CN113317413B - Processing method and application of fermented feed for aquatic products based on hermetia illucens - Google Patents
Processing method and application of fermented feed for aquatic products based on hermetia illucens Download PDFInfo
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- CN113317413B CN113317413B CN202110627082.2A CN202110627082A CN113317413B CN 113317413 B CN113317413 B CN 113317413B CN 202110627082 A CN202110627082 A CN 202110627082A CN 113317413 B CN113317413 B CN 113317413B
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Classifications
-
- 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
- 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
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—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
- A23K10/30—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
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Abstract
The invention belongs to the field of biological feeds, and particularly relates to a processing method and application of a fermented feed for aquatic products based on hermetia illucens; the method comprises the following steps: activating the strain to obtain bacillus, saccharomycete and lactobacillus fermentation seed liquid; mincing fresh hermetia illucens larva, homogenizing, mixing with bran, rice bran and secondary powder, adding acid protease, mixing to obtain fermentation medium, inoculating bacillus fermentation seed liquid, saccharomycete fermentation seed liquid and lactobacillus fermentation seed liquid, and performing anaerobic fermentation under sealed condition to obtain the final product; the invention adopts the hermetia illucens fresh insects to carry out the synergistic fermentation of bacteria and enzymes, the fermentation is thorough, the finished product has high small peptide content, is rich in organic acid and has antibacterial activity; meanwhile, the storage time is long, the use is convenient, and the economic benefit is good.
Description
Technical Field
The invention belongs to the field of biological feeds, and particularly relates to a processing method and application of a fermented feed for aquatic products based on hermetia illucens.
Background
The aquatic feed accounts for 40% -60% of the aquaculture cost, fish meal is one of the most important feed protein sources in the aquatic feed, and more than 2/3 of the fish meal is internationally used as the aquatic feed, and reaches about 60% in China. With the rapid development of the aquaculture industry, the problem of global fish meal resource shortage is becoming more serious, and the aquaculture cost is increased due to the fact that the aquaculture feed relies on fish meal and fish oil extracted from bait fish, which severely restricts the sustainable development of the aquaculture industry. The hermetia illucens is an insect of hermetia illucens of the diptera, is widely distributed, and larvae thereof contain rich essential amino acids, wherein leucine, valine and lysine are high in content; in addition, the hermetia illucens has a nutrition enrichment function, can degrade kitchen waste and animal manure, enrich beneficial nutritional ingredients in the kitchen waste and animal manure, and convert the kitchen waste and animal manure into insect protein with high added value, and has good application prospect. Therefore, the black soldier fly resource is reasonably developed and utilized, and the black soldier fly resource is converted into high-quality biological feed by utilizing a microbial fermentation technology, so that the method has important significance.
At present, hermetia illucens feed conversion has been studied to a certain extent, such as patent CN 112544783a, which adopts hermetia illucens larva powder to replace fish meal; patent CN 112544813A is prepared by drying and pulverizing hermetia illucens, degreasing, mixing with fowl feed, and making into fowl feed. The equipment investment of the dry and crushing treatment of the hermetia illucens powder is large, the energy consumption is high, the protein can be destroyed by high-temperature treatment, the ash content is high, and the digestion and absorption rate of the organism is reduced; in addition, the whole or chopped feeding of the fresh black soldier flies can influence the digestion and absorption rate, reduce the growth performance of the organism, and partial small aquatic animals cannot directly eat the black soldier flies, meanwhile, the black soldier flies have the risk of carrying viruses and bacteria, and the proper addition amounts of the fresh black soldier flies are different for different aquatic animals; the hermetia illucens has different life cycles, and can be used for aquatic feeding, and the hermetia illucens pupa or adult is difficult to utilize, so that the storage and transportation of the hermetia illucens are difficult.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a processing method and application of a fermented feed for aquatic products based on hermetia illucens, which is used for fermenting the hermetia illucens larvae to convert protein macromolecules into small peptides, is rich in probiotics, has acid fragrance and antibacterial effect, and is convenient to store.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
(1) Respectively inoculating bacillus, saccharomycetes and lactobacillus into corresponding liquid culture mediums for activation culture to respectively obtain bacillus fermentation seed liquid, saccharomycetes fermentation seed liquid and lactobacillus fermentation seed liquid;
(2) Preparing a fermentation medium: mincing fresh hermetia illucens larva, homogenizing to obtain hermetia illucens homogenate, mixing with bran, rice bran and wheat middling, adding acid protease, and mixing to obtain a fermentation medium;
(3) Inoculating the bacillus fermentation seed liquid, the saccharomycete fermentation seed liquid and the lactobacillus fermentation seed liquid obtained in the step (1) into the fermentation culture medium obtained in the step (2), regulating the fermentation temperature and the water content of the culture medium, uniformly mixing to obtain a fermentation mixture, and carrying out anaerobic fermentation under a closed condition to obtain the fermented feed for the aquatic products.
Preferably, the bacillus in the step (1) is bacillus natto or bacillus subtilis; the saccharomycete is saccharomyces cerevisiae; the lactobacillus is one or more of lactobacillus plantarum, lactobacillus fermentum, lactobacillus paracasei, lactobacillus rhamnosus or bacillus coagulans.
Preferably, the weight percentages of the components of the fermentation medium in the step (2) are respectively as follows:
homogenizing 45% -60% of hermetia illucens;
15% -30% of bran;
15% -30% of rice bran;
secondary powder 9.95-18%;
0.05 to 1.5 percent of acid proteinase.
Preferably, the water content of the hermetia illucens homogenate in the step (2) is 60-70%.
Preferably, the acid protease in step (2) has an enzyme activity of 6 ten thousand u/g.
Preferably, the inoculum size of the bacillus fermentation seed liquid and the saccharomycete fermentation seed liquid in the step (3) is 0.1% -9% of the weight of the fermentation medium; live bacteria total of the bacillus and the saccharomycetesNumber average 1.0X10 8 ～2.8×10 9 cfu/mL; the yeast fermentation seed liquid is Saccharomyces cerevisiae fermentation seed liquid.
Preferably, the lactobacillus fermentation seed liquid in the step (3) is lactobacillus plantarum fermentation seed liquid, and the total number of viable bacteria is 1.5X10 8 ～3.5×10 9 cfu/mL, the inoculation amount is 0.8% -5.7% of the weight of the fermentation medium.
Preferably, the lactobacillus fermentation seed liquid in the step (3) further comprises one or more of lactobacillus fermentum fermentation seed liquid, lactobacillus paracasei fermentation seed liquid, bacillus coagulans fermentation seed liquid or lactobacillus rhamnosus fermentation seed liquid; the total number of viable bacteria of the lactobacillus fermentum fermentation seed liquid, lactobacillus paracasei fermentation seed liquid, bacillus coagulans fermentation seed liquid or lactobacillus rhamnosus fermentation seed liquid is 1.0X10 8 ～6.1×10 9 cfu/mL, the inoculation amount is 0.5% -6.5% of the weight of the fermentation medium.
Preferably, the water content of the fermentation mixture in step (3) is 40% to 60%.
Preferably, the fermentation time in the step (3) is 5-20 days, and the fermentation temperature is 24-38 ℃.
Among them, acid protease is purchased from the biotechnology limited company of the state of the art Cheng Mei and is a common commercial enzyme preparation.
Strain sources: (1) bacillus: bacillus natto (Bacillus natto) CGMCC1.1086 and Bacillus subtilis (Bacillus subtilis) CGMCC1.921
(2) Yeast: saccharomyces cerevisiae (Saccharomyces cerevisiae) CGMCC2.1527
(3) Lactic acid bacteria: lactobacillus plantarum (Lactobacillus plantarum) CGMCC1.557, lactobacillus fermentum (Lactobacillus fermentum) CGMCC1.15608, lactobacillus paracasei (Lactobacillus paracasei) CGMCC1.12731, lactobacillus rhamnosus (Lactobacillus rhamnosus) CGMCC1.577 and bacillus coagulans (Bacillus coagulans) CGMCC1.10823;
the strains are purchased from China general microbiological culture Collection center.
The beneficial effects of the invention are as follows:
(1) The invention adopts the mode of synergistic fermentation of bacterial enzymes and one-step anaerobic fermentation, and has thorough fermentation, high content of small peptide, acid fragrance, good palatability and high animal feeding quantity.
(2) The invention adopts a plurality of probiotics, and the used saccharomycetes increase nutrient substances such as mycoprotein, vitamins, a plurality of digestive enzymes and the like of a fermentation medium through self-propagation; the bacillus can secrete abundant extracellular enzyme systems, and during the propagation process, a large amount of protease, amylase, cellulase, lipase and the like are secreted, so that the degradation of protein, polysaccharide, fat and other organic matters in feed raw materials is facilitated, and the feed digestibility is improved; lactic acid bacteria produce organic acid, free amino acid, vitamins, functional bioactive peptide, unknown growth factors and other probiotics through self metabolism, and have antibacterial effect, improve intestinal flora growth environment and improve organism immunity.
(3) The product obtained by the method is convenient to store and transport; can be mixed with other feeds in a convenient proportion, can partially replace fish meal without affecting animal growth performance, has good feed effect and outstanding economic benefit.
Drawings
FIG. 1 is a graph showing the molecular weight distribution of the fermented product and the fermented material protein; wherein M is a protein Marker, F1 refers to a protein band of the fermentation raw material of example 1, and F2 is a protein band of the fermentation finished product.
FIG. 2 is a graph showing in vitro bacteriostatic activity of a fermented feed for aquatic products; wherein PC is a positive control; NC is negative control; SF is an experimental group for controlling Shigella flexneri by fermented feed; SA is an experimental group for controlling staphylococcus aureus by fermented feed; AH is an experimental group for controlling aeromonas hydrophila by fermenting feed; o157 is the experimental group of the fermented feed controlling Escherichia coli O157:H27; ET is an experimental group of fermented feed controlling edwardsiella tarda.
Detailed description of the preferred embodiments
The invention is further described below in connection with specific embodiments.
Example 1:
(1) Activation culture of bacillus, saccharomycetes and lactic acid bacteria:
A. firstly, preparing a culture medium of bacillus, saccharomycetes and lactobacillus;
bacillus liquid and solid media: 10g of tryptone, 10g of sodium chloride, 5g of yeast powder, 1L of distilled water, natural pH, sterilization at 121 ℃ for 20min, and the components of the bacillus solid culture medium are the same as those of the liquid culture medium; except that 15g of agar was added.
Yeast liquid and solid medium: filtering with 8 layers of gauze after 200g of potato is boiled and soft, filtering with 20g of glucose, 3g of potassium dihydrogen phosphate, 1.5g of magnesium sulfate, fixing the volume to 1L with distilled water, naturally sterilizing at 121 ℃ for 20min; the components of the saccharomycete solid culture medium are the same as those of the liquid culture medium, except that 15g of agar is added;
lactic acid bacteria liquid and solid medium: 10g of beef extract, 10g of tryptone, 20g of glucose, 5g of yeast powder, 2g of dipotassium hydrogen phosphate, 2g of diammonium hydrogen citrate, 5g of anhydrous sodium acetate, 1mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 1L of distilled water, pH7.2 and sterilizing at 121 ℃ for 20min; the components of the lactobacillus solid culture medium are the same as those of the liquid culture medium, except that 15g of agar is added;
B. the method for activating and culturing bacillus, saccharomycetes and lactobacillus comprises the following steps:
culturing bacillus liquid strain: inoculating glycerol bacteria preserved at-80deg.C to bacillus solid culture medium, culturing at 30deg.C for 18 hr for resuscitating strain, selecting single colony, inoculating into shake flask (250 mL) containing 100mL bacillus liquid culture medium, culturing at 180r/min at 30deg.C for 12 hr to obtain bacillus fermentation seed liquid; inoculating the strain into a liquid culture medium according to the actual requirement in an inoculum size of 2 percent for expansion culture; the bacillus comprises bacillus natto CGMCC1.1086 and bacillus subtilis CGMCC1.921.
Culturing yeast liquid strain: inoculating glycerol bacteria preserved at-80deg.C to yeast solid culture medium, culturing at 28deg.C for 20 hr to recover strain, then picking single colony, inoculating into shake flask (250 mL) containing 100mL yeast liquid culture medium, culturing at 180r/min at 28deg.C for 12 hr to obtain yeast fermentation seed liquid; inoculating the strain into a liquid culture medium according to the actual requirement in an inoculum size of 2 percent for expansion culture; the microzyme is Saccharomyces cerevisiae CGMCC 2.1527.
Culturing lactobacillus liquid strain: inoculating glycerol bacteria preserved at-80 ℃ to a lactobacillus solid culture medium respectively under aseptic conditions, culturing at 37 ℃ for 24 hours to recover strains, picking single bacterial colonies, inoculating into an anaerobic bottle (50 mL) filled with 50mL of lactobacillus liquid culture medium, standing at 37 ℃ for 18 hours to obtain lactobacillus fermentation seed liquid, and inoculating into the liquid culture medium according to actual needs by an inoculum size of 2% for expansion culture; the lactobacillus comprises lactobacillus plantarum CGMCC1.557, lactobacillus fermentum CGMCC1.15608, lactobacillus rhamnosus CGMCC1.577, bacillus coagulans 1.10823 and lactobacillus paracasei CGMCC1.12731.
(2) Preparing anaerobic fermentation culture medium, wherein the weight percentages of the components are as follows: the hermetia illucens homogenate is 50 percent (the water content is 70 percent); 17.5% of bran; 16.5% of rice bran; secondary powder 15%; 1% of acid protease, and uniformly mixing to obtain the anaerobic fermentation medium.
(3) Inoculating the bacillus subtilis fermentation seed liquid, the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the lactobacillus fermentum fermentation seed liquid, the lactobacillus paracasei fermentation seed liquid, the bacillus coagulans fermentation seed liquid and the lactobacillus rhamnosus fermentation seed liquid which are subjected to the activation culture in the step (1) into a fermentation culture medium, wherein the inoculum size of the bacillus subtilis and the saccharomyces cerevisiae is 1.5 percent of the weight of the fermentation culture medium, and the total number of the active bacteria of the bacillus subtilis and the saccharomyces cerevisiae is 1.7x10 percent 9 cfu/mL, lactobacillus plantarum inoculation amount is 2% of the weight of the fermentation medium, and the total number of viable bacteria is 2.3X10 9 cfu/mL, the inoculum size of the lactobacillus fermentum, the lactobacillus paracasei, the bacillus coagulans and the lactobacillus rhamnosus is 1 percent of the weight of the fermentation medium, and the total number of viable bacteria is 1.8x10 9 cfu/mL, uniformly mixing, and adjusting the water content to 54% to obtain an anaerobic fermentation mixture; anaerobic fermentation is carried out in a fermentation bag under the sealed condition, the fermentation temperature is 30 ℃, the finished product is obtained after 7 days of fermentation, the fermented feed for aquatic products is obtained, various indexes are detected to verify the quality of the feed, and the results are shown in the table 1.
Table 1 shows the results of the detection indexes of the fermented product of example 1
| Detecting items | Content of |
| Small peptide (%) | 54.83 |
| Cellulose | 15.17 |
| Hemicellulose | 19.65 |
| pH | 4.51 |
| Viable count of lactic acid bacteria | 1.29×10 9 cfu/g |
Example 2:
(1) The bacillus, saccharomycete and lactobacillus are activated and cultured, and the specific operation is the same as that of example 1;
(2) Preparing anaerobic fermentation culture medium, wherein the weight percentages of the components are as follows: the hermetia illucens homogenate is 55 percent (the water content is 70 percent); bran 11.2%; rice bran 20%; 13% of secondary powder; and (3) acid protease 0.8%, and uniformly mixing to obtain the anaerobic fermentation medium.
(3) Inoculating the bacillus natto fermentation seed liquid, the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the lactobacillus fermentum seed liquid and the lactobacillus rhamnosus seed liquid after the activation culture in the step (1) to a fermentation culture medium, wherein the inoculum size of the bacillus natto and the saccharomyces cerevisiae is 1.2 percent of the weight of the fermentation culture medium, and the inoculum size of the bacillus natto and the living bacteria of the saccharomyces cerevisiae are respectively equal to the inoculum size of the bacillus natto and the lactobacillus rhamnosusThe total number is 2.3X10 9 cfu/mL, lactobacillus plantarum inoculation amount is 3% of the weight of the fermentation medium, and the total number of viable bacteria is 2.8X10 9 cfu/mL, the inoculum size of lactobacillus fermentum and lactobacillus rhamnosus is 2% of the weight of the fermentation medium, and the viable count of the two bacteria is 4.2X10 9 cfu/mL, uniformly mixing, and adjusting the water content to 50% to obtain an anaerobic fermentation mixture; anaerobic fermentation is carried out in a fermentation bag under the sealed condition, the fermentation temperature is 33 ℃, the finished product is obtained after fermentation for 9 days, namely the fermented feed for aquatic products is obtained, and various indexes are detected to verify the quality of the feed, and the results are shown in the table 2.
Table 2 shows the results of the respective detection indexes of the fermented product of example 2
Example 3:
(1) The bacillus, saccharomycete and lactobacillus are activated and cultured, and the specific operation is the same as that of example 1;
(2) Preparing anaerobic fermentation culture medium, wherein the weight percentages of the components are as follows: the hermetia illucens homogenate is 48 percent (the water content is 70 percent); bran 20%; 14.7% of rice bran; secondary powder 16%; and (3) acid protease 1.3%, and uniformly mixing to obtain the anaerobic fermentation medium.
(3) Inoculating the bacillus natto fermentation seed liquid, the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the bacillus coagulans seed liquid and the lactobacillus paracasei seed liquid after the activation culture in the step (1) to a fermentation culture medium, wherein the inoculating amount of bacillus natto and the saccharomyces cerevisiae is 1% of the weight of the fermentation culture medium, and the total number of viable bacteria of the bacillus natto and the saccharomyces cerevisiae is 2.6x10 9 cfu/mL, lactobacillus plantarum inoculation amount is 4.2% of the weight of the fermentation medium, and the total number of viable bacteria is 1.9X10 9 cfu/mL, the inoculation amount of the bacillus coagulans and the lactobacillus paracasei is the weight of the fermentation medium1.5% of the total number of viable bacteria of both bacteria was 5.7X10% 9 cfu/mL, uniformly mixing, and adjusting the water content to 45% to obtain an anaerobic fermentation mixture; anaerobic fermentation is carried out in a fermentation bag under the sealed condition, the fermentation temperature is 36 ℃, the finished product is obtained after fermentation for 11 days, the fermented feed for aquatic products is obtained, various indexes are detected to verify the quality of the feed, and the results are shown in the table 3.
Table 3 shows the results of the respective detection indexes of the fermented product of example 3
| Detecting items | Content of |
| Small peptide (%) | 48.31％ |
| Cellulose | 17.41％ |
| Hemicellulose | 20.48％ |
| pH | 4.59 |
| Viable count of lactic acid bacteria | 6.92×10 8 cfu/g |
Tables 1-3 show the results of various measurements of the finished fermentation product, wherein the measured small peptide content refers to the proportion of small peptide to the crude protein content in the measured sample; the measured substances comprise small peptides, oligopeptides, a small amount of polypeptides, free amino acids and the like, and the results show that the invention has thorough fermentation and high content of the small peptides.
Example 4:
and (5) measuring the in-vitro antibacterial activity of the product.
Accurately weighing 1g of the aquatic fermented feed prepared in the example 1, adding 10mL of 50% methanol extract, leaching for 1h, filtering to obtain filtrate for later use, and filtering the extract through a 0.22 μm membrane filter before use; the antibacterial effect of the aquatic fermented feed on escherichia coli O157:H27 (Escherichia coli O157:H27) NCTC 12900, shigella flexneri (Shigella flexneri) CMCC (B) 51572, staphylococcus aureus (Staphylococcus aureus) ATCC 25923, aeromonas hydrophila (Aeromonas hydrophila) CGMCC 1.1801 and Edwardsiella tarda (Edwardsiella tarda) ATCC 1597is obtained by adopting an agar filter paper sheet diffusion method and taking 50% methanol solution as a negative control and taking kanamycin (0.05 g/mL) as a positive control.
Table 4 shows the inhibitory effect of hermetia illucens feed extract on various harmful bacteria
Note that: ++ means that the diameter of the inhibition zone is 10-13mm; + represents a bacteriostasis ring diameter of 7-10mm; -no zone of inhibition.
As can be seen from fig. 2 and table 4, the fermented feed extract for aquatic products prepared in example 1 has a relatively obvious inhibition effect on aeromonas hydrophila, and the diameter of the inhibition zone is greater than 10mm; the inhibition effect on staphylococcus aureus and shigella flexneri is secondary; has no inhibiting effect on Escherichia coli O157H 7 and Edwardsiella tarda.
Example 5:
feeding effect experiment of the product;
the fermented feed for aquatic products prepared by the invention is added into common aquatic feed for feeding special aquatic animals, and the addition amount is 5% -25% (by weight) of the common aquatic feed.
Selecting the aquatic fermented feed prepared in the embodiment 1, and carrying out a feeding experiment by taking Chinese soft-shelled turtles as experimental objects; the test point is Zhejiang Kangxing biotechnology Co. 60 Chinese soft-shelled turtles in the adult stage are taken, the average weight of the Chinese soft-shelled turtles is about 700g, the Chinese soft-shelled turtles are randomly combined into 2 groups, 3 parallel feeding groups of each feed are arranged, the feed is continuously fed for 60 days, the feed formula is shown in table 5, and the experimental results are shown in table 6.
The 10% by weight of the black soldier fly fermented feed prepared in the embodiment 1 replaces 10% of fish meal in the common feed, and the result shows that the weight gain rate and the feed coefficient of the Chinese soft-shelled turtle are not obviously different from those of a control group (P is more than 0.05), so that the feed prepared by the invention has good effect and outstanding economic benefit.
Table 5 shows the feed formulation of the Chinese soft-shelled turtle for test
TABLE 6 results of the finished feed feeding test prepared in example 1
| Feed variety | Control group | Test group |
| Initial mass (g) | 741.4±1.31 | 730.2±2.07 |
| Terminal quality (g) | 805.1±8.31 | 793.7±9.46 |
| Weight gain Rate (%) | 8.6±0.01 a | 8.7±0.06 a |
| Feed coefficient | 1.68±0.05 b | 1.66±0.03 b |
Note that: the same row of data marked with the same letter indicates that the difference was not significant (P > 0.05) and marked with a different letter indicates that the difference was significant (P < 0.05).
Description: the above embodiments are only for illustrating the present invention and not for limiting the technical solution described in the present invention; thus, while the invention has been described in detail with reference to the various embodiments described above, it will be understood by those skilled in the art that the invention may be modified or equivalents; all technical solutions and modifications thereof that do not depart from the spirit and scope of the present invention are intended to be included in the scope of the appended claims.
Claims (1)
1. The application of the fermented feed for aquatic products based on hermetia illucens in inhibiting harmful bacteria and special aquatic animal cultivation is characterized by comprising the following preparation steps of:
(1) Inoculating bacillus subtilis, saccharomyces cerevisiae and lactobacillus to corresponding liquid culture mediums respectively for activation culture to obtain bacillus subtilis fermentation seed liquid, saccharomyces cerevisiae fermentation seed liquid and lactobacillus fermentation seed liquid respectively; wherein the lactobacillus comprises Lactobacillus plantarum, lactobacillus fermentum, lactobacillus paracasei, bacillus coagulans and Lactobacillus rhamnosus;
(2) Preparing anaerobic fermentation culture medium, wherein the weight percentages of the components are as follows: homogenizing hermetia illucens to obtain a homogenate with water content of 70%; 17.5% of bran; 16.5% of rice bran; secondary powder 15%; 1% of acid protease, and uniformly mixing to obtain an anaerobic fermentation culture medium;
(3) Inoculating the bacillus subtilis fermentation seed liquid, the saccharomyces cerevisiae fermentation seed liquid, the lactobacillus plantarum fermentation seed liquid, the lactobacillus fermentum fermentation seed liquid, the lactobacillus paracasei fermentation seed liquid, the bacillus coagulans fermentation seed liquid and the lactobacillus rhamnosus fermentation seed liquid which are subjected to the activation culture in the step (1) into a fermentation culture medium, wherein the inoculum size of the bacillus subtilis and the saccharomyces cerevisiae is 1.5 percent of the weight of the fermentation culture medium, and the total number of the active bacteria of the bacillus subtilis and the saccharomyces cerevisiae is 1.7x10 percent 9 cfu/mL, lactobacillus plantarum inoculation amount is 2% of the weight of the fermentation medium, and the total number of viable bacteria is 2.3X10 9 cfu/mL, the inoculum size of the lactobacillus fermentum, the lactobacillus paracasei, the bacillus coagulans and the lactobacillus rhamnosus is 1 percent of the weight of the fermentation medium, and the total number of viable bacteria is 1.8x10 9 cfu/mL, uniformly mixing, and adjusting the water content to 54% to obtain an anaerobic fermentation mixture; anaerobic fermentation is carried out in a fermentation bag under the sealing condition, the fermentation temperature is 30 ℃, and the finished product is obtained after 7 days of fermentation, namely the fermented feed for aquatic products;
the harmful bacteria are as follows: aeromonas hydrophila, staphylococcus aureus and shigella flexneri;
the special aquatic animal is a Chinese soft shell turtle and is operated as follows: the aquatic fermented feed is added into the aquatic feed, and the addition amount is 10% -25% by weight.
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