CN105475624B - Microbial fermentation feed and production method and application thereof - Google Patents
Microbial fermentation feed and production method and application thereof Download PDFInfo
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Abstract
The invention discloses a micro-meterBiological fermentation feed and a production method and application thereof. The method for preparing the microbial fermentation feed comprises the following steps: 1) uniformly mixing lactobacillus, clostridium butyricum Cb-2, saccharomycetes, spore bacteria, a fermentation base material and water to obtain a fermentation raw material; 2) sequentially carrying out aerobic fermentation and anaerobic fermentation on the fermentation raw materials obtained in the step 1) to obtain the fermented product. The microbial fermented feed contains live microbial bacteria and beneficial bacteria of which the total number is more than 5 × 10
9cfu/g, the quantity of the mould and the content of mycotoxin are lower than the national limit standard, the fermentation liquor is rich in organic acid, the content of lactic acid reaches 2.5%, the pH value is lower than 4.5, the titer of the antibacterial peptide is more than 1000u/g, the neutral protease is 150u/g, the α -amylase is 300u/g, the dry weight is higher than that of the neutral protease, the total protease is more than 600u/g, the whole fermentation process is controlled within 8-24h, the fermentation time is saved, and the energy consumption is controlled within 3%.
Description
Technical Field
The invention belongs to the field of feeds, and relates to a microbial fermentation feed and a production method and application thereof.
Background
With the improvement of living standard of people, people pay more attention to the safety, nutrition and health of animal food, which promotes the healthy and large-scale development of the breeding industry. In recent years, the number of large-scale livestock and poultry farms in China is continuously increased, the scale is continuously enlarged, and hundreds of millions of tons of feed are needed every year; however, the current microbial fermented feed is not ideal in application condition and poor in application effect, mainly due to the lagging strain (single yeast or spore bacteria are adopted), the poor process (simple aerobic fermentation), high energy consumption and increased cost, and the like. The following problems mainly exist in the production and circulation process: 1) the problem of large energy consumption of fermentation is as follows: the common fermentation process adopts aerobic bacteria for fermentation, and the energy consumption of the whole fermentation process is up to 20 percent, so that the conversion rate of the fermented feed is reduced; if the energy of the feed is increased, carbon sources such as sugar and the like need to be additionally added, so that the production cost is increased. 2) The problems of 'keep-alive' of beneficial bacteria and 'preservation' of bioactive components are as follows: through the production, storage and transportation and circulation processes of the fermented feed, the activity of beneficial bacteria and the 'keep-alive and preservation' of bioactive components in the product are difficult to reach expected values, and some of the beneficial bacteria and the bioactive components are even kept for about a week.
Disclosure of Invention
The invention aims to provide a microbial fermentation feed and a production method and application thereof.
The method for preparing the microbial fermentation feed comprises the following steps:
1) uniformly mixing lactobacillus, clostridium butyricum Cb-2, saccharomycetes, spore bacteria, a fermentation base material and water to obtain a fermentation raw material;
wherein the lactobacillus consists of lactobacillus plantarum Lp and pediococcus pentosaceus PP with equal mass;
the spore bacteria consist of spore bacteria B7348 and spore bacteria N9 with equal mass;
2) and (2) sequentially carrying out aerobic fermentation and anaerobic fermentation on the fermentation raw materials obtained in the step 1), and obtaining the microbial fermentation feed after the fermentation is finished.
In step 1) of the above method, the content of viable bacteria in the lactic acid bacteria is 2 × 10
6cfu/g; wherein the lactobacillus plantarum Lp and the pediococcus pentosaceus PP are both available from Shandong Baolaili bioengineering GmbH;
the content of the viable bacteria in the clostridium butyricum Cb-2 is 10
6cfu/g；
The yeast is selected from Candida utilis; the content of viable bacteria in the yeast is 10
6cfu/g; wherein the candida utilis is available from Shandong Baolaili bioengineering GmbH;
the content of viable bacteria in the spore bacteria is 2 × 10
6cfu/g; wherein the bacillus B7348 is available from biological engineering GmbH of Shandong Bao Lai;
the spore bacteria N9 can be obtained from biological engineering GmbH of Shandong Bao;
the fermentation base material consists of bran, brewer's grains, insect manure and montmorillonite;
wherein the bran accounts for 20-30 parts by mass, more specifically 25 parts by mass;
the mass portion of the beer lees is specifically 20-30, more specifically 25;
the mass part of the insect manure is specifically 20-30 parts, more specifically 25 parts;
the mass part of the montmorillonite is specifically 20-30 parts, more specifically 25 parts.
The dosage of the lactobacillus is that 50g of lactobacillus is added into each ton of fermentation base material;
the dosage of the clostridium butyricum Cb-2 is that 20g of clostridium butyricum Cb-2 is added into each ton of fermentation base material;
the using amount of the microzyme is that 20g of microzyme is added into each ton of fermentation base material;
the dosage of the spore bacteria is 20g of the spore bacteria added into each ton of fermentation base material;
the amount of water is 500kg per ton of fermentation base material.
In the aerobic fermentation step in the step 2), the fermentation time is 8-12 h, specifically 10 h;
the temperature is 15-30 ℃, specifically 25 ℃;
in the anaerobic fermentation step, the water content of the system is 30-45 percent, specifically 38 percent;
the fermentation time is 24-36 h, specifically 24 h;
the temperature is 15-25 ℃, in particular 20 ℃.
In addition, the microbial fermented feed prepared by the method also belongs to the protection scope of the invention.
The microbial fermented feed is a natural in-vitro predigested feed containing a microbial preparation, an immune promoter, an acidifier, an enzyme preparation and a mildew remover, which is fermented by adopting a globally leading microbial feed fermentation technology under the reasonable compatibility condition of 6 anaerobic bacteria and aerobic bacteria, and is a concentrated feed with complete nutrition and high-efficiency utilization. The lactobacillus strain used in the invention is the strain with the same-period latest technical performance in the United states, and other complex bacteria are also the strains bred by space breeding, and a novel complex flora with more remarkable performance advantage is formed by reasonable compatibility, and the best fermentation effect is ensured by utilizing the synergistic effect of the novel complex flora; breaks through the phenomenon that the performance of a single strain or a plurality of strains is unstable or the strains are simply applied in the conventional fermentation process in the market. The fermentation method is a global leading TMF biotechnology and a two-state two-step fermentation process, and the fermentation energy consumption is greatly reduced; by the anaerobic fermentation of the lactic acid bacteria, the growth of harmful mixed bacteria is inhibited, the excellent fermentation environment is ensured, and the functionality of the produced fermented feed is further ensured, namely the functional components have higher content and stronger activity. Compared with the traditional single microecological preparation, an enzyme preparation, an acidifier, a growth promoter and other products, the biological enzyme preparation has more complete functions, and simultaneously has the functions of producing enzyme, acid and bacteriocin, removing mildew and toxin, enhancing immunity, promoting growth and the like.
The microbial fermentation feed provided by the invention has the following characteristics:
1. the fermented feed product contains live microbial bacteria and beneficial bacteria of which the total number is more than 5 × 10
9cfu/g；
2. The quantity of the mould and the content of mycotoxin in the fermented feed are lower than the national limit standard;
3. the produced fermented feed product is rich in acetic acid, propionic acid, butyric acid, lactic acid and other organic acids, the content of the lactic acid reaches 2.5 percent, and the pH value is lower than 4.5;
4. the antibacterial peptide titer is more than 1000u/g, the neutral protease is 150u/g, the α -amylase is 300u/g, the dry weight is more than that, and the total protease is more than 600 u/g;
5. the whole fermentation process of the fermented feed is controlled within 8-24h, so that the fermentation time is saved, the energy consumption is reduced, the energy consumption is controlled within 3 percent, and the fermentation cost is reduced;
6. the method adopts the international leading production technology and packaging technology to reach the optimal state of bioactive components, greatly prolongs the service life of the fermented feed by more than one year, and ensures the strong functionality of the fermented feed.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.
The bran used in the following examples was purchased from magnetic kiln flour mills, and the brewery grain used was purchased from Taian Shengliyuan wine industry; the used insect manure is purchased from a jujube village insect farm; the montmorillonite is obtained from Taian Dakang animal husbandry.
The following bacteria were purchased from bio-engineering gmbh, Shandong Bao Li, and the deposit numbers of each of the bacteria are shown below:
lactobacillus plantarum Lp: CCTCC NO: m2010150;
pediococcus pentosaceus PP: CCTCC NO: m2012029;
clostridium butyricum Cb-2: CCTCC NO: m2011384;
bacillus B7348: CCTCC NO: m2010260;
spore bacterium N9: CCTCC NO: m2011301;
candida utilis: CCTCC NO: m2010090.
The Lactobacillus plantarum Lp and Pediococcus pentosaceus PP were purchased from bioengineering GmbH, Bay, Shandong.
Wherein, the preservation number is CCTCC NO: the lactobacillus plantarum LP of M2010150 has been described in another invention patent, whose information is: the patent numbers are: 201010240725.X, publication No. CN101914475A, grant No. CN101914475B, the name of invention is "a lactic acid bacteria for biological preservation and its application", the patent of the invention has been granted and disclosed, become the prior art, so this application no longer provides the proof of preservation of Lactobacillus plantarum LP;
the preservation number is CCTCC NO: the pediococcus pentosaceus PP of M2012029 has been described in another invention patent, whose information is: the patent numbers are: 20121008224.6, publication No. CN102599348A, and publication No. CN102599348B, entitled "a lactic acid bacteria semisolid fermentation product and process for improving animal growth performance", the patent of the invention has been granted and published, and becomes the prior art, so the application no longer provides the preservation evidence of the Pediococcus pentosaceus PP;
the preservation number is CCTCC NO: clostridium butyricum Cb-2 of M2011384 has been described in another invention patent, which is informative: the patent numbers are: 20111045528.5, publication No. CN102517238A, and publication No. CN102517238B, entitled "A Bacillus cereus producing acid and its application", the patent of this invention has been granted and published, become the prior art, so this application no longer provides the preservation evidence of this Clostridium butyricum Cb-2;
the preservation number is CCTCC NO: the bacillus B7348 of M2010260 has been described in another invention patent, whose information is: the patent numbers are: 201010589074.5, publication No. CN102120975A, and publication No. CN102120975B, entitled "a Bacillus subtilis with strong bacteriostatic action and its application", the patent of this invention has been granted and published, become the prior art, so this application no longer provides the preservation evidence of this Bacillus B7348;
the preservation number is CCTCC NO: the bacillus N9 of M2011301 is already described in another invention patent, and the information of the invention patent is as follows: the patent numbers are: 20111027806.4, publication No. CN102329749A, and publication No. CN102329749B, entitled "a Bacillus subtilis strain bred by aerospace breeding technology and its application", the patent of the invention has been granted and published, and becomes the prior art, so the application no longer provides the preservation proof of the Bacillus subtilis N9;
the preservation number is CCTCC NO: the candida utilis of M2010090 is already described in another invention patent, and the information of the invention patent is as follows: the patent numbers are: 20101018508.3, publication No. CN101864369A, and publication No. CN101864369B, entitled "a yeast strain with strong organic selenium tolerance, enrichment and transformation capability and its application", the patent of the invention has been granted and disclosed, and becomes the prior art, so the application no longer provides the preservation proof of the Candida utilis;
the specific compositions of the feeds used in the following examples 2-5 are shown in table 21:
TABLE 21 concrete composition of the feed used in examples 2 to 5
| Lactating sows | Corn (corn) | Bean pulp | Cotton seed dregs | Bean curd skin | Puffed soybean | Premix compound | Total up to | |
| 61.3 | 17 | 3 | 12 | 1.7 | 5 | 100 | ||
| Laying hen | Corn (corn) | Bean pulp | Cotton seed dregs | Bran | Stone powder | Premix compound | Total up to | |
| 60 | 21 | 2 | 4 | 8 | 5 | 100 | ||
| Corn (corn) | Bean pulp | Cotton seed dregs | Bran | Bean curd skin | Glucose | Premix compound | Total up to | |
| Weaned pig | 59 | 19.2 | 5 | 11 | 0.8 | 5 | 100 | |
| Fattening pig | 52.5 | 10 | 6.5 | 17.5 | 8.5 | 5 | 100 |
DDGS: distiller's dried grain and solubles, pig premix: purchased from bio-engineering, Inc. of Baolinlai, Shandong; chicken premix: purchased from bio-engineering, Inc. of Baolinlai, Shandong;
the laying hen feed, the weaned pig feed, the fattening pig feed and the lactating sow feed are purchased from Shandong Baolaili biological engineering Limited company, and the product numbers are Lu feeding pre-character (2015)139003, Lu feeding pre-character (2015)139006, Lu feeding pre-character (2015)139007 and Lu feeding pre-character (2015)139010 in sequence.
Example 1 preparation of microbial fermented feed
1) Uniformly mixing bran, brewer's grains, insect manure and montmorillonite according to equal mass to obtain a fermentation base material;
then uniformly mixing lactobacillus, clostridium butyricum Cb-2, candida utilis, spore bacteria, the fermentation base material and water to obtain a fermentation raw material;
wherein the lactobacillus consists of lactobacillus plantarum Lp and pediococcus pentosaceus PP with equal mass;
the spore bacteria consist of spore bacteria B7348 and spore bacteria N9 with equal mass;
the viable count of the lactobacillus plantarum Lp, pediococcus pentosaceus PP, clostridium butyricum Cb-2, candida utilis, spore bacteria B7348 and spore bacteria N9 is 10
6cfu/g；
The addition amount of the lactic acid bacteria is 50g of lactic acid bacteria added into each ton of fermentation base material;
the dosage of the clostridium butyricum Cb-2 is that 20g of clostridium butyricum Cb-2 is added into each ton of fermentation base material;
the usage amount of the yeast is that 20g of yeast is added into each ton of fermentation base material;
the dosage of the spore bacteria is 20g of the spore bacteria added into each ton of the fermentation base material;
the amount of water is 500kg per ton of fermentation substrate.
2) Carrying out aerobic fermentation on the fermentation raw material obtained in the step 1), adjusting the water content of a system to be 38% after the fermentation is finished, and then carrying out anaerobic fermentation to obtain the microbial fermentation feed provided by the invention after the fermentation is finished;
wherein, the aerobic fermentation is 'solid state' aerobic fermentation, spore bacteria and yeast fermentation are started, and fermentation is carried out for 10 hours at the temperature of 25 ℃;
the anaerobic fermentation is carried out by vacuumizing or packaging with a respiratory membrane, and clostridium butyricum and lactobacillus are started to ferment for 24 hours at 20 ℃.
And after the fermentation is finished, the obtained microbial fermented feed can be packaged according to the corresponding specification (such as 20 kg-40 kg per bag) according to the actual requirement.
The obtained fermented feed product is detected according to national standards GB478935-2010 and GB/T26428-2010, and the total number of the live micro-ecological bacteria and beneficial bacteria contained in the fermented feed product is more than 5 multiplied by 10
9cfu/g, total beneficial bacteria 5.07 x 10
9cfu/g
According to the national standard GB/T13092-2006, the obtained fermented feed product is detected by an ELISA detection kit, and the quantity of the mould and the mycotoxin content in the fermented feed product are lower than the national limit standard and are 3.1 multiplied by 10
4cfu/kg; 12ppb of aflatoxin, 37ppb of ochratoxin and 0.6ppb of T-2 toxin;
according to GB/T22142-2008, the obtained fermented feed product is detected, so that the fermented feed product is rich in acetic acid, propionic acid, butyric acid, lactic acid and other organic acids, the content of the lactic acid reaches 2.5 percent, and the content of the lactic acid reaches 3.2 percent; the pH was less than 4.5, and the pH was 3.7 as measured by a pH meter;
according to GB/T23881-2009, the antibacterial peptide titer in the fermented feed product is more than 1000u/g, the neutral protease is 150u/g, the α -amylase is 300u/g, the dry weight is more than 300u/g, and the total protease is more than 600 u/g.
Example 2 application of microbial fermentation feed in commercial layer chicken breeding
1.1.1 Experimental design
300 feathers of 375-day-old healthy Hailan brown laying hens are selected and randomly divided into 2 groups, namely a control group and an experimental group, wherein each group is provided with 3 parallel tests, and each parallel test has 50 feathers. The experimental group was fed with fermented feed at a fixed ratio (4% of the microbial fermented feed obtained in example 1 was added to the complete feed, i.e., the mass ratio of the complete feed to the microbial fermented feed obtained in example 1 was 100:4), and the other conditions were identical to those of the control group. The basic daily ration of the experimental group and the basic daily ration of the control group are completely consistent, and the experimental group and the control group can freely take food and drink water.
1.1.2 measurement index
Egg laying performance: daily average laying rate, egg breaking rate, average egg weight, daily average feed intake, feed-egg ratio and the like;
egg shape index: the egg-shaped index is equal to the transverse diameter/longitudinal diameter multiplied by 100 percent;
eggshell is relatively heavy: the relative weight of eggshell is equal to the weight of eggshell/weight of egg x 100%;
half unit: hu-100 lg (H-1.7W0.37+0.76),
wherein H is the concentrated protein height/mm, and W is the eggshell weight/g.
1.1.3 results and analysis
1.1.3.1 Effect on egg quality
The experimental group and the control group of laying hens are fed for 30 days according to the experimental design, the related indexes of the egg quality are detected, and the experimental results are shown in table 1.
TABLE 1 egg quality test results
As can be seen from table 1, the average daily laying rate of the experimental group is significantly increased, the feed-egg ratio is significantly decreased, and the egg breaking rate and the death and culling rate are also significantly decreased, compared with the control group. Research results show that the microbial fermented feed can obviously improve the solar energy rate, reduce the material-egg ratio and greatly reduce the egg breaking rate and the death rate.
1.1.3.2 Effect on egg laying Performance
The thicker the eggshell thickness is, the harder the eggshell quality of the egg is, and the egg is not easy to break; the hough unit is an important index representing the quality of eggs, which represents the freshness and protein quality of eggs. The larger the Hough unit, the higher the concentrated protein, and the fresher the egg. After the laying hens in the experimental group and the control group are fed for 30 days according to the experimental design, the relevant indexes of the egg laying performance are tested, and the experimental results are shown in table 2.
Table 2 egg laying performance test results
As can be seen from Table 2, the microbial fermented feed can obviously improve the quality of the eggshells, thicken the eggshells and reduce the breakage rate; the half unit of the egg is improved, and the protein quality and the freshness of the egg are ensured.
1.1.4 nodules
Application experiment results show that the microbial fermentation feed can obviously improve the daily average laying rate, reduce the feed-egg ratio and obviously reduce the egg breaking rate and the death rate; in addition, the product quality can be obviously improved, the appearance is better, the eggshell is thickened, and the freshness and the protein quality of the egg are improved.
The research result proves that the safe and efficient microbial fermented feed can promote the production performance of the laying hens, improve the egg quality and increase the disease resistance. The microbial fermented feed contains rich beneficial bacteria, namely lactobacillus with high biological activity and a large quantity, and other high-efficiency fresh bioactive substances, and plays a powerful probiotic role. A large amount of lactic acid bacteria rapidly colonize after entering the intestinal tract to form dominant flora, so that a large amount of organic acid is generated, the pH value in the intestinal tract is reduced, the propagation of intestinal pathogenic bacteria is inhibited, the immunity of the laying hens is improved, meanwhile, the acidic environment of the intestinal tract is favorable for activating the activities of various digestive enzymes such as pepsin and the like, the digestion and absorption of organisms on nutrient substances are promoted, and the production performance and the product quality of the laying hens are improved.
Example 3 application of microbial fermented feed in weaned pig breeding
2.1.1 Experimental design
90 healthy Taishan black weaned piglets with the body weight of about 8.5kg and the age of 35 days are selected for carrying out feeding experiments.
According to the principle of similar weight and consistent sex proportion, the weight is randomly divided into two groups, namely an experimental group and a control group, wherein each group is provided with three parallels, and each parallels 15 heads. The experimental group was fed with basal ration and fermented feed (2% of the microbial fermented feed obtained in example 1 was added to the complete feed, i.e., the mass ratio of the complete feed to the microbial fermented feed obtained in example 1 was 100:2), and the control group was fed with basal ration alone for 30 days of the experimental period. The basic daily ration of the weaned piglets of the experimental group and the control group is completely consistent, the weaned piglets are fed in the conventional modes of free feeding, drinking water and the like, and the disease prevention and the feeding management are carried out according to the conventional immunization program.
2.1.2 measurement of indices
Measurement of growth Performance: and respectively carrying out fasting weighing at the beginning and the end of the experiment, recording the initial weight, the final weight and the parallel feed consumption, and calculating the average daily gain, the average feed consumption and the feed conversion rate of each group.
Determination of incidence and diarrhea rate: during the test period, the health condition of the swinery is observed and recorded at regular time every day, and the morbidity and diarrhea quantity of each parallel experiment of each group are recorded, wherein the morbidity symptoms comprise piglets suffering from cough, diarrhea and leg pain.
And (3) fecal flora determination: and selecting the most representative lactic acid bacteria, escherichia coli and enterococcus faecalis in intestinal tracts of the piglets for culture and counting. Fresh excrement of 9 piglets is randomly selected in parallel from each group of 10 days, 20 days and 30 days respectively for counting strains.
Determination of total serum protein and serum urea content: the total protein content of serum is measured by a biuret method, and the urea content of serum is measured by a spectrophotometry method.
2.1.3 results and analysis
2.1.3.1 Effect on growth Performance
The effect of microbial fermented feed on growth performance, namely weight gain and feed-weight ratio, of weaned piglets is studied experimentally, and the results are shown in table 3.
TABLE 3 Effect of growth Performance results
From table 3, it was found that the addition of the microbial fermented feed to the basal diet improved the growth performance and feed conversion rate of piglets to some extent, compared to the control group. Compared with a control group, the daily average weight gain of an experimental group is improved by 13.4%, the feed conversion rate is obviously improved by 7.04%, the daily feed intake is improved by 5.4%, and the difference is obvious (P is less than 0.05); feed gain was reduced with significant differences (P < 0.05).
2.1.3.2 Effect on morbidity and diarrhea
The effect of microbial fermented feed on the morbidity and diarrhea of weaned piglets was studied experimentally, and the results are shown in table 4.
TABLE 4 Effect of onset and diarrhea
As can be seen from Table 4, no death cases were found in the control group of the experimental group during the experimental period. Compared with a control group, the morbidity of the experimental group is reduced by 44.1%, the diarrhea rate is greatly reduced by 75%, and the difference is obvious (P is less than 0.05). And the observation in the experimental period shows that the condition of the weaned piglets in the control group and the experimental group at the early stage of the experiment is not very different from that of the diarrhea, but the diarrhea of the experimental group is reduced along with the feeding of the fermented feed at the middle and later stages. Research results show that the feeding of the microbial fermentation feed can effectively reduce diarrhea and morbidity of piglets and promote the growth of the piglets.
2.1.3.3 Effect on fecal microflora
The effect of microbial fermented feed on microbial flora in feces was investigated, and the results of the experiment after feeding 14d and 21d are shown in table 5.
TABLE 5 results of the Effect of the microbial flora
As can be seen from Table 5, in experiment 14d, compared with the control group, both Lactobacillus and enterococcus faecalis were slightly increased, but the difference was not significant (P >0.05), and Escherichia coli was reduced by 5.9% and the difference was significant (P < 0.05); in experiment 21d, the lactobacillus and the enterococcus faecalis were increased by 7.0% and 5.4%, respectively, compared to the control group, and the difference was significant (P <0.05), and the escherichia coli was decreased by 8.3%, and the difference was significant (P < 0.05).
2.1.3.4 Effect on serum Total protein and serum Urea content
The effect of microbial fermented feed on serum total protein and serum urea of weaned pigs was studied experimentally, and the results are shown in table 6.
TABLE 6 Effect of serum Total protein and serum Urea content
As can be seen from Table 6, the microbial fermented feed can obviously increase the total protein content of serum from 50.1g/L to 55.7g/L, which is increased by 11.2%, and the difference is obvious (P < 0.05); serum urea was significantly reduced by 25.6% and the difference was significant (P < 0.05).
2.1.4 nodules
Research results show that the microbial fermented feed can obviously improve the growth performance and the feed conversion rate of weaned pigs, and has good promotion effect on weight gain and improvement of the feed utilization rate; obviously reducing the morbidity and diarrhea rate of piglets, and showing that the feed can enhance the body resistance of organisms and maintain the balance and coordination of intestinal environment; the intestinal microbial flora is reasonably regulated, so that the number of beneficial bacteria such as lactic acid bacteria and the like is greatly increased, the growth of escherichia coli is inhibited, and the number is reduced; can increase the total protein content of serum and reduce the urea content of serum, which shows that the protein and amino acid utilization of piglets can be effectively improved.
Due to the fact that microbial fermentation feed can improve the micro-ecological environment of the gastrointestinal tract of piglets, the feed is pre-digested, and good digestion and utilization rate of animals on the feed is guaranteed; fresh probiotic bacteria entering the intestinal tract maintain the advantages of beneficial bacteria in the digestive tract through field planting, bacteriocin in the feed can effectively inhibit the growth of harmful bacteria, and the anti-stress capability of the organism is improved; the dominant lactic acid bacteria in the intestinal tract generate a large amount of organic acid substances, so that the acidic environment of the intestinal tract is ensured, the growth of harmful bacteria can be inhibited, the growth and the propagation of other anaerobic bacteria in the intestinal tract can be promoted, the acidic environment is favorable for the digestion and the absorption of nutrient substances, and the utilization rate of the feed is further improved.
Example 4 application of microbial fermented feed in fattening pig breeding
2.2.1 Experimental design
90 healthy and disease-free live pigs growing uniformly and weighing about 35kg in the same pigsty are selected and randomly divided into two groups, the two groups are respectively an experimental combination control group, 45 heads in each group are respectively made into three parallel groups, each group is 15 heads in parallel, the experiment period is indefinite, and the experiment is finished when all the experimental group live pigs are fed to the weight of about 100 kg. During the experiment, the experimental groups are fed with a certain amount of fermented feed according to the specified proportion (4% of the microbial fermented feed obtained in the example 1 is added on the basis of the complete feed, namely the mass ratio of the complete feed to the microbial fermented feed obtained in the example 1 is 100:4), the parallel feeding management conditions of other groups are kept consistent, and the sanitation, immunization and disinfection procedures are carried out according to the conventional method.
2.2.2 measurement of indices
Measurement of growth Performance: and respectively carrying out fasting weighing at the beginning and the end of the experiment, recording the initial weight, the final weight and the parallel feed consumption, and calculating the average daily gain, the average feed consumption and the feed conversion rate of each group.
Determination of carcass performance: after the experiment is finished, each group randomly samples and slaughters 5 heads to measure the carcass performance and meat quality traits, including indexes such as body weight, slaughter rate, carcass length, backfat thickness, eye muscle area, meat division rate and the like.
Meat quality characteristics determination: the muscle quality characteristic indexes include: flesh color, marbling, muscle water power, tenderness, general chemical components of muscle, antioxidant property, nutrition, health care and flavor quality characteristics.
2.2.3 results and analysis
2.2.3.1 Effect on growth Performance
The effect of the microbial fermented feed on the growth performance, i.e. weight gain and feed-weight ratio, of the fattening pigs was experimentally studied, and the results are shown in table 7.
TABLE 7 influence results on growth Performance
From table 7, it was found that the growth performance and feed conversion rate of the fattening pigs were improved to some extent by feeding the microbial fermented feed as compared with the control group. Compared with a control group, the daily average weight gain of an experimental group is increased by 10.9%, the material-to-weight ratio is reduced by 10.03%, the feed conversion rate is obviously increased, and the difference is obvious (P is less than 0.05).
2.2.3.2 Effect on carcass Performance
The influence of the microbial fermented feed on the carcass performance of the fattening pigs is experimentally researched, and the research result is shown in table 8.
TABLE 8 Effect of carcass Properties results
As can be seen from table 8, compared with the control group, the carcass weight difference of the slaughtered experimental pigs of each group is significant (P <0.05), which means that the carcass weight fed with the fermented feed is significantly higher than that of the control group, and is increased by about 7.9%; the slaughter rate difference of two groups of experimental pigs is not significant (P > 0.05); carcass length difference was significant (P <0.05), about 4.0% increase; 6-7, the thickness of the back and the rib fat is thinned, and the reduction is about 2.1%; the eye muscle area is obviously increased by 20.6 percent; the meat cutting rate is obviously reduced by 11.6 percent.
2.2.3.3 Effect of meat quality characteristics
Effects on flesh color and marbling
The influence of the microbial fermented feed on meat color and marbling in meat quality characteristics of fattening pigs is studied in an experiment, and the experimental results are shown in table 9.
TABLE 9 effect of flesh color and marbling
As can be seen from Table 9, there was no significant difference in the respective measurement indexes of flesh color and marbling between the experimental group and the control group (P > 0.05).
(2) Influence on Hydraulic and tenderness of muscular system
The influence of the microbial fermented feed on the water power and tenderness of muscle systems in the meat quality characteristics of fattening pigs is studied in an experiment, and the experimental results are shown in table 10.
TABLE 10 Effect of muscular Water power and tenderness results
As can be seen from table 10, the water loss rate, drip loss, cooking loss and tenderness difference were significant (P <0.05) compared to the experimental combination control group. Research results show that the feeding of the microbial fermented feed can improve the water power of a muscle system in the feeding stage of growing-finishing pigs.
Effects on general chemical composition of muscle
The influence of the microbial fermented feed on conventional chemical components of muscles in meat quality characteristics of fattening pigs is studied in an experiment, and the experimental result is shown in table 11.
TABLE 11 results of the Effect of conventional chemical composition of muscle
As can be seen from table 11, there was no significant difference in dry matter, intramuscular fat, crude protein and crude ash content of the chicken in the two experimental groups (P > 0.05).
Effect on muscle antioxidant Properties
The influence of the microbial fermented feed on the muscle oxidation resistance in the meat quality characteristics of the fattening pigs is studied in an experiment, and the experimental result is shown in table 12.
TABLE 12 Effect of muscle antioxidant Performance 24h after slaughter
As can be seen from table 12, compared with the control group, the muscle SOD activity of the experimental group was significantly higher than that of the control group; the muscle MDA content difference is not significant (P >0.05), but the muscle MDA content of the experimental group is lower than that of the control group in general.
Influence on nutrition, health care and flavor quality characteristics
The influence of the microbial fermented feed on the nutrition, health care and flavor quality characteristics of the meat quality characteristics of the fattening pigs is studied in an experiment, and the experimental results are shown in table 13.
TABLE 13 Effect of muscle Nutrition and health and flavor quality Properties results
As can be seen from table 13, the content differences of total cholesterol, soluble collagen, unsaturated fatty acid, saturated fatty acid, and monounsaturated fatty acid were not significant (P >0.05) compared to the control group; the content of inosinic acid, total collagen and polyunsaturated fatty acid is obviously higher than that of the control group. The microbial fermented feed can increase the contents of muscle flavor substances and health-care active substances, and has good functional health-care effect.
2.2.4 nodules
The microbial fermented feed with a certain proportion is fed in the raising process of the live pigs in the growing and fattening period, so that the growing performance and carcass performance of the growing and fattening pigs can be obviously improved, the meat quality characteristic quality (pork series waterpower, tenderness and oxidation resistance are obviously improved), the cholesterol content in the meat products can be reduced to different degrees, and the content and the composition of functional active substances such as inosinic acid, collagen, polyunsaturated fatty acid and the like are improved.
The microbial fermented feed can improve the production performance and increase the breeding income, can be used as a feed concentrate supplement for improving the quality of commercial pork, has unique development and utilization values and wide economic benefits in the aspects of improving the physical and chemical properties of the pork, improving the flavor and taste of the pork, nourishing, health care and the like, and is worthy of large-scale popularization, application and demonstration.
Example 5 application of microbial fermented feed in sow farming
2.3.1 Experimental design
The pregnant sow variety is the existing sow variety in a farm, 40 healthy and active pregnant sows to 70-day-old pregnant sows with the same or similar gestation times are selected, the sows are randomly divided into two groups according to the principle that the weight and the gestation ratio are similar, the two groups are respectively an experimental group and a control group, and 20 sows in each group are not provided with repeated tests. The experimental group was fed with a certain proportion of microbial fermented feed (7% of the microbial fermented feed obtained in example 1 was added to the complete feed, i.e. the mass ratio of the complete feed to the microbial fermented feed obtained in example 1 was 100: 7), the other proportion was normal ration, the control group was fed with normal basal ration, the nutritional levels of all experimental rations were kept consistent, the other epidemic prevention, management and feeding technical levels were kept consistent, and the experimental period was 21 days after delivery.
2.3.2 measurement of indices
The growth performance of the sow is as follows: can reflect the indicators of postnatal feed intake of sow growth performance, sow constipation in a farrowing house and the like.
Number of dead births, number of weak births: the number of dead piglets, weak piglets, live piglets and weak piglets of the same litter size.
The lactation performance of the sow is as follows: the total average litter size, the number of live born piglets, the lactation capacity of the sows, the daily lactation amount, the average individual birth weight, the weaning individual weight and other indexes of each sow.
The health condition of the piglets is as follows: the indexes of the sow trigeminal morbidity, the retarded productivity, the early productivity, the piglet diarrhea rate, the weaning healthy piglet number, the piglet mortality and the like.
2.3.3 results and analysis
2.3.3.1 sow Performance Effect
The experiment researches the influence of the microbial fermented feed on the growth performance of the sows, and the experimental result is shown in table 14.
TABLE 14 sow growth Performance Effect results
As can be seen from Table 14, the growth performance of the sows fed with the microbial fermented feed is obviously superior to that of the sows fed with the common ration, the daily feed intake is obviously increased, and the constipation of the sows is obviously reduced. Therefore, the microbial fermentation feed can promote the optimization and improvement of the growth performance of the sows.
2.3.3.2 influence of dead and weak litter size
The experiment researches the influence of the microbial fermented feed on the number of stillbirth and weak litter size of sows, and the experimental results are shown in table 15.
TABLE 15 influence of number of dead and weak beats
Note: weak number refers to the number of piglets with weight less than 0.9 kg; weak litter size means the ratio of weak litter size to live litter size
As can be seen from Table 15, the feeding of the microbial fermented feed did not significantly affect the lactation period and the weak litter size, but significantly improved the litter size and decreased the weak litter size.
2.3.3.3 sow lactation Performance Effect
The experiment researches the influence of the microbial fermented feed on the lactation performance of the sows, and the experimental result is shown in table 16.
TABLE 16 influence of the lactation Performance of sows
As can be seen from table 16, when the sows are fed with the microbial fermented feed, the lactation yield and daily lactation yield of the sows can be obviously improved, and the weight of weaned piglets is obviously improved; however, there was no significant effect on the average litter size and the number of live litter size of sows.
2.3.3.4 effects on piglet health status
The effect of microbial fermented feed on the health condition of piglets was studied in this experiment, and the experimental results are shown in table 17.
TABLE 17 influence of piglet health status results
As can be seen from Table 17, no sow triple combination symptom is found in sows fed with the microbial fermented feed, the slow-release rate, the early yield and the diarrhea rate of the sows are remarkably reduced, and the weaned healthy piglet rate is obviously higher than that of the control group.
2.3.4 nodules
The microbial fermented feed for the sows can obviously improve the growth performance of the sows, reduce the number of dead births and weak births, improve the lactation capacity of the sows, obviously reduce the morbidity of piglets and greatly ensure the health condition of the piglets. The microbial fermented feed can quickly regulate and supplement beneficial bacteria in intestinal tracts, colonize in the intestinal tracts, and produce various nutrient substances and functional active substances such as lactic acid, bacteriocin, vitamins, amino acid and the like through propagation to play a role, so that the growth performance and the health condition of sows and piglets are ensured, the lactation performance of sows and the immunity of the sows are improved, and relevant performance indexes of the piglets are improved.
Claims (9)
1. A method for preparing microbial fermented feed, comprising the following steps:
1) uniformly mixing lactobacillus, clostridium butyricum Cb-2, candida utilis, spore bacteria, a fermentation base material and water to obtain a fermentation raw material;
wherein the lactobacillus consists of lactobacillus plantarum Lp and pediococcus pentosaceus PP with equal mass;
the spore bacteria consist of spore bacteria B7348 and spore bacteria N9 with equal mass;
the fermentation base material consists of bran, brewer's grains, insect manure and montmorillonite; wherein the bran accounts for 20-30 parts by mass; the mass portion of the beer lees is specifically 20-30; the mass part of the insect manure is specifically 20-30 parts; the mass part of the montmorillonite is specifically 20-30 parts;
2) sequentially carrying out aerobic fermentation and anaerobic fermentation on the fermentation raw materials obtained in the step 1), and obtaining the microbial fermentation feed after the fermentation is finished;
wherein in the step 1), the using amount of the lactic acid bacteria is 50g of the lactic acid bacteria added into each ton of the fermentation base material; the dosage of the clostridium butyricum Cb-2 is that 20g of clostridium butyricum Cb-2 is added into each ton of fermentation base material; the dosage of the candida utilis yeast is 20g of yeast added into each ton of fermentation base material; the dosage of the spore bacteria is 20g of the spore bacteria added into each ton of fermentation base material; the amount of the water is 500kg per ton of the fermentation base material;
wherein in the aerobic fermentation step in the step 2), the fermentation time is 8-12 h; the temperature is 15-30 ℃;
in the anaerobic fermentation step, the water content of the system is 30-45%; the fermentation time is 24-36 h; the temperature is 15-25 ℃.
2. The method of claim 1, wherein: the content of viable bacteria in the lactic acid bacteria in the step 1) is 2 multiplied by 10
6cfu/g；
The content of the viable bacteria in the clostridium butyricum Cb-2 is 10
6cfu/g；
The content of viable bacteria in the yeast is 10
6cfu/g; the content of viable bacteria in the spore bacteria is 2 × 10
6cfu/g。
3. The method according to claim 1 or 2, characterized in that: in the step 1), the mass part of the bran in the fermentation base stock is specifically 25 parts;
the mass portion of the beer lees is specifically 25 portions;
the mass part of the insect manure is specifically 25 parts;
the mass part of the montmorillonite is specifically 25 parts.
4. The method of claim 1, wherein: in the step 2) of aerobic fermentation, the fermentation time is 10 h.
5. The method of claim 1, wherein: in the step 2) of aerobic fermentation, the fermentation temperature is 25 ℃.
6. The method of claim 1, wherein: in the step 2) of anaerobic fermentation, the water content of the system is 38%.
7. The method of claim 1, wherein: in the step 2) of anaerobic fermentation, the fermentation time is 24 hours.
8. The method of claim 1, wherein: in the step 2) of anaerobic fermentation, the fermentation temperature is 20 ℃.
9. A microbial fermented feed prepared by the method of any one of claims 1 to 8.
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