CN117546959A - Deep fermentation cured corn for compound strain culture crabs and preparation method thereof - Google Patents

Deep fermentation cured corn for compound strain culture crabs and preparation method thereof Download PDF

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CN117546959A
CN117546959A CN202311718168.1A CN202311718168A CN117546959A CN 117546959 A CN117546959 A CN 117546959A CN 202311718168 A CN202311718168 A CN 202311718168A CN 117546959 A CN117546959 A CN 117546959A
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corn
preparation
culture
astaxanthin
feed
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王道胜
李娟�
李其松
刘艳玲
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Anhui Baojie Biotechnology Co ltd
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    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures

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Abstract

The invention discloses a compound crab feed prepared from deeply fermented corn and strain cultures. The invention discloses a preparation method of the feed, which comprises the steps of curing corn particles, and tabletting to obtain tabletting corn; uniformly mixing the pressed corn, gelatin and water, continuously adding glutaraldehyde under stirring, stirring at 40-50 ℃ for 1-2h, cooling to-10 to-30 ℃, preserving heat for 10-30min, freeze-drying, and washing to obtain pretreated pressed corn; adding water, bacillus subtilis bacterial liquid, a saccharomyces cerevisiae preparation and a lactobacillus plantarum preparation into the pretreated tabletting corn, uniformly mixing, and carrying out solid fermentation to obtain fermented corn; drying corncob to constant weight, grinding, sieving, dripping tetraethyl silicate, washing, drying, calcining at 300-350deg.C under nitrogen protection for 10-30min, cooling, and adding astaxanthin to obtain compound astaxanthin; adding compound astaxanthin, clostridium butyricum culture and Phaffia rhodozyma culture into the fermented corn, mixing uniformly, sealing and packaging.

Description

Deep fermentation cured corn for compound strain culture crabs and preparation method thereof
Technical Field
The invention relates to the technical field of deep processing of grains, in particular to a feed for deeply fermented corn and strain culture compound crabs and a preparation method thereof.
Background
Corn is a common aquaculture feed raw material, has the main advantages of rich energy and protein content, is a feed which is easy to digest and cheap, and is suitable for eating various aquatic products. However, the proportion of fatty acid in the feed is unbalanced when corn is used as aquatic feed, so that the fat content and the fatty acid content of the aquatic feed are affected. Meanwhile, excessive corn is added into the feed to reduce the immunity of aquatic animals, influence the resistance of the aquatic animals to pathogens and the digestion and absorption capacity of the aquatic animals to other nutritional ingredients. If the corn feed can be developed and utilized in a deeper level, not only the economic benefit of corn processing enterprises can be greatly brought, but also the defects of the original corn feed can be improved.
The corn is used for crab culture, contains a large amount of carbohydrate and grease, and can provide a large amount of energy sources for the young crabs in the later growth period. At present, the corn is often subjected to feed pre-digestion treatment before being used for crab culture. The feed predigestion is to convert macromolecules in feed raw materials into micromolecules which are easier to be absorbed by cultured animals through the actions of microorganisms, enzymes and the like, so that the feed utilization rate is improved. High temperature steam curing and microbial fermentation are very effective pre-digestion techniques.
In the crab culturing process, the antibiotic feed plays a great promoting role in preventing crab diseases and creating economic benefits. However, with the improvement of living standard, people pay more attention to the quality of consumer products, and the safety problem of crab products is becoming a focus of attention, so that the use of antibiotics is forbidden in the European Union. Development of efficient, residue-free, nuisanceless green feeds is an urgent and inevitable consequence of social development.
Studies have shown that: the clostridium butyricum is added into the feed, so that the total antioxidant capacity, superoxide dismutase activity and digestive enzyme activity of the hepatopancreas of the young crabs can be improved. And clostridium butyricum is bacillus capable of producing acid and has strong stress resistance. The rhodotorula is one of natural yeasts capable of producing astaxanthin, has the advantages of mild culture conditions, astaxanthin as a main component in pigments and the like, and becomes a preferred microorganism for industrial production of astaxanthin. And a proper amount of astaxanthin is added into the young crab feed, so that not only is the important promotion effect on the raising survival rate of young crabs improved, but also the quality of the adult crabs can be improved, and the raising yield and benefit are improved.
At present, the clostridium butyricum and the rhodozyma are compounded, so that the young crab immunity is enhanced, the crab production performance is improved, and the method has a very good prospect.
Disclosure of Invention
The invention aims to solve the defects in the prior art, and provides a feed for deeply fermenting corn and strain culture compound crabs and a preparation method thereof.
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, curing the corn particles by adopting high-temperature steam, and tabletting to obtain tabletting corn;
s2, uniformly mixing the pressed corn and the gelatin, adding water, continuously adding glutaraldehyde under the stirring state, adjusting the temperature to 40-50 ℃, continuously stirring for 2-4min, cooling to-10 to-30 ℃, preserving heat for 10-30min, freeze-drying, and washing to obtain the pretreated pressed corn;
s3, adding water, bacillus subtilis bacterial liquid, a saccharomyces cerevisiae preparation and a lactobacillus plantarum preparation into the pretreated tabletting corn, uniformly mixing, carrying out heat dissipation and oxygen introduction treatment when the solid fermentation is carried out to 45-50 ℃, and obtaining the fermented corn when the pH of the system is reduced to 4.0-4.5;
s4, drying corncob to constant weight, grinding, sieving, adding into ethanol solution, adding ammonia water until the pH value of the system is 8.0-8.5, dropwise adding tetraethyl silicate in a stirring state, continuously stirring for 1-2h, washing, drying, calcining for 10-30min at 300-350 ℃ under the protection of nitrogen, cooling, and adding astaxanthin for ultrasonic treatment to obtain compound astaxanthin;
s5, adding the compound astaxanthin, the clostridium butyricum culture and the phaffia rhodozyma culture into the fermented corn, uniformly mixing, sealing and packaging.
Preferably, in S1, the gelatinization degree of the curing treatment to the corn reaches 60-80%.
Preferably, in S2, the mass ratio of the corn flakes, the gelatin and the glutaraldehyde is 100:3-5:0.1-1.
Preferably, in S3, the mass ratio of the pretreated tabletting corn to the bacillus subtilis bacterial liquid to the saccharomyces cerevisiae preparation to the lactobacillus plantarum preparation is 100-120:1-2:4-6:1-2.
Preferably, in S2, absolute ethyl alcohol and water are used for alternately washing for 2-3 times, so that residual glutaraldehyde can be effectively removed, and the biosafety is improved.
Preferably, in the step S3, the bacillus subtilis bacterial liquid is obtained by adopting a bacillus subtilis culture medium for aerobic fermentation for 22-26 hours; the bacillus subtilis culture medium comprises: 2-3g/100g of beef extract, 2-3g/100g of peptone, 4-6g/100g of brown sugar and 4-6g/100g of NaCl, and the pH value of the system is maintained to be 7.0+/-0.5 in the aerobic fermentation process.
Preferably, in S3, the Saccharomyces cerevisiae preparation is obtained by aerobic fermentation of Saccharomyces cerevisiae culture medium for 22-26 hours; the saccharomyces cerevisiae culture medium comprises: 0.5-1.5g/100g yeast extract, 0.5-1.5g/100g peptone and 4-6g/100g brown sugar.
Preferably, in S3, the lactobacillus plantarum preparation is obtained by adopting lactobacillus plantarum culture medium for aerobic fermentation for 16-20 hours; the lactobacillus plantarum culture medium comprises: 3-5g/100g of soybean peptone, 5-7g/100g of glucose, 0.05-0.15g/100g of tween 80, 0.1-0.3g/100g of diammonium citrate, 0.4-0.6g/100g of sodium acetate and 0.1-0.3g/100g of dipotassium hydrogen phosphate.
Preferably, in S4, the mass ratio of the corncob to the tetraethyl silicate to the astaxanthin is 5-15:1-5:1-3.
Preferably, in S4, the ultrasonic treatment time is 1-2h, and the ultrasonic frequency is 5-12kHz.
Preferably, in S5, the mass ratio of the fermented corn, the complex astaxanthin, the clostridium butyricum culture and the rhodozyma culture is 110-130:10-20:2-8:8-12.
Preferably, in S5, the clostridium butyricum culture is prepared by the following steps: inoculating the inclined plane of activated clostridium butyricum into an RCM liquid culture medium, and standing and culturing at 37 ℃ until the inclined plane reaches a logarithmic phase, wherein the inclined plane is used as seed liquid for standby; inoculating the seed liquid into a fermentation tank according to the inoculation amount of 3-5%, wherein the culture medium in the fermentation tank comprises the following components: glucose 30.0g/L, yeast powder 5.0g/L, fish meal 30.0g/L, K 2 HPO 4 1.05g/L、MgSO 4 ·7H 2 O 1.21g/L、NaCl 0.3g/L、CaCO 3 1.5g/L, glucose solution was used for feeding during fermentation.
Preferably, in S5, the rhodozyma culture is prepared by the following steps: inoculating the rhodotorula bevel into a liquid culture medium (glucose 25g/L and yeast extract 8 g/L), and performing shake culture at 22 ℃ for 80 hours at 160 r/min; then, the culture was inoculated into a fermentation medium (glucose 2.5%, fructose 2%, yeast extract 0.5%) at an inoculum size of 10%, and the culture was performed under shaking for 168 hours at an initial pH=8.0 of the fermentation broth, an inoculum size of 20%, and a shaking speed of 160 r/min.
The feed for the deeply fermented corn and strain culture compound crabs is prepared by adopting the preparation method of the feed for the deeply fermented corn and strain culture compound crabs.
The beneficial effects are that:
according to the invention, corn is cured and sterilized by high-temperature steam, then fermentation and pre-digestion are carried out under the combined action of a plurality of microorganisms, and clostridium butyricum and rhodozyma cultures with the effects of improving the production performance and immunity of the young crabs are compounded, so that the feed mainly containing the corn for the young crabs is prepared, the nutrition is rich, and the yield of the crabs can be greatly improved.
According to the invention, after the corn kernels are cured at high temperature, the corn kernels are dispersed with gelatin, the gelatin effectively permeates into a pressed corn structure, and after cross-linking, the gelatin is frozen at low temperature, so that the internal porosity of the gelatin can be greatly increased, and the pressed corn is coated and permeated with a porous gelatin structure, so that the subsequent inoculation and fermentation of bacillus subtilis, saccharomyces cerevisiae and lactobacillus plantarum are facilitated, and the heat dissipation and the smooth entering of oxygen are facilitated in the fermentation process, so that the whole fermentation process is more thorough. The fermented corn obtained by the invention can provide needed nutrition for young crabs, is rich in nutrition, and can provide rich load structures for clostridium butyricum cultures and rhodozyma cultures by matching the fermented corn with the compound astaxanthin, so that the biological activities of clostridium butyricum and rhodozyma are effectively ensured, and the immunity and the growth speed of crabs are further improved.
The corncob tissues are uniform, the corncob tissues are dispersed in ethanol solution after being dried and ground, silicon dioxide is deposited on the surfaces of particles of the corncob tissues, a structure which takes the silicon dioxide as a shell layer and takes the interior of the corncob tissues as a uniform carbon hole core layer is formed after carbonization, the astaxanthin is effectively adsorbed, meanwhile, the core layer structure has high stability, the influence of air oxidation on the astaxanthin is reduced, the astaxanthin loss caused by unsealing of a packaging bag is avoided, the problem of low astaxanthin content caused by long-time placement is avoided, the utilization efficiency of composite astaxanthin is effectively ensured, the dosage of astaxanthin is reduced, the cost is saved, and a load carrier is further provided for clostridium butyricum cultures and rhodotorula cultures.
The applicant found through experiments that: compared with the common activated carbon added as a load carrier, the astaxanthin content in the juvenile crab shells fed by the method is obviously improved, and the average value of three immune indexes of superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) in juvenile crab serum is also obviously improved.
Drawings
FIG. 1 is a graph showing the comparison of the growth rates of the group of example 5, the group of comparative example 1 and the group of comparative example 2.
FIG. 2 is a SOD, CAT, T-AOC comparison plot of example 5, comparative example 1, comparative example 2, and blank.
FIG. 3 is a graph showing comparison of enzyme contents of the example 5 group, the comparative example 1 group, the comparative example 2 group and the blank group.
FIG. 4 is a graph comparing the SOD, CAT, T-AOC increase rates of the group of example 5, the group of comparative example 1, and the group of comparative example 2.
FIG. 5 is a graph showing the comparison of the increase rates of the enzyme contents in the group of example 5, the group of comparative example 1 and the group of comparative example 2.
FIG. 6 is a graph showing comparison of the mRNA expression levels of the EcR genes in the group of example 5, the group of comparative example 1, the group of comparative example 2 and the blank group.
Detailed Description
The invention is further illustrated below in connection with specific embodiments.
The following strains were purchased from China center for type culture Collection of microorganisms, including Bacillus subtilis (strain No. CICC 10023), saccharomyces cerevisiae (strain No. CICC 1296), and Lactobacillus plantarum (strain No. CICC 22200); the rhodotorula maritima strain is purchased from bioengineering stock, inc., baolaili, shandong, and Clostridium butyricum is purchased from Beijing ocean technology development Co.
The bacillus subtilis liquid is obtained by adopting a bacillus subtilis culture medium to perform aerobic fermentation for 24 hours; the bacillus subtilis culture medium comprises: beef extract 2.5g/100g, peptone 2.5g/100g, brown sugar 5g/100g and NaCl 5g/100g, and the pH of the system is maintained to be 7.0+/-0.5 in the aerobic fermentation process.
The following Saccharomyces cerevisiae preparation is obtained by aerobic fermentation of Saccharomyces cerevisiae culture medium for 22-26 hr; the saccharomyces cerevisiae culture medium comprises: 1g/100g of yeast extract, 1g/100g of peptone and 5g/100g of brown sugar.
The lactobacillus plantarum preparation is obtained by adopting lactobacillus plantarum culture medium for aerobic fermentation for 16-20 hours; the lactobacillus plantarum culture medium comprises: 4g/100g of soybean peptone, 6g/100g of glucose, 0.1g/100g of Tween 80, 0.2g/100g of diammonium citrate, 0.5g/100g of sodium acetate and 0.2g/100g of dipotassium hydrogen phosphate.
The following clostridium butyricum cultures were prepared using the following steps: inoculating the inclined plane of activated clostridium butyricum into an RCM liquid culture medium, and standing and culturing at 37 ℃ until the inclined plane reaches a logarithmic phase, wherein the inclined plane is used as seed liquid for standby; inoculating the seed liquid into a fermentation tank according to the inoculation amount of 3-5%, wherein the culture medium in the fermentation tank comprises the following components: glucose 30.0g/L, yeast powder 5.0g/L, fish meal 30.0g/L, K 2 HPO 4 1.05g/L、MgSO 4 ·7H 2 O 1.21g/L、NaCl 0.3g/L、CaCO 3 1.5g/L, M total 5% of continuously variable feed.
The following rhodozyma cultures were prepared by the following steps: inoculating the rhodotorula bevel into a liquid culture medium (glucose 25g/L and yeast extract 8 g/L), and performing shake culture at 22 ℃ for 80 hours at 160 r/min; then, the culture was inoculated into a fermentation medium (glucose 2.5%, fructose 2%, yeast extract 0.5%) at an inoculum size of 10%, and the culture was performed under shaking for 168 hours at an initial pH=8.0 of the fermentation broth, an inoculum size of 20%, and a shaking speed of 160 r/min.
Example 1
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles for 30min by using high-temperature steam, ensuring the gelatinization degree of the corn to reach 60%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 3kg of gelatin, adding 200kg of water into the mixture, adding 0.1kg of glutaraldehyde into the mixture under stirring at a stirring speed of 300r/min, continuously stirring at a temperature of 40 ℃ for 2min, cooling to-10 ℃ at a speed of 5 ℃/min, preserving heat for 10min, and alternately washing with absolute ethyl alcohol and water for 2 times after freeze drying to obtain pretreated pressed corn;
s3, adding 40kg of water, 1kg of bacillus subtilis bacterial liquid, 4kg of saccharomyces cerevisiae preparation and 1kg of lactobacillus plantarum preparation into 100kg of pretreated pressed corn, stirring for 1h in a mixer at a stirring speed of 50r/min, fermenting in a trough type solid fermentation mode, wherein the stacking height of materials is 80cm, and when the temperature reaches 45 ℃, carrying out heat dissipation and oxygen introduction treatment on the materials until the pH value of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 5kg of corncobs to constant weight at 50 ℃, grinding, sieving with a 100-mesh sieve, adding into 40kg of ethanol water solution with the mass fraction of 40%, adding ammonia water to the pH value of a system of 8.0-8.5, dropwise adding 1kg of tetraethyl silicate in a stirring state, continuously stirring for 1h, alternately washing with water and ethanol, drying, calcining at 300 ℃ for 10min under the protection of nitrogen, cooling, adding 1kg of astaxanthin, and carrying out ultrasonic treatment for 1h at the ultrasonic frequency of 5kHz to obtain the compound astaxanthin;
s5, adding 10kg of composite astaxanthin, 2kg of clostridium butyricum culture and 8kg of rhodozyma culture into 110kg of fermented corn, uniformly mixing, sealing and packaging.
Example 2
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 50min, ensuring the gelatinization degree of the corn to reach 80%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 5kg of gelatin, adding 300kg of water into the mixture, adding 1kg of glutaraldehyde into the mixture under stirring, continuously stirring at the temperature of 50 ℃ for 4min, cooling to-30 ℃ at the speed of 10 ℃/min, preserving heat for 30min, and alternately washing with absolute ethyl alcohol and water for 2 times after freeze drying to obtain pretreated pressed corn;
s3, adding 60kg of water, 2kg of bacillus subtilis bacterial liquid, 6kg of saccharomyces cerevisiae preparation and 2kg of lactobacillus plantarum preparation into 120kg of pretreated pressed corn, stirring for 2 hours in a mixer at a stirring speed of 200r/min, fermenting in a trough type solid fermentation mode, wherein the stacking height of materials is 100cm, and when the temperature reaches 50 ℃, carrying out heat dissipation and oxygen introduction treatment on the materials until the pH value of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 15kg of corncobs to constant weight at the temperature of 60 ℃, grinding, sieving with a 100-mesh sieve, adding into 100kg of ethanol water solution with the mass fraction of 50%, adding ammonia water to the pH value of a system of 8.0-8.5, dropwise adding 5kg of tetraethyl silicate in a stirring state, continuously stirring for 2 hours, alternately washing with water and ethanol, drying, calcining for 30 minutes at the temperature of 350 ℃ under the protection of nitrogen, cooling, adding 3kg of astaxanthin, and carrying out ultrasonic treatment for 2 hours at the ultrasonic frequency of 12kHz to obtain the compound astaxanthin;
s5, adding 20kg of composite astaxanthin, 8kg of clostridium butyricum culture and 12kg of rhodozyma culture into 130kg of fermented corn, uniformly mixing, sealing and packaging.
Example 3
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 20min, ensuring the gelatinization degree of the corn to reach 65%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 4.5kg of gelatin, adding 220kg of water into the mixture, adding 0.8kg of glutaraldehyde into the mixture under stirring at a stirring speed of 350r/min, continuously stirring at a temperature of 47 ℃ for 3min, cooling to-15 ℃ at a speed of 9 ℃/min, preserving heat for 25min, and alternately washing with absolute ethyl alcohol and water for 3 times after freeze drying to obtain pretreated pressed corn;
s3, adding 55kg of water, 1.2kg of bacillus subtilis bacterial liquid, 5.5kg of saccharomyces cerevisiae preparation and 1.3kg of lactobacillus plantarum preparation into 105kg of pretreated pressed corn, stirring for 110min in a mixer at a stirring speed of 80r/min, fermenting in a trough type solid fermentation mode, wherein the stacking height of materials is 95cm, radiating heat and introducing oxygen to the materials when the temperature reaches 47 ℃, until the pH of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 12kg of corncobs to constant weight at 53 ℃, grinding, sieving with a 100-mesh sieve, adding into 80kg of ethanol water solution with mass fraction of 42%, adding ammonia water to the pH value of the system of 8.0-8.5, dropwise adding 4kg of tetraethyl silicate in a stirring state, continuously stirring for 80min, alternately washing with water and ethanol, drying, calcining for 15min at 340 ℃ under the protection of nitrogen, cooling, adding 2.5kg of astaxanthin, and carrying out ultrasonic treatment for 70min at ultrasonic frequency of 10kHz to obtain compound astaxanthin;
s5, adding 18kg of composite astaxanthin, 4kg of clostridium butyricum culture and 11kg of rhodozyma culture into 115kg of fermented corn, uniformly mixing, sealing and packaging.
Example 4
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 60min, ensuring the gelatinization degree of the corn to be 75%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 3.5kg of gelatin, adding 280kg of water into the mixture, adding 0.2kg of glutaraldehyde into the mixture under stirring at the stirring speed of 450r/min, continuously stirring at the temperature of 43 ℃ for 3min, cooling to-25 ℃ at the speed of 7 ℃/min, preserving heat for 15min, and alternately washing with absolute ethyl alcohol and water for 3 times after freeze drying to obtain pretreated pressed corn;
s3, adding 45kg of water, 1.8kg of bacillus subtilis bacterial liquid, 4.5kg of saccharomyces cerevisiae preparation and 1.7kg of lactobacillus plantarum preparation into 115kg of pretreated pressed corn, stirring for 70min in a mixer at a stirring speed of 160r/min, fermenting in a trough type solid fermentation mode, wherein the stacking height of materials is 85cm, radiating heat and introducing oxygen to the materials when the temperature reaches 49 ℃, until the pH of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 8kg of corncobs to constant weight at 57 ℃, grinding, sieving with a 100-mesh sieve, adding into 60kg of ethanol water solution with the mass fraction of 48%, adding ammonia water to the pH value of a system of 8.0-8.5, dropwise adding 2kg of tetraethyl silicate in a stirring state, continuously stirring for 100min, alternately washing with water and ethanol, drying, calcining for 25min at 320 ℃ under the protection of nitrogen, cooling, adding 1.5kg of astaxanthin, and carrying out ultrasonic treatment for 110min at the ultrasonic frequency of 6kHz to obtain the compound astaxanthin;
s5, adding 12kg of compound astaxanthin, 6kg of clostridium butyricum culture and 9kg of rhodozyma culture into 125kg of fermented corn, uniformly mixing, sealing and packaging.
Example 5
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 40min, ensuring the gelatinization degree of the corn to reach 70%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 4kg of gelatin, adding 250kg of water into the mixture, adding 0.5kg of glutaraldehyde into the mixture under stirring at a stirring speed of 400r/min, continuously stirring at a temperature of 45 ℃ for 3min, cooling to-20 ℃ at a speed of 8 ℃/min, preserving heat for 20min, and alternately washing with absolute ethyl alcohol and water for 3 times after freeze drying to obtain pretreated pressed corn;
s3, adding 50kg of water, 1.5kg of bacillus subtilis bacterial liquid, 5kg of saccharomyces cerevisiae preparation and 1.5kg of lactobacillus plantarum preparation into 110kg of pretreated pressed corn, stirring for 90min in a mixer at a stirring speed of 120r/min, fermenting in a trough solid fermentation mode, wherein the stacking height of materials is 90cm, radiating heat and introducing oxygen to the materials when the temperature reaches 48 ℃, until the pH of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 10kg of corncobs to constant weight at the temperature of 55 ℃, grinding, sieving with a 100-mesh sieve, adding into 70kg of ethanol water solution with the mass fraction of 45%, adding ammonia water to the pH value of a system of 8.0-8.5, dropwise adding 3kg of tetraethyl silicate in a stirring state, continuously stirring for 90min, alternately washing with water and ethanol, drying, calcining for 20min at the temperature of 330 ℃ under the protection of nitrogen, cooling, adding 2kg of astaxanthin, and carrying out ultrasonic treatment for 90min at the ultrasonic frequency of 8kHz to obtain the compound astaxanthin;
s5, adding 15kg of compound astaxanthin, 5kg of clostridium butyricum culture and 10kg of rhodozyma culture into 120kg of fermented corn, uniformly mixing, sealing and packaging.
Comparative example 1
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 40min, ensuring the gelatinization degree of the corn to reach 70%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 4kg of gelatin, adding 250kg of water into the mixture, adding 0.5kg of glutaraldehyde into the mixture under stirring at a stirring speed of 400r/min, continuously stirring at a temperature of 45 ℃ for 3min, cooling to-20 ℃ at a speed of 8 ℃/min, preserving heat for 20min, and alternately washing with absolute ethyl alcohol and water for 3 times after freeze drying to obtain pretreated pressed corn;
s3, adding 50kg of water, 1.5kg of bacillus subtilis bacterial liquid, 5kg of saccharomyces cerevisiae preparation and 1.5kg of lactobacillus plantarum preparation into 110kg of pretreated pressed corn, stirring for 90min in a mixer at a stirring speed of 120r/min, fermenting in a trough solid fermentation mode, wherein the stacking height of materials is 90cm, radiating heat and introducing oxygen to the materials when the temperature reaches 48 ℃, until the pH of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, carrying out ultrasonic treatment on 10kg of activated carbon and 2kg of astaxanthin for 90min, wherein the ultrasonic frequency is 8kHz, so as to obtain the compound astaxanthin;
s5, adding 15kg of compound astaxanthin, 5kg of clostridium butyricum culture and 10kg of rhodozyma culture into 120kg of fermented corn, uniformly mixing, sealing and packaging.
Comparative example 2
A preparation method of a deeply fermented corn and strain culture compound crab feed comprises the following steps:
s1, sieving large corn particles to remove powder and impurities, filling the large corn particles into a pressure cooker, introducing steam, sterilizing the corn particles by using high-temperature steam for 40min, ensuring the gelatinization degree of the corn to reach 70%, and processing the corn particles into 2.5+/-0.5 mm of pressed corn by a corn tabletting machine;
s2, uniformly mixing 100kg of pressed corn and 4kg of gelatin, adding 250kg of water into the mixture, uniformly stirring, and alternately washing the mixture with absolute ethyl alcohol and water for 3 times after freeze drying to obtain pretreated pressed corn;
s3, adding 50kg of water, 1.5kg of bacillus subtilis bacterial liquid, 5kg of saccharomyces cerevisiae preparation and 1.5kg of lactobacillus plantarum preparation into 110kg of pretreated pressed corn, stirring for 90min in a mixer at a stirring speed of 120r/min, fermenting in a trough solid fermentation mode, wherein the stacking height of materials is 90cm, radiating heat and introducing oxygen to the materials when the temperature reaches 48 ℃, until the pH of the materials is reduced to 4.0-4.5, and stopping fermenting to obtain fermented corn;
s4, drying 10kg of corncobs to constant weight at the temperature of 55 ℃, grinding, sieving with a 100-mesh sieve, adding into 70kg of ethanol water solution with the mass fraction of 45%, adding ammonia water to the pH value of a system of 8.0-8.5, dropwise adding 3kg of tetraethyl silicate in a stirring state, continuously stirring for 90min, alternately washing with water and ethanol, drying, calcining for 20min at the temperature of 330 ℃ under the protection of nitrogen, cooling, adding 2kg of astaxanthin, and carrying out ultrasonic treatment for 90min at the ultrasonic frequency of 8kHz to obtain the compound astaxanthin;
s5, adding 15kg of compound astaxanthin, 5kg of clostridium butyricum culture and 10kg of rhodozyma culture into 120kg of fermented corn, uniformly mixing, sealing and packaging.
The feeds obtained in example 5 and comparative examples 1 to 2 were subjected to physical and chemical index detection, and the results were as follows:
example 5 Comparative example 1 Comparative example 2
Moisture content 40.70 41.52 40.33
pH 4.21 4.03 4.63
Clostridium butyricum content 1.80×10 7 CFU/g 1.42×10 7 CFU/g 1.35×10 7 CFU/g
Astaxanthin content 10.64μg/g 1.54μg/g 8.46μg/g
After the feeds obtained in example 5 and comparative examples 1 to 2 were left at room temperature for 30 days, physical and chemical index detection was performed again, and the results were as follows:
example 5 Comparative example 1 Comparative example 2
Moisture content 40.97 42.06 41.59
pH 3.89 3.68 3.92
Clostridium butyricum content 1.92×10 7 CFU/g 1.87×10 7 CFU/g 1.51×10 7 CFU/g
Astaxanthin content 10.53μg/g 0.98μg/g 8.07μg/g
The above results illustrate: according to the invention, after corncob is dried and ground, silicon dioxide is deposited on the particle surfaces of the corncob, and a structure with the silicon dioxide as a shell layer and a uniform carbon hole core layer inside is formed after carbonization, so that the astaxanthin is effectively adsorbed, and the initial astaxanthin content of the feed can be effectively improved; meanwhile, as the stability of the core layer structure is high and the sealing package is adopted, the influence of air oxidation on astaxanthin is effectively reduced, and the problem of low astaxanthin content caused by long-time placement is avoided.
The experimental aquariums (100 cm multiplied by 40cm multiplied by 60 cm) are adopted for cultivation, the water level is set at 30cm, the water temperature is 25 ℃, and micro-aeration oxygenation equipment is adopted. Eriocheir sinensis seedlings were used as test subjects and divided into 5 groups (3 replicates per group, 15 crab seedlings per replicate), which are example 5 group, comparative example 1 group, comparative example 2 group and blank group, respectively. Wherein, the feed obtained in the embodiment 5 is used for feeding in the embodiment 5, the feed obtained in the comparative example 1 is used for feeding in the comparative example 1, the feed obtained in the comparative example 2 is used for feeding in the comparative example 2, and the corn flour is used for feeding in the blank group.
Food is fed every evening, the experimental period is 21 days, and the growth rate of each group of crab seedlings is measured every 3 days.
Growth rate = (total mass of the group on the day-total mass of the blank group on the day)/(total mass of the blank group on the day x 100%)
As shown in fig. 1, the weight gain was evident in each of the example 5 group, the comparative example 1 group, and the comparative example 2 group, as compared with the blank group. The weight gain of the group of the example 5 and the group of the comparative example 1 is obviously better than that of the group of the comparative example 2.
The applicant believes that: the corn kernels are dispersed with the gelatin after being cured at high temperature, the gelatin effectively permeates into a pressed corn structure, and after being frozen at low temperature after being crosslinked, the internal porosity of the gelatin can be greatly increased, and the pressed corn is coated and permeated into a porous gelatin structure, so that the subsequent inoculation and fermentation of bacillus subtilis, saccharomyces cerevisiae and lactobacillus plantarum are facilitated, and the smooth entering of heat dissipation and oxygen is facilitated in the fermentation process, so that the whole fermentation process is more thorough. The fermented corn obtained by the invention can provide needed nutrition for young crabs, is rich in nutrition, and can provide rich load structures for clostridium butyricum cultures and rhodozyma cultures by matching the fermented corn with the compound astaxanthin, so that the biological activities of clostridium butyricum and rhodozyma are effectively ensured, and the immunity and the growth speed of crabs are further improved.
On the day of the end of the experiment, 2, i.e. 6, from each of the repeated crab fries were randomly selected, dissected under anesthesia on ice, the hepatopancreas was rapidly isolated, and the hepatopancreas was homogenized (10% w/v) in ice physiological saline (0.85% NaCl). The mixture was centrifuged at 2500rpm at 4℃for 10min, and the supernatant was collected to determine the enzyme activity. And superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) are measured by adopting corresponding kits.
The results of superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) are shown in figure 2, and the results of enzyme activity are shown in figure 3. Because some of the indices were too small, it was difficult to observe the phase difference in the figures, the applicant turned the example 5 group, the comparative example 1 group, and the comparative example 2 group into the increase rate as compared with the blank group, as shown in fig. 4 and 5.
Growth rate = (a certain group a index value-blank group a index value)/(blank group a index value x 100%)
As can be seen from fig. 2 to 5: the immune index and enzyme activity of the example 5 group and the comparative example 2 group are significantly better than those of the comparative example 1 group. The applicant believes that: according to the invention, corn cob is dried and ground and then dispersed in ethanol solution, silicon dioxide is deposited on the particle surfaces of the corn cob, a structure which takes the silicon dioxide as a shell layer and takes the interior as a uniform carbon hole core layer is formed after carbonization, the astaxanthin is effectively adsorbed, meanwhile, the stability of the core layer structure is high, the influence of air oxidation on astaxanthin is reduced, the utilization efficiency of composite astaxanthin is effectively ensured, and a load carrier is further provided for clostridium butyricum cultures and Phaffia rhodozyma cultures; the compound astaxanthin, clostridium butyricum culture and rhodozyma culture can effectively enhance the immunity of young crabs and improve the immunity index of the young crabs.
Extracting juvenile crab total RNA by using Trizol Reagent according to a specification method. Using PrimeScript TM The RT Reagent Kit (Perfect Real Time) synthesized cDNA according to the instructions. And (3) determining the relative expression quantity of the mRNA of the ecdysone receptor (EcR) gene by adopting a fluorescent quantitative PCR technology.
As shown in FIG. 6, the relative expression levels of mRNA of the EcR gene in the group of example 5, the group of comparative example 1, and the group of comparative example 2 were higher than those in the blank group, and the relative expression level in the group of example 5 was the highest.
In the nutritional study of crustaceans, the Molting Frequency (MF) is another important growth indicator. And as a targeting receptor for ecdysone, ecR plays a key role in crustacean shelling. Fig. 6 shows that: the invention effectively improves the expression of the juvenile crab ec R gene mRNA by multiple means of enhancing the fermentation degree, adopting the combination of clostridium butyricum culture and rhodozyma culture, improving the astaxanthin content and the utilization efficiency and the like, thereby improving the molting frequency and finally promoting the growth of the juvenile crab ec R gene mRNA.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.

Claims (10)

1. The preparation method of the feed for the deeply fermented corn and strain culture compound crabs is characterized by comprising the following steps:
s1, curing the corn particles by adopting high-temperature steam, and tabletting to obtain tabletting corn;
s2, uniformly mixing the pressed corn and the gelatin, adding water, continuously adding glutaraldehyde under the stirring state, adjusting the temperature to 40-50 ℃, continuously stirring for 2-4min, cooling to-10 to-30 ℃, preserving heat for 10-30min, freeze-drying, and washing to obtain the pretreated pressed corn;
s3, adding water, bacillus subtilis bacterial liquid, a saccharomyces cerevisiae preparation and a lactobacillus plantarum preparation into the pretreated tabletting corn, uniformly mixing, carrying out heat dissipation and oxygen introduction treatment when the solid fermentation is carried out to 45-50 ℃, and obtaining the fermented corn when the pH of the system is reduced to 4.0-4.5;
s4, drying corncob to constant weight, grinding, sieving, adding into ethanol solution, adding ammonia water until the pH value of the system is 8.0-8.5, dropwise adding tetraethyl silicate in a stirring state, continuously stirring for 1-2h, washing, drying, calcining for 10-30min at 300-350 ℃ under the protection of nitrogen, cooling, and adding astaxanthin for ultrasonic treatment to obtain compound astaxanthin;
s5, adding the compound astaxanthin, the clostridium butyricum culture and the phaffia rhodozyma culture into the fermented corn, uniformly mixing, sealing and packaging.
2. The method of producing a feed for crabs according to claim 1, characterized in that in S1, the gelatinization degree of the cured corn is 60 to 80%.
3. The preparation method of the crab feed according to claim 1, wherein in the S2, the mass ratio of the pressed corn to the gelatin to the glutaraldehyde is 100:3-5:0.1-1.
4. The preparation method of the crab feed according to claim 1, wherein in the S3, the mass ratio of the pretreated pressed corn, the bacillus subtilis bacterial liquid, the saccharomyces cerevisiae preparation and the lactobacillus plantarum preparation is 100-120:1-2:4-6:1-2.
5. The method for preparing the feed for crabs according to claim 1, wherein in S3, the bacillus subtilis liquid is obtained by adopting a bacillus subtilis culture medium for aerobic fermentation for 22-26 hours; the bacillus subtilis culture medium comprises: 2-3g/100g of beef extract, 2-3g/100g of peptone, 4-6g/100g of brown sugar and 4-6g/100g of NaCl, and the pH value of the system is maintained to be 7.0+/-0.5 in the aerobic fermentation process.
6. The method for preparing crab feed according to claim 1, wherein in S3, the saccharomyces cerevisiae preparation is obtained by aerobic fermentation of a saccharomyces cerevisiae culture medium for 22-26 hours; the saccharomyces cerevisiae culture medium comprises: 0.5-1.5g/100g yeast extract, 0.5-1.5g/100g peptone and 4-6g/100g brown sugar.
7. The method for preparing feed for crabs according to claim 1, wherein in S3, the lactobacillus plantarum preparation is obtained by adopting lactobacillus plantarum culture medium for aerobic fermentation for 16-20 hours; the lactobacillus plantarum culture medium comprises: 3-5g/100g of soybean peptone, 5-7g/100g of glucose, 0.05-0.15g/100g of tween 80, 0.1-0.3g/100g of diammonium citrate, 0.4-0.6g/100g of sodium acetate and 0.1-0.3g/100g of dipotassium hydrogen phosphate.
8. The preparation method of the crab feed according to claim 1, wherein in the S4, the mass ratio of the corncob to the tetraethyl silicate to the astaxanthin is 5-15:1-5:1-3.
9. The method for preparing crab feed according to claim 1, wherein in S5, the mass ratio of the fermented corn, the complex astaxanthin, the clostridium butyricum culture and the rhodozyma culture is 110-130:10-20:2-8:8-12.
10. A feed for deeply fermenting corn and strain culture compound crabs, which is characterized by being prepared by the method for preparing the feed for deeply fermenting corn and strain culture compound crabs according to any one of claims 1-9.
CN202311718168.1A 2023-12-14 2023-12-14 Deep fermentation cured corn for compound strain culture crabs and preparation method thereof Pending CN117546959A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117958355A (en) * 2024-04-02 2024-05-03 内蒙古海邻科技发展有限公司 Method for producing biological feed by using corn byproducts

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117958355A (en) * 2024-04-02 2024-05-03 内蒙古海邻科技发展有限公司 Method for producing biological feed by using corn byproducts
CN117958355B (en) * 2024-04-02 2024-06-11 内蒙古海邻科技发展有限公司 Method for producing biological feed by using corn byproducts

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