CN109874751B - Growth substrate and preparation method and application thereof - Google Patents
Growth substrate and preparation method and application thereof Download PDFInfo
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- CN109874751B CN109874751B CN201910243110.3A CN201910243110A CN109874751B CN 109874751 B CN109874751 B CN 109874751B CN 201910243110 A CN201910243110 A CN 201910243110A CN 109874751 B CN109874751 B CN 109874751B
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Abstract
The invention provides a growth substrate and a preparation method and application thereof, wherein the method comprises the following steps: pretreatment of raw materials: mixing and pretreating the flammulina velutipes fungus dregs and the vinegar dregs in proportion; fermenting raw materials: inoculating a single strain or a mixture of multiple strains with a certain inoculation amount to a fermentation raw material, and fermenting for a period of time under a certain fermentation condition to obtain a growth substrate. The fermentation raw materials of the growth substrate prepared by the method are the flammulina velutipes mushroom dregs and the vinegar dregs which are mixed according to a proportion, are low in price and easy to obtain, have simple components, can be used for raising earthworms, have better effect than the direct feeding of the flammulina velutipes mushroom dregs and the traditional feeding method, and can solve the problem of environmental pollution caused by agricultural wastes.
Description
Technical Field
The invention belongs to the field of biological fermentation, and particularly relates to a growth substrate, and a preparation method and application thereof.
Background
Earthworms, commonly known as earthworms and ricefield eel, are omnivorous animals living in soil, and have the habit of going out in the daytime and night, preferring high-protein food, and mainly feed on animal wastes and organic waste garbage. According to determination, the earthworm mainly comprises mineral elements such as protein, unsaturated fatty acid, carbohydrate, vitamin, calcium, phosphorus and the like, has the functions of clearing heat, calming the liver, relieving asthma and dredging collaterals in medicine, can be used as protein feed for breeding livestock, poultry, fish and the like in agricultural production, loosens soil, improves soil fertility, can also treat organic garbage in production and life in cities and rural areas, changes waste into fertilizer, improves environment, and has extremely wide application and higher economic value. Therefore, earthworm breeding has become one of emerging industries.
At present, the problem of environmental pollution caused by agricultural wastes such as edible fungus residues, vinegar residues and the like is very serious. China is a large edible fungus producing country, and according to the incomplete statistics of edible fungus society in China, the total edible fungus production amount in China in 2011 reaches 2570 million tons, which accounts for more than 70% of the total edible fungus production amount in the world, and the output value exceeds 1400 billion yuan. With the development of the edible fungus industry, the amount of waste solid culture medium-mushroom dregs after harvesting edible fungus sporocarp is increasing (wei zhitao, zhou national english, huqing, etc.. edible fungus dregs utilization state research [ a ]. chinese edible fungus 2010.29 (5)). According to statistics, the production amount of the dutch mushroom dregs reaches 2.1 x 106m3 every year, and the production amount of the American mushroom dregs reaches 3.4 x 106m3 every year. And as the country with the largest edible fungus production capacity in China, the production amount of fungus residues in 2011 is 8360 ten thousand tons, and the quantity is huge. At present, the method for treating the edible fungus dregs is mainly to discard or burn the edible fungus dregs at will (Dong Xue Mei, Wang Yan Feng, Jing Sun Xuan, etc.. comprehensive utilization and development research of the edible fungus dregs [ A ]. Chinese edible fungus. 2013.32(6)), and a small amount of the edible fungus dregs are directly used as fertilizer to return to the field or used as secondary cultivation base material of the edible fungus, but the edible fungus dregs are discarded or burned, so that not only is the waste of resources caused, but also the environment is polluted. The waste edible fungus residues are converted into the culture medium required by the growth of the earthworms by a solid fermentation treatment method, so that the problem of environmental pollution can be effectively solved, and waste is changed into valuable.
At present, the substrates for breeding earthworms mainly comprise animal wastes, municipal excess sludge, fermented mushroom dregs and a mixture thereof required by the growth of the earthworms. The common animal manure for earthworm cultivation mainly comprises pig manure, chicken manure, sheep manure, cow manure and the like, and the base material prepared by the appropriate proportion has the best effect of earthworm cultivation; fresh urban excess sludge can be directly used for raising earthworms, and the earthworms grow normally; the fermented mushroom dregs can also be used for earthworm cultivation, which mainly comprises stack retting fermented straw mushroom residue, pleurotus cornucopiae, agaricus bisporus and the like, and can obviously improve the growth and reproductive capacity of earthworms; the mixture of the fungus dregs and earthworm needed for growth mainly comprises kitchen waste, additives (urea, ammonium sulfate and the like) and the like. The research provides a theoretical basis for the utilization of the product of the treated flammulina velutipes mushroom dregs for earthworm cultivation, the vinegar dregs serving as the earthworm cultivation substrate are not reported, and the process for obtaining the substrate suitable for earthworm cultivation by multi-bacterium fermentation of the flammulina velutipes mushroom dregs and the vinegar dregs according to a certain proportion is not reported.
With the development of society and economy, the substrates for earthworm cultivation such as animal wastes are not easy to obtain, and some substrates have the defects of complex components, numerous additives and the like, so that the earthworm cultivation effect is influenced.
Disclosure of Invention
The invention provides a growth substrate and a preparation method and application thereof, the growth substrate is used for breeding earthworms, the invention aims at the problems that the resource utilization of agricultural wastes such as flammulina velutipes mushroom dregs and vinegar residues is low, the development and utilization of high added value are less, the environment pollution is easy to cause and the like, the substrate suitable for the growth of the earthworms is obtained by adopting the microorganism solid state fermentation technology of the agricultural wastes such as the flammulina velutipes mushroom dregs and the vinegar residues, a new way is provided for the resource high-efficiency utilization of the agricultural wastes, and the raw material of the growth substrate is the mixture of the flammulina velutipes mushroom dregs and the vinegar residues according to a proportion, is cheap and easy to obtain, has simple components, and can solve the problem of the environment pollution caused by the agricultural wastes.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method of preparing a growth substrate comprising the steps of:
pretreatment of raw materials: mixing and pretreating the flammulina velutipes fungus dregs and the vinegar dregs in proportion;
fermenting raw materials: inoculating a single strain or a mixture of multiple strains with a certain inoculation amount to a fermentation raw material, and fermenting for a period of time under a certain fermentation condition to obtain a growth substrate.
In the scheme, the flammulina velutipes mushroom dregs and the vinegar dregs are mixed according to the weight ratio of 1: 7-1: 7.
Preferably, the flammulina velutipes mushroom dregs and the vinegar dregs are mixed according to the weight ratio of 7: 3.
In the scheme, the strain is one or a mixture of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium.
Further, the strain is a mixed bacterial liquid prepared by mixing any two of bacillus subtilis, candida tropicalis and phanerochaete in a ratio of 1: 1.
Preferably, the strain is a mixed bacterial liquid with the ratio of candida tropicalis, bacillus subtilis and phanerochaete chrysosporium of 1: 1.
In the scheme, the inoculation amount of the strain is 1-9%, the fermentation condition is that the fermentation time is 1-5 days, the fermentation temperature is 20-32 ℃, and the fermentation pH value is 5.5-7.5.
Preferably, the inoculation amount of the strain is 7%, the fermentation condition is 3 days, the fermentation temperature is 26 ℃, and the fermentation pH value is 7.5.
The growth substrate is prepared by the preparation method of the growth substrate.
Use of the growth substrate for earthworm cultivation.
Compared with the prior art, the invention has the beneficial effects that:
1. the fermentation raw materials of the growth substrate prepared by the method are the flammulina velutipes mushroom dregs and the vinegar dregs which are mixed in proportion, and the growth substrate is low in price, easy to obtain and simple in components.
2. The fermented growth substrate obtained by the method is used for feeding earthworms, the daily gain multiple of the earthworms is found to be obviously higher than that of unfermented earthworms and that of blank earthworms, and the feeding effect is superior to that of direct feeding of flammulina velutipes mushroom residues and the traditional feeding method.
3. The method for fermenting agricultural production wastes by using multiple fermentation bacteria in cooperation converts the waste needle mushroom dregs and vinegar residues into the growth matrix required by earthworm growth through solid state fermentation treatment, has simple process, is suitable for production, and can solve the problem of environmental pollution caused by the agricultural wastes.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a graph showing the change in crude protein content in a fermentation substrate of a single-strain fermentation according to the present invention.
FIG. 2 is a graph showing the change in crude protein content in the fermentation substrate of the multi-bacterial fermentation of the present invention.
FIG. 3 is a graph showing the effect of the ratio of fermentation raw materials on the crude protein content of the fermentation substrate according to the present invention.
FIG. 4 is a graph showing the effect of the inoculum size of the mixed strain on the crude protein content of the fermentation substrate.
FIG. 5 is a graph showing the effect of fermentation temperature on crude protein content of the fermentation substrate according to the present invention.
FIG. 6 is a graph showing the effect of pH on crude protein content in fermentation media according to the present invention.
Detailed Description
The invention will be described in detail below with reference to the drawings, but the scope of the invention is not limited thereto.
The preparation method of the growth substrate comprises the following steps:
pretreatment of raw materials: mixing and pretreating the flammulina velutipes fungus dregs and the vinegar dregs in proportion;
fermenting raw materials: inoculating a single strain or a mixture of multiple strains with a certain inoculation amount to a fermentation raw material, and fermenting for a period of time under a certain fermentation condition to obtain a growth substrate.
The needle mushroom fungus residues and the vinegar residues are mixed according to the weight ratio of 1: 7-1: 7. Preferably, the flammulina velutipes mushroom dregs and the vinegar dregs are mixed according to the weight ratio of 7: 3.
The strain is one or a mixture of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium. Further, the strains are mixed bacteria liquid with the ratio of the bacillus subtilis to the candida tropicalis being 1:1, mixed bacteria liquid with the ratio of the bacillus subtilis to the phanerochaete chrysosporium being 1:1, mixed bacteria liquid with the ratio of the candida tropicalis to the phanerochaete chrysosporium being 1:1 or mixed bacteria liquid with the ratio of the candida tropicalis to the phanerochaete chrysosporium being 1: 1. Preferably, the strain is a mixed bacterial liquid with the ratio of candida tropicalis, bacillus subtilis and phanerochaete chrysosporium of 1: 1.
The inoculation amount of the strain is 1-9%, the fermentation condition is that the fermentation time is 1-5 days, the fermentation temperature is 20-32 ℃, and the fermentation pH value is 5.5-7.5. Preferably, the inoculation amount of the strain is 7%, the fermentation condition is 3 days, the fermentation temperature is 26 ℃, and the fermentation pH value is 7.5.
The growth substrate is prepared by the preparation method of the growth substrate.
Use of the growth substrate for earthworm cultivation.
The growth substrate for culturing the earthworms mainly comprises flammulina velutipes mushroom dregs and vinegar dregs, and the solid fermentation bacteria are one or a mixture of bacillus subtilis, candida tropicalis or phanerochaete chrysosporium. The specific fermentation pretreatment, component analysis, strain culture and protein content determination method comprises the following steps:
1. the pretreatment method of the flammulina velutipes fungus residue and the vinegar residue comprises the following steps: the dried needle mushroom dregs and vinegar dregs are respectively weighed by 50g precisely, crushed by a crusher and placed in a ventilated and dry place for later use.
2. The method for analyzing the main components of the flammulina velutipes fungus residues and the vinegar residues can be carried out according to the following steps:
2.1. determination of ash content
The total ash content of the samples was determined by the burning method (GB 5009.4-2016).
2.2 determination of fat content
The fat content of the samples was determined by Soxhlet extraction (GB 5009.6-2016).
2.3 determination of the cellulose content
The content of cellulose in the samples was determined by titration (GB 5009.88-2014).
2.4 elemental content analysis
The content of C, N, H and P was determined using an elemental analyzer and an inductively coupled plasma mass spectrometer.
2.5 determination of protein content
The content of crude protein in the flammulina velutipes mushroom dregs and the vinegar dregs is measured by adopting a Kjeldahl method (GB 5009.4-2016).
The analysis results of the main components of the flammulina velutipes fungus residue and the vinegar residue are as follows:
the water content in the mushroom dregs is 44.82%; the crude protein content is 7.32%; crude fat 1.71%; ash content 17.87%; the cellulose content was 26.53%. The water content in the vinegar residue is 50.26%; crude protein content 5.35%; the crude fat content was 1.87%; ash content 9.27%; the cellulose content was 35.86%. The content of C, N, H and P in the mushroom dregs is respectively 24.40%, 2.16%, 3.21% and 3.2%; the content of C, N, H and P in the vinegar residue is 43.67%, 1.88%, 5.87% and 6.36% respectively. Through analysis of main components of the flammulina velutipes fungus dregs and the vinegar dregs, the fungus dregs and the vinegar dregs are found to be rich in nutrient substances required by the growth of earthworms and energy required by activities.
3. The method for culturing the bacillus subtilis, the candida tropicalis and the phanerochaete chrysosporium can be carried out according to the following steps:
beef extract peptone medium: accurately weighing 3g of beef extract, 10g of peptone and 5g of NaCl, completely dissolving in 1000ml of double distilled water, adjusting pH to 7.4-7.6 with 0.1M NaOH solution, and sterilizing in an autoclave at 121 ℃ for 30min for later use.
Potato glucose medium (PDA): taking 46g of potato glucose culture medium, dissolving with 1000ml of double distilled water completely, adjusting pH to natural, sterilizing with 121 deg.C autoclave for 30min, and keeping.
Taking 500ml conical flask, and autoclaving at 121 deg.C for 30min in autoclave for use. Transferring 100mL of culture medium into a conical flask under aseptic conditions, inoculating, holding the test tube with the left hand during inoculation, extending the burned inoculating loop into a strain tube with the right hand, cooling, taking a strain liquid loop, transferring the strain liquid loop into a culture medium tube, slightly grinding the tube wall close to the liquid level, slightly tilting the test tube, dipping a little liquid for blending, and mixing the strain liquid with the culture medium. And (3) placing the inoculated bacteria in a constant-temperature oscillator at 37 ℃ and 200r, culturing for 24h, and taking out for later use. The bacillus subtilis is cultured by a beef extract peptone culture medium, and the candida tropicalis and the phanerochaete chrysosporium are cultured by a PDA culture medium.
4. The protein content was measured by the method 2.5.
The function of each component in the fermentation product is as follows:
needle mushroom fungus dreg: is the main raw material for fermentation and the main substrate for earthworm cultivation. It contains protein, fat, cellulose, and substances such as C, N, H and P necessary for earthworm to live, and can provide nutrient for earthworm.
Vinegar residue: is the main raw material for fermentation and the main substrate for earthworm cultivation. It contains protein, fat, water, C, N, H and P, which are essential for earthworm, and contains a lot of cellulose to provide energy for earthworm and regulate pH.
Bacillus subtilis, candida tropicalis and phanerochaete chrysosporium: is the main fermentation strain.
Drugs and reagents:
bacillus subtilis, Candida tropicalis and Phanerochaete chrysosporium are all purchased from China center for industrial microorganism culture preservation management. Potassium sulfate, copper sulfate, concentrated sulfuric acid, sodium hydroxide, boric acid, methylene blue indicator and the like are purchased from national medicine group chemical reagent limited, and the purity is chemical purity.
The statistical method comprises the following steps:
data were processed using SAS6.12 statistical software and results were used
Showing that the significance of the difference between each administered group and the control group was compared by t-test between groups. P < 0.05, indicating that the group was significantly different from the control group.
The experimental method comprises the following steps:
the fermentation strain comprises bacillus subtilis, candida tropicalis and phanerochaete chrysosporium, the fermentation substrate comprises flammulina velutipes mushroom dregs and vinegar dregs, the total weight of the fermentation substrate is 100g, and the component proportion and the fermentation method of the fermentation substrate are shown in examples 1-6.
Example 1
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 1: 1;
bacillus subtilis, Candida tropicalis or Phanerochaete chrysosporium;
the total mass of fermentation raw materials is 100g, the inoculation amount is 5%, the initial pH value is adjusted to 7, the fermentation time is 5 days, the fermentation is carried out in an illumination incubator at the temperature of 25 ℃, samples are taken in 0, 1, 2, 3, 4 and 5 days, the crude protein content is measured, and the optimal single-bacterium fermentation strain is determined by recording.
FIG. 1 shows that the effect of single-strain fermentation experiments of Bacillus subtilis, Candida tropicalis and Phanerochaete chrysosporium on the crude protein content of a fermentation substrate is studied when the inoculation amount is 5%, the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 1:1, the fermentation time is 5 days, the pH value is 7, and the fermentation temperature is 25 ℃. Results show that the crude protein content of the three different strains after fermentation is in a trend of increasing firstly and then decreasing, the crude protein content of the three different strains reaches a peak value on the 3 rd day of fermentation, and the crude protein content of the single strains of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium on the 3 rd day is 14.1%, 13.9% and 14.9% respectively. Wherein the content of crude protein in the Phanerochaete chrysosporium fermentation substrate is the highest. Therefore, the crude protein content was used as the evaluation criterion for the earthworm diet, and the fermentation end point was selected as day 3.
Example 2
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 1: 1;
fermentation strain: a mixed bacterial liquid with the ratio of the bacillus subtilis to the candida tropicalis being 1:1, a mixed bacterial liquid with the ratio of the bacillus subtilis to the phanerochaete chrysosporium being 1:1, a mixed bacterial liquid with the ratio of the candida tropicalis to the phanerochaete chrysosporium being 1:1 or a mixed bacterial liquid with the ratio of the candida tropicalis, the bacillus subtilis and the phanerochaete chrysosporium being 1: 1. Preferably, the strain is a mixed bacterial liquid with the ratio of candida tropicalis, bacillus subtilis and phanerochaete chrysosporium of 1: 1.
The total mass of fermentation raw materials is 100g, the fermentation time is 5 days, the inoculation amount is 5%, the initial pH value is adjusted to 7, the fermentation is carried out in an illumination incubator at the temperature of 25 ℃, samples are taken in 0, 1, 2, 3, 4 and 5 days, the crude protein content is measured, and the optimal multi-bacterium fermentation strain is determined by recording.
As shown in figure 2, the effect of the multi-bacteria fermentation experiment of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium on the crude protein content of the fermentation substrate is studied when the inoculation amount is 5%, the weight ratio of the flammulina velutipes residues to the vinegar residues is 1:1, the pH value is 7, the fermentation time is 5 days, and the fermentation temperature is 25 ℃. The results show that the change of the crude protein content in the mixed fermentation process has a tendency of rising first and then falling, and the crude protein content reaches a peak value at 3 days of fermentation, and in fig. 2, it can be seen that the crude protein content in the fermented product of the mixed bacteria liquid with the proportion of three bacteria, namely phanerochaete chrysosporium, candida tropicalis and bacillus subtilis, of 1:1 is 18.5% which is the highest, while the mixed bacteria liquid with the proportion of candida tropicalis and bacillus subtilis is 1:1, the mixed bacteria liquid with the proportion of phanerochaete chrysosporium and candida tropicalis of 1:1 and the mixed bacteria liquid with the proportion of phanerochaete chrysosporium and bacillus subtilis of 1:1 are 16.4%, 16.7% and 16.5% respectively after fermentation. Therefore, the optimal zymocyte is determined to be mixed bacterium liquid with the proportion of Phanerochaete chrysosporium, candida tropicalis and bacillus subtilis being 1: 1.
Example 3
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 10: 0, 7:3, 1:1, 3: 7 and 0: 10;
fermentation strain: a mixed bacterial liquid with the proportion of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium of 1: 1;
the total mass of fermentation raw materials is 100g, the fermentation time is 5 days, the inoculation amount is 5%, the initial pH value is adjusted to 7, the fermentation raw materials are placed in an illumination incubator at the temperature of 25 ℃ for fermentation, samples are taken in 0, 1, 2, 3, 4 and 5 days, the crude protein content of the samples is measured, the records are recorded, and the optimal fermentation ratio of the flammulina velutipes mushroom dregs to the vinegar dregs is determined.
FIG. 3 shows that the inoculation amount is 5%, the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 10: 0, 7:3, 1:1, 3: 7, the fermentation time is 0: 10 for 5 days, the pH value is 7, and the fermentation temperature is 25 ℃, so that the influence of different proportions of fermentation raw materials on the crude protein content of a fermentation substrate in a multi-fungus fermentation experiment is researched. The result shows that with the gradual reduction of the content of the needle mushroom dregs in the fermentation proportion, the content of the vinegar residues is gradually increased, the content of crude protein in the fermentation product is in a trend of slightly increasing and then continuously decreasing, and the content of the crude protein reaches a peak value when the proportion of the dregs to the vinegar residues is 7:3, and the content of the crude protein at the moment is 16.7 percent, so the optimal proportion of the fermentation raw materials is selected as the dregs: the ratio of the vinegar residue is 7: 3.
Example 4
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 7: 3;
fermentation strain: a mixed bacterial liquid with the proportion of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium of 1: 1;
the total mass of fermentation raw materials is 100g, the fermentation time is 5 days, the inoculation amount is 1, 3, 5, 7 or 9 percent, the initial pH value is adjusted to 7, the fermentation is carried out in an illumination incubator at the temperature of 25 ℃, samples are taken in 0, 1, 2, 3, 4 and 5 days, the crude protein content is measured, and the optimal multi-bacterium inoculation ratio is determined by recording.
FIG. 4 shows the results of the experiments on the effect of different inoculum sizes on the crude protein content of the fermentation substrate in the fermentation experiments, wherein the weight ratio of the flammulina velutipes residue to the vinegar residue is 7:3, the fermentation time is 5 days, the fermentation temperature is 25 ℃, the pH value is 7, and the inoculum sizes are 1%, 3%, 5%, 7% and 9%. The results show that the protein content in the fermentation substrate increases and then decreases with increasing inoculum size, reaching a peak at an inoculum size of 7% and a crude protein content of 16.6%. Therefore, the optimal inoculum size was determined to be 7%.
Example 5
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 7: 3;
fermentation strain: a mixed bacterial liquid with the proportion of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium of 1: 1;
the total mass of fermentation raw materials is 100g, the fermentation time is 5 days, the inoculation amount is 5%, the initial pH value is adjusted to 7, the fermentation is carried out in an illumination incubator at the temperature of 20, 23, 26, 29 or 32 ℃, samples are taken at 0, 1, 2, 3, 4 and 5 days, the crude protein content is measured, and the optimal fermentation temperature is determined by recording.
FIG. 5 shows the weight ratio of the needle mushroom dregs to the vinegar dregs being 7:3, the fermentation time being 5 days, the inoculation amount of the mixed fungus being 7%, the pH value being 7, and the fermentation temperature being controlled at 20, 23, 26, 29 and 32 ℃, and the influence of different temperatures on the crude protein content in the fermentation substrate in the fermentation experiment was studied. The results show that the crude protein content in the fermentation substrate tends to increase and then decrease with increasing fermentation temperature and peaks at 26 ℃ where the crude protein content is 17.0%. The optimum fermentation temperature was thus determined to be 26 ℃.
Example 6
Fermentation raw materials: the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 7: 3;
fermentation strain: a mixed bacterial liquid with the proportion of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium of 1: 1;
the total mass of fermentation raw materials is 100g, the fermentation time is 5 days, the inoculation amount is 5 percent, the initial pH is adjusted to be 5.5, 6, 6.5, 7 or 7.5, the mixture is placed in an illumination incubator at the temperature of 25 ℃ for fermentation, samples are taken at 0, 1, 2, 3, 4 and 5 days, the crude protein content is measured, and the optimal fermentation pH value is determined by recording.
FIG. 6 shows that the weight ratio of the flammulina velutipes fungus residue to the vinegar residue is 7:3, the fermentation time is 5 days, the inoculation amount of the mixed fungus is 7%, the fermentation temperature is controlled at 26 ℃, and the influence of different pH values on the crude protein content in the fermentation substrate in the fermentation experiment is studied when the pH values are adjusted to 5.5, 6, 6.5, 7 and 7.5. The results show that the crude protein content in the fermentation substrate increases with increasing pH and levels off at pH 7 to 7.5, and that the crude protein content of the fermentation product is up to 16.8% at pH 7.5. Therefore, the optimum fermentation pH was selected to be 7.5.
Example 7
The fermentation substrate is used for earthworm breeding experiments:
in this example 7, with regard to the # 2 da earthworms as an experimental subject, 10 earthworms are put into a cultivation vessel, wherein about 1/3 fresh and wet soil is placed in advance, and the fermentation products of the above experiment, that is, the growth substrate, the unfermented raw material and the common soil prepared by the method of the present invention are added to the soil in advance as an earthworm feed with a dry weight of 50g, 3 groups are parallel, the humidity is maintained at 60%, the cultivation temperature is 25 ℃, the mouth of the cultivation vessel is sealed with gauze with a pore diameter of less than 0.5mm, so as to prevent the earthworms from escaping, the earthworms are cultivated for 9 days, the influences of different fermentation products on the daily weight gain and the daily breeding number of the earthworms are examined, and the number of grown earthworms, the number of young earthworms and the number of earthworm cocoons are recorded every 3 d. Meanwhile, the peel and the leaf and stem of green vegetables which are commonly used earthworm breeding feed in the earthworm breeding industry are used as positive controls.
According to the experimental result, the daily gain times of the earthworms in the fermentation substrate group are obviously higher than those of the earthworms in the unfermented group and those of the earthworms in the blank group (p is less than 0.05), the daily gain times of the earthworms in the fermentation substrate group are 0.0463 +/-0.0040, the daily gain times of the earthworms in the unfermented group are 0.0264 +/-0.0020, and the daily gain times of the earthworms in the blank group are 0.0301 +/-0.0005. Therefore, the fermented needle mushroom fungus residues and vinegar residues can be used for raising earthworms, and the effect is superior to that of direct needle mushroom fungus residue raising and traditional needle mushroom fungus residue raising methods.
And (4) conclusion:
the multi-bacteria fermentation experiments and the experiments with different fermentation ratios show that the optimal fermentation strain is the mixture of three bacteria liquids of Phanerochaete chrysosporium, candida tropicalis and bacillus subtilis in the ratio of 1:1, the optimal ratio of the fermentation raw materials of flammulina velutipes residues to vinegar residues is 7:3, and the optimal fermentation time is 3 days.
The optimal conditions for obtaining solid state fermentation by the single factor test are as follows: the inoculum size is 7 percent, the temperature is 26 ℃, and the pH value is 7.5.
The fermented product with higher crude protein and the growth substrate are obtained under the optimal conditions and are used for feeding the earthworms, the daily gain multiple of the earthworms is found to be obviously higher than that of unfermented earthworms and that of blank earthworms, and the feeding effect is better than that of the direct feeding of the flammulina velutipes mushroom residues and the traditional feeding method.
It should be understood that although the present description has been described in terms of various embodiments, not every embodiment includes only a single embodiment, and such description is for clarity purposes only, and those skilled in the art will recognize that the embodiments described herein may be combined as suitable to form other embodiments, as will be appreciated by those skilled in the art.
The above-listed detailed description is only a specific description of a possible embodiment of the present invention, and they are not intended to limit the scope of the present invention, and equivalent embodiments or modifications made without departing from the technical spirit of the present invention should be included in the scope of the present invention.
Claims (4)
1. A method of preparing a growth substrate, comprising the steps of:
pretreatment of raw materials: mixing and pretreating the flammulina velutipes fungus dregs and the vinegar dregs according to the weight ratio of 7: 3;
fermenting raw materials: inoculating a certain inoculation amount of mixed bacterial liquid of multiple strains on a fermentation raw material, and fermenting for a period of time under a certain fermentation condition to obtain a growth substrate; the strain is a mixed bacterial liquid prepared by mixing any two of bacillus subtilis, candida tropicalis and phanerochaete chrysosporium in a ratio of 1:1, or the strain is a mixed bacterial liquid prepared by mixing candida tropicalis, bacillus subtilis and phanerochaete chrysosporium in a ratio of 1:1, the inoculation amount of the strain is 1-9%, the fermentation time is 1-5 days, the fermentation temperature is 20-32 ℃, and the fermentation pH value is 5.5-7.5; the growth substrate is used for breeding earthworms.
2. The method of claim 1, wherein the inoculum size of the strain is 7%, the fermentation time is 3 days, the fermentation temperature is 26 ℃, and the fermentation pH is 7.5.
3. A growth substrate, characterized in that it is produced by a method for producing a growth substrate according to any one of claims 1 to 2.
4. Use of a growth substrate according to claim 3, for the cultivation of earthworms.
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| CN101811911A (en) * | 2010-04-23 | 2010-08-25 | 镇江恒欣肥料科技有限公司 | Matrix produced by using vinegar residue for greening and production process thereof |
| CN102058031A (en) * | 2010-11-24 | 2011-05-18 | 天津大学 | Earthworm bait prepared from fungus bran through mixed culture fermentation and preparation method thereof |
| CN103371125A (en) * | 2012-04-12 | 2013-10-30 | 天津市绿色坐标生态科技有限公司 | Method for breeding earthworm by waste fungus chaff generated in edible fungus cultivation |
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| CN102058031A (en) * | 2010-11-24 | 2011-05-18 | 天津大学 | Earthworm bait prepared from fungus bran through mixed culture fermentation and preparation method thereof |
| CN103371125A (en) * | 2012-04-12 | 2013-10-30 | 天津市绿色坐标生态科技有限公司 | Method for breeding earthworm by waste fungus chaff generated in edible fungus cultivation |
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