U.S. patent application number 09/862389 was filed with the patent office on 2002-05-23 for procedure for preparing a food additive, and an additive and its use.
Invention is credited to Apajalahti, Juha, Jatila, Hanna, Jukola, Elias, Lauraeus, Marko, Nurminen, Paivi, Virkki, Markku, Vuorenmaa, Juhani.
Application Number | 20020061345 09/862389 |
Document ID | / |
Family ID | 26160285 |
Filed Date | 2002-05-23 |
United States Patent
Application |
20020061345 |
Kind Code |
A1 |
Vuorenmaa, Juhani ; et
al. |
May 23, 2002 |
PROCEDURE FOR PREPARING A FOOD ADDITIVE, AND AN ADDITIVE AND ITS
USE
Abstract
The invention relates to a procedure for preparing a food
additive, in which a brewing yeast raw material containing
oligosaccharides and/or polysaccharides is filtered and treated
hydrolytically so that the cell wall structure is opened.
Furthermore, the invention relates to a food additive prepared by
hydrolytically treating a filtered brewing yeast raw material
containing oligosaccharides and/or polysaccharides so that the cell
structure is opened. Moreover, the invention relates to the use of
the food additive in question for the prevention of gastric
disorders and intestinal diseases, and to a preparation containing
such additive.
Inventors: |
Vuorenmaa, Juhani; (Tampere,
FI) ; Virkki, Markku; (Espoo, FI) ; Jukola,
Elias; (Helsinki, FI) ; Lauraeus, Marko;
(Helsinki, FI) ; Jatila, Hanna; (Helsinki, FI)
; Apajalahti, Juha; (Helsinki, FI) ; Nurminen,
Paivi; (Masala, FI) |
Correspondence
Address: |
ABELMAN FRAYNE & SCHWAB
Attorneys at Law
150 East 42nd Street
New York
NY
10017
US
|
Family ID: |
26160285 |
Appl. No.: |
09/862389 |
Filed: |
May 21, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09862389 |
May 21, 2001 |
|
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09331708 |
Aug 18, 1999 |
|
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Current U.S.
Class: |
426/62 ;
426/807 |
Current CPC
Class: |
A23K 10/24 20160501;
A23L 33/21 20160801; Y02P 60/87 20151101; A23K 10/14 20160501; A23K
10/32 20160501; A23K 10/10 20160501; Y02P 60/877 20151101; A23K
10/37 20160501; Y10S 426/807 20130101 |
Class at
Publication: |
426/62 ;
426/807 |
International
Class: |
A23J 001/18; A23K
001/18 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 23, 1996 |
FI |
965192 |
May 14, 2001 |
FI |
N/A |
Claims
What we claim is:
1. A process for preparing a feed additive for use in the
prevention of gastric disorders, intestinal diseases and the
promotion of growth in animals, which comprises filtering a brewing
yeast raw material and treating the filtered brewing yeast raw
material hydrolytically to open the cell wall structure to increase
the amount of free oligosaccharides and free polysaccharides and
oligosaccharides and polysaccharides on the surface of the cell
wall.
2. The process as defined in claim 1, wherein the brewing yeast is
filtered mechanically.
3. The process as defined in claim 1, wherein the dry matter of the
filtered brewing yeast is at least 15 w-%.
4. The process as defined in claim 1, wherein the dry matter of the
filtered brewing yeast is 18-20 w-%.
5. The process as defined in claim 1, wherein the filtered raw
material is treated with an acid and/or an alkali.
6. The process as defined in claim 1, wherein the filtered raw
material is treated enzymatically.
7. The process as defined in claim 1, wherein the hydrolysis
product obtained in the hydrolysis is non-fractionated.
8. The process as defined in claim 1, wherein the raw material is
further treated mechanically, hydrostatically, pneumatically,
thermally or combinations thereof, whereby the cell structure is
broken up.
9. The process as defined in claim 1, wherein the product is
treated with a detergent.
10. The process as defined in claim 1, wherein the oligosaccharide
and/or polysaccharide product obtained is added to feed in an
amount of about 0.05 to about 1.5 w-%, calculated in terms of dry
matter.
11. The process as defined in claim 10, wherein the oligosaccharide
and/or polysaccharide product obtained is added to feed in an
amount of about 0.1 to about 1 w-%, calculated in terms of dry
matter.
12. A feed additive for the prevention of intestinal diseases and
promotion of growth, prepared in accordance with the process of
claim 1.
13. The additive as defined in claim 12, wherein the raw material
has been filtered mechanically.
14. The additive as defined in claim 12, wherein the additive has
been prepared by treating the filtered raw material with an acid
and/or an alkali.
15. The additive as defined in claim 12, wherein the additive has
been prepared by treating the filtered raw material
enzymatically.
16. The additive as defined in claim 12, for use in conjunction
with feed for animals and humans for the prevention of gastric
disorders and intestinal diseases and for the promotion of
growth.
17. The additive as defined in claim 16, wherein the amount of
additive used is about 0.05 to about 1.5 w-% of the amount of the
feed.
18. Use of the additive as defined in claim 12 wherein the amount
of the additive used is 0.05-1.5 w-% of the amount of feed.
19. Use of the additive as defined in claim 17 for animals.
20. Use of the additive as defined in claim 17 for humans.
21. A preparation containing a feed additive, designed for the
prevention of intestinal diseases and for the promotion of growth
and intended to be given to an animal to be fed, wherein the
preparation contains an additive according to claim 12 in an amount
of 0.01-0.6 g/kg, calculated from the daily ration of feed stuff as
dry matter per kilogram of living weight.
22. The preparation as defined in claim 21, wherein the amount of
additive contained in the preparation is about 0.05 to about 1.5
w-% of the daily ration of feed stuff.
Description
[0001] The present invention relates to a procedure for preparing a
food additive as defined in the preamble of claim 1. Moreover, the
invention relates to a food additive, its use and a preparation
containing the additive.
[0002] A balance of intestinal microbes is a condition for the
health and well-being of animals and for their productivity.
Disturbances of this balance appear as diarrhoea and other
intestinal health problems and may even lead to death.
[0003] The commonest nutritional method used to avoid the effects
of noxious microbes on the health of single-stomach animals is to
add various antibiotic and chemotherapeutic substances inhibiting
microbial growth to the fodder used to feed the animal. To maintain
intestinal balance and to avoid the use of antibiotics, it is also
possible to use fodders containing added probiotic products, such
as various microbes, acids and yeasts.
[0004] Another method used to prevent intestinal diseases is to
inhibit the adherence of noxious microbes on the wall of the
intestine. A method used to achieve this is to add to the fodder
mixtures various oligosaccharides, which adhere to the receptors on
the intestinal wall or to microbial receptors, thus preventing
noxious microbes from settling on the intestinal wall. Moreover, it
has been established that certain oligosaccharides, e.g.
fructo-oligosaccharides promote the growth of bifidomicrobes
beneficial to animals.
[0005] A problem with the use of antibiotics is that it promotes
the development of microbial strains immune to antibiotics and thus
leads to health risks to humans. A problem with probiotic products
is that they have a variable and generally low efficacy and are
quite expensive to use. Likewise, a problem with fodders containing
pure oligosaccharides is that they have a variable and generally
low efficacy in preventing intestinal diseases. In addition, the
price of pure oligosaccharides is high.
[0006] The object of the present invention is to eliminate the
problems described above.
[0007] A specific object of the present invention is to disclose a
procedure for preparing a food additive having a more efficient
effect on intestinal microbes that promotes the health and/or
growth of animals.
[0008] A further object of the invention is to disclose a food
additive that makes it possible to reduce more effectively
intestinal diseases in animals.
[0009] A further object of the invention is to disclose the use of
a new additive prepared according to the present invention and a
preparation containing such an additive.
[0010] As for the features characteristic of the invention,
reference is made to the claims.
[0011] In the procedure of the invention for preparing a food
additive, a filtered brewing yeast raw material containing
oligosaccharides and/or polysaccharides is treated so that its cell
structure is changed and the amount of free oligosaccharides and/or
polysaccharides and/or the amount of oligosaccharides and/or
polysaccharides on the surface of the cell structures are/is
increased, i.e. e.g. the cell structure breaks up, to release the
oligosaccharides and/or polysaccharides for use to prevent
intestinal diseases. This treatment can also be used to release
said components.
[0012] The invention also discloses products prepared by the method
of the invention, their use and preparations containing additives
according to the claims.
[0013] In a brewing yeast raw material containing oligosaccharides
and/or polysaccharides, the oligosaccharides and/or polysaccharides
are fixedly bound to the cell walls and other insoluble structures
in the raw material. Investigations carried out in conjunction with
the present invention revealed that adding such raw material
directly to fodder does not produce any favourable effects of
oligosaccharides; the animal (and human) digestive system is
generally unable to decompose e.g. the cell wall of a yeast cell
and release the desired oligosaccharides and/or polysaccharides. It
was further established in the investigations that by treating the
raw materials so that the amount of free oligosaccharides and/or
polysaccharides and/or the amount of oligosaccharides and/or
polysaccharides on the surface of the cell structures are/is
increased, e.g. the cell structure of the raw material breaks up, a
product is obtained that, when given to an animal together with
fodder, substantially reduces intestinal diseases in the
animal.
[0014] The mechanism of action of the product obtained in
preventing intestinal diseases has not been fully elucidated in the
investigations carried out, so it is based on various assumptions.
According to one model, using the products obtained by the present
method in conjunction with fodders inhibits microbial adherence to
the intestine, in other words, the oligosaccharides and/or
polysaccharides and/or other substances released in conjunction
with the break-up of the cell structure of the raw material are
assumed to act as analogues to the receptors of noxious microbes,
such as E.coli, in the intestine and to inhibit the ability of the
microbes to attach to the wall of the intestine.
[0015] According to another model, the products obtained via
break-up of the raw material cell structure affect the growth of
noxious microbes in both the small and the large intestines, in
other words, beneficial intestinal microbes, such as lactic acid
bacteria and bifidobacteria, are able to utilise the
oligosaccharides and/or polysaccharides for their nutrition whereas
noxious microbes, such as E.coli and salmonella, are not. This
favours the growth of beneficial microbes at the expense of noxious
ones.
[0016] According to a third model, the decomposition products
obtained via hydrolytic treatment of the raw material are assumed
to have an effect on the immune reaction of the animal, i.e.
certain raw material components, e.g. saccharine structures
containing phosphor in yeast may improve the animal's immune
reaction, thereby inhibiting intestinal diseases. Further the
hydrolytic treatment of filtered yeast effects on the type and the
strength of immune response.
[0017] Further, the components formed via hydrolytic treatment of
the raw material may affect the adsorption of toxins; i.e. the
components may bind and neutralise microbial toxins, thus
inhibiting intestinal diseases. The assumed mechanisms of action
described above may also work in combination, inhibiting intestinal
diseases in animals.
[0018] Brewing yeast is produced as a side product of brewery
industry. Normally, a brewing yeast mixture is transferred after
the brewing to storage tanks in which the brewing yeast becomes
precipitated on the tank base. After this, the beer remaining on
the surface is separated. The dry matter of a brewing yeast
produced in this way usually varies between 7-13% by weight.
[0019] In the method in accordance with the invention, the brewing
yeast that became precipitated on the bottom of the tank is
filtered mechanically and/or pneumatically. The brewing yeast may
be filtered by any known filtering method. To prevent the filters,
e.g. filtering discs, from getting blocked up, a vibrator such as a
micro vibrator may be used at a high frequency and/or some other
corresponding technique which prevents the filter from getting
blocked up. The density of the filter is chosen based on the
particle size.
[0020] The dry matter of a brewing yeast filtered in accordance
with the invention is 15% by weight at the minimum, preferably
18-20% by weight.
[0021] In the filtered brewing yeast raw material used, the
oligosaccharides and/or polysaccharides are bound to the cell
structures of the raw material. oligo- and polysaccharides can be
released from the raw material by breaking up the cell structure of
the raw material hydrolytically using an acid and/or an alkali,
and/or enzymatically. Acids usable in the hydrolysis are e.g.
ordinary mineral acids, such as hydrochloric acid, sulphuric acid,
phosphoric acid, nitric acid, etc., as well as strong organic
acids, such as formic acid, acetic acid, propionic acid, etc. The
pH range used in acid hydrolysis may be below 4, e.g. about 2. In
alkali hydrolysis, the alkalis used may be e.g. ordinary alkaline
hydroxides such as sodium hydroxide, caustic potash etc., ammonium
hydroxide or other alkalis releasing oligosaccharides and/or
polysaccharides.
[0022] Among the enzymes usable in enzymatic hydrolysis are various
cellulolytic and proteolytic enzymes, e.g. cellulases, acid or
alkaline proteases, which may be selected according to the
properties of the raw material to be used. In hydrolysis of yeast,
e.g. Torula yeast, the culture solution of a derivative of Trametes
sanguinea can be used. Furthermore, other added enzymes, proteases,
ribonucleases and deaminases can be used in the hydrolysis. The
enzyme treatment can also be implemented using a combination of
several enzymes, simultaneously or in succession; e.g. protease
treatment or protease treatment followed by ribonuclease treatment
and deaminase treatment, in which process the protease breaks down
the RNA, and when the amino acids are released, the ribonuclease
breaks down the RNA into various nucleotides and the deaminase
converts the adenosine mononucleotide into inosine mononucleotide.
The protease treatment can be implemented using any known protease.
Generally, the procedure can be implemented using enzymes mentioned
in the specifications referred to below and/or other known enzymes
having the desired effect of breaking up the cell structure,
together and/or separately, e.g. as described in the specifications
referred to below.
[0023] In a hydrolysis, the yeast can be heated to a temperature of
over 40.degree. C., in autolysis and enzymatic hydrolysis e.g. to
40-65.degree. C. and in acid and alkalic hydrolysis e.g.
70-90.degree. C. The duration of the heating may vary depending on
the temperature, e.g. between 1-12 hours.
[0024] Both the soluble and the insoluble fraction obtained via
filtration and hydrolysis contain certain amounts of the desired
oligosaccharides and/or poly-saccharides. The soluble and insoluble
fraction can expressly be used e.g. in the production of a feed
stuff or foodstuff; on the other hand, the soluble fraction or the
insoluble fraction can be used.
[0025] Hydrolytic decomposition of yeasts is described in the
following patent specifications and applications: U.S. Pat. No.
3,914,450, U.S. Pat. No. 3,443,969, U.S. Pat. No. 5,288,509, EP 299
078, JP 57-219695 and PCT/FI/96/00326. These and other prior-art
methods can be used in conjunction with the present invention, the
usable fraction being expressly the fraction containing
oligosaccharides and/or polysaccharides or the non-fractionated
product obtained as such. Thus, specification PCT/FI96/00326
describes the recovery of flavours, such as nucleotides, peptides
and amino acids e.g. from brewing yeast and baking yeast, whereas
in the present invention it is primarily the other components that
are recovered, and, if desired, the flavours can be separated e.g.
as described in the specification referred to. On the other hand,
the separation of flavours can be omitted; in other words, part or
all of the flavours can be included in the product prepared
according to the invention.
[0026] Besides hydrolytic treatment of filtered brewing yeast, it
is also possible to apply a treatment of the raw material with a
detergent and/or a treatment that breaks up the cell structure of
the raw material, e.g. by subjecting the cell structure to a
mechanical, hydrostatic and/or pneumatic force, and/or to a heat
treatment. Moreover, it is possible to use combinations of the
above-mentioned methods, e.g. a treatment breaking up the cell
and/or a heat treatment combined with an enzymatic or other
hydrolytic treatment. If desired, the product obtained via
hydrolytic treatment and/or via a treatment breaking up the cell
and/or via heat treatment can be treated with a detergent to wash
it.
[0027] If desired, the food additive produced according to the
invention can be processed further, e.g. by fractionating or
concentrating the saccharine structures obtained in the treatment.
The further treatment, such as fractionation or concentration, can
be implemented by any method known in itself. The fractionated
and/or concentrated products obtained can be used as such for
fodder or food, or they can be mixed with feed stuffs and/or
foodstuffs known in themselves.
[0028] The product prepared by the method of the invention can be
added to a fodder or foodstuff as such, moisturised or dried, and
it can generally be treated as desired.
[0029] The food additive prepared by the method of the invention
can be used in fodders for single-stomach animals, e.g. pigs,
poultry, calves, fur animals such as foxes and minks, pets such as
dogs and cats, horses, especially foals, fish and so on, to prevent
intestinal diseases. The food additive can be used in fodders/foods
for single-stomach animals in amounts of approx. 0.05-1.5w-%,
preferably about 0.1-1 w-% of the total amount of fodder,
calculated in terms of dry matter and depending on the degree of
hydrolysation; the percentages have been calculated based on a
degree of hydrolysation of 50%; the percentages depend on the
degree of hydrolysation. The additive can be used together with
fodder/food or as such. The additive is preferably so used that the
amount of additive used is 0.1-0.6 g/kg, calculated from the daily
ration of foodstuff and/or feed stuff in terms of dry matter per
kilogram of the animal's living weight.
[0030] The food additive of the invention can also be used in food
for people, e.g. in food products for children or adults or as a
preparation served separately to promote health, to balance
intestinal microbes and to inhibit intestinal diseases.
[0031] The feed additive prepared by the method of the invention,
when added to fodder intended for animals, effectively inhibits the
growth of harmful micro-organisms and promotes the growth of
beneficial microbes. At the same time, the growth of the animals,
utilisation of fodder and the overall economy of production are
improved. The feed additive according to the invention is
approximately 30% more efficient in preventing the attachment and
the growth of noxious microbes than a product prepared from an
unfiltered brewing yeast. The invention improves the economy of the
production of the feed additive and the preparation according to
the invention. Further, the environmental emissions caused by the
production are reduced because the animal is able to utilise the
fodder more effectively. In addition, the use of the products of
the invention, i.e. organic feed products, in the fodder for
animals makes it possible to stop using antibiotics in fodder. The
risk for the development of microbial strains immune to antibiotics
is reduced and the health risks they cause for humans are also
reduced.
[0032] In the following, the invention will be described in detail
by the aid of embodiment examples by referring to the attached
drawings, in which
[0033] FIG. 1 illustrates the adherence of bacteria in the mucus of
an intestine treated using products prepared from yeast by the
method of the invention.
EXAMPLE 1
[0034] In a laboratory test, food additive was prepared from baking
yeast (PCT/FI96/00326). In the test, the effect of a processed
yeast fraction on the adherence of E.coli bacteria to the mucous
membranes in the intestine of a pig was tested using micro-titre
plates; the test is described in the publication Conway, P. L.,
(1990) Infection and Immunity, 58, 3178-3182. Presence of
K88-specific receptors in preine ileal mucus is age dependent.
[0035] It was established that the additive in a 1-% solution
inhibits microbial adherence by 70-90%, depending on the coli
strain. The results are shown in Table 1.
1 TABLE 1 E. coli strains Inhibition strain 1 77% strain 2 83%
strain 3 70% strain 4 82% strain 5 74% strain 6 90% strain 7
80%
EXAMPLE 2
[0036] In a laboratory test, food additive was prepared from dried
blood by treating it with a detergent enzyme.
[0037] It was established that the additive inhibits bacterial
adherence by 90-95%, depending on the coli strain. The results are
shown in Table 2.
2 TABLE 2 E. coli strains Inhibition strain 1 93% strain 2 strain 3
89% strain 4 94% strain 5 strain 6 96% strain 7 90%
EXAMPLE 3
[0038] In a laboratory test, food additive was prepared from sugar
beet cuts by acid hydrolysis. Inhibition of bacterial adherence was
determined as in Example 1. The results are shown in Table 3.
[0039] It was established that the additive inhibits bacterial
adherence by 92-96% (Table 3).
3 TABLE 3 E. coli strains Inhibition strain 1 92% strain 2 96%
EXAMPLE 4
[0040] Additive was prepared from larch by hydrostatic heat
treatment. Inhibition of bacterial adherence was determined as
above. The results are shown in Table 4.
[0041] It was established that the additive inhibits bacterial
adherence by 96-98%.
4 TABLE 4 E. coli strains Inhibition strain 1 96% strain 2 98%
EXAMPLE 5
[0042] In this test, four equal groups of pigs were fed with the
following fodders:
[0043] Group 1: basic fodder (reference)
[0044] Group 2: basic fodder+40 ppm Avilamysine
[0045] Group 3: basic fodder+product prepared from yeast by
hydrolytic treatment, in an amount of 0.5 w-% (of dry matter)
[0046] Group 4: basic fodder+product prepared from yeast by
hydrolytic treatment, in an amount of 1.0 w-% (of dry matter).
[0047] The results are shown in Table 5.
5TABLE 5 Effect of yeast addition on development of piglets Group 1
2 3 4 Yeast % 0 0 0.5 1.0 Avilamysine - + - - Piglets 72 72 72 72
Starting weight, kg 9.5 10.6 10.7 9.0 Final weight, kg 23.0 24.1
24.2 22.4 ADG, g/d 456 502 512 433 FCR kg fodder/kg 1.97 1.81 1.77
1.96 Weight increase
[0048] The analysed fodder composition did not differ from the
calculated composition for any one of the groups. Both Avilamysine
and the 0.5 w-% addition of yeast preparation increased the growth
and fodder consumption effectively as compared with the reference
group (group 1, Table 5). The yeast preparation and Avilamysine
were substantially equal in effectiveness. The 1 w-% addition of
yeast preparation had a slightly negative effect on the growth of
the piglets; it clearly reduced the fodder consumption, which may
have been the cause for the lower result. The test result indicates
that the amount of the yeast product in fodder/foodstuff may
preferably be under 1 w-%, e.g. up to 0.9 w-%.--The use of yeast,
e.g. brewing yeast as protein raw material, in fodders is known in
prior art. The amounts of yeast used are 2-10 w-% of the fodder,
and yeast has been used to replace other protein raw materials,
such as crushed soy, without any harmful effects on growth.
EXAMPLE 6
[0049] An amount of a product obtained from baking yeast by
hydrolytic treatment, was added to the fodder of growing piglets.
The fodder for the reference group contained Olaqvindox
chemotherapeutic substance, 50 mg/kg. In the fodder for the yeast
group, instead of Olaqvindox, yeast fraction was added in an amount
of 0.5%. The results are shown in Table 6.
[0050] The yeast fraction addition clearly reduced diarrhoea in the
piglets; the average diarrhoea index was 1.5 for the yeast group
and 2.5 for the Olaqvindox group. In addition, 100% of the farrows
in the olaqvindox group had to be treated with an antibiotic or
with zinc oxide because of diarrhoea. For the yeast group, the
corresponding need was 12.5%.
6 TABLE 6 Olaqvindox 50 Yeast frac- Group ppm tion 0.5% Pigs 79 87
Initial weight, kg 7.10 7.50 Final weight, kg 12.47 12.96
Additional growth, 255 261 g/day Fodder efficiency kg/kg 1.61 1.60
Diarrhoea index 2.5 1.5 Treatments for diar- 100 12.5 rhoea, % of
farrows Diarrhoea index graduation: 1 = normal faeces, 2 = loose
faeces, 3 = watery diarrhoea
EXAMPLE 7
[0051] An amount of a product obtained from baking yeast by
hydrolytic treatment, was added to the fodder of growing pigs. The
measurement was implemented as in the preceding example. The
results are shown in Table 7.
7 TABLE 7 Olagvindox Yeast fraction Group 50 ppm 0.5 % Number of
pigs 150 140 Initial weight, kg 21.7 21.3 Test days, d 33 33
Additional growth, g/day .sup. 777.sup.a .sup. 847.sup.a Fodder
efficiency kg/kg 2.07 1.87
EXAMPLE 8
[0052] An amount of a product obtained from baking yeast by
hydrolytic treatment, was added to the fodder of growing piglets to
investigate its effect on the growth and health of piglets and on
fodder utilisation. Each test group comprised 6.times.4 piglets.
The test groups were divided as shown in Table 8.
8 TABLE 8 Olaqvindox Avilamysine Additive 0 50 ppm 40 ppm Yeast
fraction + - + - + -
[0053] In the fodder for the yeast groups, additive, i.e. yeast
fraction, prepared from yeast by hydrolytic treatment was added in
an amount of 0.5%. The results are shown in Table 9.
[0054] The addition of yeast fraction somewhat improved the growth
of the piglets and the fodder utilisation (Table 9). The effect of
yeast is particularly evident in the case of fodders without
additives, in which the addition of the yeast fraction increased
pig growth to the same level as for fodders with additives.
9TABLE 9 Effect of yeast fraction on the growth of piglets and
fodder utilisation no addi- no addi- Addition of yeast tive tive
fraction - + - + Number of piglets 72 72 24 24 Initial weight, kg
13.42 13.62 13.50 13.50 Final weight, kg 23.73 24.80 22.38 24.55
Additional growth, 493 535 .sup. 429.sup.a .sup. 526.sup.b g/day
Fodder efficiency, 1.94 1.73 2.16 1.78 kg of fodder/kg of
additional growth .sup.a,b (p < 0.05)
EXAMPLE 9
[0055] In this test, yeast fractions were prepared for
microbiological tests. The raw materials used were baking yeast and
brewing yeast, which were treated with an acid, enzymatically or
autolysed with salt.
[0056] In the acid hydrolysis, the pH of the yeast suspensions was
maintained at the value 4.0 by using a strong HCl solution (10 h),
and the temperature was maintained at 60.degree. C. The next day,
the pH was lowered to the value 2.0 (11 h). Finally, the
temperature was raised to 68.degree. C. (12 h). The reaction
mixture obtained was neutralised (pH 6.2) and centrifuged (4000
rpm, 20 min). From the soluble (supernatant) fraction and the cell
residue, the dry matter content and adhesion were determined as in
Example 1. Table 10 presents the dry matter content values.
[0057] In the enzymatic hydrolysis, the yeast suspensions were
subjected to a heat treatment (95.degree. C. for about 10 min.),
whereupon they were transferred into a fermentor, pH 5.8,
temperature 65.degree. C. The proteolytic enzyme used was papain
(Promod 144 L). In the final enzymatic treatment with ribonuclease,
the nucleotides of RNA were split and deamizyme GMP was converted
into IMP. The reaction mixtures were centrifuged (4000 rpm, 20
min). From the soluble fraction and the cell residue, the dry
matter content and adhesion were determined as described above. The
dry matter content values are presented in Table 10.
[0058] In the autolysis, the yeast was autolysed in a fermentor,
temperature 50.degree. C., with 0.5% NaCl added, mixing speed 100
rpm and reaction time 24 h. The reaction mixture was centrifuged
(4000 rpm, 20 min). From the soluble fraction and the cell residue,
the dry matter content and adhesion were determined as described
above. The dry matter content values are presented in Table 10.
[0059] The brewing yeast used in this example was processed in the
same way as the baking yeast (above) except that it was centrifuged
(4000 rpm, 20 min.) before the processing to remove most of the
soluble beer components from it. After this, hydrolyses and
autolysis were carried out as described above.
10 TABLE 10 Sample Dry matter, w-% Brewing yeast 12.0 Baking yeast
19.3 Brewing yeast 19.0 Baking yeast, Autol., supern. 19.3 Brewing
yeast, Autol., supern. 9.4 Brewing yeast, Autol., total fraction
20.9 Baking yeast, Autol., total fraction 18.1 Brewing yeast,
Autol., cells 25.7 Baking yeast, Autol., cells 30.0 Baking yeast,
Acid hydr., total frac- 17.8 tion Brewing yeast, Acid hydr., total
frac- 18.7 tion Baking yeast, Acid hydr., cells 32.7 Brewing yeast,
Acid hydr., cells 24.4 Brewing yeast, Acid hydr., supern. 19.1
Baking yeast, Acid hydr., supern. 10.9 Baking yeast, Enz., cells
20.3 Brewing yeast, Enz., cells 18.5 Baking yeast, Enz., supern.
14.4 Brewing yeast, Enz., supern. 10.6
[0060] When the dry matter content (supernatant) of the yeast
extracts is considered, it can be seen that, in a comparison of
different process types, the dry matter content of the
enzymatically treated extract fractions is the highest dry matter
content value. Thus, it can be assumed that the dry matter yield
into yeast extract is highest and, conversely, that the dry matter
yield in the cell fraction is lowest. In each hydrolysis, the
extracts produced from baking yeast had a higher dry matter content
than brewing yeast (the source material dry matter content, too,
was by 0.5% higher for baking yeast than for brewing yeast). There
was no significant difference between the autolysate and the
acid-hydrolysed extract fraction. The dry matter content values for
the cell fractions corresponding to yeast extract confirm the dry
matter distribution of the enzymatically treated fractions to be in
line with what could be concluded about the dry matter content of
yeast extract, in other words, the dry matter content values of the
cell residue were correspondingly all lowest in the enzymatic
process.
[0061] When the dry matter distribution is calculated from the
extract (supernatant), which does not fully reflect the situation
as the cell residue still contains some soluble dry matter not
extracted, about 46% of the dry matter in the dry matter
distribution of the enzymatic process was in the yeast extract when
baking yeast was being processed. The corresponding value for
brewing yeast was about 28%. Accordingly, the extract yield will be
about 50% of the total dry matter. The yield values for brewing
yeast were clearly lower.
[0062] In acid hydrolysis the extract yield with baking yeast was
about 44% of the dry matter and in autolysis about 34% of the dry
matter. For brewing yeast, the corresponding figures were about 32%
(acid) and about 38% (autolysis).
EXAMPLE 10
[0063] A laboratory test was carried out to establish the ability
of processed baking yeast fractions to inhibit the adherence of
E.coli K88 bacteria to the mucus in a pig's small intestine. The
test procedure is described in Example 1. In this procedure, the
wells in a micro-titre plate are covered with mucus isolated from a
pig's intestine. Radioactively branded bacteria are added onto the
mucus either as such or together with the substance under
examination. The bacteria are incubated in the micro-titre wells
and non-adhering bacteria are washed away. The adhering bacteria
are loosened using a detergent and their number is calculated based
on their radioactivity.
[0064] Yeast was hydrolysed with enzyme and hydrochloric acid. The
enzyme used in the enzymatic hydrolysis was papain (Promod 144 L),
pH 5.8., temperature 65.degree. C. In the final enzymatic treatment
with ribonuclease, the nucleotides of RNA were split and deamizyne
GMP was converted into IMP. The reaction mixtures were centrifuged
(1400 rpm, 20 min). About 48% of the dry matter was in the yeast
extract.
[0065] In the acid hydrolysis, pH 2, temperature 68.degree. C., the
reaction mixture was centrifuged (4000 rpm, 20 min), total extract
yield about 50% of total dry matter.
[0066] In this test, fresh baking yeast and processed and
spray-dried baking yeast fractions were used as adherence
inhibitors: soluble and solid fraction of enzymatically decomposed
yeast, soluble and solid fraction of acid-hydrolysed yeast. The
concentration of all yeast fractions and fresh yeast in the
reaction mixture in the test was 0.16% (dry matter). The results
are shown in FIG. 1. For a bacterium added without yeast fraction,
the adherence to the mucus is represented by the value 100%.
EXAMPLE 11
[0067] In the test, a food additive in accordance with the
invention was prepared from a brewing yeast produced from beer
industry the concentration of dry matter of which was 9 w-%.
[0068] The brewing yeast was mechanically filtered by a fine filter
vibrating the filter discs by a micro vibrator at a high frequency.
The yeast was filtered to a concentration of dry matter of 18 w-%.
During the filtering, 3-4% of the dry matter was lost along with
the liquid, and the yield of the dry matter of the yeast was 50-60%
of the estimated amount.
[0069] The filtered brewing yeast was hydrolyzed with an acid. In
the hydrolyzing, the pH value of the yeast sludge was kept at a
value of 2-3 with a strong acid (4 h) and at a temperature of
70-85.degree. C. Then the pH value was increased to a value of 4-5
and the resulting product was cooled. The resulting end product may
be used as such or it may dried with known methods.
EXAMPLE 12
[0070] In the test there was a comparison of the effect of a
hydrolyzate as produced in example 11 and of the effect of a
hydrolyzate produced using unfiltered brewing yeast on the
attachment of E. coli bacteria on the mucous membrane of the
intestine of a piglet by micro titer discs; the test has been
described in the publication Conway, P. L., (1990) Infection and
Immunity, 58, 1378-3182. Presence of K88-specific receptors in
porcine ileal mucus is age dependent.
[0071] The results are shown in Table 11.
11TABLE 11 Concen- tration of the test mat- The attachment of E.
coli % ter in Hydroly- Filtered % the zate, un- Hydroly- from an
analysis filtered zate, fil- unfiltered % Control yeast tered yeast
yeast 100 0.16 21.4 16.2 76 0.08 45.6 32.4 71 0.016 82.3 60.7
74
[0072] From the results it was obvious that the food additive in
accordance with the invention was approximately 25-30% more
efficient in preventing the attachment of E.coli than a product
prepared from an unfiltered brewing yeast.
EXAMPLE 13
[0073] In the test, the effect of a yeast hydrolyzate as produced
in example 1 and a yeast hydrolyzate 2 according to the invention
was examined on the immunity of the intestine of rats. The immunity
was determined by measuring the concentration of IgA in the
gastrointestinal tract and by determining the proportional parts of
the immune cells in the tissue samples of the intestine. The
comparison definitions were made by a control and by betaglucan
products.
[0074] In each test, 6 rats were used. The tissue samples from the
duodenum and ileum were taken after 28 eight days of the beginning
of the feeding test. They were diluted and the IgA was determined
by using a new ELISA method, which is used to measure the immune
response at intestine level. The previous methods have been used to
measure the immunity indirectly by means of cellular cultures and
blood samples. The methods now used may be used to measure both the
cell-mediated and the antibody-mediated immunity in the intestine
and thereby directly measure the inhibiting effect of different
products against the intestinal diseases. In the definitions,
monoclonal antibodies specific to the immune cells of rats were
used.
[0075] The product according to the invention (0.3%) and betaglucan
did not have a major effect on the concentration of IgA in the
gastrointestinal tract. Both of them slightly increased the
concentration of IgA as compared to the control. With smaller
portions the product of the present invention increased the
concentration of IgA as compared to the control.
[0076] The frequencies (+cells/0,5 cm villus) of the makrophages
and CD8 positive cells are shown in Table 12.
12 TABLE 12 Hydrolyza- Hydroly Control Betaglucan te, Ex. 1 zate 2
Macrophages 8,2 5,8 15,7 13,0 CD8+ 8,5 13,0 19,5 15,0
[0077] Table 12 shows that the product in accordance with the
invention significantly increased the frequency of the macrophage
cells and clearly increased the frequency of the CD8 positive
cells. The product in accordance with the invention stimulates in
the gastrointestinal tract those types of immune response that have
specific influence on the intracellular infection (viruses,
parasites and intracellularly reproducing bacteria), thus also
increasing the protection against infections caused by other than
E.coli, whereas the betaglucan product was more or less
ineffective.
[0078] The invention is not restricted to the examples of its
embodiments described above, but different variations of it are
possible within the framework of the inventive idea defined by the
claims.
* * * * *