US20020061345A1 - Procedure for preparing a food additive, and an additive and its use - Google Patents
Procedure for preparing a food additive, and an additive and its use Download PDFInfo
- Publication number
- US20020061345A1 US20020061345A1 US09/862,389 US86238901A US2002061345A1 US 20020061345 A1 US20020061345 A1 US 20020061345A1 US 86238901 A US86238901 A US 86238901A US 2002061345 A1 US2002061345 A1 US 2002061345A1
- Authority
- US
- United States
- Prior art keywords
- additive
- yeast
- raw material
- filtered
- feed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 41
- 239000000654 additive Substances 0.000 title claims abstract description 32
- 230000000996 additive effect Effects 0.000 title claims abstract description 29
- 235000013373 food additive Nutrition 0.000 title abstract description 18
- 239000002778 food additive Substances 0.000 title abstract description 18
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 119
- 210000004027 cell Anatomy 0.000 claims abstract description 33
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- 229920001542 oligosaccharide Polymers 0.000 claims abstract description 26
- 150000004676 glycans Chemical class 0.000 claims abstract description 22
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- 229920001282 polysaccharide Polymers 0.000 claims abstract description 21
- 208000028774 intestinal disease Diseases 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 13
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- 239000002253 acid Substances 0.000 claims description 18
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- 230000007062 hydrolysis Effects 0.000 claims description 10
- 238000006460 hydrolysis reaction Methods 0.000 claims description 10
- 239000003674 animal food additive Substances 0.000 claims description 6
- 239000003599 detergent Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
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- 239000000047 product Substances 0.000 description 32
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- 239000003242 anti bacterial agent Substances 0.000 description 6
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- 208000035404 Autolysis Diseases 0.000 description 5
- 206010057248 Cell death Diseases 0.000 description 5
- 206010012735 Diarrhoea Diseases 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
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- 230000028043 self proteolysis Effects 0.000 description 5
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 4
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- 102000035195 Peptidases Human genes 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- 241000700159 Rattus Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 3
- 235000013405 beer Nutrition 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
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- 230000028993 immune response Effects 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 108090000526 Papain Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 230000000973 chemotherapeutic effect Effects 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 230000009931 harmful effect Effects 0.000 description 2
- 230000002706 hydrostatic effect Effects 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000008105 immune reaction Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical group C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 108091005658 Basic proteases Proteins 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 241000282421 Canidae Species 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical class OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 241000218652 Larix Species 0.000 description 1
- 235000005590 Larix decidua Nutrition 0.000 description 1
- 231100000757 Microbial toxin Toxicity 0.000 description 1
- 241000282339 Mustela Species 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 241000006364 Torula Species 0.000 description 1
- 241000042002 Trametes sanguinea Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
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- 230000001461 cytolytic effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 210000001198 duodenum Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
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- 230000010534 mechanism of action Effects 0.000 description 1
- 238000012009 microbiological test Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000003359 percent control normalization Methods 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 235000011118 potassium hydroxide Nutrition 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
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- 150000003839 salts Chemical class 0.000 description 1
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- 239000011780 sodium chloride Substances 0.000 description 1
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- 238000003860 storage Methods 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
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- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
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- -1 various microbes Substances 0.000 description 1
- 230000036642 wellbeing Effects 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/24—Animal feeding-stuffs from material of animal origin from blood
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/32—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from hydrolysates of wood or straw
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S426/00—Food or edible material: processes, compositions, and products
- Y10S426/807—Poultry or ruminant feed
Definitions
- 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.
- 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.
- 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.
- various antibiotic and chemotherapeutic substances inhibiting microbial growth are added to the fodder used to feed the animal.
- fodders containing added probiotic products such as various microbes, acids and yeasts.
- 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.
- certain oligosaccharides e.g. fructo-oligosaccharides promote the growth of bifidomicrobes beneficial to animals.
- 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.
- a problem with fodders containing pure oligosaccharides is that they have a variable and generally low efficacy in preventing intestinal diseases.
- the price of pure oligosaccharides is high.
- the object of the present invention is to eliminate the problems described above.
- 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.
- a further object of the invention is to disclose a food additive that makes it possible to reduce more effectively intestinal diseases in animals.
- 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.
- 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.
- the invention also discloses products prepared by the method of the invention, their use and preparations containing additives according to the claims.
- 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.
- beneficial intestinal microbes such as lactic acid bacteria and bifidobacteria
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- cellulases e.g. cellulases, acid or alkaline proteases
- the culture solution of a derivative of Trametes sanguinea can be used.
- 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.
- the yeast in a hydrolysis, can be heated to a temperature of over 40° C., in autolysis and enzymatic hydrolysis e.g. to 40-65° C. and in acid and alkalic hydrolysis e.g. 70-90° C.
- the duration of the heating may vary depending on the temperature, e.g. between 1-12 hours.
- 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.
- flavours can be separated e.g. as described in the specification referred to.
- 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.
- 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.
- a treatment breaking up the cell and/or a heat treatment combined with an enzymatic or other hydrolytic treatment.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- Additive was prepared from larch by hydrostatic heat treatment. Inhibition of bacterial adherence was determined as above. The results are shown in Table 4.
- Group 1 basic fodder (reference)
- Group 2 basic fodder+40 ppm Avilamysine
- Group 3 basic fodder+product prepared from yeast by hydrolytic treatment, in an amount of 0.5 w-% (of dry matter)
- Group 4 basic fodder+product prepared from yeast by hydrolytic treatment, in an amount of 1.0 w-% (of dry matter).
- 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.
- 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.
- 100% of the farrows in the olaqvindox group had to be treated with an antibiotic or with zinc oxide because of diarrhoea.
- the corresponding need was 12.5%.
- 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.
- 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.
- the yeast suspensions were subjected to a heat treatment (95° C. for about 10 min.), whereupon they were transferred into a fermentor, pH 5.8, temperature 65° C.
- the proteolytic enzyme used was papain (Promod 144 L).
- 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.
- the yeast was autolysed in a fermentor, temperature 50° 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.
- 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. 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.
- 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.
- 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.
- 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.
- 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.
- 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° C.
- 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.
- 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-%.
- 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.
- the filtered brewing yeast was hydrolyzed with an acid.
- 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° 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.
- 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.
- 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.
- 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.
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Abstract
Description
- 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.
- 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.
- 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.
- 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.
- 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.
- The object of the present invention is to eliminate the problems described above.
- 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.
- A further object of the invention is to disclose a food additive that makes it possible to reduce more effectively intestinal diseases in animals.
- 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.
- As for the features characteristic of the invention, reference is made to the claims.
- 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.
- The invention also discloses products prepared by the method of the invention, their use and preparations containing additives according to the claims.
- 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.
- 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 asE.coli, in the intestine and to inhibit the ability of the microbes to attach to the wall of the intestine.
- 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 asE.coli and salmonella, are not. This favours the growth of beneficial microbes at the expense of noxious ones.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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 ofTrametes 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.
- In a hydrolysis, the yeast can be heated to a temperature of over 40° C., in autolysis and enzymatic hydrolysis e.g. to 40-65° C. and in acid and alkalic hydrolysis e.g. 70-90° C. The duration of the heating may vary depending on the temperature, e.g. between 1-12 hours.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- 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.
- In the following, the invention will be described in detail by the aid of embodiment examples by referring to the attached drawings, in which
- 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.
- 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 ofE.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.
- 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.
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% - In a laboratory test, food additive was prepared from dried blood by treating it with a detergent enzyme.
- It was established that the additive inhibits bacterial adherence by 90-95%, depending on the coli strain. The results are shown in Table 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% - 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.
- It was established that the additive inhibits bacterial adherence by 92-96% (Table 3).
TABLE 3 E. coli strains Inhibition strain 1 92% strain 2 96% - Additive was prepared from larch by hydrostatic heat treatment. Inhibition of bacterial adherence was determined as above. The results are shown in Table 4.
- It was established that the additive inhibits bacterial adherence by 96-98%.
TABLE 4 E. coli strains Inhibition strain 1 96% strain 2 98% - In this test, four equal groups of pigs were fed with the following fodders:
- Group 1: basic fodder (reference)
- Group 2: basic fodder+40 ppm Avilamysine
- Group 3: basic fodder+product prepared from yeast by hydrolytic treatment, in an amount of 0.5 w-% (of dry matter)
- Group 4: basic fodder+product prepared from yeast by hydrolytic treatment, in an amount of 1.0 w-% (of dry matter).
- The results are shown in Table 5.
TABLE 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 - 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.
- 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.
- 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%.
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 - 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.
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 777a 847a Fodder efficiency kg/kg 2.07 1.87 - 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 6×4 piglets. The test groups were divided as shown in Table 8.
TABLE 8 Olaqvindox Avilamysine Additive 0 50 ppm 40 ppm Yeast fraction + − + − + − - 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.
- 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.
TABLE 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 429a 526b g/day Fodder efficiency, 1.94 1.73 2.16 1.78 kg of fodder/kg of additional growth - 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.
- 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° C. The next day, the pH was lowered to the value 2.0 (11 h). Finally, the temperature was raised to 68° 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.
- In the enzymatic hydrolysis, the yeast suspensions were subjected to a heat treatment (95° C. for about 10 min.), whereupon they were transferred into a fermentor, pH 5.8, temperature 65° 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.
- In the autolysis, the yeast was autolysed in a fermentor, temperature 50° 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.
- 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.
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 - 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.
- 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.
- 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).
- A laboratory test was carried out to establish the ability of processed baking yeast fractions to inhibit the adherence ofE.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.
- 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° 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.
- In the acid hydrolysis, pH 2, temperature 68° C., the reaction mixture was centrifuged (4000 rpm, 20 min), total extract yield about 50% of total dry matter.
- 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%.
- 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-%.
- 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.
- 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° 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.
- 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 ofE. 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.
- The results are shown in Table 11.
TABLE 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 - 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 ofE.coli than a product prepared from an unfiltered brewing yeast.
- 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.
- 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.
- 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.
- The frequencies (+cells/0,5 cm villus) of the makrophages and CD8 positive cells are shown in Table 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 - 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 thanE.coli, whereas the betaglucan product was more or less ineffective.
- 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.
Claims (22)
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Cited By (4)
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WO2008152207A1 (en) | 2007-06-13 | 2008-12-18 | Glykos Finland Oy | Nutritional compositions |
WO2011005982A1 (en) * | 2009-07-08 | 2011-01-13 | Alltech, Inc. | Bacterial adherence and anti-adherence to mucus, epithelial cells and other cells |
WO2017219106A1 (en) * | 2016-06-24 | 2017-12-28 | Yessinergy Holding S/A | Immunomodulating and growth-promoting composition controlling the population of undesirable bacteria in the intestinal microbiota, and use thereof |
WO2019046919A1 (en) * | 2017-09-06 | 2019-03-14 | Yessinergy Holding S/A | Composition of growth-promoting prebiotic additives for animal feedstuffs and use thereof |
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WO2005016024A1 (en) * | 2003-08-15 | 2005-02-24 | Grain Processing Corporation | Method for dissociation of cells |
WO2012007590A1 (en) | 2010-07-16 | 2012-01-19 | Nutreco Nederland B.V. | Improved drying of components |
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Cited By (10)
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WO2008152207A1 (en) | 2007-06-13 | 2008-12-18 | Glykos Finland Oy | Nutritional compositions |
CN101778947A (en) * | 2007-06-13 | 2010-07-14 | 格莱科斯芬兰公司 | nutritional compositions |
US20100184150A1 (en) * | 2007-06-13 | 2010-07-22 | Glykos Finland Oy | Nutritional compositions |
EA024620B1 (en) * | 2007-06-13 | 2016-10-31 | Ханккия-Мааталоус Ой | Saccharide comprising product and method for producing the same |
WO2011005982A1 (en) * | 2009-07-08 | 2011-01-13 | Alltech, Inc. | Bacterial adherence and anti-adherence to mucus, epithelial cells and other cells |
US20110034400A1 (en) * | 2009-07-08 | 2011-02-10 | Alltech, Inc. | Compositions and methods for analyzing bacterial adherence and anti-adherence to mucus, epithelial cells and other cells |
CN102639707A (en) * | 2009-07-08 | 2012-08-15 | 全技术公司 | Bacterial adherence and anti-adherence to mucus, epithelial cells and other cells |
WO2017219106A1 (en) * | 2016-06-24 | 2017-12-28 | Yessinergy Holding S/A | Immunomodulating and growth-promoting composition controlling the population of undesirable bacteria in the intestinal microbiota, and use thereof |
US10973245B2 (en) | 2016-06-24 | 2021-04-13 | Yessinergy Holding S/A | Immunomodulatory and growth promoting and controlling composition of intestinal microbiota undesirable bacteria and its use |
WO2019046919A1 (en) * | 2017-09-06 | 2019-03-14 | Yessinergy Holding S/A | Composition of growth-promoting prebiotic additives for animal feedstuffs and use thereof |
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