CN115843919B - Preparation method and application of edible fungus concentrated feed - Google Patents

Abstract

The invention belongs to the technical field of fermented feeds and the field of poultry aquaculture, and particularly relates to a preparation method and application of an edible fungus concentrated feed. Adding edible fungus strains into a compound fermentation medium, fermenting under the fermentation condition of combining the stepwise regulation and control of rotation speed and pH, separating solid from liquid after fermentation to obtain mycelia, and performing dispersion treatment and alkali treatment on the mycelia to obtain the edible fungus concentrated feed. According to the invention, by analyzing the influence of different carbon sources, nitrogen sources, phosphorus source concentrations and effectors on the growth of the thalli, the optimal compound fermentation medium is obtained through compounding and verification, and the production cost is reduced while the yield of the thalli is improved. The preparation of the edible fungi with high digestibility, which is applied to the field of feed, is realized by combining a compound fermentation culture medium with a fermentation regulation and post-treatment process, so that the effect of feeding the feed is improved.

Description

Preparation method and application of edible fungus concentrated feed

Technical Field

The invention belongs to the technical field of fermented feeds and the field of poultry aquaculture, and particularly relates to a preparation method and application of an edible fungus concentrated feed.

Background

In the current production practice for preparing the concentrated feed of the edible fungi, the fermentation medium for preparing the fermented edible fungi is used by enterprises all the time: 25g/L of glucose, 10g/L of peptone, 10g/L of yeast extract, 2g/L of ammonium sulfate, 0.4g/L of magnesium sulfate, 0.5g/L of potassium dihydrogen phosphate and pH value of 6.0. However, the prices of glucose, peptone and yeast extract are higher, and the whole production cost of enterprises is further increased along with the rising price of raw materials.

The edible fungi is a high-quality biological resource, low in fat and energy, rich in protein, triterpene, vitamin and other nutrient substances, and has wide application in the aquiculture of poultry. However, edible fungi are processed as feed, and there is always a low digestibility in feeding poultry and aquatic products, which results in waste of resources.

Therefore, under the condition of ensuring the yield of the thalli, the culture medium components which are more in line with the interests of enterprises are sought, the production cost is reduced, and meanwhile, the preparation of the edible fungi with high digestibility in the feed field is improved, so that the feed feeding effect is further improved.

Disclosure of Invention

The invention aims to solve the technical problem of providing a preparation method of edible fungi concentrated feed aiming at the defects of the prior art.

The invention also solves the technical problem of providing the application of the edible fungi concentrated feed.

In order to solve the technical problems, the invention adopts the following technical scheme:

a process for preparing concentrated feed of edible fungus includes such steps as adding edible fungus strain to compound fermentation medium, fermenting under the condition of regulating rotational speed and pH, solid-liquid separation to obtain mycelia, dispersing and alkali treating.

The compound fermentation medium is characterized in that the influence of different carbon sources, nitrogen sources, phosphorus source concentrations and effectors (bran, zinc sulfate, nicotinamide and biotin) on the growth of the thalli is analyzed through a single factor test, and a proper amount of carbon-nitrogen ratio and effectors are found to have an obvious promotion effect on the growth of the thalli. And then carrying out compound verification on the basis of the optimal concentration.

Specifically, the carbon source of the compound fermentation medium is corn sugar, the nitrogen source is corn steep liquor, the phosphorus source is phosphoric acid, and the effector is bran, zinc sulfate, nicotinamide and biotin. Effectors are related to enzyme synthesis or activity and bacterial growth, for example, cellulose and vitamins in bran have promotion effect on enzyme induction and bacterial growth; zinc ions can promote the improvement of enzyme activity similar to cofactors, nicotinamide has the effect similar to vitamins, and biotin can promote the growth of thalli.

Specifically, the compound fermentation medium comprises the following components: 10-35g/L of corn sugar, 10-30g/L of corn steep liquor, 4-12g/L of ammonium sulfate, 0.5-1.5g/L of phosphoric acid, 0.1-1g/L of magnesium sulfate, 0.2-1.5g/L of bran, 0.02-0.2g/L of zinc sulfate, 0.002-0.2g/L of nicotinamide, 0.0005-0.05g/L of biotin and pH=6.0.

Preferably, the compound fermentation medium is: 26g/L of corn sugar, 25g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of phosphoric acid, 0.4g/L of magnesium sulfate, 0.6g/L of bran, 0.05g/L of zinc sulfate, 0.002g/L of nicotinamide, 0.005g/L of biotin and pH=6.0.

Wherein the edible fungi comprise any one or a combination of a plurality of pleurotus geesteranus, beefsteak mushroom and velvet mushroom.

The preferred edible fungi are Pleurotus geesteranus and beefsteak mushroom, and the most preferred edible fungi are beefsteak mushroom.

Wherein, the staged regulation takes the thallus quantity reaching 7-15g/L as a regulation node and is divided into the fermentation early-stage regulation and the fermentation middle-stage and later-stage regulation. The preferred stepwise regulation takes the cell quantity reaching 7g/L as a regulating node.

Specifically, the fermentation early-stage regulation and control is carried out, the rotating speed is 300-500r/min, and the pH value is 4-6; the fermentation is regulated in the middle and later stages, the rotating speed is 100-300r/min, and the pH value is 7-8.

Preferably, the fermentation is regulated and controlled in the earlier stage, the rotating speed is 400r/min, and the pH value is 5; and (3) regulating and controlling the middle and later fermentation period, wherein the rotating speed is 250r/min, and the pH value is 7.2.

Wherein the temperature of the fermentation is 25-29 ℃, the ventilation is 0.8-1.2vvm, and the fermentation culture time is 2-5 days.

Preferably, the fermentation temperature is 27 ℃, the ventilation is 0.8-1.2vvm, and the fermentation culture time is 3 days.

Wherein the dispersion treatment is to mechanically break the wall of mycelium for 30-120s.

Specifically, after adding 2 times of volume of water into mycelium obtained by solid-liquid separation, mechanically breaking the cell wall for 30-120s by using a wall breaking machine (Jiuyang wall breaking machine). The preferred mechanical wall breaking treatment time is 30s.

Wherein, the alkali treatment is to treat the mycelium after the dispersion treatment for 8 hours at the temperature of between pH7.2 and 14 and the temperature of 55 ℃, then adjust the pH value to between 6 and 7 and analyze the change of digestibility under different pH values.

The preferred alkali treatment is to treat the dispersion-treated mycelium at pH12, 55℃for 8 hours.

Specifically, the preparation method of the edible fungi concentrated feed comprises the following steps:

(1) Inoculating the activated edible fungus strain into seed liquid, and culturing until the wet weight concentration of the strain reaches 14g/L;

(2) Inoculating the seed solution containing edible fungus strains in the step (1) into a compound fermentation culture medium according to the inoculation amount of 10% v/v, wherein the aeration rate is 0.8-1.2vvm, the tank pressure is 0.1Mpa, the minimum dissolved oxygen concentration is controlled to be 30-40%, and the fermentation culture is carried out for 2-4 days under the conditions of stepwise regulation of the rotating speed of 100-500r/min and the pH value of 4-8;

(3) Performing dispersion treatment on mycelium obtained by solid-liquid separation on thalli obtained by fermentation culture in the step (2);

(4) And (3) performing alkali treatment on the mycelium subjected to the dispersion treatment in the step (3), and directly drying and crushing the mycelium to prepare mycelium powder.

The edible fungi concentrated feed prepared by the preparation method is also within the scope of the invention.

The use of the edible fungus concentrate in ruminant or aquaculture or poultry animal feeding is also within the scope of the present invention.

Specifically, in the assessment of milk yield of cows, cowshed of Xie 1-1, 1-2 and 2-1 is selected, the number of lactation cows is 3-4, the number of lactation is 200-220 days, and the daily milk yield is 30-36kg, and the cows are divided into 3 groups, each group comprises 110 heads, and the groups are respectively a control group, a test group 1 and a test group 2. The control group was fed with normal basal diet (mainly alfalfa 18%, corn silage 25%, corn meal 12%, soybean meal 12%, oat 4%, concentrated feed 5%, other 24%), the test group 1 added mycelium powder without post-treatment to 150g per cow on the basal diet (i.e. the bacterial liquid after fermentation under the condition of pH5/400rpm was separated by example 6, the bacterial liquid was directly dried and crushed, and sieved by a 100 mesh sieve), the test group 2 added mycelium powder with post-treatment to 150g per cow on the basal diet (i.e. the treatment mode of test group d in example 8 was adopted, and the bacterial liquid was dried and crushed and sieved by a 100 mesh sieve). The experimental period was 3 months, and the average daily milk yield of three groups was counted every half month. The experimental results are as follows, the effect on milk yield is not obvious in one month of feeding, the milk yield of the experimental group 2 is improved by nearly 2kg compared with that of the control group from the 6 th week, and the feeding effect of the experimental group 1 is improved by a relatively small amplitude. At week 10 of feeding, milk production was increased by 1.3kg and 3.2kg for test groups 1 and 2, respectively. The results show that the treated mycelium powder has remarkable promotion effect on milk yield of cows.

Specifically, in the evaluation of the weever cultivation effect, 300 weever tails (about 8 cm) with basically consistent length are selected and divided into 3 groups, the groups are carried out in a cultivation net cage, the original formula feed is fed by a control group, fish meal and yeast powder in the original weever raw feed are replaced by mycelium powder by a test group, wherein the fish meal and the yeast powder are replaced by the mycelium powder on the basis of the original formula in the test group 1 (bacterial liquid separation after fermentation is carried out under the condition of pH5/400rpm in the example 6, the bacterial liquid is directly dried and crushed, and the mycelium is sieved by a 100-mesh sieve); test group 2 replaced the fish meal and yeast meal with treated mycelium powder based on the original formulation (using the treatment method of test group d in example 8, oven dried, crushed and sieved with a 100 mesh sieve). The feeding amount of the groups 3 is 2% of the fish mass every day, and the groups are fed in the morning and evening. The experimental conditions of the 3 groups are consistent, and the cultivation period is 2 months. Compared with a control group, the weever weight gain rates of the test group 1 and the test group 2 are respectively improved by 11.4 percent and 23.8 percent, the survival rate is improved by 7.8 and 21.3 percent, the growth performance of the weever can be obviously improved, and the culture benefit of the weever can be improved.

Specifically, in the evaluation of broiler breeding, 400 broilers of 3 days old were randomly divided into 4 groups, the control group was fed with basic ration, and the test group 1 was fed with basic ration +70kg ^a Per ton of ration (mycelium powder obtained by directly drying, crushing and sieving with 100-mesh sieve of the fungus body by separating fungus liquid after fermentation in the fermentation tank 3 in the example 5), and feeding the basic ration +70kg in the test group 2 ^b Per ton of ration (bacterial liquid separation after fermentation under the condition of pH5/400rpm in example 6 is adopted), and bacterial cells are directly driedMycelium powder obtained by crushing and sieving with a 100-mesh sieve), and feeding basic ration +70kg in test 3 ^c Per ton of daily ration (mycelium powder obtained by sieving with 100 mesh sieve by adopting the treatment mode of the test group d of the example 8), and the daily feed intake and daily gain and feed-meat ratio of 50 days old are measured. Results average daily gain was increased by 11.2%, 16.2% and 23.3% for test groups 1, 2 and 3, respectively, relative to the control group. Daily feed intake is also improved obviously.

The beneficial effects are that:

(1) The optimal compound fermentation medium is obtained through analysis of the influence of different carbon sources, nitrogen sources, phosphorus source concentrations and effectors (bran, zinc sulfate, nicotinamide and biotin) on the growth of the thalli, and the preparation method is verified, so that the thalli yield is improved, and meanwhile, the production cost is reduced.

(2) The preparation of the edible fungi with high digestibility, which is applied to the field of feed, is realized by combining a compound fermentation culture medium with fermentation regulation (combination of stepwise regulation of rotation speed and pH) and a post-treatment process (dispersion treatment and alkali treatment), so that the effect of feeding the feed is improved.

Drawings

The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.

FIG. 1 shows the morphological changes of the cells at different rotational speeds.

FIG. 2 is a graph showing the dispersion treatment of cells having different diameters.

Detailed Description

The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available.

The pleurotus geesteranus and beefsteak mushrooms described in the examples below were purchased from a Taobao agricultural product store at day 12 of 2021.

In the current production practice, the fermentation medium used for preparing the fermented edible fungus body is: 25g/L of glucose, 10g/L of peptone, 10g/L of yeast extract, 2g/L of ammonium sulfate, 0.4g/L of magnesium sulfate, 0.5g/L of monopotassium phosphate and pH6.0 (as a control group, no. 0). But the prices of glucose, peptone and yeast extract are higher, and the overall production cost of enterprises is increased. Therefore, under the condition of ensuring the yield of the thalli, the culture medium components which are more beneficial to enterprises are sought, and the production cost is reduced.

EXAMPLE 1 Effect of different Components on the fermentation yield of the thallus

Transferring Pleurotus geesteranus strain into PDA slant culture medium, culturing in constant temperature incubator at 25deg.C for 3-4 days, and activating strain.

Inoculating the activated strain into seed solution, and culturing at 26deg.C and 200r/min for 3 days until the wet weight concentration of thallus reaches 14g/L. Then, the culture was inoculated into 150mL of a liquid fermentation medium containing different components according to an inoculum size of 10% v/v, and cultured at 27℃for 4 days under 220 r/min. After the fermentation, the solid-liquid separated cells were washed twice with pure water, the dry weight of the cells was measured, and the dry weight concentration (yield) was calculated and filled into the following table.

PDA slant culture medium comprises the following components: cleaning potato, peeling, weighing 200g, cutting into small pieces, adding water, boiling for 20-30 min, filtering with eight layers of gauze, heating, adding 15-20g agar and 5-10g glucose, continuously heating, stirring, adding water to 1000mL, packaging into test tube or conical flask after agar is dissolved, adding plug, wrapping, sterilizing at 115deg.C for 20 min, cooling, and storing.

The seed liquid comprises the following components: glucose 10g/L, molasses (converted sugar content) 15g/L, mgSO4 0.6g/L, feSO4.7H2O 0.08g/L, KH2PO4 0.6g/L, and soybean juice. ( The soybean juice is used for replacing water ingredients, and the soybean juice treatment mode comprises the following steps: taking 25g of soybeans, adding 800mL of water, steaming for 30 minutes, filtering with a filter cloth to remove residues, adding the components into soybean juice, quantifying to 1L with water, and regulating the pH to 5.5. )

The fermentation medium composition is shown in table 1 below, wherein corn sugar is expressed as total sugar of corn hydrolysate, and corn hydrolysate is prepared by the following method: corn flour and water in a mass ratio of 1:2.2, mixing, adjusting pH to 5.2, adding 16U/g alpha amylase based on corn flour mass, liquefying at 95 ℃ for 2 hours, then cooling to normal temperature for standby, measuring about 225g/L total sugar, diluting to required sugar concentration during dosing, and adding 200U/g saccharifying enzyme based on corn flour mass during fermentation. Wherein the carbon source is corn sugar, the nitrogen source is corn steep liquor, the phosphorus source is phosphoric acid, and the effector is bran, zinc sulfate, nicotinamide and biotin. Effectors are related to enzyme synthesis or activity and bacterial growth, for example, cellulose and vitamins in bran have promotion effect on enzyme induction and bacterial growth; zinc ions are similar to cofactors, can promote the improvement of enzyme activity, nicotinamide has the effect similar to vitamins, and biotin can promote the growth of thalli.

TABLE 1 Effect of different fermentation Medium Components on the yield of fermentation broths

Note that: the units are g/L; each numbered group is an average of 3 replicates.

Based on the research foundation, the influence of different carbon sources, nitrogen sources, phosphorus source concentrations and effectors (bran, zinc sulfate, nicotinamide and biotin) on the growth of the thalli is analyzed through a single factor test, and a proper amount of carbon-nitrogen ratio and effectors are found to have obvious promotion effect on the growth of the thalli. Therefore, the compound verification is carried out on the basis of the optimal concentration, when the fermentation medium is 26g/L of corn sugar, 25g/L of corn steep liquor, 8g/L of ammonium sulfate, 1g/L of phosphoric acid, 0.4g/L of magnesium sulfate, 0.6g/L of bran, 0.05g/L of zinc sulfate, 0.002g/L of nicotinamide, 0.005g/L of biotin and pH=6.0, the concentration of the fermentation thalli can reach 16.78g/L, which is obviously higher than the level of a control group of an organic nitrogen source, and the economic feasibility is higher.

Example 2 Effect of different rotational speeds on cell morphology and cell concentration

The seed solution cultured in example 1 was inoculated into 4 10L stainless steel fermenters containing 6L according to an inoculum size of 10% v/v, respectively, using the fermentation medium compounded in example 1. Culturing at 27 deg.C under 0.8-1.2vvm and 0.1Mpa under the lowest dissolved oxygen concentration of 30-40%. After fermentation, the solid-liquid separated thalli are washed twice by pure water, the diameter of the thalli is measured, dried, crushed, weighed and calculated dry weight concentration is measured, and the protein content (total nitrogen conversion measured by national standard method, the same applies below) and the dry matter digestibility are detected, wherein the detection method is measured by referring to GB/T6435-2006. The results are shown in fig. 1 and table 2.

TABLE 2 influence of different rotational speeds on fermentation

Note that: each fermenter was repeated 3 times.

As can be seen from the fermentation data in Table 2, the growth rate of the cells and the protein content were increased with the increase of the rotation speed, but the growth rate of the cells was not significantly increased and the protein content was decreased with the increase of 650 rpm. The high rotation speed has a remarkable influence on the size and the digestibility of the thallus balls, and the digestion rate is lower than 50% when the rotation speed is higher than 500rpm and the diameter is lower than 2 mm. In terms of yield, a high rotation speed is beneficial to the production of thalli, but in terms of application end, the digestibility of thalli with high yield is too low to be beneficial to the application of downstream products.

EXAMPLE 3 periodic control of rotational speed to achieve high yield and high digestibility of bacterial cells

The fermentation medium compounded in the example 1 is adopted, seed liquid cultured in the example 1 is respectively inoculated into 4 10L stainless steel fermentation tanks filled with 6L according to the inoculation amount of 10% v/v, but the fermentation level and the product performance are improved by adopting the step rotation speed regulation measures. The rotation speed is set to 400rpm in the early stage of fermentation, 200rpm is adopted in the middle and later stages of fermentation, the time point of rotation speed adjustment is determined to be limited by the limit that the thallus quantity reaches about 7-15g/L, the ventilation quantity is 0.8-1.2vvm, the tank pressure is 0.1Mpa, the lowest dissolved oxygen concentration is controlled to be 30-40%, and the fermentation time is 3 days. The results are shown in Table 3.

TABLE 3 periodic control of rotational speed bacterial fermentation results

	Fermentation tank 1	Fermentation tank 2	Fermentation tank 3	Fermentation tank 4
					Cell mass g/L at the time of rotation speed adjustment	8.3	11.2	13.6	15.2
Mycelium pellet diameter mm	2.51±0.08	2.24±0.14	2.01±0.12	1.62±0.07
					Cell mass g/L at the end of fermentation	17.42±1.22	18.02±1.24	18.65±1.02	17.35±1.45
Protein content%	45.21±3.83	46.88±1.62	46.16±1.13	47.05±2.43
					Dry matter digestibility%	50.85±1.51	51.67±0.82	48.56±1.87	47.22±1.48

In the logarithmic phase (7-9 g/L) of the thalli, the rotating speed is regulated down, and the influence on the growth of mycelium pellets is remarkable. In the middle and later stages of fermentation, the method has no promotion effect on the improvement of yield, protein content and digestibility.

EXAMPLE 4 Effect of different pH on cell morphology and cell concentration

The seed solution cultured in example 1 was inoculated into 5 10L stainless steel fermenters containing 6L according to an inoculum size of 10% v/v, respectively, using the fermentation medium compounded in example 1. Culturing at 27 deg.C and 250rpm under different pH values (constant pH value regulated by ammonia water, the same applies below) for 3 days, and controlling the lowest dissolved oxygen concentration at 30-40% under the pressure of 0.1Mpa and aeration rate of 0.8-1.2 vvm. After the fermentation, the solid-liquid separated cells were washed twice with pure water, and the cell diameter, dry weight concentration and protein content, and dry matter digestibility were measured, and the results are shown in Table 4.

TABLE 4 influence of different pH on fermentation

Note that: uniformity of ^# : a large number of +indicates good uniformity.

As can be seen from table 4, the pH has a significant effect on the mycelium morphology, the morphology size has a significant effect on the dry matter digestibility, and the fermentation pH is preferably controlled to about 7 in combination with the yield and final product quality characteristics.

Example 5 effect of staged pH control on fermentation and product Properties

The fermentation medium compounded in the example 1 is adopted, seed liquid cultured in the example 1 is respectively inoculated into 4 10L stainless steel fermentation tanks filled with 6L according to the inoculation amount of 10% v/v, but the fermentation level and the product performance are improved by adopting the step-by-step pH regulation measures. The pH5 is set in the early stage of fermentation, the pH7 is adopted in the middle and later stages of fermentation, the time point of pH adjustment is determined by taking the thallus quantity reaching 7-15g/L as a limit, the rotating speed is 250rpm, the ventilation quantity is 0.8-1.2vvm, the tank pressure is 0.1Mpa, the lowest dissolved oxygen concentration is controlled to be 30-40%, and the fermentation time is 3 days, and the result is shown in the table 5.

TABLE 5 staged pH control of thallus fermentation results

	Fermentation tank 1	Fermentation tank 2	Fermentation tank 3	Fermentation tank 4
					Cell amount g/L at pH adjustment	7.1	9.4	12.0	14.4
Mycelium pellet diameter mm	3.52±0.12	2.84±0.14	2.21±0.23	2.02±0.17
					Mycelium pellet homogeneity	++++	+++	+	+
Cell mass g/L at the end of fermentation	14.09±1.01	15.26±1.52	15.69±1.13	15.32±1.45
					Dry matter digestibility%	64.82±3.23	62.67±2.82	57.56±2.26	54.22±3.57

When the concentration of the thallus reaches about 7-9g/L during pH adjustment, the mycelium pellet has larger form, better uniformity and higher digestibility level after fermentation.

Example 6 effects of combination of rotational speed and pH control on yield and digestibility

And (3) strain: beefsteak mushroom

Culture medium and culture conditions: reference is made to example 1.

The seed liquid cultured in the example 1 is respectively inoculated into 3 10L stainless steel fermentation tanks with 6L according to the inoculation amount of 10 percent v/v, and the fermentation level and the product performance are improved by adopting the combined measures of stepwise regulating and controlling the stirring rotation speed and the pH. Setting pH5 and rotation speed of 400rpm at the early stage of fermentation, regulating pH7.2 with ammonia water and rotation speed of 250rpm when the thallus amount reaches about 7g/L, controlling ventilation amount of 0.8-1.2vvm, controlling tank pressure of 0.1Mpa, controlling minimum dissolved oxygen at about 30%, and fermenting until residual sugar is about 1 g/L. As shown in Table 6, the growth rate and the digestibility of the cells can be effectively improved by stepwise control of the rotation speed and pH.

TABLE 6 bacterial growth and digestibility changes under different conditions

	Fermentation cylinder 1 (comparison 1)	Fermentation tank 2	Fermentation tank 3 (comparative 2)
				Conditions (conditions)	pH5/400rpm	pH5/400rpm-pH7.2/250rpm	pH7.2/250rpm
Mycelium pellet diameter a mm	1.82±0.24	2.64±0.31	3.11±0.20
				Mycelium pellet homogeneity	++	+++	++++
Amount of thallus g/L	14.29	17.76	18.97
				Yield g/L/h	0.24	0.44	0.26
Conversion rate of thallus%	54.33	67.52	72.13
				Dry matter digestibility b%	54.85±2.25	63.43±2.55	67.54±3.21

Note that: ^a mycelium pellet diameter is an average of three lower pot samples taken, ^b the dry matter digestibility is the average of 3 measurements.

EXAMPLE 7 Effect of cell dispersion treatment on digestibility

The cells in example 6 were further subjected to dispersion treatment, and the digestibility was measured. As shown in table 7 below, after adding 2 volumes of water to each mycelium, mechanical wall breaking treatment was performed on the mycelium by using a wall breaking machine (jiuyang wall breaking machine), the smaller the mycelium pellet diameter was, the less likely to have a dispersion effect, the larger the mycelium pellet diameter was, the fewer the mycelium pellets were present after mechanical wall breaking, the more favorable it was for improving the digestibility, and the more remarkable the effect was (fig. 2).

TABLE 7 variation of digestibility of different samples under wall breaking conditions

Experimental group	1	2	3	4	5
						The diameter of the thallus is mm	1.82	2.64	3.11	3.11	3.11
Processing time s	30	30	30	60	120
						Percentage of bacterial loss%	10.22	12.42	14.24	17.61	22.52
Dry matter digestibility%	56.42±4.46	70.23±1.79	73.62±2.21	75.90±3.21	77.82±2.78

Example 8 Effect of alkali treatment on digestibility

Referring to the procedure of experimental group 3 in example 7, 5 groups of samples were prepared, two replicates each, and the change in digestibility at different pH conditions was analyzed. The results are shown in Table 8 below, the pH was controlled to 12, and the digestion rate was significantly improved by hot water bath for 8 hours, and the cell lysis was severe by continuing the up-regulation. The samples of experimental groups 3, 4 and 5 were adjusted back to ph6.5 and concentrated and dried to find that the three ash results were: 12.3%, 18.8% and 37.5%, so a treatment with pH12 is preferred.

TABLE 8 influence of pH on digestibility

Experimental group	a	b	c	d	e
						pH	6	8	10	12	14
Temperature (DEG C)	55	55	55	55	55
						Percentage of bacterial loss% a	11.26	11.59	14.24	21.61	42.52
Dry matter digestibility% b	71.42±2.40	74.23±2.15	80.62±1.29	91.90±2.25	95.32±4.12

Note that: percentage of bacterial loss% ^a And dry matter digestibility% ^b The results of (2) are the average of two replicates.

Example 9 cow milk yield assessment

Cow houses of Xuyi pastures 1-1, 1-2 and 2-1 are selected, lactating cows with 3-4 times of gestation, 200-220 days of lactation and 30-36kg of daily milk yield are divided into 3 groups, and about 110 heads of each group are respectively a control group, a test group 1 and a test group 2. The control group was fed with normal basal diet (mainly alfalfa 18%, corn silage 25%, corn meal 12%, soybean meal 12%, oat 4%, concentrated feed 5%, other 24%), the test group 1 added mycelium powder without post-treatment to 150g per cow on the basal diet (i.e. the bacterial liquid after fermentation under the condition of pH5/400rpm was separated by example 6, the bacterial liquid was directly dried and crushed, and sieved by a 100 mesh sieve), the test group 2 added mycelium powder with post-treatment to 150g per cow on the basal diet (i.e. the treatment mode of test group d in example 8 was adopted, and the bacterial liquid was dried and crushed and sieved by a 100 mesh sieve). The experimental period was 3 months, and the average daily milk yield of three groups was counted every half month, and the results are shown in table 9.

TABLE 9 statistics of milk yield of lactating cows

Week 2

Week 4

Week 6

Week 8

Week 10

Week 12

Control group

33.42±1.02

34.10±0.87

33.82±0.63

33.18±0.75

33.52±1.41

34.02±1.13

Test group 1

33.53±1.25

33.72±0.42

34.72±1.07

35.22±1.04

34.82±1.15

35.82±0.84

Test group 2

34.66±0.34

35.32±1.23

35.72±0.68

36.58±1.16

37.74±0.61

37.10±1.07

Within one month of feeding, the effect on milk yield is not obvious, from week 6, the milk yield of the test group 2 is improved by nearly 2kg compared with that of the control group, and the feeding effect of the test group 1 is improved by a relatively small range. At week 10 of feeding, milk production was increased by 1.3kg and 3.2kg for test groups 1 and 2, respectively. The results show that the treated mycelium powder has remarkable promotion effect on milk yield of cows.

Example 10 evaluation of Perch culture Effect

Mixing the ingredients, pulverizing to 80 mesh, adding 2% gelatin, and making into granule feed with particle diameter of 3mm, drying and preserving. Granulating according to the formula of table 10:

table 10 weever cultivation ingredient list (unit percentage)

Selecting 300 tails (about 8 cm) of weever with basically consistent length, dividing into 3 groups, feeding the groups in a culture net cage, feeding a control group with the raw formula feed, and replacing fish meal and yeast powder in the raw formula feed by mycelium powder in a test group, wherein the fish meal and the yeast powder are replaced by mycelium powder in the test group 1 on the basis of the raw formula (bacterial liquid separation after fermentation under the condition of pH5/400rpm in example 6 is adopted, directly drying and crushing thalli, and sieving by a 100-mesh sieve); test group 2 replaced the fish meal and yeast meal with treated mycelium powder based on the original formulation (using the treatment method of test group d in example 8, oven dried, crushed and sieved with a 100 mesh sieve). The feeding amount of the groups 3 is 2% of the fish mass every day, and the groups are fed in the morning and evening. The experimental conditions of the 3 groups are consistent, and the cultivation period is 2 months. Compared with a control group, the weever weight gain rates of the test group 1 and the test group 2 are respectively improved by 11.4 percent and 23.8 percent, the survival rate is improved by 7.8 and 21.3 percent, the growth performance of the weever can be obviously improved, and the culture benefit of the weever can be improved.

Example 11 Effect on broiler chickens

400 3-day-old broilers were randomly divided into 4 groups, and the diet design for each group of feed feeding is shown in table 11 below. The production management and epidemic prevention conditions are the same. Daily feed intake and daily gain and feed conversion ratio were measured at 50 days of age (Table 12).

Table 11 table for designing daily ration for raising broiler chickens

Group of	Proportioning of
		Control group	Basic ration
Test group 1	Basic ration plus 70kg mycelium powder a Daily ration per ton
		Test group 2	Basic ration plus 70kg mycelium powder b Daily ration per ton
Test group 3	Basic ration plus 70kg mycelium powder c Daily ration per ton

Wherein, the basic ration comprises the following components: 55% of corn, 35% of bean pulp, 5% of peanut meal, 2% of calcium hydrophosphate, 0.5% of salt and 2.5% of other materials. Mycelium powder ^a For the separation of the bacterial liquid after fermentation in the fermentation tank 3 in example 5, the bacterial cells were directly dried and pulverized to obtain mycelium powder (dried and pulverized to pass through a 100 mesh sieve), the mycelium powder ^b For the separation of the fermented bacterial liquid under the condition of pH5/400rpm in example 6, the mycelia powder obtained by directly drying and pulverizing the bacterial cells (by drying and pulverizing and sieving with 100 mesh sieve) was obtained ^c Mycelium powder (oven dried, pulverized and sieved through 100 mesh) obtained in the manner of treatment of test group d of example 8.

Table 12 daily feed intake, daily gain, and feed to meat ratio results statistics for broiler farming

As can be seen from the above table, the average daily gain of test groups 1, 2, 3 was increased by 11.2%, 16.2% and 23.3%, respectively, relative to the control group. Daily feed intake is also improved obviously. In addition, beefsteak mushroom has better effect than pleurotus geesteranus, and the mycelium powder has heavier fishy smell and possibly has a certain food calling effect.

The invention provides a preparation method of edible fungi concentrated feed and an application thought and method thereof, and the method and the way for realizing the technical scheme are more specific, the above is only a preferred embodiment of the invention, and it should be pointed out that a plurality of improvements and modifications can be made by those skilled in the art without departing from the principle of the invention, and the improvements and modifications are also considered as the protection scope of the invention. The components not explicitly described in this embodiment can be implemented by using the prior art.

Claims (5)

1. A preparation method of edible fungus concentrated feed is characterized in that edible fungus strains are added into a compound fermentation medium, fermentation is carried out by utilizing fermentation conditions of combination of stepwise regulation and control of rotation speed and pH, mycelium is obtained through solid-liquid separation after fermentation is finished, and then the mycelium is subjected to dispersion treatment and alkali treatment to prepare the edible fungus concentrated feed;

wherein, the compound fermentation culture medium comprises the following components: 10-35g/L of corn sugar, 10-30g/L of corn steep liquor, 4-12g/L of ammonium sulfate, 0.5-1.5g/L of phosphoric acid, 0.1-1g/L of magnesium sulfate, 0.2-1.5g/L of bran, 0.02-0.2g/L of zinc sulfate, 0.002-0.2g/L of nicotinamide, 0.0005-0.05g/L of biotin and pH=6.0;

the stepwise regulation takes the thallus quantity reaching 7-15g/L as a regulation node and is divided into fermentation early regulation and fermentation middle-late regulation, specifically, the fermentation early regulation and the fermentation rotating speed is 300-500r/min, and the pH value is 4-6; the fermentation is regulated and controlled in the middle and later period, the rotating speed is 100-300r/min, and the pH value is 7-8;

wherein the temperature of the fermentation is 25-29 ℃, the ventilation is 0.8-1.2vvm, and the fermentation culture time is 2-5 days;

wherein the dispersion treatment is to mechanically break the wall of mycelium for 30-120s;

wherein, the alkali treatment is to treat the mycelium after the dispersion treatment for 8 hours at the temperature of between pH7.2 and 14 and the temperature of 55 ℃, and then to adjust the pH value to between 6 and 7.

2. The method for preparing concentrated feed for edible fungi according to claim 1, wherein the edible fungi comprises any one or a combination of a plurality of pleurotus geesteranus, beefsteak mushroom and velvet mushroom.

3. The method for preparing concentrated feed of edible fungi according to claim 1, comprising the steps of:

(1) Inoculating the activated edible fungus strain into seed liquid, and culturing until the wet weight concentration of the strain reaches 14g/L;

(2) Inoculating the seed solution containing edible fungus strains in the step (1) into a compound fermentation culture medium according to the inoculation amount of 10% v/v, wherein the aeration rate is 0.8-1.2vvm, the tank pressure is 0.1Mpa, the minimum dissolved oxygen concentration is controlled to be 30-40%, and the fermentation culture is carried out for 2-4 days under the conditions of stepwise regulation of the rotating speed of 100-500r/min and the pH value of 4-8 until the residual sugar is 1g/L;

(3) Performing dispersion treatment on mycelium obtained by solid-liquid separation on thalli obtained by fermentation culture in the step (2);

(4) And (3) performing alkali treatment on the mycelium subjected to the dispersion treatment in the step (3), and directly drying and crushing the mycelium to prepare mycelium powder.

4. The method for preparing an edible fungus concentrate according to any one of claims 1 to 3.

5. Use of the edible fungus concentrate according to claim 4 for the preparation of ruminant or aquaculture or poultry animal feed.