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
  • A23K10/30—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
  • A23K40/00—Shaping or working-up of animal feeding-stuffs
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/14—Hydrolases (3)
  • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
  • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
  • C12N9/2405—Glucanases
  • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
  • C12N9/244—Endo-1,3(4)-beta-glucanase (3.2.1.6)
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/14—Hydrolases (3)
  • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
  • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
  • C12N9/2477—Hemicellulases not provided in a preceding group
  • C12N9/248—Xylanases
  • C12N9/2482—Endo-1,4-beta-xylanase (3.2.1.8)
• • 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

Definitions

• the present invention relates to the agricultural field of animal nutrition and relates to a method of improving forage degradation.
• Wheat straw represents an inexpensive resource that could contribute to strengthening the forage and food autonomy of certain farms.
• fodder represents a qualitatively very important resource that could advantageously be valued in animal nutrition.
• wheat straw is limited in ruminants with a high production level, such as cattle. Indeed, despite its high fiber content, wheat straw has a low nutritional value due to poor digestion by ruminants. Indeed, from a chemical point of view, the presence of bonds between the constituents of the plant wall leads to a more or less strong resistance to the attack of the digestive microbiota rumen. This resistance results in a low degradation of the feed in the animal.
• the publication by Y. Wang et al shows the effects of alkaline pretreatment on the efficiency of exogenous enzymes to increase the digestibility of wheat straw.
• the wheat straw is pretreated with an alkaline solution and then with an enzyme solution by spraying.
• the results thus show that the combination of alkaline chemical pretreatment and enzymatic treatment improves the digestibility of wheat straw.
• the enzymatic treatment used is applied to the wheat straw upstream of the feed of the animal in order to respect a sufficient incubation time and to allow an effect on the digestibility.
• the enzymatic treatment can therefore be qualified as pretreatment in that it is a preliminary treatment of the straw. wheat so that the latter, once the incubation time is respected, can be used later in animal feed and that the digestion is improved.
• a first subject of the invention relates to a method for improving the ruminal degradation of wheat straw in a ruminant comprising the following steps:
• step b) feeding the ruminant with the wheat straw pretreated chemically in step b),
• said method being characterized in that it further comprises a step d) of direct administration to the ruminant of an enzymatic rumen additive, which means that said enzymatic additive is not used in pretreatment on the straw.
• a second subject of the invention relates to the direct use in a ruminant of an enzymatic rumen additive, before, during or after feeding said ruminant with wheat straw having undergone chemical pretreatment.
• the "direct" use in a ruminant means that the enzymatic additive is administered directly to the ruminant, in particular by the oral route, without going through a pretreatment of the wheat straw with the enzymatic additive.
• a first subject of the invention relates to a method for improving the ruminal degradation of wheat straw in a ruminant comprising the following steps:
• step b) feeding the ruminant with the wheat straw pretreated chemically in step b),
• said method being characterized in that it further comprises a step d) of direct administration to the ruminant of an enzymatic rumen additive, which means that said enzymatic additive is not used in pretreatment on wheat straw.
• Degradation is defined as the degradation of food by the action of reticulo-rumen microorganisms.
• pretreatment denotes a treatment carried out on wheat straw upstream of the administration to the animal, said treatment being carried out by placing a chemical solution in contact with the wheat straw.
• the process according to the invention thus makes it possible to improve the degradation of wheat straw when it is used to feed a ruminant.
• the process developed makes it possible to make the parietal constituents of wheat straw more accessible to the degradation enzymes of the rumen microorganisms and to improve their degradation.
• Digestibility is a criterion that defines the degree to which an organic matter is digested by an animal. The higher the digestibility, the better the digestion.
• the process according to the invention also makes it more suitable for animal feed because at present it is underutilized because of the difficulties animals have in digesting it. compared to other foods.
• the first step of the process according to the invention therefore consists of providing wheat straw.
• Wheat straw is the agricultural product represented by the part of the stalk or stubble of wheat.
• the wheat straw provided is that conventionally used by those skilled in the animal feed industry and can be in various forms, both in the form of straw bales and in bulk form.
• the straw may first undergo a mechanical treatment step whose objective is to reduce the size of the strands and to allow both a better accessibility of the fibers to subsequent treatments and better degradation.
• the wheat straw may previously be minced, lacerated or defibrated, or a grinding.
• the second step of the process according to the invention consists in carrying out a chemical pretreatment of the wheat straw.
• the chemical pretreatment used in the process of the invention is a pretreatment carried out by an alkaline solution as conventionally used by those skilled in the art.
• the chemical pretreatment can be carried out with a solution of sodium hydroxide (NaOH), sodium hydroxide, lime, potassium hydroxide (KOH), anhydrous ammonia (NH 3 ), aqueous ammonia (NH 4 OH), urea or a mixture of these compounds.
• the chemical pretreatment carried out in step b) is carried out with a solution of sodium hydroxide (NaOH).
• the chemical pretreatment can be carried out in different ways according to the techniques known to those skilled in the art, whether industrial or not.
• the chemical pretreatment can be carried out wet, semi-wet, semi-dry, or dry.
• the pretreatment when carried out wet or semi-wet, it can be applied by spraying or by dipping.
• said solution can be concentrated from 1.6 to 5% and then mixed at a rate of 1 to 3 L / kg of wheat straw.
• the wheat straw can then be distributed to the animals 24 to 48 hours after mixing.
• the sodium hydroxide solution is concentrated between the semi-wet and dry channels at 12% and then mixed in the machine with the straw at a rate of 0.4 L / kg of straw.
• the action time of soda is about 8 days.
• the straw can be dried in the open air.
• the soda solution is more concentrated (16%).
• the solution is then mixed with the chopped straw at a rate of about 0.3 L / kg of wheat straw.
• the latter is then passed through a die press.
• the improving action of the soda is very fast (from 20 seconds to about one minute) thanks to the high temperature and pressure prevailing in the die.
• the chemical pretreatment is carried out semi-wet way.
• the method according to the invention comprises a step of feeding an animal with the straw which has been pretreated chemically.
• This feeding step is a conventional step well known to those skilled in the breeding industry.
• the animals are then fed with the pretreated wheat straw.
• the animals are ruminants, such as, for example, cows or cattle.
• the administration step consists in administering the ruminant enzymatic additive to the ruminant so that the ruminant is found in the reticulo-rumen of said ruminant.
• An example of administration of the enzymatic additive is the oral route.
• This administration step can be performed before, simultaneously with, or after the feeding step of the ruminant with the chemically pre-treated wheat straw.
• the feeding step means a maximum of 24 hours before.
• the enzyme additive will never be administered more than 24 hours before the administration of the chemically pre-treated wheat straw.
• the enzymatic rumen additive will be administered from 2 hours to 8 hours prior to administration of the chemically pretreated wheat straw.
• the feeding step means at most 24 hours later.
• the enzyme additive will never be administered more than 24 hours after the administration of the chemically pre-treated wheat straw.
• the enzymatic rumen additive will be administered from 2 hours to 8 hours after administration of the chemically pre-treated wheat straw.
• the administration step is performed simultaneously with the feeding step.
• a step of direct administration in the ruminant, in particular orally, of an enzymatic ruminate additive made it possible to significantly improve the degradation, or even , to create a synergy on the degradation with the chemical pretreatment of wheat straw.
• the method according to the invention can be applied on a large scale on the whole of a farm and inexpensive way since the enzyme is not used in pretreatment on straw but is administered directly to the animal as an enzyme additive.
• the ruminant enzymatic additive is directly administered to the ruminant before, during feeding or after feeding with chemically pre-treated wheat straw, and it is no longer necessary to spray it on the whole stock of straw. wheat or wheat straw to eat soon.
• the process according to the invention makes it possible to dispense with the incubation times normally required to allow the enzyme to act when it is used in pretreatment of wheat straw.
• the enzymatic rumen additive is a mixture comprising at least two enzymes. Enzymes used in the enzymatic rumen additive may be glucanases, xylanases, esterases, or carboxymethylcellulases.
• the enzymatic rumen additive is a mixture of glucanase and xylanase, and in particular endo-1,3 (4) -beta-glucanase and endo-beta-1,4-xylanase. .
• the endo-1, 3 (4) -3-glucanase enzyme is a highly concentrated preparation of ⁇ -glucanase, obtained by submerged fermentation of a selected strain of Trichoderma reesei.
• the endo-b-1, 4-xylanase enzyme is a highly concentrated xylanase preparation obtained by submerged fermentation of a selected strain of Trichoderma reesei. According to the EC nomenclature of Enzymes Commission number:
• the endo-1, 3 (4) - glucanase enzyme has the EC code 3.2.1.6,
• the endo-1,4-xylanase enzyme has the EC code 3.2.1.8.
• the dose of glucanase can be between 200 units / g of straw and 7000 units / g of straw, preferably 4000 units / g of straw at 6000 units / g of straw, for example about 4670 units / g of straw.
• the xylanase dose may be from 200 units / g of straw to 7000 units / g of straw, preferably from 4000 units / g of straw to 6000 units / g of straw, for example about 4670 units / g of straw.
• the enzymatic units correspond to the amount of enzyme necessary for:
• the glucanase may be an endo-1,3 (4) -glucanase and the xylanase may be an endo-1, 4-xylanase.
• the glucanase / xylanase ratio is at least 0.5, preferably at least 0.75, and most preferably about 1.
• the enzymatic rumen additive is in a form suitable for animal administration.
• the enzymatic rumen additive can be in the form of capsules, bolus, powder, granules or liquid, such as in ampoules.
• the enzymatic rumen additive is in powder or liquid form, preferably in the form of a powder.
• a capsule or capsule is a solid preparation, consisting of a hard or soft envelope of varying shape and capacity.
• a tablet or bolus is a solid preparation containing a unit for taking one or more active ingredients (or preparation). They are obtained by compression of a constant volume of particles. Destined for cattle, they become bigger and they are called boluses.
• said enzymatic ruminate additive when the enzymatic rumen additive is administered simultaneously with the chemically pretreated wheat straw, said enzymatic ruminate additive may be in the form of a powder which will be mixed with the pretreated wheat straw. chemically. The mixture is then used to feed the ruminant.
• the ruminant belongs to the family Bovidae.
• the family Bovidae includes several sub-families, including cattle (including cattle) and goats (including sheep and goats).
• the improvement in the degradation of wheat straw can be determined by the techniques known to those skilled in the art, for example by measuring the loss of dry matter by incubation in ruminal liquid. The details of the measurements are presented in the example section below.
• the process according to the invention is particularly advantageous because it surprisingly allows a synergistic action between the enzymatic rumen additive and the chemical pretreatment of the wheat straw on the degradation of the wheat straw.
• the effect on ruminal degradation is greater than the sum of the combined effects of chemical pretreatment and enzymatic ruminate additive alone.
• the wheat straw is better valued and it is simpler for the farms to tend towards food self-sufficiency without generating significant additional cost.
• a second subject of the invention relates to the direct use in a ruminant of an enzymatic rumen additive before, after or simultaneously with feeding said ruminant with wheat straw having previously undergone alkaline pretreatment, and preferably simultaneously with feeding said ruminant, which means that said enzymatic additive is not used in pretreatment on wheat straw.
• the enzymatic rumen additive is an enzyme mixture comprising at least two enzymes. Enzymes used in the enzymatic rumen additive may be glucanases, xylanases, esterases, or carboxymethylcellulases.
• the enzymatic rumen additive is a mixture of glucanase and xylanase and in particular endo-1,3 (4) -beta-glucanase and endo-beta-1,4-xylanase.
• the dose of endo-1, 3 (4) -3-glucanase can be between 200 units / g of straw and 7000 units / g of straw, preferably of 4000 units / g of straw at 6000 units / g of straw, for example about 4670 units / g of straw.
• the dose of endo-3-1, 4-xylanase can be between 200 units / g of straw and 7000 units / g of straw, preferably of 4000 units / g of straw to 6000 units / g of straw as for example about 4670 units / g of straw.
• the ratio endo-1, 3 (4) -3-glucanase / endo-3-1, 4-xylanase is at least 0.5, preferably at least 0.75, and especially about 1.
• the enzymatic rumen additive is in a form suitable for animal administration.
• the enzymatic rumen additive can be in the form of capsules, bolus, powder, granules or liquid, such as in ampoules.
• the enzymatic rumen additive is in the form of powder or liquid, preferably in powder form.
• the use according to the invention is particularly advantageous because it makes it possible to improve the degradation of wheat straw by animals.
• the simultaneous use makes it possible to create a synergistic effect on the degradation of wheat straw.
• FIG. 1 Comparison of the effect of an aqueous pretreatment alone (H 2 0) or alkaline alone (NaOH 5%), of the use of an enzymatic ruminate additive alone (represented hereinafter by (GP + XP) x10) or the combination of an alkaline pretreatment and an enzymatic ruminate additive (hereinafter represented by (5% + NaOH + (GP + XP) x10)) on the degradation of the dry matter of wheat straw ( percentage of disappearance / degradation of the dry matter of wheat straw).
• the two dashed lines respectively represent the value of the disappearance / degradation of the dry matter of wheat straw pretreated with soda (5% NaOH) (upper line) and not pretreated (control (C)) (lower line).
• Wheat straw is milled on a 2 cm mesh (Electra) and then the fine particles are removed through a sieve of 1. 18 mm (Penn State Separator).
• Pretreatment is applied semi-wet for 24 hours on straw before incubation with rumen juice. Two types of pretreatment are applied. Pretreatment with water (control) and chemical pretreatment.
• Pretreatment with water is a negative control. Indeed, the pre-treatment is made by a soda solution in water.
• a negative control pre-treated with water only and (ii) a negative control without pre-treatment, and then dissociating the water pre-treatment effect and soda, and therefore to be able to conclude specifically about the effect of soda as a pre-treatment.
• the chemical pretreatment is applied by contacting the wheat straw with a 5% sodium hydroxide (NaOH) solution. 9 ml of solution are used for 3 g of wheat straw.
• NaOH sodium hydroxide
• the additive is added in the modules at the time of incubation with the ruminal liquid.
• Enzyme ruminate additive is a mixture of two enzymes in a 1: 1 ratio:
• an endo-1,3 (4) -beta-glucanase hereinafter GP, having an activity of 70000 units / g and an endo beta 1-4 xylanase, hereinafter XP, having an activity of 70000 units / g.
• the enzymatic units correspond to the amount of enzyme necessary for:
• Ten doses of the enzymatic mixture corresponds to 0.4 g of the enzyme mixture for 3 g of straw, ie 0.2 g of GP and 0.2 g of XP for 3 g of straw, or 4670 units (rounded to the tens) of GP and XP for 1 g of straw. 4.
• Ruminai content is collected approximately 2 hours post-meal in dry dairy cows, carrying a ruminal cannula and fed with a ration based on wheat straw, corn silage and a mineral and vitamin supplement.
• the ruminai content is filtered through a 1.6 mm mesh metal screen to obtain rumen fluid.
• the latter is bubbled with C0 2 oxygen-free.
• Test 2 Unpretreated wheat straw and enzymatic rumen additive (see point 3. above).
• Trial 3 duplicate of Trial 2 to validate and confirm the results.
• Test 4 Pretreated wheat straw (see point 2.) and enzymatic rumen additive (see point 3.)
• the untreated straw corresponds to pretreated straw according to the same protocol as point 2. but replacing the alkaline solution with water.
• 3 white modules are made. They contain only incubation medium as described in point 4 of this example.
• the incubation flasks receive 200 ml of incubation medium each and then are saturated with oxygen-free CO 2 .
• the quantity of straw, pretreated or not, within each module is 3 grams.
• the flasks are incubated at 39 ° C for 96 hours to allow fermentation.
• the modules After 96 hours of incubation, the modules are placed in the ice to stop the fermentation. The amount of non-degraded dry matter is then measured by filtering and drying the incubation residue at 60 ° C for 4 days (hereinafter referred to as residue).
• the incubated substrate corresponds to pretreated or unpretreated wheat straw.
• the dry matter degradation results were subjected to an analysis of variance using the linear general univariate model of the statistical data processing software SPSS® (IBM®, SPSS®, version 22).
• the model integrates the effects of the covariate and the products, the covariate being the degradation of the control dry matter (in percentage%).
• An Iog10 transformation of the percent values was performed for the analysis.
• FIG. 1 shows a significant effect on the ruminal degradation of the dry matter by the use of an enzymatic rumen additive on wheat straw pretreated with sodium hydroxide (5% NaOH + (GP + XP) x10 ).
• the results show that pretreatment with sodium hydroxide (5%) improves the ruminal degradation of straw with respect to (i) no pre-treatment and (ii) pre-treatment with water.
• Pretreatment with sodium hydroxide makes it possible to improve rumen degradation of the dry matter by 22.3 points compared with untreated wheat straw (control).
• the synergistic effect is understood to mean that ruminal degradation in animals of chemically pretreated wheat straw and in the presence of enzymatic rumen additive is greater than the ruminal degradation achieved by chemical pretreatment alone, by the administration. of the enzymatic additive alone or by the two combined but working independently.
• the process according to the invention is therefore particularly advantageous in that it makes it possible to significantly improve the ruminal degradation of wheat straw when the latter is chemically pretreated.

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• Life Sciences & Earth Sciences (AREA)
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• Polymers & Plastics (AREA)
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• 2018
  • 2018-06-05 FR FR1854868A patent/FR3081682B1/fr active Active
• 2019
  • 2019-06-04 US US16/972,216 patent/US20210307360A1/en not_active Abandoned
  • 2019-06-04 AU AU2019281142A patent/AU2019281142A1/en active Pending
  • 2019-06-04 EP EP19737839.1A patent/EP3801045A1/de active Pending
  • 2019-06-04 WO PCT/FR2019/051325 patent/WO2019234347A1/fr unknown
  • 2019-06-04 BR BR112020024781-2A patent/BR112020024781A2/pt unknown
  • 2019-06-04 CA CA3101868A patent/CA3101868A1/fr active Pending
  • 2019-06-04 MX MX2020013246A patent/MX2020013246A/es unknown
  • 2019-06-04 CN CN201980037921.6A patent/CN112272521A/zh active Pending
  • 2019-06-05 AR ARP190101532A patent/AR115483A1/es active IP Right Grant

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