; Mendel, 1906).
Pancreatic lipase.

Keene and Hewer (1929) found that

pancreatic lipase was usually present after the thirtysecond week of fetal development in the human.

The lipase

levels have been found to be high at birth in the human
(Zweifel, 1874-; Schmidt, 1914-).

Kitts et al. (1950) have

shown that the level of lipase is high at birth in the pig
and it remained high throughout the 50-day period of the
study.
Supplementation of Diets with Enzymes and Hydrolysates
This section will deal with two aspects of the use of
supplemental enzymes and hydrolysates in the diet.

These

are the studies of experimentally produced enzyme defi­
ciencies and the use of enzymes and hydrolysates to im­
prove the utilization of foods by young animals.

.Vhile a

great number of studies have been conducted using hydroly­
sates with older animals, these studies will not be covered
here because it is felt that they do not apply directly to
the problem being considered.

17
Studies of enzyme deficiencies.

Cruickshank (1915)

was one of the first investigators to use substitution
therapy in a study with depancreatized dogs.***

When a meat

diet was fed to these dogs the coefficients of digestibil­
ity for protein and fat were 76 and 33%* respectively.
Feeding raw pancreas increased these values to 92 and 79%)
respectively.

Similar findings have been reported by

Coffey et al* (194-Oa, 194-Ob), but the effects were somewhat
inconsistant in the case of fat.

They also reported that

the loss of carbohydrate in the feces was reduced consider­
ably when fresh pancreas was fed.
Several studies have been reported in 'which pancreatin
was administered to depancreatized dogs.

Selle (1937)

found that 3 £• of pancreatin per 100 g. of a meat diet re­
duced the nitrogen loss in the feces by 60% but failed to
check the loss of fat.

Larger amounts of pancreatin fur­

ther reduced the nitrogen loss but were not more effective
in checking the fat loss.

Enteric-coating of the pancrea­

tin to prevent destruction in the stomach gave no improve­
ment.

Schmidt et al. (1937) obtained similar reductions

in nitrogen losses when using 25 g* of pancreatin per day
with a beef diet.

In addition, fecel fat loss was reduced

^For convenience, the term "depancreatized" will be
used whenever all of the external secretion of the pancreas
was excluded from the intestines, whether by depancreatization, ligation of the pancreatic ducts, etc.

18
by a similar amount.

These reductions occurred regardless

of the level of food intake.

Coffey et al. (194-Ob) re­

ported that 2 g. of pancreatin per day was not effective
in reducing fecal nitrogen and fat losses but was effec­
tive in reducing fecal carbohydrate losses.

In four cases

of achylia pancreatica in humans, Beazell et al. (194-1)
found that oral administration of pancreatin resulted in
60% reductions in fecal fat and nitrogen losses.
In depancreatized dogs fed a 62% starch diet, Beazell
et al. (1937) found that supplementing the diet with vari­
ous amylases decreased the fecal starch loss by 30 to 50%.
In general amylases of vegetable origin which are inacti­
vated at a lower pH were more effective than pancreatic
amylase.

Enteric-coating of the pancreatic amylase im­

proved its effectiveness.

Even with very high levels of

amylases it was not possible to reduce the fecal starch
loss to its pre-operative level.
Ivy et al. (1937) have shown that as much as 20% of
the starch in the diet may be digested in the stomach when
supplemental amylases were used.
Use of enzymes and hydrolysates in feeding young ani­
mals.

The use of enzymes in feeding young animals is not

a new idea.

Kellner (1924-) credited Liebig with the for­

mulation of a milk substitute for calves which contained
starch that had been saccharified with malt,

holler (1953)

19
has reveiwed several studies on the use of malt-treated
starch in calf feeding.

Jhile satisfactory results were

obtained, it is difficult to interpret these studies be­
cause of the large amount of lactose which was available
from the skim milk in the diets.
The results of the few studies on the use of supple­
mental enzymes are somewhat conflicting.

Johnson (194-9)

found that sucrase added to the diet did not prevent the
deleterious effects from feeding sucrose to 2-day old pigs.
However, since Becker et ad. (1954-b) obtained similar ef­
fects from feeding fructose, this failure may not have been
due to the lack of activity of sucrase.
Williams and Knodt (1951) found that supplementation
of a milk replacer with papain or pancreatin resulted in
decreased growth and feed consumption by calves.

The milk

replacer used was of the skim milk type.
Studies involving the use of supplemental proteolytic
enzymes with soybean-type diets for pigs have been reported
by Lewis et al. (1955a, 1955b).

The rate of gain was in­

creased as much as 29% from 1 through 4- weeks of age.

Pep­

sin provided a greater response than proteolytic enzymes of
plant or fungus origin.
Jorpes et al. (194-6) reported increased weight gains
in premature infants when a pancreatic digest of casein was
added to the diet.

This response was not obtained when

20

undigested casein was used.

Feeding an enzymatic digest

of bovine plasma to infants from 1.5 to 9 months of age
produced growth rates and nitrogen retention as great as
evaporated milk (Albanese et al. , 1951)Summary of the Literature Review
The most important points of the literature review as
they apply to the current problem may be briefly summarized
as follows.

Calves fed a corn-soybean flour milk replacer

do not begin to grow at the normal rate until they are about
30 days of age.

Results of digestion trials indicate that

this lack of growth is due to the young calf's inability
to digest these milk replacers.

It would appear that

young animals are not able to utilize nutrients which are
not present in milk and which require enzymatic degrada­
tion before absorption.

The apparent deficiencies of a

number of digestive enzymes in the young animal would tend
to support this conclusion.

There is some evidence that

supplemental enzymes will improve food utilization in
cases of enzyme deficiencies.

However, the results with

the use of supplemental enzymes have not been uniformly
successful•

EXPERIMENTAL PROCEDURE
The first of the two experiments was carried out to
determine if predigesting a soybean, flour-corn milk re­
placer with malt diastase and/or papain would improve its
utilization by calves.

A 2 x 2 factorial design using 24-

Holstein calves was employed in this experiment.
trol group received Milk Replacer 1 (Table 1).

The con­
The chemi­

cal analysis of this replacer is presented in Table 2.

The

remaining groups received Milk Replacer 1 that had been
predigested with the following enzymes; 1.0% malt diastase,^
0.2papain, 2 and a combination of these treatments.
The selection of treatment levels was based on the
strength of the enzyme preparations and the calculated
amount of substrate present in the milk replacer.

The re­

placer to be predigested was mixed with four parts of water
and the digestion was carried out at 40° C. for 12 hours.
The digestion was carried out daily in amounts sufficient
to meet the needs of the following day.
The calves used in the experiment were obtained from
the University herd and from local dairymen.

They were

^Dry malt syrup (60° L), produced by Standard Brands,
Inc., New York.
o
No. 1 powder, produced by Paul-Lewis Laboratories,
Inc., Milwaukee.
- 21 -

22

Table 1*

Composition of the experimental milk replacers

1

Milk Repl;acer
2

3

(%)

c%)

(%)

33.4

41.9

Ingredient

Soybean flour
Dried skim milk

-

Finely ground corn

48.0

Cerelose

-

—

-

—

57.9
—

39.5

23.5

10.0

10.0

10.0

Dried whey

3.0

5.0

5.0

Aurofac

1.0

1.0

1.0

Dicalcium phosphate

2.0

2.0

2.0

Salt

0.5

0.5

0.5

0.1

0.1

0.1

Corn distillers dried solubles

Vitamin and mineral mixture

2

^Contained 5*0 gm. Aureomycin and appreciable Vitamin
B^2 P er
o
Mixture contained:
20 gm.
Vitamin A concentrate 20,000 U.S.P • units/gm.
Irradiated yeast, 9,000 I.U. Vitamin D/gm.
Cobalt Chloride
Cupric sulfate
Ferrous sulfate

5 gm.
1.5 gm.
1.25 gm.
6 gm.

25
Table 2.

Chemical composition of the feeds used in the two
experiments-*-

Crude hitrogenAsh
free
fiber
extract

Dry
matter

Crude
protein

Ether
extract

(%)

(%)

(%)

(%)

(%)

(%)

Beplacer 1

84-.9

27-6

1*5

2.1

47.3

6.4

Replacer 2

90.7

26.8

3.9

1.3

52.6

6.1

Replacer 5

94-.1

24-.8

1.3

0.4

59.2

8.4

Starter

88.0

18.4

3.5

4.7

56.1

5.3

Alfalfa :
hay

86.8

16.0

1.4

29.8

32.5

7.1

Feed

^"The feeds were analyzed using accepted A.O.A.C. (1950)
procedures.
placed on experiment as they became available throughout
the year with, the exception that no calves were on experi­
ment during the summer months.

The calves were randomly

assigned to the various experimental groups.

Any animal

that was apparently sick or abnormal during the first days
of the experiment was removed and replaced by the next ani­
mal of the same sex and relative weight.
The calves were left with their dams for the first
4-8 hours.

They were then weighed and placed into indi­

vidual pens bedded with wood shavings.

The milk replacers

were fed as a gruel using the amounts of milk and/or water

24

indicated in the feeding schedule (Table 3)•

The treated

milk replacers were fed through 40 days of age in order
Table 3*

Age
(days)

Daily feeding schedule

Water
(lb.")

Milk replacer

(lb.)

(lb.)

0-2

with dam

-

-

3-5

6

0.2

—

6-9

4

0*5

4

10-20

2

0.8

8

21-40

-

1.0

12

-P
H
1
O''
o

vVhole milk

-

-

ad libitum

to be certain that all calves were past the critical period
when milk replacer feeding was terminated*

The calves

were kept on experiment an additional 20 days to determine
if there were any carry-over effects from the treatments.
Milk Replacer 1 in dry form was fed ad libitum to all
calves after 7 days of age and was the only feed used dur­
ing the last 20 days of the experiment.
Body weights were obtained at the beginning of the
experiment, at 40 and 60 days of age, and at weekly inter­
vals throughout the experiment.

The weights presented in

the results for 20 days of age were obtained by interpola­
tion*

The amounts of feed fed to the individual calves

25
were recorded daily.

Refusals of milk and milk replacer

were recorded at the time of feeding.

Unconsumed dry feed

was weighed and discarded at the same times that body
weights were taken.
The feed consumption and growth data for the various
20-day periods, the period from 3 to 40 days, and the
period from 3 to 60 days were subjected to the usual
methods of analysis of variance (Snedecor, 1956).

In the

few cases in which a calf died before the end of the ex­
periment, a missing value was calculated following the
method outlined by Snedecor (1950).

In the event that

significant differences were found, the individual differ­
ences between means were tested by means of a multiple
range test (Duncan, 1955)*
Because of the inconclusive (in the case of malt
diastase) and unsatisfactory (in the case of papain) re­
sults of the first experiment, a second experiment was
carried out in order to further evaluate the use of sup­
plemental enzymes with milk replacers.

Three milk replac­

ers (Table 1) were fed with and without 0.5% pepsin.^

Pep­

sin was chosen because (1) it normally occurs in the di­
gestive tract and, therefore, would not be toxic and (2)

***1:3000 N. P., produced by Cudahy Laboratories, Omaha,
Neb.

26
it was the enzyme that showed the most promise in the pig
studies (Lewis e_t al. , 1955a)*

The level of pepsin used

was the one which had heen recommended for use with pigs
(Lewis et al., 1955a).

The milk replacers were not pre­

digested in order to eliminate another possible cause of
the unsatisfactory results obtained from papain.
If malt diastase was effective in the first experi­
ment one would expect as great or greater effect from the
use of the end-product of carbohydrate digestion.

There­

fore, Milk Repla,cer 2 (Table 1) was formulated in which the
corn was replaced by cerelose.

Milk Replacer 5 utilized

skim milk as the primary source of protein.

Its purpose

was to serve as a positive control.
A starter 2 and second-cutting alfalfa hay were fed ad
libitum to all calves after 7 days of age.

This change

from the first experiment was made in order to conform
more closely with the usual methods of feeding calves.
The body weights were taken at five day intervals
throughout this experiment.
With the above exceptions the second experiment was
conducted in the same manner as the first experiment.

^Composed of 42.8% ground corn, 51.3% crimped oats,
13.3% soybean oil meal, 10.0% distillers solubles, 1.0%
dicalcium phosphate, 1.0% salt, 0.5% Aurofac 2A, and 0.1%
vitamin A and D mix.

RESULTS
The results of the two experiments are presented in
Tables 4 and 5* respectively.

Although data had been col­

lected at 5-hay intervals, it was not practical to use
these detailed data because of relatively large errors in­
herent in the measurement of body weight.

The use of 20-

day intervals for the presentation of the data was arbi­
trarily adopted because the relative error in body weight
measurement was reduced to a reasonable size and because
these intervals coincided with the most radical changes in
the feeding regime.

It is recognized that this arbitrary

treatment of the data may tend to cover up some of the
trends that were present in the more detailed data.

There­

fore these detailed data will be discussed in those cases
where it is felt that it would make important contribu­
tions to the interpretation of the data.
Milk and Milk Replacer Consumption
The consumption of whole milk and milk replacer was
similar for all groups in both experiments (Tables 4 and 5).
This would be expected because of the controlled feeding
regime that was used.

The small variations were caused by

occasional feed refusals by a few calves.

- 27 -

In general,

28

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ft p d CM CM CM
P CO 0 0 0
d ft d d d d d
0 0 0 0 0
d 0
o o ft o O o o
ft 0 0 0 0
ft 0
r~HKA•H i
—Ii-1ft ft
0 P ft P P P P
0 o 0 0
d 0 ft
cd eft p (ft [ftP (ft
§
d A A A <A
0 A
—1o
0 i
d NO •H NO NO ft ft
CO ft
ft
d d d d
cd d d
0 0 d 0 0 0 0
o o tiOO o o o
0 cd 0 •H 0 0 0 0
-1
—1i
—1ft 1
CO ft ft 0 i

0 a,
ft OJ ft ft ft ft ft ft

f
t
f
tf
tf
tf
t
ft00O0000

CCS f t

O ft

0 <H fc O ft

31
acceptability of the milk replacers was rot a problem in
these experiments.
Starter Consumption^
In both experiments the level of starter consumption
was low for all calves through 20 days of age (Tables 4and 5)*

The differences among the various groups were not

statistically significant in either experiment.
In the first experiment the use of papain decreased
starter consumption to a considerable degree.

This de­

crease was statistically significant during the period 21
through 4-0 days and for the total consumption through 4-0
days.

Following 4-0 days the consumption of starter fol­

lowed a trend similar to that of the preceding period.
However, the difference was of smaller magnitude and was
not significant.

For the entire experiment the calves re­

ceiving papain consumed significantly less starter than the
other calves.

^For convenience when referring to the first experi­
ment, the term "milk replacer" refers to the treated milk
replacer that was fed as a gruel and the term "starter" re­
fers to the untreated milk replacer that was fed in the dry
form. From a functional standpoint, the untreated milk re­
placer was used as a starter in this experiment. The com­
position of Milk Replacer 1 was the same as the starter
used in the second experiment with the exception of the use
of finely ground corn and soybean flour in place of coarsely
ground corn, crimped oats, and soybean oil meal.

32
Malt diastase had no effect on starter consumption
during the periods in which it was used (through 4-0 days
of age).

However, during the period from 4-1 through 60

days, the calves that had received malt diastase consumed
significantly more starter than the other calves.

The in­

creased starter consumption by these calves in the complete
experiment was completely accounted for by the increase in
the last period and was not significant.
In the second experiment the calves receiving pepsin
consumed approximately the same amount of starter as the
control calves.

The small differences were not significant

in any of the periods considered.
The rate of starter consumption from 21 through 4-0
days of age appeared to be inversely related to the level
of cerelose in the milk replacer.

The results of the sta­

tistical analysis revealed that only the difference between
the calves receiving Milk Replacer 1 and the calves receiv­
ing Milk Replacer 2 was significant.

Similar results were

obtained when the total consumption through 4-0 days was
considered.

From 4-1 through 60 days the rate of starter

consumption by the calves that had received Milk Replacers
1 and 3 was approximately the same and was somewhat lower
for the calves that had received Milk Replacer 2.
this difference was not significant.

However,

Similar relationships

were obtained when the starter consumption for the entire

33
experiment was considered and these differences were like­
wise not significant.
Hay Consumption
Hay consumption by the individual calves of the
second experiment varied from 0 to 35 lbs. and did not ap­
pear to be related to starter consumption or weight gain
(Table 5)*

The differences among the various groups were

not significant in any of the periods.
Weight Gain
In general, the rate of gain by the various groups in
the first experiment tended to follow trends similar to
the rates of starter consumption (Table 4-).

In all periods

the calves that had received papain exhibited a lower rate
of gain than the calves that had not received papain and
the calves that had received malt diastase exhibited a
higher rate of gain than the calves that had not received
malt diastase.

These differences were not statistically

significant with the exceptions of the differences due to
malt diastase from 41 to 60 days and for the complete ex­
periment .
In the second experiment the use of pepsin produced
no significant effect on the rate of gain during the first
and last periods of the experiment.

However, from 21

34
through 40 days the rate of gain for the calves receiving
pepsin was significantly greater than the rate for the
control calves.

A large part of this difference was ac­

counted for by two calves in the control group that died
shortly after the end of this period.

The differences

for the total gain through 40 days and through 60 days
were not significant.
The calves that had received skim milk (Replacer 3)
exhibited a rate of gain approximately equal to the normal
growth rate for Holstein calves (Matthews and Fohrman,
1954).

The calves receiving the two vegetable replacers

exhibited the typical critical period.

From 3 to 20 days

the calves receiving Replacer 3 gained significantly more
weight than the other replacer groups.

From 21 to 40 days

the difference between Replacers 1 and 3 was not so pro­
nounced and was not significant.

The rate of gain by the

Replacer 2 group was significantly lower than the rates
for the other groups in this period.

When the data for

these periods was combined it was found that the rate of
gain for the Replacer 3 group was significantly greater
than the rates for the other groups.
The rates of gain for the three milk replacer groups
did not vary greatly in the last period and the differences
among the groups were not significant.

When the entire

35
experiment was considered the only significant difference
among the groups was the difference between Replacers 2
and 3 •
Mortality
In the first experiment two calves from the group that
had received papain alone and one calf from the group that
had received the combination of enzymes died after 40 days
of age.

The history of all cases was similar.

These

calves consumed little or no starter up to 40 days of age.
After the feeding of milk replacer was terminated at 40
days, the calves still refused to consume significant
amounts of starter and all efforts to get them to do so
failed.

The calves became extremely emaciated and died

within 3 to 10 days.

At necropsy the adrenal cortex of

all three calves showed hemorrhage and/or necrosis.

Mild

hyperemia and some hemorrhages were noted in the digestive
tract.

At the present time these lesions should be con­

sidered to be of a non-specific nature.

They were probably

caused by the long period of reduced nutrient intake pre­
ceding death (Follis, 1948).
In the second experiment two calves fed Milk Replacer
2 without pepsin died under circumstances similar to those
in the first experiment.
lesions were found.

However, at necropsy no gross

DISCUSSION
Most of the increased gain by the calves that had re­
ceived malt diastase occured after 40 days of age when the
calves no longer received the enzymatically treated millc
replacer.

This period of increased gain coincided with a

period of significantly increased starter intake.

»Vhile

it is possible that the increased starter consumption and
weight gain was due to some indirect effect of the use of
malt diastase, this possibility should be discounted until
there is more evidence to support it.

This is particularly

true in view of the studies which have been reported since
the completion of this experiment which indicate that malt
diastase could not be expected to be effective under con­
ditions of this experiment.
Ratcliff et al. (1937) found no growth response from
feeding animal diastase to calves.

However, in this study

the enzyme was not used until the calves were 28 days of
age at which time they should have been past the critical
period.

In studies more comparable to the present one,

Dollar and Porter (1937) found that predigestion of oat
flour and flaked corn with alpha- and beta-amylase did not
improve these feeds for calves through three weeks of age.
Chromatographic examination of the digestion mixture showed

- 36 -

37
that the primary end-product was maltose*
using the blood sugar response method,

From studies

Dollar and Porter

(1957) concluded that glucose and lactose are the only
carbohydrates that can be utilized by the young calf.
ability to utilize lactose decreased with age.

The

Ingestion

of sucrose, maltose, dextrin and soluble starch gave no
blood sugar response.

They also examined the amylolytic

activity of the pancreas and the maltase, lactase and su­
crase activities of the intestinal mucosa.

The results

gave good correlation with the blood sugar findings:

lac­

tase activity was high in the very young calf, but that of
amylase and maltase was very low; the activity of amylase
and maltase increased with age while that of lactase de­
creased; no sucrase activity was detected.

Larson et al.

(1956) found that maltose is readily hydrolysed and absorbed
when introduced posterior to the rumen of a nine-monthsold bovine, but they were unable to demonstrate the diges­
tion of starch.

It may be possible that the amylolytic

enzyme systems of the bovine never become well developed
because with a normally functioning rumen very little
starch is presented to the lower portions of the digestive
tract•
In studies on the carbohydrate utilization in the
young pig using the blood sugar response method, Dollar et
al. (1957) obtained results identical to those obtained in

38

calves.

In contrast, Cunningham and Brisson (1957a, 1957c)

reported that starch and maltose digestion in the two—dayold pig was practically complete.

However, as Dollar and

Porter (1957) have pointed out, "(digestibility studies)
are clearly misleading as they ignore the contribution of
the bacterial flora of the large gut to the degradation
of carbohydrate— a process that does not necessarily con­
tribute to the nutrition of the host."
It follows from the above findings that predigestion
of cereal starches by amylases must be complemented by
the action of maltase to complete the degradation to glu­
cose before they can be utilized by the young calf.

There­

fore, it would appear that the improved gain obtained from
the calves that had received malt diastase was not due to
the action of the enzyme.
Since cerelose is composed largely of glucose, its
use in the second experiment should have overcome the de­
ficiency of a utilizable carbohydrate in the diet.

The

lack of significant improvement in gain through 20 days of
age indicates that the lack of an available energy source
is not the sole deficiency in the corn-soybean flour milk
replacer.

If a milk replacer composed of an animal pro­

tein (i.e., casein) and corn had been used, it would have
been expected to provide some information on whether avail­
able protein is the sole deficiency.

However, Dollar and

39
Porter (1957) have employed this type of diet with unsatis­
factory results which indicate that both protein and energy
are lacking in the corn-soybean flour milk replacer.
The results from 21 to 40 days of age indicate that
the use of high levels of cerelose with plant protein is
unsatisfactory.

The large depression of weight gain by

the Replacer 2 can be largely accounted for by the de­
pressed starter intake compared to the Replacer 1 group.
While the consumption of starter by the calves receiving
Replacer 3 was somewhat depressed when compared to the
calves receiving Replacer 1, this depression was not impor­
tant because of the superior ingredients contained in the
milk replacer.

The recovery of appetite by the calves that

had received the high level of cerelose after the termina­
tion of milk replacer feeding lends further support to the
conclusion that high levels of cerelose depress appetite.
The glucostatic theory of the regulation of food in­
take (Mayer, 1953) provides a means of explaining this ap­
petite depression.

The glucose in Replacer 2 would be

readily absorbed and would cause elevated blood sugar levels
for some time after feeding.

In Replacer 3 a considerable

portion of the carbohydrate was lactose which does not give
as great an increase in blood sugar as glucose in calves
of this age.

Most of the carbohydrate in Replacer 1 was

starch which should give no increase in blood sugar

40
level*

It follows that one would expect the greatest

starter consumption by the calves that received Replacer 1,
an intermediate consumption by the calves that received
Replacer

and the least consumption by the calves that

received Replacer 2.

This is in agreement with the results

of the experiment.
The use of papain in the first experiment appears to
have produced deleterious effects.

This is indicated by

the depressed growth rate, the significantly depressed
starter consumption, and the higher mortality rate.

Fur­

ther support for this conclusion is provided by the recovery
of appetite and the relative improvement in rate of gain
by the surviving calves after the termination of papain
use.

These results are in agreement with those of /i/illiams

and Knodt (1952).
The cause of these deleterious effects cannot be de­
termined from the results of this experiment or from the
literature.

Lewis et al. (1955a) used papain at the rate

of 1% of the diet of young pigs without the production of
adverse effects.

Uo toxic effects were found in laboratory

animals when it was fed at much higher rates than were
used in this study (rfeiner, 1955).

It may be possible

that the deleterious effects are dependent upon the specie

^Results of blood glucose determinations which are
not included in this thesis are in agreement with this.

4-1

used.

Another possibility is that these effects were in

some way associated with the predigestion process*

In

this regard the results of the study by Cunningham and
Brisson (1957b) on the use of pepsin in baby pig diets is
of some interest*

They found that soybean protein predi­

gested with pepsin resulted in severe diarrhea and death
of 2-day-old pigs.

When the diet was not predigested there

was no adverse effect from the use of pepsin.
The effect of pepsin in the second experiment cannot
be evaluated with certainty on the basis of the available
data.

Although the rate of gain was increased signifi­

cantly from 21 to 40 days, most of this increase can be
attributed to the low rate of gain of the two calves in the
control group which died shortly after the end of this
period.

The rate of gain of these calves was the lowest

of all calves during this period.

In addition, starter

consumption by these calves was very low which would indi­
cate that they were not normal.

At the present time it is

not possible to attribute these deaths to the diet or to
suggest a mechanism by which pepsin could have prevented
the deaths.

In addition to these considerations, if pepsin

were effective in improving weight gains, one would have
expected as much or more improvement in the period preced­
ing 20 days of age.

This improvement was not obtained.

Therefore, the most reasonable conclusion at the present

42

time is that pepsin was not; effective in improving the rate
of gain*
Tliis conclusion is in agreement with the more recently
reported work with calves (Ratcliff et al., 1956) and pigs
(Alsmeyer e_t al^., 1957; Conrad and Beeson, 1957; Cunning­
ham and Brisson, 1957b; Hanson, 1957)*^

It is not in

agreement with the earlier pig work reported by the Iowa
group (Baker et al., 1956; Lewis et al.* 1955a; Lewis et
al., 1955b).
The low production of hydrochloric acid and the spe­
cificity of the various proteolytic enzymes provide two
possible explanations for the ineffectiveness of pepsin in
most studies.

In the case of the pig it has been reported

that the pH of the stomach contents was never lower than
3*5 and it was usually nearer 4.0 (Lewis et al., 1955a).
Since the pH optimum for pepsin activity is from 1.5 to
2.0 and since pepsin is relatively inactive at pH 4 (Fruton
and Simmonds, 1955)* it would not appear likely that pepsin
could improve food utilization in the young pig through
its proteolytic activity.
There is no information available concerning the
hydrochloric acid production by the abomasum of the young

^In addition to the negative results cited, a number
of other studies yielding essentially the same results
have come to the author's attention, either by personal
communication or by the popular press.

4-3
calf*

However, since a number of other species have

yielded essentially the same results as the pig it is
possible that a similar situation exists in the calf*
A second explanation is analogous to that discussed
in regard to the ineffectiveness of supplemental amylases*
From the specificity of the various proteolytic enzymes
(Fruton and Simmonds, 1953)» it follows that the endproducts of digestion by an individual enzyme would be a
mixture of peptides.

These peptides would not be utilized

unless the animal possessed the other enzymes necessary to
complete the degradation of the peptides to a utilizable
form.

Therefore, the use of supplemental pepsin could be

expected to be effective only if it or an enzyme of simi­
lar specificity was the only enzyme deficient.
Evidence supporting this possibility is lacking be­
cause the studies concerning enzyme production in the
young have not dealt with specific enzymes and because
little is known about the amino acid sequence of different
proteins.

Since the proteins of milk are almost completely

digested by the young animal, it must follow that the young
animal has a sufficient quantity of all enzymes required
for the complete degradation of milk proteins.

It is pos­

sible that some of the specific enzymes required for com­
plete degradation of plant proteins are different than
those required for milk proteins and that these enzymes

44

are deficient in the suckling animal.

In this regard it

has been shown that pancreatic and peptic digests of plant
proteins yield end-products which differ considerably from
those obtained from animal proteins (Birk and Bondi, 1956;
Bondi and Birk, 1956).

In the plant protein digests a

considerable quantity of peptides were precipitated by
trichloroacetic acid.

These peptides were not obtained

from animal protein digests.
In conclusion, it would appear from a considerable
amount of direct and circumstantial evidence that animals
at birth are deficient in many if not all of the digestive
enzymes except those required for the complete degradation
of the various components of milk.

Therefore, any attempt

to develop a feeding regime excluding milk would necessi­
tate the use of ingredients which can be completely de­
graded by the enzymes present or the provision of all of
the additional enzymes required for the degradation of the
given ingredient together with the necessary conditions for
the action of these enzymes (pH, etc.).

It would appear

that most studies including the present ones have not met
these conditions.

SUMMARY
Two experiments were carried out to determine the
effect of supplementing milk replacers with, enzymes.

In

the first experiment a corn-soybean flour milk replacer
was predigested with malt diastase, papain, and a combina­
tion of these enzymes.

Fifty-five pounds of milk were fed

through 20 days of age and 50.5 lh. of the treated milk re­
placer were fed through 40 days of age.
terminated at 60 days.

The experiment was

Decreased starter consumption and

growth from 20 to 40 days and an increased mortality rate
indicated that papain produced deleterious effects.

Malt

diastase produced no improvement in gain during the periods
it was used.
In the second experiment, soybean flour-corn, soybean
flour-cerelose, and skimmilk-cerelose milk replacers were
supplemented with pepsin but were not predigested.

All

other aspects of the experiment were similar to the first
experiment.

It was concluded that pepsin produced no im­

provement in growth with any of the replacers.

The skiiu-

milk-cerelose replacer produced significantly greater
weight gains than the other replacers during the first 20day period.

In the second 20-day period the soybean flour-

cerelose replacer produced significantly smaller gains than

- 45 -

46
the other replacers.

Starter consumption by this group

was also significantly depressed.
From considerations of the literature and the results
of the present experiments it appears that young animals
possess only those enzymes which are necessary for the de­
gradation of the components of milk.

Future use of enzymes

in diets replacing milk will require a greater considera­
tion of enzyme specificity and optimum conditions for en­
zyme activity than has been used in previous experiments.

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50
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51
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(Vet. Bui. 13:222. 1941)
Langendorf, 0. 1879* Ueber die Entstehung der Verdauungsfermente beim Embryo.
(The appearance of digestive
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Physiol. Abthlg. 3:93.
Langstein, L. and M. Soldin.
(1909).

1908.

Cited by Ibrahim

Larsen, H. J., G. E. Stoddard, N. L. Jacobson and R. S.
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Mayer, J. 1953. Genetic, traumatic and environmental
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Am. J. Physiol.

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Moriggia.

1876.

Cited by Langendorf (1879).

Noller, C. H. 1955- Corn-soybean flour rations in the
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Ph.D. thesis, Michigan
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52

Noller, C. H. , C. F. Huffman, G. M. Ward and C. W. Duncan,
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6th ed.

Iowa

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(Nutr. Abs. Rev. 21:110. 1951)

53
Seiner, 8, 1955* Papain. A Review of the Literature.
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6.
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Wolffhugel.
Zweifel.

1876.

1874-•

Cited by Langendorf (1879)*

Cited by Langendorf (1879)*

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