CN117256731A - Black soldier fly larva fermentation liquor, preparation method and application thereof, and shrimp feed - Google Patents
Black soldier fly larva fermentation liquor, preparation method and application thereof, and shrimp feed Download PDFInfo
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- CN117256731A CN117256731A CN202311184508.7A CN202311184508A CN117256731A CN 117256731 A CN117256731 A CN 117256731A CN 202311184508 A CN202311184508 A CN 202311184508A CN 117256731 A CN117256731 A CN 117256731A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 64
- 241000709785 Hermetia illucens Species 0.000 title claims abstract description 63
- 230000004151 fermentation Effects 0.000 title claims abstract description 53
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 241000238557 Decapoda Species 0.000 title claims abstract description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 38
- 244000005700 microbiome Species 0.000 claims abstract description 23
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 17
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 10
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 10
- 241000190950 Rhodopseudomonas palustris Species 0.000 claims abstract description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 10
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 10
- 235000015099 wheat brans Nutrition 0.000 claims abstract description 10
- 240000007817 Olea europaea Species 0.000 claims abstract description 9
- 235000013312 flour Nutrition 0.000 claims abstract description 9
- 241000607528 Aeromonas hydrophila Species 0.000 claims abstract description 8
- 244000062241 Kaempferia galanga Species 0.000 claims abstract description 8
- 235000013421 Kaempferia galanga Nutrition 0.000 claims abstract description 8
- 208000035240 Disease Resistance Diseases 0.000 claims abstract description 6
- 239000002131 composite material Substances 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 241000143060 Americamysis bahia Species 0.000 claims abstract 3
- 239000000463 material Substances 0.000 claims description 22
- 238000002156 mixing Methods 0.000 claims description 17
- 238000010564 aerobic fermentation Methods 0.000 claims description 13
- 238000002791 soaking Methods 0.000 claims description 13
- 241000894006 Bacteria Species 0.000 claims description 7
- 230000003078 antioxidant effect Effects 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
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- 238000000034 method Methods 0.000 claims description 6
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- 239000002245 particle Substances 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
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- 238000005119 centrifugation Methods 0.000 claims description 3
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- 230000000694 effects Effects 0.000 abstract description 6
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- 241000255925 Diptera Species 0.000 description 1
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- 241000709334 Hermetia Species 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
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- 210000001015 abdomen Anatomy 0.000 description 1
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- 230000032683 aging Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
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- 230000002147 killing effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
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- 238000003756 stirring Methods 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
Classifications
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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
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- 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
-
- 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
- A23K20/163—Sugars; Polysaccharides
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
- A61K35/64—Insects, e.g. bees, wasps or fleas
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Insects & Arthropods (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
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Abstract
The invention provides a hermetia illucens larva fermentation liquid, a preparation method and application thereof, and shrimp feed, and belongs to the technical field of feed preparation. Is prepared from bacillus subtilis and composite microorganism fermentation mixture; the mixture comprises cured black soldier fly larvae, flour, wheat bran, brown sugar, water and mixed alcohol extract; the preparation method of the cured black soldier fly larvae comprises the following steps: scalding the hermetia illucens larvae for 10-20 s at the temperature of 80-100 ℃; the raw materials of the mixed alcohol extract comprise galangal leaves, olive leaves and betel nut pulp; the compound microorganism comprises fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum. The hermetia illucens larva fermentation liquid provided by the invention improves the antioxidation activity and disease resistance to aeromonas hydrophila of the shrimps, and improves the weight and survival rate of the shrimps.
Description
Technical Field
The invention relates to the technical field of feed preparation, in particular to a hermetia illucens larva fermentation liquid, a preparation method and application thereof, and shrimp feed.
Background
The black soldier fly is a saprophytic insect of the genus hermetia of the order diptera, the hermetia illucens, the larvae of the black soldier fly are rich in protein, essential amino acid, unsaturated fatty acid, mineral substances and other nutrient substances, the content of the essential amino acid such as methionine, lysine and arginine is similar to that of fish meal, and the black soldier fly is an ideal feed raw material; meanwhile, the hermetia illucens larvae are rich in active substances such as antibacterial peptide, chitin, chitosan and the like, and can improve the immunity and disease resistance of cultured animals as baits. The hermetia illucens larva is pretreated by means of fermentation, so that the proteins of the larva can be decomposed into small peptides, amino acids and other nutrient substances and antimicrobial peptides and other active substances are fully released, and the absorption and utilization rate and the functionality are improved; meanwhile, the fermentation treatment damages and decomposes stimulative enzymes, mucus and the like of the hermetia illucens larvae, so that the stimulative damage to intestinal tracts of aquatic animals is avoided, and the palatability is enhanced. However, the bad smell of rancid smell generated by animal proteins during fermentation can greatly reduce the palatability of the ferment. Therefore, a proper fermentation method needs to be established, which can quickly and fully decompose the hermetia illucens larvae, release nutrition and active substances, and inhibit the generation of bad odor of foul smell at the same time, so that the fermentation liquor has good palatability.
Disclosure of Invention
In order to solve the problems, the invention provides the hermetia illucens larva fermentation liquor, the preparation method and application thereof, and the shrimp feed.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a black soldier fly larva fermentation broth which is prepared from bacillus subtilis and a composite microorganism anaerobic fermentation mixture;
the mixture comprises the following components in parts by weight: 20-30 parts of cured black soldier fly larvae, 3-8 parts of flour, 5-10 parts of wheat bran, 1-3 parts of brown sugar, 50-60 parts of water and 0.05-0.1 part of mixed alcohol extract;
the preparation method of the cured hermetia illucens larvae comprises the following steps: scalding the hermetia illucens larvae for 10-20 s at the temperature of 80-100 ℃;
the raw materials of the mixed alcohol extract comprise galangal leaves and olives She Hebin Lang Guorou;
the compound microorganism comprises fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum.
Preferably, the mass ratio of the bacillus subtilis to the mixture is 0.5-1.5:100; the effective viable count of the bacillus subtilis is 1 to 5 multiplied by 10 7 CFU/ml。
Preferably, the mass ratio of the compound microorganism to the mixture is 2-5:100.
Preferably, the fermenting yeast in the compound microorganism has a bacterial content of 1 to 5×10 6 CFU/ml, rhodopseudomonas palustris with a bacterial content of 1-5×10 7 CFU/ml, lactobacillus plantarum with a bacterial content of 1-5×10 7 CFU/ml。
Preferably, the preparation method of the mixed alcohol extract comprises the following steps:
pulverizing and mixing the rhizoma Alpiniae Officinarum leaves and fructus Canarii albi She Hebin Lang Guorou to obtain pulverized materials;
mixing the obtained crushed material with ethanol solution, and soaking to obtain soaking solution;
and (3) centrifuging the obtained soaking solution to obtain a supernatant, and concentrating the supernatant until the volume is reduced by 70-80% to obtain the mixed alcohol extract.
Preferably, the mass ratio of the galangal leaves to the olive She Hebin Lang pulp is 1:1:1;
the particle size of the crushed material is smaller than 80-100 meshes;
the mass ratio of the crushed material to the ethanol solution is 1:10-20, and the mass concentration of the ethanol solution is 75%;
the soaking time is 1d;
the centrifugation conditions include: the time is 20-30 min, and the rotating speed is 5000-8000 rpm.
The invention also provides a preparation method of the hermetia illucens larva fermentation liquid, which comprises the following steps:
1) Mixing the cured black soldier fly larvae, flour, wheat bran, brown sugar, water, mixed alcohol extract and bacillus subtilis, and performing aerobic fermentation for 2d to obtain an aerobic fermentation material;
2) And (2) mixing the aerobic fermentation material obtained in the step (1) with the compound microorganism, and performing anaerobic fermentation for 4-6 d to obtain the black soldier fly larva fermentation liquor.
The invention also provides application of the black soldier fly larva fermentation liquor in preparation of feed for improving the antioxidant activity of shrimps.
The invention also provides application of the black soldier fly larva fermentation liquor in preparation of the feed for improving disease resistance of shrimps to aeromonas hydrophila.
The invention also provides a shrimp feed, which comprises a basic feed and the hermetia illucens larva fermentation liquid according to the technical scheme;
the mass ratio of the basic feed to the hermetia illucens larva fermentation liquid is 100:3-5.
The invention carries out proper curing on the larva body of the hermetia illucens larva, and aims at:
(1) After the insect body is properly cured, the body fluid is changed from liquid into solid, and the insect body sterilizing enzymes and mucus are destroyed, so that the method is more suitable for the growth of fermentation type microorganisms, and improves the fermentation speed and degradation effect;
(2) Killing the microorganisms on the body surface of the insect body to a certain extent, and reducing the pollution of mixed bacteria;
(3) The protein of the insect body is denatured, so that the fermentation speed is improved.
According to the invention, the mixed extract of the alpinia galanga leaves, the olive leaves and the betel nut pulp is added, and the active substances of the mixed extract can effectively inhibit the growth of harmful miscellaneous bacteria, so that the fermentation speed is improved, the generation of bad smell is reduced, and the safety of fermentation liquor is ensured; the volatile smell of the plants can neutralize the bad smell generated by animal protein fermentation.
Detailed Description
The invention discloses a black soldier fly larva fermentation broth which is prepared from bacillus subtilis and a composite microorganism anaerobic fermentation mixture;
the mixture comprises the following components in parts by weight: 20-30 parts of cured black soldier fly larvae, 3-8 parts of flour, 5-10 parts of wheat bran, 1-3 parts of brown sugar, 50-60 parts of water and 0.05-0.1 part of mixed alcohol extract;
the preparation method of the cured hermetia illucens larvae comprises the following steps: scalding the hermetia illucens larvae for 10-20 s at the temperature of 80-100 ℃;
the raw materials of the mixed alcohol extract comprise galangal leaves and olives She Hebin Lang Guorou;
the compound microorganism comprises fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum.
In the invention, the mass ratio of the bacillus subtilis to the mixture is preferably 0.5-1.5:100; the bacterial content of the bacillus subtilis is preferably 1 to 5 multiplied by 10 7 CFU/ml. The source and the culture method of the bacillus subtilis are not particularly limited, and conventional methods are adopted. In the present invention, the cakeThe bacillus subtilis has the functions of promoting the conversion of organic matters in fermentation liquor, secreting antibacterial substances and inhibiting the growth and reproduction of pathogenic bacteria by decomposing macromolecular organic matters through aerobic fermentation.
In the invention, the mass ratio of the compound microorganism to the mixture is preferably 2-5:100. In the present invention, the microbial content of the fermenting yeast in the composite microorganism is preferably 1 to 5X 10 6 CFU/ml, the rhodopseudomonas palustris preferably 1 to 5X 10 in bacterial content 7 CFU/ml, the bacterium content of Lactobacillus plantarum is preferably 1-5×10 7 CFU/ml. The sources and the culture methods of the fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum are not particularly limited, and the fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum can be obtained by adopting conventional methods. In the invention, the effect of the compound microorganism is to further promote the decomposition of organic matters through anaerobic fermentation, inhibit the proliferation of harmful bacteria in the intestinal tract after entering the intestinal tract, improve the balance of intestinal flora, purify the intestinal environment and improve the immunity of cultured organisms.
In the present invention, the method for preparing the mixed alcohol extract preferably comprises the steps of: pulverizing and mixing the rhizoma Alpiniae Officinarum leaves and fructus Canarii albi She Hebin Lang Guorou to obtain pulverized materials; mixing the obtained crushed material with ethanol solution, and soaking to obtain soaking solution; and (3) centrifuging the obtained soaking solution to obtain a supernatant, and concentrating the supernatant until the volume is reduced by 70-80% to obtain the mixed alcohol extract. In the invention, the mass ratio of the galangal leaves to the olive She Hebin Lang pulp is preferably 1:1:1. In the present invention, the particle size of the pulverized material is preferably less than 80 to 100 mesh. In the present invention, the mass ratio of the pulverized material to the ethanol solution is preferably 1:10 to 20, and the mass concentration of the ethanol solution is preferably 75%. In the present invention, the soaking time is preferably 1d. In the present invention, the conditions of the centrifugation preferably include: the time is 20-30 min, and the rotating speed is 5000-8000 rpm.
The invention also provides a preparation method of the hermetia illucens larva fermentation liquid, which comprises the following steps:
1) Mixing the cured black soldier fly larvae, flour, wheat bran, brown sugar, water, mixed alcohol extract and bacillus subtilis, and performing aerobic fermentation for 2d to obtain an aerobic fermentation material;
2) And (2) mixing the aerobic fermentation material obtained in the step (1) with the compound microorganism, and performing anaerobic fermentation for 4-6 d to obtain the black soldier fly larva fermentation liquor.
And mixing the cured black soldier fly larvae, flour, wheat bran, brown sugar, water, mixed alcohol extract and bacillus subtilis, and performing aerobic fermentation for 2d to obtain an aerobic fermentation material. In the present invention, the aerobic fermentation conditions include: the temperature is 35-37 ℃ and the pH value is 6.5-7.0.
The invention mixes the obtained aerobic fermentation material with the compound microorganism and performs anaerobic fermentation for 4-6 d to obtain the black soldier fly larva fermentation liquor. In the present invention, the anaerobic fermentation conditions include: the temperature is 30-35 ℃, and the pH value is 6.5-6.8.
The invention also provides application of the black soldier fly larva fermentation liquor in preparation of feed for improving the antioxidant activity of shrimps. In the present invention, the antioxidant activity preferably includes total antioxidant activity, superoxide dismutase activity and/or malondialdehyde content.
The invention also provides application of the black soldier fly larva fermentation liquor in preparation of the feed for improving disease resistance of shrimps to aeromonas hydrophila.
The invention also provides a shrimp feed, which comprises a basic feed and the hermetia illucens larva fermentation liquid according to the technical scheme; the mass ratio of the basic feed to the hermetia illucens larva fermentation liquid is 100:3-5. The invention has no special limitation on the basic feed, and the basic feed is prepared by adopting conventional commercial feed, wherein the crude protein content is 32%, and the crude fat content is 7.8%. The application method of the shrimp feed is not particularly limited, and a person skilled in the art can routinely feed the shrimp feed, for example, the shrimp feed is fed 2 times per day according to 6% of the total weight of the shrimps.
The present invention will be described in detail with reference to examples for further illustration of the invention, but they should not be construed as limiting the scope of the invention.
Example 1
The preparation method of the hermetia illucens larva fermentation liquid comprises the following steps:
(1) Scalding hermetia illucens larva at 100deg.C for 20S, naturally airing surface moisture, weighing, and crushing (aging the larva into solid small particles after crushing, reducing liquid viscosity, facilitating attachment of fermentation strain, and improving protein denaturation and fermentation speed);
(2) The weight ratio is as follows: worm 20: flour 5: wheat bran 5: brown sugar 1: water 50, mixing well; (wheat bran is crushed, 80 meshes is passed, and brown sugar is crushed and dissolved)
(3) The preparation method of the mixed extract of the alpinia galanga leaves, olive leaves and betel nut pulp comprises the following steps: cleaning and sun-drying rhizoma Alpiniae Officinarum leaf, olive leaf and betel nut pulp, pulverizing, sieving with 80 mesh sieve, weighing powder, and weighing according to weight ratio of 1:1:1, adding ethanol solution with the mass concentration of 75% 10 times (weight ratio) into the mixed powder, soaking for 1 day, stirring for several times, centrifuging at 5000r/min for 30min, collecting supernatant, concentrating under reduced pressure to 80% of volume, and obtaining concentrated mixed extract of rhizoma Alpiniae Officinarum leaf, olive leaf and betel nut pulp.
(4) Adding 0.05 part (weight ratio) of concentrated mixed extract of galangal leaves, olive leaves and betel nut pulp into the mixture in the step (2), and uniformly mixing to obtain a material to be fermented; (effective in inhibiting the growth of harmful microorganisms, inhibiting and neutralizing the unpleasant odor produced by fermentation of animal proteins)
(5) Adding bacillus subtilis accounting for 1 percent of the mass percentage of the material to be fermented, wherein the effective viable bacteria content is 5 multiplied by 10 7 CFU/ml, mixing uniformly, and aerobically fermenting for 2 days;
(6) Then adding 1% of compound microorganism which is 5 x 10 of fermenting yeast 6 CFU/mL, rhodopseudomonas palustris 5×10 7 CFU/mL, lactobacillus plantarum 5 x 10 7 CFU/mL, mixing uniformly;
(7) And performing sealed anaerobic fermentation for 5 days to obtain the black soldier fly larva fermentation liquor.
Experimental example 1
Commercial feed (crude protein 32.0%, crude fat 7.8%) was used as basal feed.
Control group: and feeding basic feed.
Experimental group: before feeding, 1%, 3% and 5% of black soldier fly larva fermentation liquor are respectively added into the feed according to the mass percentage, and the mixture is uniformly stirred and air-dried for use.
Young red-chelating crayfish with intact appendages, healthy and non-diseased and in the molting interval were selected for the culture trial, randomly divided into 4 groups of 3 replicates each, 30 replicates each. The cultivation test is carried out in a net cage of an indoor circulating filtration cultivation system, the specification of each net cage is 100cm multiplied by 100cm, the water level is 70cm, and 30 PVC pipes are placed at the bottom of each cultivation box to serve as shielding objects. The water temperature is 26-30 ℃ and the pH is 7.8 during the cultivation. The feeding amount of the feed per day is 6% of the weight of the shrimp, and the feed is adjusted according to the actual feeding condition. Feeding is carried out twice a day, the time is 07:00 and 18:00 respectively, and the feeding period is 8 weeks. After 8 weeks of cultivation, 10 shrimps were each repeatedly taken and subjected to aeromonas hydrophila challenge experiments, and 1 x 10 were injected from the abdomen of the crayfish 8 CFU/ml aeromonas hydrophila fluid, 15 μl per tail injection, and survival rate after 72h was recorded. The results of different feeds on the growth, hepatopancreatic immune index and antibacterial ability of the red swamp crayfish are shown in table 1:
TABLE 1 Effect of different feeds on growth of Procambrus clarkii, hepatopancreatic Immunity index and antibacterial Capacity
The results in the table 1 show that when the addition amount of the black soldier fly larva fermentation liquor reaches 3% and 5%, the growth and survival rate of the red swamp crayfish are obviously increased; when the addition amount reaches more than 1%, the total antioxidant activity of the hepatopancreas and the superoxide dismutase activity are obviously increased, and when the addition amount reaches more than 3%, the malondialdehyde content is obviously reduced, which indicates that when the addition amount reaches more than 3%, the antioxidant activity of the crayfish body can be obviously improved; the result of the virus attack by the aeromonas hydrophila shows that when the addition amount reaches more than 3 percent, the disease resistance of the body of the crayfish to the aeromonas hydrophila can be obviously improved.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (10)
1. The hermetia illucens larva fermentation liquid is characterized by being prepared from bacillus subtilis and a compound microorganism anaerobic fermentation mixture;
the mixture comprises the following components in parts by weight: 20-30 parts of cured black soldier fly larvae, 3-8 parts of flour, 5-10 parts of wheat bran, 1-3 parts of brown sugar, 50-60 parts of water and 0.05-0.1 part of mixed alcohol extract;
the preparation method of the cured hermetia illucens larvae comprises the following steps: scalding the hermetia illucens larvae for 10-20 s at the temperature of 80-100 ℃;
the raw materials of the mixed alcohol extract comprise galangal leaves and olives She Hebin Lang Guorou;
the compound microorganism comprises fermenting yeast, rhodopseudomonas palustris and lactobacillus plantarum.
2. The hermetia illucens larva fermentation broth according to claim 1, wherein the mass ratio of the bacillus subtilis to the mixture is 0.5-1.5:100; the effective viable count of the bacillus subtilis is 1 to 5 multiplied by 10 7 CFU/ml。
3. The hermetia illucens larva fermentation broth of claim 1, wherein the mass ratio of the compound microorganism to the mixture is 2-5:100.
4. A hermetia illucens larva fermentation broth according to claim 1 or 3, wherein the effective viable bacteria content of the fermenting yeast in the composite microorganism is 1-5 x 10 6 CFU/ml, rhodopseudomonas palustris with a bacterial content of 1-5×10 7 CFU/ml, the effective viable bacteria content of the lactobacillus plantarum is 1 to 5 multiplied by 10 7 CFU/ml。
5. The hermetia illucens larva fermentation broth of claim 1, wherein the method of preparing the mixed alcohol extract comprises the steps of:
pulverizing and mixing the rhizoma Alpiniae Officinarum leaves and fructus Canarii albi She Hebin Lang Guorou to obtain pulverized materials;
mixing the obtained crushed material with ethanol solution, and soaking to obtain soaking solution;
and (3) centrifuging the obtained soaking solution to obtain a supernatant, and concentrating the supernatant until the volume is reduced by 70-80% to obtain the mixed alcohol extract.
6. The hermetia illucens larva fermentation broth of claim 5, wherein the mass ratio of galangal leaves to olive She Hebin boy pulp is 1:1:1;
the particle size of the crushed material is smaller than 80-100 meshes;
the mass ratio of the crushed material to the ethanol solution is 1:10-20, and the mass concentration of the ethanol solution is 75%;
the soaking time is 1d;
the centrifugation conditions include: the time is 20-30 min, and the rotating speed is 5000-8000 rpm.
7. A method for preparing the black soldier fly larva fermentation broth according to any one of claims 1 to 6, comprising the following steps:
1) Mixing the cured black soldier fly larvae, flour, wheat bran, brown sugar, water, mixed alcohol extract and bacillus subtilis, and performing aerobic fermentation for 2d to obtain an aerobic fermentation material;
2) And (2) mixing the anaerobic fermentation material obtained in the step (1) with the compound microorganism, and performing anaerobic fermentation for 4-6 d to obtain the black soldier fly larva fermentation liquor.
8. Use of the black soldier fly larva fermentation broth according to any one of claims 1-6 for preparing feed for improving the antioxidant activity of shrimps.
9. Use of the black soldier fly larva fermentation broth according to any one of claims 1-6 for preparing feed for improving disease resistance of shrimp against aeromonas hydrophila.
10. A shrimp feed, comprising a basal feed and the hermetia illucens larva fermentation broth of any one of claims 1-6;
the mass ratio of the basic feed to the hermetia illucens larva fermentation liquid is 100:3-5.
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