Note: Descriptions are shown in the official language in which they were submitted.
~0~9Z3
Ihis invention relates to a novel feed composition
suitable for feeding to young calves, and more particularly
to such a compositon which comprises a milk replacer.
It has been a common practice Eor many years to raise
calves by means other than allowing them to nurse their
mothers. Normally, a young ealf is complet:ely dependent
upon its mother,s milk for its nutritional requirements ~or
a period up to 42 days after its birth, until such time as
it is able to obtain all the nutrien-ts required for normal
and healthy growth from other sources, par-ticularly plant
matter. According to conventional methods of raising calves
apart from their mothers, ealves are fed after weaning (i.e.
after -the first three to four days of their lives) and for a
period up to and until -they are abou-t seven weeks old on a
so-ealled "millc replacer".
Ihe conventional millc replacers consist rnainly oE millc
produets in the form of slci~n milk and whey -together with
oils, fats, vitamins and traee minerals giving a balanced
blend of proteins, fat, carbohydrates, vitamins and minerals
meeting the ealves' nu-tritional requirements.
The major problem with eonventional millc replacers is
one oE cost. The high, and rising, costs of milk produc-ts
malce eonventional millc replaeers expensive, and result in
higher eosts Eor raising calves, which are reflec-ted in higher
meat prices -to the purchasers of meat produets and high cos-ts
o-E raising replaeement stoek.
The high costs assoeiated with the use of eonventional
milk replaeers containing milk protein have led to the
consideration oE the inclusion of non~milk protein in milk
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replacers. By blending together non-milk proteins~ e.g.
proteinaceous plant materials, in the appropriate amounts,
and with the addition of synthetic essential amino acids
if necessary, it is possible to obtain a mixture containing
the essential amino acid~ in proportions substantially
correspondiny to those o milk protein, and which could
theoretically be substituted, at least partially, for
milk protein in milk replacers. However, replacement of
any substantial amount of milk protein source by a non-
10~ milk protein source has~ to date, been unsuccessful.
For the early part of a calf's life, its rumen isinsufficeintly developed to allow the proper and re~uired
digestion of oods containing any substantial amount of
non-milk protein. In order to digest plant proteins ade-
quately, a normal flora of bacteria and protozoa needs to be
established in the calf's rumen~ ~hese bacteria and
protozoa then produce and secrete the enzymes which are
capable of digesting plant proteins. The development of
~his flora of bacteria and protozoa in the rumen is a
gradual one, and although the calf i9 usually producing
enzymes by about age 2 weeks~ the development is only fully
completed after the calf reaches the age o 6-8 weeks.
Notwithstanding these problems associated with the
use o~ non-milk protein in milk replacers, attempts have
been made to reduce the costs of conventional milk replacers
by using a variety of products as substitutes for a portion
of the milk protein in the milk replacer. while some
manufacturer~ of milk replacers containing substantial
amounts o non-milk protein claim e~ficienk utilization of
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of the non-milk protein, it is generally considered that the
better quality milk replacers are those consisting entirely oE milk
protein. The use of low-quality milk replacers, having a higher
proportion of non~milk protein, is felt to have contributed to
: .
the many nutritional, health and environmental problems associated
with the raising of calves, and the resulting deaths of calves
represent a serious financial loss to calf raisers. Thus while
some initial savings on the cost of the milk replacer may be
obtained by replacing some of the milk protein by non-milk
protein, these savings are often e~ualled or exceeded by the
losses occurring at the farm level when these product~ are Eed
to young calves.
We have now found that young calves may be raised on
a milk replacer containing non-milk protein without the aore-
mentio~ed nutritional, health and environmental problems resulting~
and without unacceptable digestive disturbances occurring, if the
feed is supplemented with a protease material which i~ an extra-
cellular metabolite of a Sora qLium sp.(isolate 495).
The said ~s~}~yl__m sp.(isolate 495) is maintained in
and is available rom the Chemistry and Biolo~y Research Insti~u~e,
Agriculture - Canada (ormerly known as the Microbiology Research
Institute, Canada Department of Agricultuxe), Oktawa. It is
described in United States Patent Mo. 3,515,641 dated June 2, 1970
assigned to Canadian Patents and Development Limited~ to which
referenc~ should be made for details of procedures for culturing
and growing the bacterium. Details o~ compositions o:E ~uitable ..
growth media and o other growth conditions are disclosl3d in
Example 1 of the pa~ent. The protease material i~ present in
~7~g~3
the liquid portion of the fermentation broth.
As a result of taxonomic studies conducted on the ~ -
said bacterium, we believe it should more proparly be
classified as hYSobaCter enzymoqenes. However, since such
is the name under which the bacterium is available and
under which it is discussed in the literatura, we shall
refer to it herein as a Soranqium sp.(isolate 495) for
the sake of clarity and to avoid confusion.
In the preferred practice o~ the present invention,
we use the whole fermentation broth, or we separate the
liquid portion from the solids portion using conventionaL
methods of separation ~uch a~ centri~ugation or il~ration,
and the supernatant or liquid portlon can then be concon-
trated under vacuum and dried by ~reeze-drying or spray-
drying to yield the protease material in concentrated
solid foxm.
As described in the a~ove-mentioned paten~, two distinct
enzymes can be isolated from the fermentation broth, and
the trivial name3 a- and ~-lytic protease have been assigned
20 to them. These two en~ymes are reasponsible ~or mo~t of
the proteolytic activity which is believed to account or
the beneficial result~ obtained with our feed supplement,
but since entirely satis~actory results can be obtainecl
employing a crude mixture of the enzymes which are present
in the liquid portion of the fermantation brokh, t~ere i~ no
need to isolate individual enzymes for the purposes of the
present invention. Further, the whole fermentation broth
contains many valuable nutrients, so there is l:ittle
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advantage in separating the protease mater.ial in the lic~id
portion from the cellular portion of the broth.
Empl~ying our protease material feed supplement, we
have found that calves can be raised satisfactorily on
milk replacers containing hi~h proportions of non-milk
protein. By way of example, the content of non-milk protein
may be in excess o 50% of the total protein in the milk
replacer. The formulation of milk replacers containing
substantial contents of mon-milk protein, is well within
the capabilities of those skilled in the art, and forms
no part o~ the essence of our invention. It is necessary
to note merely that n~ilk protein contains all the amino
acids required by the calf and it is desirable that the
non-milk protein milk replacer should contc~n at least the
m.inimum amounts of the amino acids believed necessary for
adequate nutrition in appropriately balanced proportions.
It can be mentioned by way o~ example that milk replacers
can be formulated from blends of soy protein meal and corn
gluten meal, which latter has a high content of the aromatic
amino acids and valine tha* are lacking from soy protein,
together with additions of methionine~
In the practice of the present invention, the protease
material feed supplement and the milk replacer may be mixed
with warm water and fed from a pail or a nipple, generally ~:
following the conventional procedures ~or raising calves
on milk replacers. Conventional feed additives, including
vitamins, trace minerals and antibiotics, may be included
in the ~eed composition.
Any amount o~ the protease material addecl to a mi.lk
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replacer containing non-milk pro-tein ~ill assist, to some
exkent, in the digestion of the non-milk ~rotein. More-
over, a large excess of protease material added to -the milk
replacer would not be harmful to the calf) but would merely
be wasteful, as more oE the protease material would be used
-than would be required to enable the calf to fully digest
the non-milk protein.
The optimal amount of protease material to be added
to the milk replacer is dependent upon several Eactors,
including the nature and composition of the milk replacer,
the proportion of mills protein to non-milk protein in the
milk replacer, and the state of development of the digestive
sy~tem of the indiviclual calves to which the milk replacer
is to he ~ed.
~ n estimate o the minimum amount of the protease
material requlred can be made in vitro by determining the
arnount of protease material required to hydrolyze a given
amount of the milk replacer in the laboratory. EIowever, the ~
estimate has to take into accounk the fact that not all ;
the enzyme material will be utilised efficiently, and that
some will be destroyed.
In practice~ we prefer to employ a somewhat yreater
~uantity of protease material than would optimally seem to ~;
be re~uired; a substantial increase in the amount of protease
material present would not substan-tially reduce the savings
realized through the use of a non-milk protein in the
preparation of the milk replacer.
~ enerally, we have found that satisfactory results can
be obtained when the protease material is present in an
~L~7~9Z3
amount exhibiting a protease activity in the range 500 to
3,000 D.U. (Delft Units) per gram of non-milk protein in the
milk replacer, and more preferably about 2,000 D.U. per gram
of non-milk protein in the milk replacer.
A Delft Unit is an arbitrary unit used to indicate
the protease activity of an enzyme preparation, and is
determined by following the assay procedure described in the
Maxatase brochure available from Royal Netherlands
Fermentation Industries, P.O. Box 1, Delft, Holland. Reference
to this bxochure should be made for the full details.
Briefly, the assay procedure relies on measuring the activity
of the enzyme preparation in digesting a standardised casein
substrate.
The protease material feed supplement may conveniently
be Eormulated as a unit dosage containing an amount of the
protease material exhibiting an appropriate protease activity,
which can then be fed to the calves along with the milk
replacer.
Advantageously, the unit dosage will contain protease
material in an amount exhibiting a protease activity of from
10,000 D.U. to 200,000 D.U. At the time when the protease
material supplement has the greatest influence of the calf's
digestion o non-milk protein, i.e. during the irst two
weeks of the calf's life, the calf will typically be
receiving about 500 g of milk replacer per day, with a
protein content of about 20%, in two equal daily feedings
of 250g. With a non-milk protein content of at least 50
in the milk replacer, it would be appropriate to employ
a dosage e~hibiting at least 12,500 D.U., preferably about
~ trade ~n~r,~!
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50,000 D.U. protease activity to be added to each -feeding.
With higher non-milk protein contents, dosages of as much
as 2007 000 D.U. may be used.
The following non-limitin~ Examples illustrate the
practice of the present invention.
EXAMPhE
..... _ _
Thirty-two Holstein bull cc~es, purchased at
approximately 6 days o~ age, were randomly allotted Oll the
basis of initial weight and age to a control group and two
experimental groups~ Two experimental replacers, with 50%
(50~P) and 80~ (80%P) of the protein in the milk replacer
supplied by plant (P) protein, were Eormulated and compared
to a control all-milk protein replacer (O~P). The
composition oE khese milk replacers is shown in Table 1, where-
in~ as throughout this speci~ication, all parts and per-
centages are by weight.
Table 1
~ O~P 50~0P 80%P
20 _ _ _ __ __
Ingredients (~0):
Dried skim milk 40 22.5 11.5
Spray dried whey 10 27.5 13.5
Dried buttermilk 15 - -
Tallow & cocoanut oil (1) 21 21 21
Dextrose & corn starch (2) 13 13.9 28.4
Promine F (3) - 8.5 16.0
Corn gluten meal - 5.5
Vitamins & Minerals (4)
Methionine - 0~1 0.1
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Table 1 cont
.
~/P 50~0P 80~P
Composition (~ dry matter basis~
Pxotein 20.9 22.4 25.4
Fat 21. 021~. O 21~ 0
Ca 1.0 1.0 1.0
- Phosphorus 0.750.75 0.75
(1) Mixed in 3:1 proportion. Fancy grade tallow, supplied by
St. Lawrence Rendering Co., Montreal, Que. Unre~ined
cocoanut oil supplied by Drew Brown Ltd., Montreal, Que.
(2) Mixed in 3:1 proportion. Both supplied by Canada Starch
Ltd., Cardinal, ont.
(3) Isolated soy protein (90~) supplied by Central Soya Co.,
Inc., Chicago, Illinois.
(4) Commercial mix supplied by Delmar Chemical Ltdo ~ Montreal,
Que.
- A ~eed supplemenk containing protease material was
prepared by growing Soranqlum sp.(isolate ~95) under the
conditions described in Example 1 (a) o~ United State~ patent
3,515,641. The whole ermentation broth was then concentrated
~y evaporation, and sufficient of the concentrated broth, ~`
~s determined by assay o~ the D~U. per gram9 was blended with
two lots of soybean meal carrier to form two mixtures, one
of higher and one of lower proteaqe material content, which
were then divided into 14 g. dosage unit3. The dosage units
o highex protease content possessed 100,000 D~U., and
those o~ lower protease content possessed 50,000 D.U.
_ g _ .
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7~9~3
The calves were fed the milk replacers allotted to th~m,
the milk replacers being mixed wi-th warm water and being
ed rom a pail or through a nipple. Each calf was given
two equal daily feeds of its allotted milk replacer. The
amounts of milk replacer and, of the water to dilute the
milk replacer fed to each calf per day were as shown in
table 2.
Tab
_ __ Week ~ _ Total
Components 1st 2nd 3rd 4th 5th 6th
Milk
Replacer (g) 500 700 900 9 9 625 31,675
Water (litres) 4 l~.5 s.o 6.o 6.o 6.o 220.5
In the case o the ~irst expe~imental group which
received the 50~ P millc replacer, one 14g dosage unit of
the 50, ooo D.U. protease material-containing feed supplement
was blended into the milk replacer before feeding, so that
20 each calf in this group received protease material o~ :
100,000 D.U. daily.
The calves in the second Pxperime~ntal group each received
one l~g dosage unit o~ ~he higher protease content with
each -feeding of the 80~P milk replacer, so.that they received
a total of 200~000 D.U. daily.
~ o protease-containing feed supplement was offered -to
the calves in the first, or control~ group.
For the first three days, each of the calves in all
three groups were injected daily with a conventional. commer-
cially available vitamin preparation and were of~ere!d 50g
-- 10 --
~ 7~9~3of electroyltes (Na Cl, K Cl and ~a HC ~) mixed with dextrose
as a carrier.
Calf s-tarter (composed of grouned corn, barley, wheat
bran, soybean meal molasses and vitamins and minerals) with
a crude protein content of 18~o~ timothy/trefoil hay, and
fresh water were offered to all calves as free choice from
day 1.
Attention was paid to balancing the proportions o~ the
essential amino acids in the 50%P and 80~oP experimental milk
replacèrs, so as to obtain, as nearly as economically feasible~
balanced proportions of amino acids substantially corresponding
to those of milk protein.
The essential amino acid contents of the milk replacers
employed i.n the presant Example are shown .in TablQ 3, as
well as the amino acid contents which are contributed by 28 g.
of the protease materia~-containing feed supplement.
: .... . . , ~ .
. .
7~323
o o~ o
~ ~ N N N
"
~ O 1` ~ ~1
.,~ ~ ~n N 1~
Id ~ O_i O a)
_I ,1 ,1
O N ~ C~~a
~ O ~ 1 ';P
.~.1 O~1 ~I O
E~ ~ ~ .~ :
1~ ~
>~ O ~ O
a~ D .~ '.
O ~Id ,~d' o ~ ,
l~ ~:S ': " .,
~ ~ O ~C~ ~
1~1 ~I ~j ~ )
~ ~'V ~ O _l O ~ '.
g ~1 '
O 0 m
.~ o o o 1~ ~ ..
U~ .'
... , .
.~ l ~ .
U o o o ~ _~ ~q . .
id h ~ ~ ,~ ~a
o ~; o o o o :~
.~ E~ ~ ,1 ,~ ,~ ~ '' .
~ ~ ,~ .... .
~ ~u~ I~
~1 ~qo ~ ~1
~ ~. ~ . .
~ C~~ ~ ~ o ~ ' .
oq ~ . ~ ~ .
~ 1
O
a a) ~ ,
a ~ 9 o .
~ S
~ o ~ o a~
~
~ . . . .
.~ ~ ~ O .~
~ ~ ~J
.~ r~ 8
O ~ O ~ O
e ~ o ~1 . . . .
~ o o ~, ~ ~ ~P ~ ~1
h ~ O ~ O ~1 1~ ~i
~I P; t~ ~`I N ~ 1
O rl ~rl ~rl
0 ~ C: ~'' ''
~U ~d ,~ OU~rl tq ,.
~1 _1 ~ ~1 ,., ~ .
~ ~ p~ o ~
~ 0 ~ o o c~ ...:
E~ ~ O In GO ~ ~ ~ ~ ~ ': ''
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~0719;~:3
As will be seen from Table l~ in addition to methionine,
corn gluten meal was employed to supplement isolated soybean
protein as it has a relatively hlgh content of aromatic amino
acids and valine which are relatively lacking in soybean
protein.
From Table 4 it will b~ seen that except for lysine and
valine, the 8 remaining essential amino acids were ofered
to the experimental calves -fed 50~P and 800~P in a higher
quantity than for the control calves fed O~P. In the ~irs-t
week, when 500 g o~ replacer was fed per cal per day (~28 g
o soybean carrier admixed with pro-tease material or
experimental calves), the lysine de~ici-t for calves fed
50~P was 0.39 g/head (4.6%) and ~or calves ~ed 80~P, 0.32
g/head (3.8~). The corresponding deicit for valine was
o.56 g (8.8~) and 0.39 g (6.1~) respectively. ~hen the
milk replacer consumption peaked with 900 g consumed/day/cal
(~28 g of soybean carrier admixed with protease material for
experimental calves), the lysine deficit was 1~33 g (8.7
and 1.2 g/head (7.9~) respectively and the val:ine deficit
was 1.25 g. (10.9~) and o.96 g/heacl (8.l~) respectively.
The eed intake was recoraed daily and calves were
weighed weekly.
All calves were weaned abruptly ater 6 weeks and kept
or an additional 2 week period on starter diet and hay.
The peroxmance of the calves up to 6 weeks of age
(weaning) is summarised in Table 4. - -
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Table ~
Characteristics O~P 50%P 80~P
.. . .. _ ~ ._ .. _ . . , . _ _ ~ __-- . . :; No. of calves 10 10 12
Ave. daily gain ~g) 617.9 639.2 710.2
~ 7
Milk replacex con-
sumption/kg of gain
(kg) 1.22 1.18 1~06
(87~o) . :
Ave. consumption of
18~ crude protein
calf starter/head/ 399 341 508
day (g) t127.3%)
Protein consumption/ 379.8 374.0 403.1
kg of gain (g) ( 106 .13~)
Plant protein o~
total protein (~) 32.5 64.2 85.8
E'eed cost/kg
o~ gain (c) 97.5 64 .1 51. 3
Age at which calves
consumed more than
500 g calf starter/
day for 3 consecutive
days 34 days 36 days 22 days
(1) ~alues in brackets relate to O~P as lOO~o.
From Table L~ it will be seen that the calves receiving
the 50~P and 80~P diets had in fact slightly higher average
daily gains than thosè fed on the O~P replacer.
Calf starter consumption per day up to weaning was
399 g ~or calves fed O~P, as agains-t 341 g and 508 g for
calves fed 50~P and 80~P respectively, the last value being
significantly greater. Calves fed 80~P consumed more than
500 g o starter/head~day for 3 consecutive days after 22
days on experiment. O~P calves consumed this guantit:y after
31~ da~s and 50~P after 36 days on experiment. Higher and ~;
:~Q~ 3
earlier intake of solid food by 80~oP calves was influenced
by earlier involvement of the rumen in ~ood digestion as
indicated by regular rumination in the course of the third
experimental week.
Hay consumption/calf/day up to weaning was 43.1~ g for
O~P, as against 65.l g and 61.5 g respectively for 50~P
and 80,bP. Again, more than 40~ higher consumption of hay
in experimental groups suggests a better functioning rumen.
Cumulative consumption of protein (including starter and
hay) /kg of gain for the same period was not significantly
different across the whole group. of the total protein
consumed, 32.5~ was of plant origin in O~P versus 64.2
and 85.8~ respectively in 50~P and 80~P groups~
No adverse e~ect was observed on the consistency o~ the
feces in the calves in the 50~0P and 80~P groups. Forty-nine
occurrences of abnormal feces were observed with the O~P
group as compared with thirty-eight occurrences in the 50~P
group and fifteen occurrences in the 80~P group. It was
concluded that no serious digestive disturbances occurred
in the calves in 50~P and 80~P groups.
EXAMPLE 2
Three groups of calves were raised up to age eight
weeks. Apart from points of dif~erence noted below, the
procedures of Example l were adopted. The first group -~
(5 calves) was ed on the O~P replacer of Example l and the
second and third groups (5 and 6 calves, respectively) received
the 80~P replacer. only the second group recei~ed a protease
material supplement in accordance with this invention.
In this Example, the whole fermentation broth~ obtained
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~L~71~Z3
as described in Example l, was concentrated by ~vaporation,
then dialyzed and reeze-dried~ The calves in the second
group received a dosage of lO0,000 D.U. freeze-dried material
with each of their twice-daily feedinys, providing them
with a daily total of 200,000 D~Ur of the protease material.
As in Example l, all ca~ves were weaned abruptly after
6 weeks and kept for an additional 2 week: period on starter
diet and hay.
At the end of the trial~ it was fourld that the average
daily gain was 480.3 g for calves in the first group, 492.8 g
Eor the second group and 450.0 g for the third group. The cal
starter consumption/calf/day was ~85.9 g for the Eirst
group, 573.8 for the second group and 560.o for the third
group.
With the calves in the third group there was a higher
occurrence of abnormality in the consistency of the feces
than in the calves in the second group (65 occurrences as
against 56).
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