CN110810661A - Application of clostridium ethanolate protein in feed of freshwater omnivorous fishes - Google Patents
Application of clostridium ethanolate protein in feed of freshwater omnivorous fishes Download PDFInfo
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- CN110810661A CN110810661A CN201910957659.9A CN201910957659A CN110810661A CN 110810661 A CN110810661 A CN 110810661A CN 201910957659 A CN201910957659 A CN 201910957659A CN 110810661 A CN110810661 A CN 110810661A
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- 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
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- 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
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
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Abstract
The invention relates to an application of clostridium ethanolate protein in feed for freshwater omnivorous fishes, belongs to the field of feed, and provides an application of clostridium ethanolate protein in feed for freshwater omnivorous fishes, wherein the preparation method of the clostridium ethanolate protein comprises the following steps: performing liquid fermentation culture by using Clostridium ethanolicum (Clostridium autoethanogenum) as a fermentation strain and using CO-containing gas as a raw material, and centrifuging and drying thalli in fermentation liquor of the liquid fermentation for the first time to obtain Clostridium ethanolicum protein; the application of the clostridium ethanolate protein in the feed for the freshwater omnivorous fishes can improve the growth performance of the freshwater omnivorous fishes, the weight gain is increased by at least 7.14%, the feed coefficient is reduced by at least 10.09%, and the protein efficiency is improved by at least 10.68%.
Description
Technical Field
The invention relates to a microbial single cell protein, in particular to an application of clostridium autoethanolate protein in a freshwater omnivorous fish feed.
Background
The freshwater omnivorous fish has the advantages of high growth speed, strong disease resistance, easiness in catching, high meat content, good meat quality and the like, is deeply loved by farmers and consumers, is cultivated nationwide, has the largest use ratio in the feed as the main protein raw material in the feed of the freshwater omnivorous fish, has the problem of feed protein raw material resource shortage more and more attracted by people along with the rapid development of livestock raising and aquaculture industry, has the trend of year-by-year increase in the situation that soybeans depend on imports, and has the soybean imports rising from 1042 million tons in 2000 to 8803 million tons in 2018.
The clostridium ethanolate protein is a microbial single-cell protein product which is prepared by taking clostridium ethanolate as a fermentation strain, carrying out liquid fermentation culture by taking CO-containing gas as a raw material, and carrying out processes such as centrifugation, drying and the like on thalli in fermentation liquor and can be used as a feed protein raw material. The clostridium ethanolate protein has the crude protein content of more than or equal to 80 parts, and the amino acid content is high and balanced. The unicellular protein serving as the protein feed of animals can relieve the current situation of shortage of protein resources in China to a certain extent, and has important significance for reducing the production cost of the breeding industry and improving the growth performance of the bred animals.
Disclosure of Invention
In view of the above problems, the present invention has been made to provide a use of clostridium autoethanolate protein in a feed for freshwater omnivorous fish that overcomes or at least partially solves the above problems.
The embodiment of the invention provides application of clostridium ethanolate protein in a feed for freshwater omnivorous fishes, wherein the preparation method of the clostridium ethanolate protein comprises the following steps:
clostridium ethanolicum (Clostridium autoethanogenum) is used as a fermentation strain, CO-containing gas is used as a raw material, liquid fermentation culture is carried out, and thalli in fermentation liquor of the liquid fermentation are subjected to centrifugation and primary drying treatment to obtain the Clostridium ethanolicum protein.
Further, the Clostridium ethanolate (Clostridium autoethanogenum) is deposited under DSM10061 and with the deposit address german collection of strains, DSMZ.
Further, the nutritional composition of the clostridium ethanolate protein comprises the following components in parts by weight: more than or equal to 80 parts of crude protein, less than or equal to 7 parts of crude ash, less than or equal to 12 parts of water and less than or equal to 1 part of ammonium salt.
Further, in the application, the preparation method of the feed for the freshwater omnivorous fishes comprises the following steps: adding clostridium ethanolate protein into the mixture, adding water, uniformly mixing, preparing into granules, and performing secondary drying to obtain the freshwater omnivorous fish feed.
Further, the mixture comprises at least one of the following: fish meal, bean pulp, rapeseed meal, cottonseed meal, wheat flour, rice bran, soybean oil, monocalcium phosphate, bentonite, mineral premix, vitamin C, choline chloride, mildew preventive and amino acid.
Further, the mixture is sieved by a 60-mesh sieve.
Further, the particle size of the particles is 2-4 mm.
Further, the fresh water omnivorous fish feed comprises the following components in parts by weight: 5-15 parts of clostridium ethanolate protein, 5 parts of fish meal, 4-29 parts of soybean meal, 14 parts of rapeseed meal, 14 parts of cottonseed meal, 21.01-29.61 parts of wheat flour, 5.30-8.2 parts of rice bran, 2.7 parts of soybean oil, 2.07-2.37 parts of monocalcium phosphate, 1.9-4.65 parts of microcrystalline cellulose, 3 parts of bentonite, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.22-0.77 part of amino acid.
One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:
1. the application of the clostridium ethanolate protein in the feed for the freshwater omnivorous fishes can improve the growth performance of the freshwater omnivorous fishes, wherein the weight gain rate is increased by at least 7.14%, the feed coefficient is reduced by at least 10.09%, and the protein efficiency is improved by at least 10.68%.
Detailed Description
The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are provided to illustrate the invention, and not to limit the invention.
Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present specification will control.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by an existing method.
In the process of breeding the freshwater omnivorous fishes, the bean pulp is usually used as a main protein raw material to be added, but the bean pulp has high anti-nutritional factor content, is not beneficial to digestion and absorption and is seriously dependent on import; the research of the invention finds that the ethanol clostridium protein can be well used as a substitute of the soybean meal, has the advantages of less anti-nutritional factors, high digestibility and the like, reduces the production cost and comprehensively improves the culture benefit.
The embodiment provides an application of clostridium ethanolate protein in a feed for freshwater omnivorous fishes, wherein a preparation method of the clostridium ethanolate protein comprises the following steps:
performing liquid fermentation culture by using Clostridium ethanolicum (Clostridium autoethanogenum) as a fermentation strain and using CO-containing gas as a raw material, and centrifuging and drying thalli in fermentation liquor of the liquid fermentation for one time to obtain Clostridium ethanolicum protein;
in this example, Clostridium ethanolicum (Clostridium autoethanogenum) was deposited under DSM10061 and was deposited under the German Collection of cultures, DSMZ.
In the liquid fermentation culture, the concentration of clostridium ethanolate is 20-25g/L, and the introduction amount of CO gas is 8000-10000m3/h；
The centrifugal rotating speed is 4000-4500r/min, and the centrifugal feeding flow is 4-6m3H; the concentration of the thalli after centrifugation is 120-150 g/L;
the primary drying temperature is 160-180 ℃ of air inlet temperature, the air outlet temperature is 75-85 ℃, instantaneous drying is carried out, the drying time is short, and the evaporation capacity of the drying tower is 5000 kg/h.
In this embodiment, the nutritional composition of the clostridium ethanolate protein comprises, in parts by weight: more than or equal to 80 parts of crude protein, less than or equal to 7 parts of crude ash, less than or equal to 12 parts of water and less than or equal to 1 part of ammonium salt.
In this embodiment, in the application, the preparation method of the feed for freshwater omnivorous fishes comprises the following steps: adding clostridium ethanolate protein into the mixture, adding water, uniformly mixing, preparing into granules, and performing secondary drying to obtain the freshwater omnivorous fish feed.
The addition amount of the water is 5-15 parts of the total weight of the clostridium ethanolate protein and the mixture.
In this embodiment, the mixture includes at least one of the following: fish meal, soybean meal, rapeseed meal, cottonseed meal, wheat flour, rice bran, soybean oil, monocalcium phosphate, bentonite, mineral premix, vitamin C, choline chloride, a mildew preventive and amino acid.
In this example, the mix was screened through a 60 mesh screen.
In this example, the particle size is 2 to 4 mm.
In this embodiment, the primary drying and the secondary drying both adopt a natural air drying method.
In the embodiment, the fresh water omnivorous fish feed comprises the following components in parts by weight: 5-15 parts of clostridium ethanolate protein, 5 parts of fish meal, 4-29 parts of soybean meal, 14 parts of rapeseed meal, 14 parts of cottonseed meal, 21.01-29.61 parts of wheat flour, 5.30-8.2 parts of rice bran, 2.7 parts of soybean oil, 2.07-2.37 parts of monocalcium phosphate, 1.9-4.65 parts of microcrystalline cellulose, 3 parts of bentonite, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.22-0.77 part of amino acid.
The following will describe in detail the application of clostridium autoethanolate protein in the present application in the feed for freshwater omnivorous fish with reference to specific examples.
Example 1
This example differs from example 1 in that:
the formula of the feed for the freshwater omnivorous fishes comprises the following components in parts by weight: 5 parts of fish meal, 20 parts of soybean meal, 5 parts of clostridium ethanolate protein, 14 parts of rapeseed meal, 14 parts of cottonseed meal, 23.12 parts of wheat flour, 6.50 parts of rice bran, 2.7 parts of soybean oil, 2.13 parts of calcium dihydrogen phosphate, 3 parts of bentonite, 2.9 parts of microcrystalline cellulose, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.49 part of amino acid.
Example 2
This example differs from example 1 in that:
the formula of the feed for the freshwater omnivorous fishes comprises the following components in parts by weight: 5 parts of fish meal, 10 parts of bean pulp, 10 parts of clostridium ethanolate protein, 14 parts of rice pulp, 14 parts of cottonseed meal, 26.61 parts of wheat flour, 7.2 parts of rice bran, 2.7 parts of soybean oil, 2.17 parts of calcium dihydrogen phosphate, 3 parts of bentonite, 3.8 parts of microcrystalline cellulose, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.27 part of amino acid.
Example 3
This example differs from example i in that:
the formula of the feed for the freshwater omnivorous fishes comprises the following components in parts by weight: 5 parts of fish meal, 4 parts of soybean meal, 15 parts of clostridium ethanolate protein, 14 parts of rapeseed meal, 10 parts of cottonseed meal, 29.61 parts of wheat flour, 8.2 parts of rice bran, 2.7 parts of soybean oil, 2.37 parts of monocalcium phosphate, 3 parts of bentonite, 4.65 parts of microcrystalline cellulose, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.22 part of amino acid.
Comparative example 1
No Clostridium ethanolicum protein was added to this comparative example.
The feed formula comprises the following components in parts by weight: 5 parts of fish meal, 29 parts of soybean meal, 14 parts of rapeseed meal, 14 parts of cottonseed meal, 21.01 parts of wheat flour, 5.30 parts of rice bran, 2.7 parts of soybean oil, 2.07 parts of monocalcium phosphate, 3 parts of bentonite, 1.9 parts of microcrystalline cellulose, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.77 part of amino acid.
The preparation method of the feed comprises the following steps: sieving feed formula with 60 mesh sieve, adding water, mixing, granulating with F-26(II) type granulator to obtain granules with particle size of 2.0mm, and drying at 65 deg.C to obtain feed.
Experimental example 1
1. Materials and methods
The test was set up with 4 treatment groups, D1 (comparative example 1-basic formula control), D2 (example 1-basic formula +5 parts clostridium ethanolate protein), D3 (example 2-basic formula +10 parts clostridium ethanolate protein) and D4 (example 3-basic formula +15 parts clostridium ethanolate protein).
The breeding test is carried out in an indoor temperature-controlled circulating water system of a test base of a stannless fishery institute of agriculture university of Nanjing, test fishes are domesticated in the system and adapted for 15 days, then the test fishes are fed with test feed 3 times a day, the test fishes are respectively fed once at 8:00-8:30, 13: 00-13: 30 and 17:00-17:30, each time, the test fishes are fed with a proper amount, and the eating condition is observed after 30min, so that the fishes are saturated without residual feed. The aeration is continuously carried out day and night, so that sufficient oxygen is ensured, the artificial interference is reduced, the silence is kept, the additional stress is prevented, and the water quality is measured once every week. The water quality during the whole test was as follows: in the feeding process, the average water temperature is 30.0 +/-1.5 ℃, the dissolved oxygen is more than or equal to 6mg/L, the ammonia nitrogen is less than or equal to 0.2mg/L, the nitrite is less than or equal to 0.2mg/L, and the pH value is 6.8-7.5. The illumination period is 12L/12D. The ingestion and death of the fish are observed every day, and the dead fish are fished out in time, weighed and checked for death reasons.
Collecting samples: after feeding for 10 weeks, stopping feeding for 24 hours, and counting the number and the total weight of the tested fishes in each breeding barrel to calculate growth performance indexes such as weight gain rate, feed coefficient and the like. Randomly taking 3 fishes from each culture barrel to collect blood, quickly taking out the fishes, immediately putting the fishes into MS-222 with the concentration of 200mg/L to perform quick deep anesthesia, wetting the tube wall of a 2mL syringe by heparin, collecting blood from tail veins, standing the blood sample in a refrigerator at 4 ℃ for 1-2h, and centrifuging at 4 ℃ and 3000rpm/min for 10min to prepare serum. Transferring the supernatant to-80 deg.C for storage and detection.
The statistical method comprises the following steps: statistical analysis was performed using SPSS12.0 software, data results were all expressed as Mean ± standard deviation (Mean ± s.d.), and after One-way ANOVA, the data were tested for significance of differences in Mean using dunnans multiple comparisons, with significance level P being 0.05.
2 results and analysis:
2.1 Effect on growth Performance of Cyprinus Carpio juvenile Fish
As can be seen from the statistical results of the tests in Table 1, the initial weight of the tested fish has no significant difference (P is more than 0.05), and compared with the control group (0 part of clostridium ethanolate protein), the weight gain rate and the protein efficiency of Cyprinus carpiovar jiangensis are significantly increased and the bait coefficient is significantly reduced (P is less than 0.05) along with the increase of the substitution amount of the clostridium ethanolate protein.
TABLE 1 influence of partial substitution of the Clostridia ethanolate protein for the Soybean meal on the growth Performance of Cyprinus carpiod
Note: the same column of shoulder marks with different letters indicate significant difference (P <0.05), the same column of shoulder marks with the same letters or without letter marks indicate insignificant difference (P >0.05), and the same is as follows.
2.2 influence of Clostridia ethanolate protein on juvenile fish antioxidant capacity to replace soybean meal in Cyprinus Carpio feed
The content of Malondialdehyde (MDA) can reflect the degree of lipid oxidation of organisms, superoxide dismutase (T-SOD) is an important antioxidant enzyme in fish bodies, can effectively eliminate free radicals in the bodies, reduces the generation of oxidative stress, has high content and low content, can reflect the anti-stress capability of the fish bodies, and the two substances are mutually matched in the bodies to jointly reflect the metabolic state of free radicals of the organisms; total antioxidant capacity (T-AOC) reflects the content of free radicals in the organism, and the antioxidant and disease-fighting capacities of the organism can be reflected by detecting various immunity and antioxidant indexes in the organism. As can be seen from Table 2: compared with the D0 group (0 part of clostridium acetobutylicum protein), the feed using 5 parts, 10 parts and 15 parts of clostridium acetobutylicum protein significantly reduces the activity of Malondialdehyde (MDA) (P <0.05), and has no significant effect on superoxide dismutase (SOD) (P > 0.05); the total antioxidant capacity (T-AOC) of group D0 was significantly lower than that of group D5 (P <0.05), with no significant difference from groups D10 and D15 (P > 0.05).
TABLE 2 influence of Clostridium ethanolate protein on the antioxidant capacity of juvenile fish by replacing soybean meal in Cyprinus carpiod feed
The clostridium ethanolate protein used in the invention can substitute for bean pulp in a formula of jian carp feed at the same protein level, so that the use amount of the bean pulp in the formula is reduced, and the utilization efficiency of the protein is improved. Meanwhile, the feed prepared by the clostridium ethanolate protein can effectively ensure the protein and energy sources of the freshwater omnivorous fish, the indexes of comprehensive growth performance, serum biochemistry, immunity and the like, and the feed has good effect on freshwater omnivorous fish juvenile fish by using 5-15 parts of clostridium ethanolate protein. Therefore, the clostridium ethanolate protein can be used as a good substitute for a protein raw material in the feed for the freshwater meat hybrid fishes.
Finally, it should also be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (8)
1. The application of clostridium ethanolate protein in the feed of freshwater omnivorous fishes is characterized in that the preparation method of the clostridium ethanolate protein comprises the following steps:
clostridium ethanolicum (Clostridium autoethanogenum) is used as a fermentation strain, CO-containing gas is used as a raw material, liquid fermentation culture is carried out, and thalli in fermentation liquor of the liquid fermentation are subjected to centrifugation and primary drying treatment to obtain Clostridium ethanolicum protein.
2. The use of a Clostridium autoethanolate protein in a feed for freshwater omnivorous fish according to claim 1, wherein Clostridium autoethanolate (Clostridium autoethanogenum) has deposit number DSM10061 and deposit address DSMZ, german collection of strains.
3. The use of clostridium autoethanolate protein in a feed for freshwater omnivorous fish according to claim 1, wherein the clostridium autoethanolate protein comprises the following nutritional components in parts by weight: more than or equal to 80 parts of crude protein, less than or equal to 7 parts of crude ash, less than or equal to 12 parts of water and less than or equal to 1 part of ammonium salt.
4. The use of clostridium autoethanolate protein in a feed for freshwater omnivorous fish according to claim 1, wherein the feed for freshwater omnivorous fish is prepared by a method comprising the following steps: adding clostridium ethanolate protein into the mixture, adding water, uniformly mixing, preparing into granules, and performing secondary drying to obtain the freshwater omnivorous fish feed.
5. The use of clostridial ethanolate protein in a feed for freshwater omnivorous fish according to any one of claim 4, wherein the mix comprises at least one of: fish meal, soybean meal, rapeseed meal, cottonseed meal, wheat flour, rice bran, soybean oil, monocalcium phosphate, bentonite, mineral premix, vitamin C, choline chloride, a mildew preventive and amino acid.
6. The use of Clostridium ethanolate protein in a feed for freshwater omnivorous fish according to claim 4, wherein the mix is passed through a 60 mesh sieve.
7. The use of clostridial ethanolate protein in a feed for freshwater omnivorous fish according to claim 4, wherein the particle size is 2-4 mm.
8. The use of clostridium autoethanolate protein in a feed for freshwater omnivorous fish according to claim 4, wherein the feed for freshwater omnivorous fish comprises, in parts by weight: 5-15 parts of clostridium ethanolate protein, 5 parts of fish meal, 4-29 parts of soybean meal, 14 parts of rapeseed meal, 10-14 parts of cottonseed meal, 21.01-29.61 parts of wheat flour, 5.30-8.2 parts of rice bran, 2.7 parts of soybean oil, 2.07-2.37 parts of monocalcium phosphate, 1.9-4.65 parts of microcrystalline cellulose, 3 parts of bentonite, 1 part of mineral premix, 0.1 part of vitamin C, 0.1 part of choline chloride, 0.05 part of mildew preventive and 0.22-0.77 part of amino acid.
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Application publication date: 20200221 |