CN115176864A - Probiotic fermented compound feed and preparation method thereof - Google Patents

Probiotic fermented compound feed and preparation method thereof Download PDF

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Publication number
CN115176864A
CN115176864A CN202210787627.0A CN202210787627A CN115176864A CN 115176864 A CN115176864 A CN 115176864A CN 202210787627 A CN202210787627 A CN 202210787627A CN 115176864 A CN115176864 A CN 115176864A
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feed
preparation
fermentation
culture medium
compound feed
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韩怀磊
李文
滕萧圪
夏雨欣
苏淇
王陶
袁珍虎
杨太平
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Xuzhou University of Technology
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    • 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
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • 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
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/22—Compounds of alkali metals
    • 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
    • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • 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

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Abstract

The invention discloses a probiotic fermented compound feed and a preparation method thereof, and the raw materials comprise 94% of soybean meal, 4% of wheat bran and 2% of ginkgo leaf residues. The manufacturing method comprises the following steps: preparing a substrate, preparing a strain culture medium, activating strains, performing solid fermentation and packaging. The invention can effectively improve the antioxidation effect of the feed, improve the contents of total phenols and total flavonoids in the feed, and improve the DPPH free radical scavenging capacity, OH scavenging capacity, iron ion reducing capacity and superoxide anion scavenging capacity. The animal nutrition supplement agent can provide rich nutrition for animals, simultaneously avoid the animals from being invaded by free radicals, reduce the inflammation and the disease risk of the animals and improve the growth quality of the animals.

Description

Probiotic fermented compound feed and preparation method thereof
Technical Field
The invention relates to the field of feeds, in particular to a probiotic fermented compound feed and a preparation method thereof.
Background
The fermented feed is prepared by fermenting coarse feed with microorganism. The coarse fodder is rich in cellulose, hemicellulose, pectin, lignin and other substances, is difficult to be directly digested and absorbed by animals, and can increase intestinal burden and cause intestinal diseases when being directly ingested. However, the microbial fermentation can effectively decompose the macromolecular substances which are difficult to digest, so that the macromolecular substances are converted into small molecules, and the digestibility of the feed is greatly improved. Meanwhile, the fermentation process can increase the number of beneficial bacteria in the feed, reduce pathogenic bacteria and effectively improve the nutritional value of the feed.
Antioxidants are a class of substances that retard or prevent the oxidation of a substrate and help trap and neutralize free radicals, thereby scavenging them. Free radicals in the animal body can be directly eliminated by the antioxidant, so that the antioxidant level of the animal body is improved. In the prior art, the feed with the antioxidant effect exists, but the antioxidant effect of the feed mostly adopts a mode of additionally adding an antioxidant, and the antioxidant effect of the feed is poor.
Disclosure of Invention
The invention aims to overcome the problems and provides a probiotic fermented compound feed and a preparation method thereof. In order to achieve the purpose, the invention adopts the following technical scheme:
a probiotic fermented compound feed and a preparation method thereof comprise the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran and 2% of ginkgo leaf residues;
the preparation method of the feed comprises the following steps:
s1, preparing a substrate
Uniformly mixing the soybean meal, the wheat bran and the ginkgo leaf residues according to a proportion, and adding water for sterilization for later use;
s2, preparing a strain culture medium
Preparing a culture medium, wherein the concentrations of raw materials are as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride, and carrying out sterilization treatment after maintaining the pH =7.0 of the culture medium;
s3, strain activation
Inoculating bacillus subtilis into the culture medium obtained in the S2, and activating strains;
s4, solid state fermentation
Adding 4% of activated strain obtained from S3 into the substrate obtained from S1, mixing, and fermenting at 37 deg.C for 60-72 hr;
s5. Packaging
After fermentation, the feed is freeze-dried, then ground into powder and packaged.
As an improvement, the material-water ratio in the step S1 is 1:1.
as an improvement, the temperature of the sterilization treatment in the step S2 is 121 ℃, and the sterilization time is 20min.
As an improvement, the conditions for activating the strains in the step S3 are as follows: culturing at 37 deg.C in 120r/min water bath shaker for 24h.
As an improvement, the activation of the strain in step S3 is performed for 2 times, and the 2 times of activation are performed by: and (3) sucking the primary activated strain, inoculating the primary activated strain into the culture medium obtained in the step S2, and culturing for 24 hours at 37 ℃ in a water bath shaker at 120 r/min.
As an improvement, during the solid state fermentation of the step S4, the fermentation material is turned intermittently, and the turning interval is 6-12h.
The invention has the advantages that:
the invention can effectively improve the antioxidation effect of the feed, improve the contents of total phenols and total flavonoids in the feed, and improve the DPPH free radical scavenging capacity, OH scavenging capacity, iron ion reducing capacity and superoxide anion scavenging capacity. The animal nutrition supplement agent can provide rich nutrition for animals, prevent the animals from being invaded by free radicals, reduce the inflammation and disease risks of the animals and improve the growth quality of the animals.
Drawings
FIG. 1 shows the change in DPPH radical scavenging ability before and after fermentation, wherein A, B, C, D represents control 1, control 2, example 1 and control 3, respectively;
FIG. 2 shows the change in OH scavenging capacity before and after fermentation, wherein A, B, C, D represents control 1, control 2, example 1 and control 3, respectively;
FIG. 3 shows the change in DPPH radical scavenging ability at each time period;
FIG. 4 shows the OH scavenging ability change at each time interval;
FIG. 5 shows the variation of the iron ion reducing power of different concentrations of extract;
FIG. 6 shows the change in superoxide anion scavenging capacity of different concentrations of extract.
Detailed Description
The present invention will be described in detail and specifically with reference to the following examples so as to facilitate the understanding of the present invention, but the following examples do not limit the scope of the present invention.
Example 1
A probiotic fermented compound feed and a preparation method thereof comprise the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran and 2% of ginkgo leaf residues.
The preparation method of the feed comprises the following steps:
s1, preparing a substrate
Uniformly mixing the soybean meal, the wheat bran and the ginkgo leaf residues in proportion, wherein the material-water ratio is 1:1 adding water for sterilization for later use;
s2, preparing a strain culture medium
Preparing a culture medium, wherein the concentration of raw materials is as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride, and carrying out sterilization treatment after maintaining the pH =7.0 of the culture medium; sterilizing at 121 deg.C for 20min;
s3, strain activation
Inoculating bacillus subtilis into 100mL of a culture medium obtained by S2, and activating strains;
the method comprises the following specific steps of primary activation: culturing at 37 deg.C in 120r/min water bath shaker for 24h;
then carrying out secondary activation: absorbing the primary activated strain, inoculating into the culture medium obtained in the step S2, and culturing for 24h at 37 ℃ in a 120r/min water bath shaking table;
s4, solid state fermentation
Adding 4% of activated strain (0.8 mL) obtained from S3 into the substrate obtained from 20g S1, mixing, fermenting at 37 ℃ for 72h, and shaking the fermentation bottle every 12 h;
s5. Packaging
After fermentation, the feed is freeze-dried, then ground into powder and packaged.
Example 2
A probiotic fermented compound feed and a preparation method thereof comprise the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran and 2% of ginkgo leaf residues.
The preparation method of the feed comprises the following steps:
s1, preparing a substrate
Uniformly mixing the soybean meal, the wheat bran and the ginkgo leaf residues in proportion, wherein the material-water ratio is 1:1 adding water for sterilization for later use;
s2, preparing a strain culture medium
Preparing a culture medium, wherein the concentrations of raw materials are as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride, and carrying out sterilization treatment after maintaining the pH =7.0 of the culture medium; sterilizing at 121 deg.C for 20min;
s3, strain activation
Inoculating bacillus subtilis into 100mL of a culture medium obtained by S2, and activating strains;
the method comprises the following specific steps of primary activation: culturing at 37 deg.C in 120r/min water bath shaker for 24h;
then carrying out secondary activation: absorbing the strains subjected to primary activation, inoculating the strains into the culture medium obtained in the step S2, and culturing for 24 hours at 37 ℃ in a 120r/min water bath shaker;
s4, solid state fermentation
Adding 4% of activated strain (0.8 mL) obtained from S3 into the substrate obtained from 20g S1, mixing, fermenting at 37 deg.C for 12-84 hr, and shaking the fermentation bottle every 12 hr;
s5. Packaging
After fermentation, the feed is freeze-dried, then ground into powder and packaged.
Comparative example 1
In the comparative example, the raw materials comprise the following components in percentage by weight:
96% of soybean meal and 4% of wheat bran.
The preparation method of the feed comprises the following steps:
s1, preparing a matrix
Uniformly mixing the soybean meal, the wheat bran and the ginkgo leaf residues in proportion, wherein the material-water ratio is 1:1 adding water for sterilization for later use;
s2, preparing a strain culture medium
Preparing a culture medium, wherein the concentration of raw materials is as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride, and carrying out sterilization treatment after maintaining the pH =7.0 of the culture medium; sterilizing at 121 deg.C for 20min;
s3, strain activation
Inoculating bacillus subtilis into 100mL of a culture medium obtained by S2, and activating strains;
the method comprises the following specific steps of primary activation: culturing at 37 deg.C in 120r/min water bath shaker for 24h;
then carrying out secondary activation: absorbing the primary activated strain, inoculating into the culture medium obtained in the step S2, and culturing for 24h at 37 ℃ in a 120r/min water bath shaking table;
s4, solid state fermentation
Adding 4% of activated strain (0.8 mL) obtained from S3 into the substrate obtained from 20g S1, mixing, fermenting at 37 ℃ for 72h, and shaking the fermentation bottle every 12 h;
s5. Packaging
After fermentation, the feed is freeze-dried, then ground into powder and packaged.
Comparative example 2
In the comparative example, the raw materials comprise the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran and 2% of garlic skin
The preparation method of the feed was the same as in comparative example 1.
Comparative example 3
In the comparative example, the raw materials comprise the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran, 1% of garlic skin and 1% of ginkgo leaf residues.
The preparation method of the feed was the same as in comparative example 1.
Experiments and conclusions
(1) Change in DPPH radical scavenging before and after fermentation
The change in DPPH radical scavenging ability before and after fermentation of the samples was measured as shown in FIG. 1 below. It can be seen that the DPPH free radical scavenging capacity of the feeds with four ratios after 72h fermentation is remarkably improved, wherein the antioxidant capacity of the feed in example 1 is the strongest, and the free radical scavenging capacity after fermentation is improved from 25.23% to 60.06% and is improved by 2.38 times.
(2) OH elimination before and after fermentation
The OH-scavenging ability of the sample before and after fermentation was measured, and the change thereof is shown in FIG. 2 below. It can be seen that the removal capacity of the feed with four proportions of OH after 72 hours of fermentation is obviously improved, wherein the removal capacity of OH in example 1 is improved to the maximum, and the removal capacity of OH is improved from 25.21% to 69.56%, which is improved by 2.76 times.
With the combination of the graphs of fig. 1 and fig. 2, the oxidation resistance of the fermented feed is improved after 72 hours, and the DPPH free radical scavenging capacity after fermentation is respectively example 1, control group 2, control group 3 and control group 1 from strong to weak; the OH scavenging ability after fermentation is from strong to weak as example 1, control group 2 and control group 3, respectively, and the antioxidant effect of example 1 is always the highest in comparison of the two antioxidant activities.
(3) Relationship between fermentation time and antioxidant effect
In the production process of example 2, samples were taken at fermentation times of 0, 12, 24, 36, 48, 54, 60, 66, 72, 78, and 84 hours, respectively, and the DPPH radical clearance and OH clearance were measured. The results are shown in fig. 3 and 4, and the antioxidant effect of the composition reaches the highest peak at 66 h.
(4) Iron ion reducing power and superoxide anion scavenging ability
Under the best measurement conditions, the change situation of the iron ion force of the sample extracting solution with different concentrations before and after fermentation is shown in the following figure 5, when the concentration of the extract is 2.5mg/mL, the reduction force of the iron ions of the extract after fermentation is equivalent to 607.68mol/L, and the reduction force of the extract after fermentation is 213.68mol/L, and as can be seen from the figure 6, along with the increase of the concentration, the superoxide anion scavenging capability of the sample extract after fermentation is obviously improved, and the total reduction force without fermentation is basically not improved.
(5) Changes of antioxidant functional components before and after fermentation
The total phenols and total flavonoids of the extract solution of example 1, which had a concentration of 2.5mg/mL before and after fermentation, were measured, and the results obtained are shown in Table 1 below. It can be seen from the table that the total phenol content in the unfermented feed extract is 5.48 μ g/mg, and the total phenol content in the fermented feed extract passing through bacillus subtilis is increased to 12.75 μ g/mg, which is 2.33 times higher than that before fermentation, because the phenolic compounds in the feed before fermentation are combined with insoluble fibers, phenols are released during the fermentation process, so that the total phenol content in the leaching solution is increased.
The content of the total flavonoids in the unfermented feed extract is 12.75 mug/mg, and after 66 hours of fermentation, the content of the total flavonoids is increased to 31.64 mug/mg, which is increased by 2.40 times, because the combined phenolic compounds and the flavonoids are released by microbial fermentation, the content of free phenolic compounds and flavonoids are obviously increased.
TABLE 1 Total phenol and Total Flavonoids content Change before and after fermentation
Figure BDA0003729327030000061
(6) EC before and after fermentation of samples 50 Value of
Antioxidant capacity and EC of samples 50 Closely related, comparison before fermentationEC of post-sample extract 50 Value, post fermentation EC 50 The more the value is decreased, the higher the degree of improvement of the antioxidant ability is. As can be seen from Table 2, EC for DPPH radical scavenging before fermentation 50 The value is 4.50mg/mL, the value is reduced to 2.23mg/mL after fermentation, and the reduction is 50.44%, which shows that the DPPH free radical scavenging capacity is improved by 2.02 times by the fermentation of the bacillus subtilis. EC for OH scavenging before fermentation 50 The value is 3.98mg/mL, the value is reduced to 1.51mg/mL after fermentation, 62.06% is reduced, and the bacillus subtilis fermentation is known to improve the OH scavenging capacity by 2.64 times. EC for superoxide anion scavenging prior to fermentation 50 The value was 2.04mg/mL, which was reduced to 0.76mg/mL after fermentation, which was reduced by 62.75%, indicating that Bacillus subtilis fermentation increased the superoxide anion capacity by 2.68 times.
TABLE 2 EC50 of antioxidant index of extract before and after fermentation
Figure BDA0003729327030000062
The embodiments of the present invention have been described in detail above, but they are merely exemplary, and the present invention is not equivalent to the embodiments described above. Any equivalent modifications and substitutions for the present invention are within the scope of the present invention for those skilled in the art. Accordingly, it is intended that all equivalent alterations and modifications be included within the scope of the invention, without departing from the spirit and scope of the invention.

Claims (6)

1. The probiotic fermented compound feed and the preparation method are characterized by comprising the following components in percentage by weight:
94% of soybean meal, 4% of wheat bran and 2% of ginkgo leaf residues;
the preparation method of the feed comprises the following steps:
s1, preparing a matrix
Uniformly mixing the soybean meal, the wheat bran and the ginkgo leaf residues in proportion, and adding water for sterilization for later use;
s2, preparing a strain culture medium
Preparing a culture medium, wherein the concentration of raw materials is as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride, and carrying out sterilization treatment after maintaining the pH =7.0 of the culture medium;
s3, strain activation
Inoculating bacillus subtilis into the culture medium obtained in the S2, and activating strains;
s4, solid state fermentation
Adding 4% of activated strain obtained from S3 into the substrate obtained from S1, mixing, and fermenting at 37 deg.C for 60-72h;
s5. Packaging
After fermentation, the feed is freeze-dried, then ground into powder and packaged.
2. The probiotic fermented compound feed and the preparation method thereof according to claim 1, wherein the feed-water ratio in the step S1 is 1:1.
3. the probiotic fermented compound feed and the preparation method thereof according to claim 1, wherein the temperature of the sterilization treatment in the step S2 is 121 ℃, and the sterilization time is 20min.
4. The probiotic fermented compound feed and the preparation method thereof according to claim 1, wherein the conditions for activating the strains in the step S3 are as follows: the cells were incubated at 37 ℃ for 24h in a 120r/min water bath shaker.
5. The probiotic fermented compound feed and the preparation method thereof according to claim 1, wherein the strain activation in step S3 is performed for 2 times, and the 2 times of activation are performed by: and (3) sucking the primary activated strain, inoculating the primary activated strain into the culture medium obtained in the step S2, and culturing for 24 hours at 37 ℃ in a water bath shaker at 120 r/min.
6. The probiotic fermented compound feed and the preparation method thereof according to claim 1, wherein during the solid state fermentation in the step S4, the fermentation material is intermittently turned over, and the turning interval is 6-12 hours.
CN202210787627.0A 2022-07-04 2022-07-04 Probiotic fermented compound feed and preparation method thereof Pending CN115176864A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103478413A (en) * 2013-09-18 2014-01-01 中国林业科学研究院林产化学工业研究所 Method for producing protein feed by mixed-strain solid-state fermentation of ginkgo leaf residues
CN110150461A (en) * 2019-07-04 2019-08-23 徐州工程学院 A kind of Composite fermentation type microbiological feed and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103478413A (en) * 2013-09-18 2014-01-01 中国林业科学研究院林产化学工业研究所 Method for producing protein feed by mixed-strain solid-state fermentation of ginkgo leaf residues
CN110150461A (en) * 2019-07-04 2019-08-23 徐州工程学院 A kind of Composite fermentation type microbiological feed and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
崔艺燕;田志梅;邓盾;刘志昌;容庭;李家洲;马现永;陈卫东;: "发酵树叶及其在鸡生产应用的研究进展", 广东农业科学 *
詹欣;辛敏;王然;唐涛;唐劲天;岳秉飞;: "不同天数银杏叶发酵液提取物的体外抗氧化活性比较", 中国实验方剂学杂志 *

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