CN111149951A

CN111149951A - Preparation method of active fermented feed for stichopus japonicus

Info

Publication number: CN111149951A
Application number: CN202010086881.9A
Authority: CN
Inventors: 赵强; 姜作真; 杨晓东; 张金浩; 高雁; 胡丽萍; 贺加贝; 柯可
Original assignee: Yantai Marine Economic Research Institute
Current assignee: Yantai Marine Economic Research Institute
Priority date: 2020-02-11
Filing date: 2020-02-11
Publication date: 2020-05-15

Abstract

The invention relates to a preparation method of an active fermented feed for stichopus japonicus. The method comprises the following steps: step 1, mixing and activating bacillus subtilis powder with the content of more than 100 hundred million per gram, glucose and water, and culturing until the content of bacillus subtilis per milliliter is more than one hundred million to obtain bacillus subtilis activation solution; step 2, mixing the bacillus subtilis activation solution with the feed raw materials, and performing primary fermentation to obtain a fermented feed raw material; and 3, activating candida, lactobacillus, streptococcus faecalis, glucose and water, mixing with the traditional Chinese medicine compound, mixing with the fermented feed raw materials for the second time, and performing secondary fermentation to obtain the active fermented feed for the stichopus japonicus. The invention controls the whole fermentation process by using bacteriocin and antibacterial peptide substances generated by high-strength beneficial bacteria to inhibit harmful bacteria and adding Chinese herbal medicines for strengthening body resistance and inhibiting pathogenic factors to regulate the phase of the fermentation bacteria.

Description

Preparation method of active fermented feed for stichopus japonicus

Technical Field

The invention relates to a preparation method of an active fermented feed for stichopus japonicus, belonging to the field of aquatic animal culture.

Background

Stichopus japonicus is the best one of more than 20 edible Stichopus japonicus in China. The natural distribution is mainly in the northern Pacific region, and is mainly distributed in China in the areas of Daliang, Qinhuang island, Jiangsu, Hongyou, Qingdao, tobacco terrace, Weihai and sunshine, etc. Stichopus japonicus is classified as one of the eight delicacies as a precious seafood. Among the stichopus japonicus families, the stichopus japonicus of the Shandong peninsula and the Liaodong peninsula has better quality. According to records in Bencao gang mu Shi Yi, the sea cucumber produced in Liaodong, which is named Liaoshen or sea cucumber, is black and brown in body color, tender in meat and thorny, has excellent quality, sweet, warm and nontoxic properties, and has the effects of tonifying kidney yin, generating blood, treating diarrhea and ulcer, and the like. The sea cucumber is named because the property of the sea cucumber is warm and tonify and is sufficient to enemy the ginseng.

In the natural environment, the stichopus japonicus mainly eats particles such as algae debris, phytoplankton corpses, microorganisms and silt mixed in the microorganisms deposited on the surface layer of the sea bottom. At present, the domestic stichopus japonicus breeding industry becomes an important supporting industry in mariculture, the scale of 'north ginseng south cultivation' and 'east ginseng west movement' is large, and the annual output value breaks through 300 billion yuan. At present, the feed used for cultivating stichopus japonicus in China is uneven, and most manufacturers only refer to the feeding mode of terrestrial cultivated animals to crush and mix various feed raw materials to prepare the feed. Because the stichopus japonicus is used as a component of a sea bottom food chain, the content of various endogenous digestive enzymes in the stichopus japonicus is lower than that of other cultured animals, so that the utilization rate of various nutrient components in the feed is far lower than that of common aquaculture animals. Therefore, huge waste of the sea cucumber feed and pollution of the culture environment caused by eutrophication are brought.

Disclosure of Invention

The invention aims to provide a method for preparing an active fermented feed for stichopus japonicus, aiming at the problems of high feed coefficient, heavy environmental load and the like in the conventional stichopus japonicus feed. The preparation method can reduce the feed coefficient of Stichopus japonicus and reduce environmental pollution.

In order to solve the technical problems, the technical scheme of the invention is as follows:

a method for preparing an active fermented feed for stichopus japonicus comprises the following steps:

step 1, mixing and activating bacillus subtilis powder with the content of more than 100 hundred million per gram, glucose and water, and culturing until the content of bacillus subtilis per milliliter is more than one hundred million to obtain bacillus subtilis activation solution;

step 2, mixing the bacillus subtilis activation solution with the feed raw materials, and performing primary fermentation to obtain a fermented feed raw material;

and 3, activating candida, lactobacillus, streptococcus faecalis, glucose and water, mixing with the traditional Chinese medicine compound, mixing with the fermented feed raw materials for the second time, and performing secondary fermentation to obtain the active fermented feed for the stichopus japonicus.

Further, in the step 1, the ratio of the bacillus subtilis powder to the glucose to the water is 10 g: 10 g: (1-1.1) L.

Further, the step 1 activation conditions are as follows: activating for 24-60h at 18-22 ℃.

Further, the weight ratio of the bacillus subtilis activation solution to the feed raw materials in the step 2 is (1-1.1): 9.

further, the primary fermentation conditions in the step 2 are as follows: fermenting for 36-48h in a closed container containing a proper amount of air at 24-26 ℃.

Further, the feed raw materials in the step 2 comprise the following components in parts by weight: 200 parts of soybean meal, 50 parts of corn protein, 200 parts of kelp powder, 200 parts of sargassum powder, 100 parts of enteromorpha powder, 100 parts of brassica rapa powder, 20 parts of schizochytrium limacinum powder, 10 parts of phospholipid, 10 parts of monocalcium phosphate and 20 parts of compound vitamin and mineral substance.

Further, in the step 3, the ratio of the candida, the lactobacillus and the streptococcus faecalis to the glucose and the water is 2 g: 40 g: 40 g: 10 g: (1-1.1) L.

Further, the weight ratio of the activated compound bacterial liquid in the step 3 to the traditional Chinese medicine compound is 3: (1-1.1).

Further, the weight ratio of the activated compound bacterial liquid mixed with the traditional Chinese medicines in the step 3 to the fermented feed raw materials is (1-1.1): 20.

further, the Chinese medicinal compound in the step 3 comprises the following raw materials in parts by weight: 30 parts of common andrographis herb, 15 parts of baical skullcap root, 30 parts of Chinese pulsatilla root, 30 parts of dandelion, 30 parts of astragalus, 30 parts of coptis root, 15 parts of Chinese angelica, 15 parts of ash bark, 30 parts of purslane and 10 parts of liquorice.

Has the advantages that:

compared with other fermented feeds, the invention controls the whole fermentation process by using bacteriocin and antibacterial peptide substances generated by high-strength beneficial bacteria to inhibit harmful bacteria and adding Chinese herbal medicines for strengthening body resistance and inhibiting pathogenic factors to regulate the phase of the fermented bacteria. The method has the characteristics of convenient production operation flow, strong operability and low production cost, and the finished product has the advantages of being capable of being stored for more than half a year and the like.

Detailed Description

Example 1

step 1, firstly mixing 1000 g of bacillus subtilis powder with the content of more than 100 hundred million per gram, 1000 g of glucose and 100 liters of water which is boiled and then cooled to room temperature, activating for 24-60 hours at the temperature of 18-22 ℃, and culturing until the content of bacillus subtilis per milliliter is more than one hundred million to obtain bacillus subtilis activation solution;

step 2, mixing 100 kg of bacillus subtilis activating solution and 900 kg of feed raw materials according to the weight ratio of 1 to 9, and then placing the mixture in a closed container (a cement pond, a glass steel tank and a stainless steel tank) containing a proper amount of air at the room temperature of 24-26 ℃ for fermentation for 36-48h to obtain a fermented feed raw material;

the feed comprises the following raw materials: 200 kg of soybean meal, 50 kg of corn protein, 200 kg of kelp powder, 200 kg of sargassum powder, 100 kg of enteromorpha powder, 100 kg of large-leaf vegetable powder, 20 kg of schizochytrium limacinum powder, 10 kg of phospholipid, 10 kg of monocalcium phosphate and 20 kg of compound vitamin and mineral substance; the mesh number of the feed raw materials is 100 meshes;

step 3, placing 100 g of candida (the content of per gram is more than 50 hundred million), 2000 g of lactobacillus (the content of per gram is more than 50 hundred million), 2000 g of streptococcus faecalis (the content of per gram is more than 50 hundred million) and 500 g of glucose in 50 liters of water which is boiled and then cooled to room temperature for activation for 24 hours to obtain activated compound bacterial liquid; mixing the activated compound bacterial liquid with 1500 g of traditional Chinese medicine compound with micro powder to 150 meshes to obtain activated compound bacterial liquid mixed with traditional Chinese medicines; mixing the activated compound bacterial liquid mixed with the traditional Chinese medicines and the fermented feed raw materials in the step 2 for the second time according to the weight ratio of 1 to 20, placing the mixture in a closed container with the temperature of 15 to 30 ℃ for 48 to 72 hours, observing the strong fermentation fragrance, and packaging the mixture;

the traditional Chinese medicine compound comprises the following components: 30 g of common andrographis herb, 15 g of baical skullcap root, 30 g of Chinese pulsatilla root, 30 g of dandelion, 30 g of astragalus, 30 g of coptis root, 15 g of Chinese angelica, 15 g of ash bark, 30 g of purslane and 10 g of liquorice.

The screening of the traditional Chinese medicine compound is obtained by creative labor of the inventor. The Chinese medicinal composition comprises coptis chinensis, common andrographis herb, Chinese pulsatilla root, baical skullcap root and dandelion, has a good inhibition effect on fungal infection, and is used as a main medicament; radix astragali and radix Angelicae sinensis promote the rapid propagation of beneficial bacteria, and are used as adjuvant drugs. The invention can effectively inhibit fungi, and has no strong inhibition effect on beneficial bacteria such as lactic acid bacteria, yeast and bacillus.

Mixing Eucommiae cortex, Coptidis rhizoma, and herba Andrographitis in the Chinese medicinal composition with fermented feed material according to weight ratio of ten-thousandth, respectively, and observing fungal contamination for 3 days, 10 days, 15 days, and 30 days, respectively, in 5 small samples per group, as shown in Table 1.

TABLE 1 fungal contamination of eucommia bark, Coptis chinensis and Andrographis paniculata

Then, five parts per million are added according to the respective formula of the coptis chinensis, the andrographis paniculata and the eucommia ulmoides, and the fungal pollution condition is observed as shown in table 2.

TABLE 2 respective compositions of Coptidis rhizoma, herba Andrographitis, and Eucommiae cortex for fungal contamination

Traditional Chinese medicine formula	Eucommia bark and coptis root	Eucommia bark and common andrographis herb	Coptidis rhizoma and herba Andrographitis
				3 days	Good effect	Good effect	Good effect
10 days	Good effect	Good effect	Good effect
				15 days	Group 1 contamination	Group 2 contamination	Good effect
30 days	Group 2 contamination	4 group contamination	Good effect

On the basis of screening out the main group of coptis and andrographis paniculata, according to the property and formula principle of the traditional Chinese medicine, a formula taking coptis, andrographis paniculata, Chinese pulsatilla root, scutellaria and dandelion as main medicines and taking astragalus, angelica, ash bark, dens equi and liquorice as auxiliary medicines is prepared, and is shown in table 3.

TABLE 3 fungal contamination of the Main and adjuvant drugs

The fermentation smell of the main and auxiliary medicines is more fragrant and pure in color. The composition is stable and reliable in use and good in effect through practical production verification. The traditional Chinese medicine compound is used for regulating the balance of various bacterial phases and effectively preventing the fungal infection. After the traditional Chinese medicine compound is added, the feed is stored for 3 months, and no mould infection batch is found as long as the bag is not opened or the package is not damaged. The proportion of the mould infection of the batch without the Chinese herbal medicines reaches more than 30 percent.

Further, the candida, lactobacillus and bifidobacterium compound bacteria liquid added for the second time in the fermentation is anaerobic fermentation flora, and the candida, lactobacillus and bifidobacterium compound bacteria liquid play a role in the secondary fermentation after the oxygen in the container is completely consumed in the bacillus subtilis flora. If the complex flora is not added for the second time, the first-time added bacillus subtilis is aerobic bacteria, and the bacillus subtilis does not have the fermentation effect after the oxygen in the container is completely consumed, so that the fermentation cannot be smoothly completed.

The cultivation example verifies that the feed prepared in the embodiment 1 is used for cultivating stichopus japonicus offspring seeds, the feeding amount is increased by 22.1 percent and the stichopus japonicus intestinal canal is thickened by 38.3 percent under the same cultivation condition, the weight is increased by 36.2 percent in the same proportion after a cultivation period of more than 70 days, the bait coefficient is reduced by 11.6 percent, the skin yield of the stichopus japonicus with the same weight is increased by 14.3 percent, and the economic benefit is remarkable.

In the same culture environment, a comparative test is carried out on the feeding effect of the fermented feed and a common compound feed (a commercially available common compound feed), as shown in table 4, stichopus japonicus is cultured in an outdoor pond, two culture areas of 100 square meters under the same conditions are respectively enclosed by purse nets, 50 kilograms of stichopus japonicus with the individual weight of 40-50 grams are respectively added, the water temperature of the culture environment is 13-18.5 ℃, feeds with different weights are added according to the ingestion condition of the stichopus japonicus, and the feeding time is 72 days (9/20 days in 2019-11/30 days in 2019).

TABLE 4 comparison of the cultivation effects of different feeds

Compared with the common feed sold in the market, the feed amount of the stichopus japonicus is increased by 22.1%, the intestinal tract is thickened by 38.3%, the weight gain rate is improved by 36.2%, the feed coefficient is reduced by 11.6%, and the carcass proportion is improved by 14.3%.

Example 1 the length of the fermentation in step 2 is important for the invention. The active fermented feed of example 1 was used for stichopus japonicus cultivation, selected stichopus japonicus individuals were relatively uniform, the average individual weight was 5.3 g, the water temperature for indoor cultivation of stichopus japonicus was 20.4-23.6 ℃, and the weight gain of stichopus japonicus at different fermentation time periods was tested during the cultivation process, as shown in table 5.

TABLE 5 comparison of duration and effect of Bacillus subtilis fermentation

Duration of fermentation (hours)	12	18	24	30	36	42	48
								15-day weight gain (%)	13.5	14.8	20.3	29.5	31.3	32.2	31.5
Weight gain (% in 30 days)	22.3	30.3	35.6	50.4	52.3	51.8	52.5
								45-day weight gain (%)	48.6	56.4	63.6	112.6	120.4	122.6	119.8

The above data show that the duration of the activated Bacillus subtilis for the first fermentation is preferably controlled to be 36-48 hours.

Comparative example 1

A method for preparing an active fermented feed for stichopus japonicus, which comprises the same steps as example 1. The difference is that the bacillus subtilis powder is directly mixed with feed raw materials without being activated.

The fermentation effects of the bacillus subtilis powder before and after activation for feed fermentation are compared, the activation environment temperature of the bacillus subtilis powder is 18-22 ℃, and after secondary fermentation, the mould infection rate is shown in table 6.

TABLE 6 comparison of duration of activation of Bacillus subtilis powder

Length of activation (h)	0	12	24	36	48	60
							Mold infection Rate (%) after Secondary fermentation	10	4	0	0	0	0

Table 6 shows that the activation time is 0h, namely the Bacillus subtilis powder in the comparative example 1 is not activated and is directly mixed with feed raw materials, and the mould infection rate after secondary fermentation is 10 percent; the activation time of the bacillus subtilis powder is more than 24 hours, namely the activation time is 24-60 hours in example 1, and the mould infection rate after secondary fermentation is 0%.

Various antibacterial substances released by high-content bacillus subtilis inhibit and kill harmful flora in the process of primary fermentation of feed raw materials. The bacillus can not only ferment feed raw materials due to huge content, but also can quickly inhibit and kill various harmful bacteria in the feed raw materials due to the capability of producing a large amount of bacteriocin and antibacterial peptide substances. According to the operation mode of the invention, 50 batches of production and processing are carried out, and no cases of fungal infection occur. If the bacillus is not activated in advance, the ratio of black mold appearing on the surface of the feed produced reaches 5 times per 50 production batches.

The invention feeds the stichopus japonicus after fermenting and degrading various raw materials suitable for being digested and absorbed by the stichopus japonicus, so that the digestion and absorption of the stichopus japonicus are easy, the feed coefficient of the stichopus japonicus is reduced, the environment in intestinal tracts of the stichopus japonicus is improved due to the fact that the stichopus japonicus is rich in various beneficial bacteria and digestive enzymes, the water quality environment of the stichopus japonicus is also improved, and the stichopus japonicus is in line with the current industry trend of green development, energy conservation and.

The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The above-described embodiments of the invention are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims, and not by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims

1. A preparation method of an active fermented feed for stichopus japonicus is characterized by comprising the following steps:

2. The method of claim 1, wherein the ratio of bacillus subtilis powder, glucose and water in step 1 is 10 g: 10 g: (1-1.1) L.

3. The method of claim 1, wherein the step 1 activation conditions are: activating for 24-60h at 18-22 ℃.

4. The method for preparing the bacillus subtilis solution of claim 1, wherein the weight ratio of the bacillus subtilis activation solution to the feed raw materials in the step 2 is (1-1.1): 9.

5. the method of claim 1, wherein the initial fermentation conditions of step 2 are: fermenting for 36-48h in a closed container containing a proper amount of air at 24-26 ℃.

6. The preparation method of claim 1, wherein the feed raw materials in the step 2 comprise the following components in parts by weight: 200 parts of soybean meal, 50 parts of corn protein, 200 parts of kelp powder, 200 parts of sargassum powder, 100 parts of enteromorpha powder, 100 parts of brassica rapa powder, 20 parts of schizochytrium limacinum powder, 10 parts of phospholipid, 10 parts of monocalcium phosphate and 20 parts of compound vitamin and mineral substance.

7. The method according to claim 1, wherein the ratio of Candida, Lactobacillus, Streptococcus faecalis, glucose and water in step 3 is 2 g: 40 g: 40 g: 10 g: (1-1.1) L.

8. The method according to claim 1, wherein the weight ratio of the activated compound bacterial liquid and the traditional Chinese medicine compound in the step 3 is 3: (1-1.1).

9. The method for preparing the fermented feed additive according to claim 1, wherein the weight ratio of the activated compound bacterial liquid mixed with the traditional Chinese medicines in the step 3 to the fermented feed raw materials is (1-1.1): 20.

10. the preparation method of claim 1, wherein the Chinese medicinal composition in step 3 comprises the following raw materials in parts by weight: 30 parts of common andrographis herb, 15 parts of baical skullcap root, 30 parts of Chinese pulsatilla root, 30 parts of dandelion, 30 parts of astragalus, 30 parts of coptis root, 15 parts of Chinese angelica, 15 parts of ash bark, 30 parts of purslane and 10 parts of liquorice.