CN115851465B - Composite fermentation inoculant, method for preparing fermented rapeseed meal for feeding and application of composite fermentation inoculant - Google Patents
Composite fermentation inoculant, method for preparing fermented rapeseed meal for feeding and application of composite fermentation inoculant Download PDFInfo
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Abstract
The invention discloses a composite fermentation microbial inoculum and a method for preparing fermented rapeseed meal for feeding and application thereof, and relates to the technical field of biological fermentation. The composite fermentation inoculant comprises rhodotorula rubra, pediococcus acidilactici, aspergillus niger, bacillus coagulans and cellulase produced by Aspergillus niger. The production process for preparing the fermented rapeseed meal for feed by adopting the composite fermentation microbial inoculum provided by the invention is simple to operate, and the prepared fermented rapeseed meal has a thick fragrance, wherein the contents of anti-nutritional factors such as thioglycoside, phytic acid, tannin, crude fiber and the like are obviously reduced, the contents of crude protein and acid soluble protein are obviously increased, the anti-nutritional factors in the rapeseed meal can be effectively removed, and the nutritional quality of the rapeseed meal is improved. The fermented rapeseed meal for feed of the invention is applied to the preparation of livestock, poultry and aquatic animal feeds.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a composite fermentation microbial inoculum, a method for preparing fermented rapeseed meal for feeding and application thereof, and particularly relates to application of the fermented rapeseed meal in replacing soybean meal or fish meal in aquatic feed.
Background
The quality protein resource for feeding aquatic animals in China is seriously dependent on import, and according to incomplete statistics, the annual import of soybean products in China is more than 5500 ten thousand tons and fish meal is more than 100 ten thousand tons. The lack of quality feed materials has become the most serious problem facing the growing aquaculture and aquaculture industry. Rape is a first large oil crop in China, and rapeseed meal is used as a byproduct after oil extraction of rapeseeds, is rich in protein content (35-42 parts), reasonable in amino acid composition and contains higher sulfur amino acid and lysine as well as microelements such as iron, manganese, zinc and selenium. However, because the anti-nutritional factors such as the sulfur-containing glycoside, the tannin, the phytic acid, the isothiocyanate and the like in the rapeseed meal have adverse effects on the growth, the development and the reproduction of animals, the great use of the rapeseed meal in animal feeds is restricted.
The microbial fermentation method is a method for directly decomposing and converting anti-nutritional factors by means of microbial life metabolic activities, has mild conditions and simple process, can efficiently degrade toxic substances such as thioglycoside, tannin, phytic acid, isothiocyanate and the like, degrade and soften crude fibers, secrete and synthesize a large amount of nutritional and hormonal substances such as active probiotics, bioactive small peptide amino acids, biochemical enzymes, growth-promoting factors and the like, thereby improving the nutritional value and palatability of the rapeseed meal, increasing the use proportion of the rapeseed meal in feed and improving the self-sufficiency rate of high-quality protein sources in China.
Therefore, development of a method for removing anti-nutritional factors in rapeseed meal by utilizing a microbial fermentation method has important significance for improving the self-supply rate of high-quality protein sources in China and promoting the sustainable development of aquaculture.
Disclosure of Invention
The invention provides a composite fermentation inoculant, a method for preparing fermented rapeseed meal for feeding and application thereof, and aims to solve the problems in the background technology.
In order to achieve the technical purpose, the invention mainly adopts the following technical scheme:
in a first aspect, the invention discloses a composite fermentation inoculant comprising rhodotorula rubra, pediococcus acidilactici, aspergillus niger, bacillus coagulans and cellulase produced by Aspergillus niger.
In some embodiments, the composite fermentation inoculant comprises the following components in parts by weight: 1-50 parts of rhodotorula rubra, 1-20 parts of Pediococcus acidilactici, 1-15 parts of Aspergillus niger, 1-30 parts of Bacillus coagulans and 1-10 parts of cellulase.
Preferably, the composite fermentation inoculant comprises the following components in parts by weight: 10-40 parts of rhodotorula rubra, 5-20 parts of Pediococcus acidilactici, 1-10 parts of Aspergillus niger, 10-25 parts of Bacillus coagulans and 5-10 parts of cellulase.
More preferably, the composite fermentation inoculant comprises the following components in parts by weight: 20-30 parts of rhodotorula rubra, 10-20 parts of Pediococcus acidilactici, 5-10 parts of Aspergillus niger, 20-25 parts of Bacillus coagulans and 5-8 parts of cellulase.
In some embodiments, the total number of viable bacteria of the rhodotorula ruber, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, and the cellulase activity is more than or equal to 2000u/g.
In some embodiments, the rhodotorula rubra, pediococcus acidilactici, aspergillus niger, bacillus coagulans and cellulases produced from aspergillus niger are derived from fermented bean curd, kimchi fermented food.
In a second aspect, the invention discloses an application of the composite fermentation inoculant in the first aspect in preparing fermented rapeseed meal for feeding.
In a third aspect, the invention discloses a method for preparing fermented rapeseed meal for feeding by using the composite fermentation inoculant in the first aspect, which comprises the following steps: crushing and sieving rapeseed meal, uniformly mixing a fermentation microbial inoculum, the rapeseed meal and water according to a certain mass ratio, placing the mixture into a breathing bag, fermenting at 25-40 ℃ for 1-3 days after stirring every 12 hours, and drying and crushing after fermentation to obtain the fermented rapeseed meal for feeding.
In some embodiments, the zymophyte agent is calculated as a mass ratio: vegetable meal: water= (0.1-0.3): 100: (50-120), fermenting at 30-40deg.C for 1-2 days.
Further, the fermentation inoculant comprises the following components in percentage by mass: vegetable meal: water= (0.1-0.2): 100: (80-100), the fermentation temperature is 30-37 ℃ and the fermentation time is 2 days.
Compared with the prior art, the invention has the following beneficial effects:
the production process for preparing the fermented rapeseed meal for feed by adopting the composite fermentation microbial inoculum provided by the invention is simple to operate, and the prepared fermented rapeseed meal has a thick fragrance, wherein the contents of anti-nutritional factors such as thioglycoside, phytic acid, tannin, crude fiber and the like are obviously reduced, the contents of crude protein and acid soluble protein are obviously increased, the anti-nutritional factors in the rapeseed meal can be effectively removed, and the nutritional value of the rapeseed meal is improved. The fermented rapeseed meal prepared by the invention can replace soybean meal or fish meal in aquatic feed, can obviously reduce feed cost and improve cultivation benefit.
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention.
Example 1
Mixing 20g of rhodotorula rubra, 10g of Pediococcus acidilactici, 10g of Aspergillus niger, 20g of Bacillus coagulans and 8g of cellulase uniformly to obtain a fermentation inoculant. Wherein the total number of viable bacteria of the rhodotorula rubra, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, and the activity of the cellulase is more than or equal to 2000u/g. The above ferment bacteria are derived from fermented food such as fermented bean curd and sauerkraut, and the above cellulase is produced from Aspergillus niger.
0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 2 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Example 2
Mixing 10g of rhodotorula rubra, 20g of Pediococcus acidilactici, 8g of Aspergillus niger, 25g of Bacillus coagulans and 5g of cellulase uniformly to obtain a fermentation inoculant. Wherein the total number of viable bacteria of the rhodotorula rubra, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, the activity of the cellulase is more than or equal to 2000u/g, the fermentation inoculants are all derived from fermented foods such as fermented bean curd, pickle and the like, and the cellulase is produced from the Aspergillus niger.
0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Example 3
Mixing 30g of rhodotorula rubra, 5g of pediococcus acidilactici, 20g of aspergillus niger, 10g of bacillus coagulans and 10g of cellulase uniformly to obtain the fermentation inoculant. Wherein the total number of viable bacteria of the rhodotorula rubra, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, the activity of the cellulase is more than or equal to 2000u/g, the fermentation inoculants are all derived from fermented foods such as fermented bean curd, pickle and the like, and the cellulase is produced from the Aspergillus niger.
0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 1 day. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Example 4
Mixing 40g of rhodotorula rubra, 2g of Pediococcus acidilactici, 1g of Aspergillus niger, 13g of Bacillus coagulans and 2g of cellulase uniformly to obtain a fermentation inoculant. Wherein the total number of viable bacteria of the rhodotorula rubra, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, and the activity of the cellulase is more than or equal to 2000u/g. The above ferment bacteria are derived from fermented food such as fermented bean curd and sauerkraut, and the above cellulase is produced from Aspergillus niger.
0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 2 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 1
Mixing 20g of rhodotorula rubra and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 2
Mixing 10g of Pediococcus acidilactici and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 3
Mixing 10g of Aspergillus niger and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 4
And mixing 20g of bacillus coagulans and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 5
50g of rhodotorula rubra, 20g of pediococcus acidilactici, 15g of aspergillus niger, 30g of bacillus coagulans and 10g of cellulase are taken and uniformly mixed to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 6
Mixing 60g of rhodotorula rubra, 30g of pediococcus acidilactici, 20g of aspergillus niger, 40g of bacillus coagulans and 15g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 7
Taking 20g of rhodotorula rubra, 10g of lactobacillus, 10g of aspergillus niger, 20g of bacillus coagulans and 8g of cellulase, and uniformly mixing to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 8
And uniformly mixing 20g of saccharomyces cerevisiae, 10g of pediococcus acidilactici, 10g of aspergillus niger, 20g of bacillus coagulans and 8g of cellulase to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 9
Mixing 20g of rhodotorula rubra, 10g of pediococcus acidilactici, 10g of spine Kong Damei, 20g of bacillus coagulans and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
Comparative example 10
Mixing 20g of rhodotorula rubra, 10g of pediococcus acidilactici, 10g of aspergillus niger, 20g of bacillus subtilis and 8g of cellulase uniformly to obtain the fermentation inoculant. 0.1g of zymophyte agent is taken and added into 100mL of water, and is stirred uniformly, inoculated into 100g of crushed rapeseed meal and mixed uniformly. Transferring the inoculated rapeseed meal into a breathing bag, sealing and fermenting, controlling the fermentation temperature to 37 ℃, turning over every 12 hours, and fermenting for 3 days. And after fermentation, drying at 65 ℃, crushing, and sieving with a 40-mesh sieve to obtain a finished product of fermented rapeseed meal.
The anti-nutritional factors in the fermented meal for feed prepared in examples 1 to 4 and comparative examples 1 to 10 were measured and the results are shown in Table 1.
TABLE 1 influence of different fermentation methods on the anti-nutritional factor content of rapeseed meal
As can be seen from Table 1, in the fermented rapeseed meal for feeding prepared by adopting the composite fermentation inoculant provided by the invention, the content of anti-nutritional factors, namely, thioglycoside, tannin and phytic acid, namely, crude fiber components is obviously reduced, and the content of the anti-nutritional factors, namely, the crude fiber components is obviously different from that of comparative examples 1-10, and P is less than 0.01.
The nutritional ingredients in the fermented meal for feed prepared in examples 1 to 4 and comparative examples 1 to 10 were measured as described above, and the results are shown in Table 2.
TABLE 2 influence of different fermentation methods on nutrient components of rapeseed meal (portions)
As can be seen from Table 2, in the fermented feed meal prepared by the composite fermentation inoculant provided by the invention, compared with the initial meal, the nutritional ingredients of the fermented feed meal are obviously increased (P is less than 0.05), which indicates that the nutritional value of the fermented feed meal prepared by the composite fermentation inoculant provided by the invention is obviously improved.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and principles of the invention are intended to be included within the scope of the invention.
And (3) testing application effects:
example 5 effect of application in Tilapia feed
In the tilapia feed (crude protein 30% and crude fat 7%), fermented rapeseed meal is used for replacing 0%, 25%, 50%, 75% and 100% of soybean meal, and the tilapia is fed for 8 weeks, and 4 tilapia feed groups are parallel. Sampling and analyzing tilapia growth and feed utilization.
TABLE 3 influence of fermented rapeseed meal on tilapia growth and feed utilization instead of soybean meal in different proportions
| Group of | 0％ | 25％ | 50％ | 75％ | 100％ |
| Initial average body weight/g | 30.20 | 30.51 | 30.26 | 30.55 | 30.42 |
| Average weight/g | 197.85 | 202.34 | 199.61 | 191.77 | 175.26 |
| Weight gain percentage% | 555.13 | 563.19 | 559.65 | 527.72 | 476.13 |
| Feed coefficient | 1.32 | 1.35 | 1.34 | 1.38 | 1.43 |
The results show that the fermented rapeseed meal can replace 75% of soybean meal in tilapia feed at most, and cannot adversely affect the growth and feed utilization of tilapia.
Example 6 application Effect in Lateolabrax japonicus feed
In the largemouth bass feed (crude protein 53% and crude fat 10%), fermented rapeseed meal is used for replacing 0%, 25%, 50%, 75% and 100% of fish meal, and the largemouth bass feed is fed for 8 weeks, and 4 largemouth bass feed groups are parallel. Sampling and analyzing the growth and feed utilization of the micropterus salmoides.
TABLE 4 influence of fermented rapeseed meal replacement with different proportions of fish meal on largemouth bass growth and feed utilization
| Group of | 0％ | 25％ | 50％ | 75％ | 100％ |
| Initial average body weight/g | 5.22 | 5.30 | 5.25 | 5.15 | 5.30 |
| Average weight/g | 25.26 | 24.65 | 20.71 | 15.83 | 10.83 |
| Weight gain percentage% | 383.91 | 365.09 | 302.14 | 198.68 | 104.34 |
| Feed coefficient | 0.75 | 0.78 | 0.84 | 0.96 | 1.22 |
The results show that the fermented rapeseed meal can replace 25% of fish meal in the micropterus salmoides feed at most, and the growth performance and feed utilization of the micropterus salmoides can not be obviously reduced.
Claims (5)
1. The application of the composite fermentation microbial inoculum in preparing the fermented rapeseed meal for feeding is that the composite fermentation microbial inoculum is used for reducing the thioglycoside, phytic acid, tannin and crude fiber anti-nutritional factors in the fermented rapeseed meal and improving the crude protein and acid soluble protein content in the fermented rapeseed meal;
the composite fermentation inoculant comprises rhodotorula rubra, pediococcus acidilactici, aspergillus niger, bacillus coagulans and cellulase produced from aspergillus niger;
the composite fermentation inoculant comprises the following components in parts by weight: 10-40 parts of rhodotorula rubra, 5-20 parts of Pediococcus acidilactici, 1-10 parts of Aspergillus niger, 10-25 parts of Bacillus coagulans and 5-10 parts of cellulase;
the total number of the viable bacteria of the rhodotorula rubra, the Pediococcus acidilactici, the Aspergillus niger and the Bacillus coagulans is 50-100 hundred million CFU/g, 100-300 hundred million CFU/g and 20-50 hundred million CFU/g respectively, and the activity of the cellulase is more than or equal to 2000u/g;
the method for preparing the fermented rapeseed meal for feeding by the composite fermentation inoculant comprises the following steps: crushing and sieving rapeseed meal, uniformly mixing a fermentation microbial inoculum, the rapeseed meal and water according to a certain mass ratio, placing the mixture into a breathing bag, fermenting at 25-40 ℃ for 1-3 days after stirring every 12 hours, and drying and crushing after fermentation to obtain the fermented rapeseed meal for feeding.
2. The use according to claim 1, characterized in that: the composite fermentation inoculant comprises the following components in parts by weight: 20-30 parts of rhodotorula rubra, 10-20 parts of Pediococcus acidilactici, 5-10 parts of Aspergillus niger, 20-25 parts of Bacillus coagulans and 5-8 parts of cellulase.
3. The use according to claim 1, characterized in that: the rhodotorula rubra, pediococcus acidilactici, aspergillus niger, bacillus coagulans and cellulase produced by the Aspergillus niger are derived from fermented bean curd and pickle fermented food.
4. The use according to claim 1, characterized in that: the fermentation inoculant is prepared from the following components in percentage by mass: vegetable meal: water= (0.1-0.3): 100: (50-120), fermenting at 30-40deg.C for 1-2 days.
5. The use according to claim 4, characterized in that: the fermentation inoculant is prepared from the following components in percentage by mass: vegetable meal: water= (0.1-0.2): 100: (80-100), the fermentation temperature is 30-37 ℃ and the fermentation time is 2 days.
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