CN112493366A - Compound microbial inoculum for solid-state fermentation weaned piglet feed, preparation method and application - Google Patents
Compound microbial inoculum for solid-state fermentation weaned piglet feed, preparation method and application Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/165—Paracasei
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/413—Acidilactici
Abstract
The invention provides a compound microbial inoculum for solid-state fermentation of weaned pig feed, a preparation method and application thereof, and relates to the technical field of fermented feed. The invention provides a complex microbial inoculum for solid state fermentation of weaned pig feed, which comprises pediococcus acidilactici ZPA017 and lactobacillus paracasei ZLP 019. The invention adopts the compound microbial inoculum of the solid-state fermentation weaned piglet feed to prepare the fermented feed, has reasonable strain compatibility, simple and efficient operation, can greatly shorten the fermentation time, effectively improve the nutritional quality of the feed, reduce the energy loss of the feed caused by oxygen consumption, heat production and the like, and reduce the use cost. The obtained feed has rich nutrient substances, reduces the content of crude fiber and anti-nutritional factors, and reduces the breeding cost; after the fermented feed is added into the feed for the weaned pigs according to a certain proportion, the feed utilization rate can be improved, the growth performance and immunity of the weaned pigs can be improved, the intestinal health of the weaned pigs can be improved, the emission of harmful bacteria in excrement is reduced, and the production environment for pig raising is facilitated.
Description
Technical Field
The invention belongs to the technical field of fermented feeds, and particularly relates to a compound microbial inoculum of a solid-state fermented weaned piglet feed, a preparation method and application.
Background
When the piglets are weaned, because the digestive system is not well developed, the digestive enzymes are not sufficiently secreted, and the nutritional ingredients in the feed are not easy to digest and absorb, the piglets are easily affected by stress factors such as environment, feed and management, the intestinal flora imbalance is caused, the diarrhea, the feed intake is reduced, the growth is hindered, and even the piglets die, and the great economic loss is caused to the production of the pigs. Currently, with the banning of feed antibiotics in animal breeding, lactobacillus fermented feed becomes a research and development hotspot of feed nutrition. The lactobacillus can be fermented in the feed to produce a large amount of metabolic products such as lactobacillus, organic acid, enzyme and the like, and can degrade anti-nutritional factors in the feed, so that the palatability of the feed and the digestion and absorption of nutrient substances are improved, the intestinal health of animals can be improved, the growth performance and the immune function of the animals are improved, and the culture environment is optimized.
At present, most of domestic lactobacillus fermented feeds adopt mixed fermentation of lactobacillus and aerobic bacteria such as yeast, bacillus, mould and the like, but the problems of random strain compatibility, complex production operation, long fermentation period (3-10 days or longer), easy infectious microbe contamination of the fermented feeds, high energy consumption and nutrition loss and the like exist. For example, in the patent application of publication No. CN110506862A, the basal materials of the fermented corn, the peeled soybean meal, the soybean hull, the soybean oil, the sucrose and the like are fermented for 3 to 6 days by using the bacillus subtilis, the bacillus licheniformis, the saccharomycetes, the lactobacillus plantarum and the lactobacillus reuteri, and the weaned pigs are fed; the application patent of publication No. CN110226671A adopts mixed bacteria of bacillus, lactic acid bacteria and yeast as fermentation strains of the complete fermentation feed, and ferments the antibiotic-free complete feed for piglets for 3-5 days; the patent with publication number CN103039699B adopts lactobacillus paracasei, bacillus subtilis and saccharomyces cerevisiae to perform liquid separate fermentation culture, then the liquid separate fermentation culture is placed in a sealed container for anaerobic fermentation, the liquid separate fermentation culture is performed for 75 to 240 hours at the temperature of 10 to 37 ℃, and then the liquid separate fermentation culture is added into weaned piglet feed according to a certain proportion. No patent of the composite fermented feed of pediococcus acidilactici and lactobacillus paracasei is found.
In order to optimize the technology of lactobacillus solid state fermentation feed, simplify the operation procedures, reduce the risk of fermented feed pollution, fully play the role of numerous metabolites generated in the lactobacillus fermentation process, and more importantly, improve the intestinal health of weaned pigs and improve the growth performance and the immune function of the weaned pigs, the invention provides the technology of the compound lactobacillus solid state fermentation weaned pig compound feed, and the verification is carried out in the breeding of the weaned pigs, so that the effects of improving the feed digestibility, promoting the healthy growth of the piglets and reducing the emission of harmful flora in excrement are achieved.
Disclosure of Invention
The invention aims to develop and develop a compound microbial inoculum for solid state fermentation weaned pig feed aiming at the actual problems and requirements in the production of weaned pigs and fermented feed. The invention also aims to provide a preparation method of the solid-state fermented weaned pig feed. The invention also aims to provide an application method of the composite microbial inoculum solid fermentation feed which is directly added into weaned pig feed in proportion.
In order to achieve the purpose, the invention adopts the following technical scheme:
a composite microbial inoculum for solid state fermentation of weaned pig feed comprises pediococcus acidilactici ZPA017 with the preservation number of CGMCC No.9040 and lactobacillus paracasei ZLP019 with the preservation number of CGMCC No. 11532.
Wherein, Lactobacillus paracasei ZLP019(Lactobacillus paracasei) and Pediococcus acidilictici ZPA017(Pediococcus acidilictici) are both preserved in the common microorganism center of China Committee for culture Collection of microorganisms (No. 3 of West Lu 1 of Beijing city, Xingyang district, Microbiol research institute of Chinese academy of sciences, postal code: 100101), and the preservation number of the Pediococcus acidilici ZPA017 ZPA017 is as follows: CGMCC No. 9040. The lactobacillus paracasei ZLP019 has a preservation number of: CGMCC No.11532, and the preservation number of the microorganism is CGMCC No. 11532.
Through a large number of experimental researches, the adopted strains of pediococcus acidilactici ZPA017 and Lactobacillus casei ZLP019 are finally selected as the pediococcus acidilactici ZPA017 (the preservation number is CGMCC No.9040) and the Lactobacillus paracasei ZLP019 (the preservation number is CGMCC No.11532) which are applied in the prior application and have the application number of 201710361900.3.
Preferably, the composite microbial inoculum is freeze-dried powder, and the effective viable bacteria content of the pediococcus acidilactici ZPA017 and the lactobacillus paracasei ZLP019 is 2.0 multiplied by 1010CFU/g and above.
The preparation method of the solid-state fermented weaned piglet feed comprises the following steps:
(1) preparing fermented weaned pig feed raw materials: preparing raw materials for fermentation according to parts by weight: 0.2 part of pediococcus acidilactici ZPA017 freeze-dried preparation, 0.2 part of lactobacillus paracasei ZLP019 freeze-dried preparation, 5 parts of glucose, 1000 parts of water and 1000 parts of weaned pig compound feed; wherein, the preservation number of the pediococcus acidilactici ZPA017 is as follows: CGMCC No.9040, the preservation number of lactobacillus paracasei ZLP019 is as follows: CGMCC No. 11532;
(2) according to the weight ratio, the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose are put into water, and after being fully dissolved, the weaned pig compound feed is added and stirred;
(3) putting the uniformly mixed feed into a clean plastic barrel, compacting, and sealing by a cover;
(4) putting the barrel into a constant-temperature, closed and clean culture device, and culturing at constant temperature to obtain the fermented feed.
The preparation method as described above, preferably, in step (1), the viable bacteria content of the lyophilized preparation of pediococcus lactis ZPA017 is 2.0 × 1010CFU/g, the viable bacteria content of the lactobacillus paracasei ZLP019 freeze-drying preparation is 2.0 multiplied by 1010CFU/g。
The production method as described above, preferably, in the step (4), the incubation is carried out at 34 ℃ for 24 hours.
The total effective viable count of lactobacillus in the prepared fermented feed reaches 3.0 multiplied by 109More than CFU/g, obviously improved contents of crude protein, acid soluble protein, alpha-D-glucose, sucrose isomaltase and the like, more than 51.39g/kg of lactic acid, less than 29mg/100g of volatile basic nitrogen, pH value of 4.29 and acid flavor.
The invention also provides application of the fermented feed obtained by the preparation method as an additive in feed.
The use as described above, preferably the fermented feed is used in a pig feed, more preferably in a feed for weaned piglets.
Furthermore, the total number of viable bacteria in the fermented feed is 3.0 multiplied by 109CFU/g and above.
Further, the fermented feed is added into basic ration, and the addition amount of the fermented feed is 1% of the basic ration.
The invention has the beneficial effects that:
the composite microbial inoculum for the solid-state fermentation weaned pig feed is used for the composite fermentation weaned pig feed, can promote the improvement of the intestinal flora environment of young animals such as weaned pigs and the like, and can digest and absorb carbohydrates in the feed, and has good application prospect in the nutrition of the piglet feed.
The preparation method of the compound lactobacillus solid state fermentation weaned pig compound feed provided by the invention can exert the probiotic advantages of pediococcus acidilactici ZPA017 and lactobacillus paracasei ZLP019, has reasonable compatibility and simple and efficient operation, can greatly shorten the fermentation time, effectively improve the nutritional quality of the feed, reduce the energy loss of the feed caused by oxygen consumption, heat production and the like, and reduce the use cost. The obtained fermented feed has viable count of lactobacillus of 3.0 × 109More than CFU/g, lactic acid content of 51.39g/kg, volatile basic nitrogen content of less than 29mg/100g, pH of 4.29, crude protein, acid soluble protein, and metabolites such as alpha-D-glucose, sucrose-isomaltase, etcObviously improves the content of crude fiber and anti-nutritional factors and reduces the content of the crude fiber and the anti-nutritional factors. After the fermented feed is fed to weaned piglets according to a certain proportion, the digestibility of the feed and the average daily gain of the piglets can be improved, the feed-weight ratio and the diarrhea rate are reduced, the immunity function of the organism is enhanced, the viable count of lactic acid bacteria in excrement is improved, and the viable count of harmful bacteria such as escherichia coli, staphylococcus aureus and the like is reduced. Therefore, the compound lactobacillus solid-state fermentation weaned pig feed can increase the nutritional value of the feed, improve the intestinal health of piglets, improve the immunity and growth performance of organisms, reduce the emission of harmful bacteria in pig manure, and is beneficial to the improvement of the breeding environment.
After the solid-state fermented feed for the weaned pigs is fed by the weaned pigs, the digestibility of the feed and the growth performance of the weaned pigs can be improved, the immunity of the weaned pigs is enhanced, the intestinal health of the weaned pigs is promoted, and the emission of harmful bacteria in excrement is reduced.
Drawings
FIG. 1 shows the results of the effect of different inoculation ratios (inoculum size) on the fermented feed at different time points.
FIG. 2 shows the effect of different fermentation temperatures on fermented feed.
FIG. 3 shows the effect of different feed water ratios on fermented feed.
FIG. 4 shows the results of conventional nutrients (xianzhi) for fermented feed.
FIG. 5 shows the results of conventional nutrient composition (dry basis) of fermented feed.
Detailed Description
The complex microbial inoculum consists of pediococcus acidilactici ZPA017 and lactobacillus paracasei ZLP 019. Both of these strains of lactic acid bacteria belong to microbial strains that are allowed for feed production in the ministry of agriculture No. 2045 (2013) bulletin, are isolated from healthy piglets by the present research laboratory, and have excellent probiotic properties: in vitro tests prove that the two strains of lactic acid bacteria have the biological characteristics of quick acid production, acid resistance, bile salt resistance, strong adhesion, good antibacterial activity and the like. Through further mass researches, genome sequencing and metabolic pathway analysis show that a perfect L-lactic acid production metabolic pathway exists in the glycolysis pathway of pediococcus acidilactici ZPA017, and alpha-D-glucose can be used for producing lactic acid; the sugar metabolism pathway of lactobacillus paracasei ZLP019 is rich in glycoside hydrolase, and can hydrolyze cellulose or complex carbohydrates derived from various plants into soluble and easily-utilized small-molecule oligosaccharides, wherein sucrose-isomaltase (EC:3.2.1.10) can hydrolyze polysaccharides such as dextrin, maltose and isomaltose, and disaccharides into alpha-D-glucose. The above studies indicate that 2 strains of lactic acid bacteria have excellent probiotic properties, can degrade indigestible cellulose or complex carbohydrates in feed, and have complementarity in the production of metabolites such as L-lactic acid and α -D-glucose.
According to the invention, by combining the physiological characteristics of weaned pigs and aiming at the problems of components, nutrition and production of the pig feed at the specific stage at present, through screening and compatibility combination of a large number of different lactic acid bacteria, 2 strains of pediococcus acidilactici ZPA017 and Lactobacillus paracasei ZLP019 have synergistic effect, the compound feed for weaned pigs is fermented together, the probiotic synergistic effect of the 2 strains can be fully exerted, the large amount of metabolites such as lactic acid bacteria, lactic acid, glucose, sucrose-isomaltase and the like in the fermented feed is promoted, the nutritional value and digestive absorption of the feed can be improved, the growth of bacteria and intestinal tract pathogenic bacteria in the feed can be quickly inhibited, the influence of mycotoxin and anti-nutritional factors is reduced, the nutritional quality of the feed for weaned pigs is improved, the breeding cost is reduced, and the healthy growth of the weaned pigs is promoted.
The following examples are intended to further illustrate the invention but should not be construed as limiting it. Modifications and substitutions may be made thereto without departing from the spirit and scope of the invention.
Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1 optimization of Process parameters of Compound Lactobacillus solid fermentation weaned piglet feed
Freeze-dried powder of Pediococcus acidilactici ZPA017 and Lactobacillus paracasei ZLP019 respectively (the viable bacteria content is 2.0 × 10 respectively)10CFU/g) was inoculated into a weaned pig compound feed (moisture content 11.71%), and the inoculation ratio (inoculation) was studiedThe amount of viable lactobacillus in the fermented feed, and the influence of fermentation parameters such as the amount of the viable lactobacillus in the fermented feed, time, temperature, feed-water ratio and the like on the contents of lactic acid, alpha-D-glucose and sucrose isomaltase in the fermented feed. The inoculation ratio (inoculation amount) is freeze-dried powder lactococcus lactis ZPA 017: lactobacillus paracasei ZLP019 is prepared and inoculated into 1000g of feed according to the weight ratio (in g: g) of 1:1, 1:3, 1:5, 3:1, 3:3, 3:5, 5:1, 5:3, 5:5, 0.5:0.5, 0.5:1, 0.5:1.5, 1:0.5, 1:1, 1:1.5, 1.5:0.5, 1.5:1, 1.5:1.5, 0.2:0.2, 0.2:0.5, 0.2:0.8, 0.5:0.2, 0.5:0.5, 0.5:0.8, 0.8:0.2, 0.8:0.5, 0.8:0.8, the ratio of material to water is 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1 respectively, the fermentation temperature is 25 ℃, 27 ℃, 30 ℃, 33 ℃, 34 ℃, 36 ℃, 37 ℃, 39 ℃, the fermentation time is 24h, 48h and 72h respectively, and the viable count, the lactic acid, the alpha-D-glucose and the sucrose isomaltase content in each fermented feed are respectively measured. Only the results of the viable count (CFU/g), lactic acid (mg/g), glucose (mg/g) and sucrose isomaltase (U/g) contents of the fermented feed under several different fermentation conditions such as inoculation ratio, fermentation time, temperature and feed-water ratio are shown. FIG. 1 shows the results of the contents of viable bacteria (CFU/g), lactic acid (mg/g), glucose (mg/g) and sucrose isomaltase (U/g) produced by the fermented feed at different inoculation ratios for 24h, 48h and 72h, respectively. FIG. 2 shows the results of the contents of viable count (CFU/g), lactic acid (mg/g), glucose (mg/g) and sucrose isomaltase (U/g) produced by fermenting the feed at different temperatures for 24 hours, and FIG. 3 shows the results of the contents of viable count (CFU/g), lactic acid (mg/g), glucose (mg/g) and sucrose isomaltase (U/g) produced by fermenting the feed at different feed-water ratios for 24 hours.
As can be seen from the graphs 1-3, the compound feed for the composite fermented weaned pigs of pediococcus acidilactici ZPA017 and Lactobacillus paracasei ZLP019 has different effects on the viable count of lactic acid bacteria and the contents of lactic acid, alpha-D-glucose and sucrose-isomaltase in the fermented feed under different inoculation ratios (inoculation amounts), time points, temperatures and feed-water ratios. And (3) analyzing the influence of the fermentation parameters on four indexes of the fermented feed by adopting a principal component analysis method to obtain the optimal fermentation parameters: the optimal inoculation ratio of the pediococcus acidilactici ZPA017 and the lactobacillus paracasei ZLP019 composite fermentation is 0.2:0.2 (w/w, 1000g feed (i.e. 0.2g of pediococcus acidilactici ZPA017 freeze-dried powder and 0.2g of lactobacillus paracasei ZLP019 freeze-dried powder are inoculated into 1000g feed)), the optimal fermentation time is 24h, the optimal culture temperature is 34 ℃, and the optimal ratio of material to water is 1:1.
Example 2 preparation of Compound Lactobacillus solid fermentation weaned piglet feed
The invention discloses an example of a preparation method of a compound lactic acid bacteria solid state fermentation weaned piglet feed, which comprises the following steps:
(1) preparing raw materials of the fermented feed in parts by weight: pediococcus acidilactici ZPA017 freeze-dried preparation (viable bacteria content is 2.0X 10)10CFU/g)0.2 portion, lactobacillus paracasei ZLP019 freeze-drying preparation (the viable bacteria content is 2.0 multiplied by 10)10CFU/g)0.2 parts, glucose 5 parts, tap water 1000 parts, and weaned pig compound feed (water content 11.71%) 1000 parts.
(2) Putting the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose into tap water, adding the weaned pig compound feed after the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose are fully dissolved, and stirring;
(3) putting the uniformly mixed feed into a clean plastic barrel, compacting, and sealing by a cover;
(4) putting the barrel into a constant-temperature, sealed and clean culture device, and culturing at 34 deg.C for 24 hr to obtain lactobacillus fermented feed.
Example 3 preparation of Compound Lactobacillus solid fermented feed for weaned piglets
The invention discloses another example of a preparation method of a compound lactic acid bacteria solid state fermentation weaned piglet feed, which comprises the following steps:
(1) preparing raw materials of the fermented feed in parts by weight: pediococcus acidilactici ZPA017 freeze-dried preparation (viable bacteria content is 2.0X 10)10CFU/g)1.5 parts, lactobacillus paracasei ZLP019 freeze-drying preparation (viable bacteria content is 2.0 multiplied by 10)10CFU/g)0.5 parts, glucose 5 parts, tap water 1000 parts, and weaned pig compound feed (water content 11.71%) 1000 parts.
(2) Putting the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose into tap water, adding the weaned pig compound feed after the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose are fully dissolved, and stirring;
(3) putting the uniformly mixed feed into a clean plastic barrel, compacting, and sealing by a cover;
(4) putting the barrel into a constant-temperature, sealed and clean culture device, and culturing at 34 deg.C for 48h to obtain lactobacillus fermented feed.
Example 4 Change of index before and after fermentation of feed
Fresh samples of the composite lactic acid bacteria fermented feeds (fermented feed 1 and fermented feed 2) were obtained by the preparation methods of examples 2 and 3, and the pH, the viable count of lactic acid bacteria, and the contents of lactic acid, volatile basic nitrogen, α -D-glucose, sucrose-isomaltase, porcine conglycinin, porcine soybean agglutinin, porcine trypsin, and the like in the fresh samples were measured. Drying the fermented feed at 60 ℃, crushing and sieving by a 60-mesh sieve to obtain a fermented feed dry sample, and measuring the contents of crude protein, crude ash, calcium, total phosphorus, acid soluble protein and crude fiber in the dry sample. The results are shown in FIGS. 4 and 5.
As can be seen from FIG. 4, according to the preparation methods of example 2 and example 3, the pH values of the fermented weaned pig compound feed (fresh base) are respectively reduced to 4.29 and 4.34, and the viable count of the lactic acid bacteria reaches 3.0 × 109And 9.5X 108CFU/g, which is obviously higher than the viable count in the unfermented feed (5.0 multiplied by 10)3CFU/g), the content of lactic acid reaches 51.39 and 36.15g/kg, the content of acid soluble protein, alpha-D-glucose and sucrose-isomaltase is also obviously increased, the content of anti-nutritional factors such as porcine conglycinin, porcine soybean agglutinin and porcine trypsin inhibitory factor is obviously reduced, which shows that the lactobacillus composite fermentation weaned piglet feed can degrade the content of the anti-nutritional factors and crude fiber in the feed, increase the content of lactobacillus, lactic acid, enzyme, protein and the like in the feed, is beneficial to the digestion and absorption of the weaned piglet intestinal tract to the nutrient substances in the feed, and improves the nutritional value of the feed. In addition, the content of volatile basic nitrogen in the fermented feed is 24.35 and 15.03mg/100g respectively, and the fermented feed is fresh and delicious, has sour flavor, and can improve palatability and promote piglet feeding.
As can be seen from FIG. 5, the fermented mixed feed (dry basis) for weaned pigs has significantly increased contents of crude protein, total phosphorus and acid soluble protein, and significantly decreased contents of crude ash, calcium and crude fiber. The results show that after the compound feed for the weaned pigs is subjected to solid-state fermentation by using the compound lactic acid bacteria, the macromolecular protein in the compound feed can be decomposed into micromolecular polypeptide or amino acid which is easier to digest and absorb by the weaned pigs, and the compound feed is beneficial to the growth of the weaned pigs.
As can be seen from the figures 4 and 5, the compound feed of the pediococcus acidilactici ZPA017 and the lactobacillus paracasei ZLP019 has great influence on the quality of the fermented feed due to the compound proportion and the fermentation parameters. The indexes of viable count of lactobacillus, metabolites and the like of the fermented feed obtained by fermentation according to the researched optimal proportion and fermentation parameters (example 2) are superior to those of the fermented feed prepared by other proportions and fermentation parameters (such as example 3). Therefore, the two lactic acid bacteria are scientifically and reasonably combined to perform compound fermentation on the feed for the weaned pigs under proper conditions, so that the optimal fermentation effect can be obtained, and the nutritional quality of the feed is improved.
Example 5 application of fermented feed in weaned pig production
1. Design of experiments
60 healthy weaned piglets (Du multiplied by length multiplied by size) with similar initial weights (9.47 +/-0.22 Kg) are selected and randomly divided into a control group and a test group of 2 groups, each group is provided with 3 repetitions, and each repetition is 10. The control group was fed with basal diet, and the test group was fed with basal diet plus 1% of the compound lactobacillus solid fermented feed prepared in example 2 (the total effective viable count of lactobacillus in the feed was 3.0 × 10)7CFU/g or more). The basal ration was formulated with reference to the us NRC (2012) standard. During the test period, piglets eat and drink water freely, and are immunized conventionally. The test period was 30 days.
2. Measurement index
At the beginning and end of the test, the individual is weighed on an empty stomach; during the test period, the growth condition of the swinery is observed every day, and the feed consumption and the diarrhea condition of the piglets are recorded; after the test is finished, randomly taking 9 parts of fresh excrement samples in each group, and measuring the viable count of lactic acid bacteria, escherichia coli, staphylococcus aureus and the like in the excrement; randomly selecting 9 pigs from each group for blood sampling, and measuring biochemical and immune indexes in serum. Data statistics single-factor analysis of variance was performed using SPSS 18.0 statistical software, and results are expressed as mean ± standard deviation.
3. Results and analysis
3.1 results of the effect of the compound lactobacillus fermented feed on the growth performance of weaned pigs are shown in Table 1.
TABLE 1 Effect of Compound Lactobacillus fermented feed on growth Performance of weaned piglets
Note: the difference between the same shoulder marks represents significant difference, and the difference between the lower case letters represents significant difference (P < 0.05). The same is as follows.
As can be seen from Table 1, the average daily gain of the test group is increased by 9.93% (P <0.05), the feed-to-weight ratio is decreased by 7.45% (P <0.05), and the diarrhea rate is decreased by 3.63% compared with the control group. Therefore, the compound lactobacillus solid fermentation feed with a certain dosage is added into the daily ration of the weaned pigs, so that the growth speed and the feed efficiency of the weaned pigs can be improved, the diarrhea rate is reduced, the feeding cost is reduced, and the pig raising benefit is improved.
3.2 digestibility of the compound lactobacillus fermented feed to the nutritional ingredients of the weaned pig feed, which is shown in table 2.
TABLE 2 influence of composite lactobacillus fermented feed on apparent digestibility of nutritional ingredients of weaned piglets (%)
As can be seen from Table 2, the apparent digestibility of the weaned piglets in the test group to the crude protein in the feed is obviously increased (P <0.05) compared with the control group, which shows that the fermented feed can promote the digestion and absorption of the weaned piglets to the protein in the feed, thereby promoting the improvement of the growth performance of the weaned piglets.
3.3 results of the effect of the composite lactobacillus fermented feed on the number of microorganisms in the feces of weaned piglets are shown in Table 3.
TABLE 3 influence of the Compound Lactobacillus fermented feed on the microbial count in weaned pig manure (CFU/g)
As can be seen from Table 3, after the fermented feed is added, compared with a control group, the number of the lactic acid bacteria in the pig manure of the test group is increased by 480.75% (P <0.05), the number of the escherichia coli is reduced by 70.98%, and the number of the staphylococcus aureus is reduced by 76.34% (P <0.05), so that the composite lactic acid bacteria fermented feed can increase the content of beneficial bacteria in the intestinal tract of the weaned pig and reduce the content of harmful bacteria, thereby improving the balance of the microecological environment of the intestinal tract of the pig and being beneficial to the growth of the piglet and the improvement of the breeding environment.
3.4 the results of the effect of the compound lactobacillus fermented feed on the serum biochemical and immune indexes of the weaned pigs are shown in Table 4.
TABLE 4 influence of the composite lactobacillus fermented feed on serum biochemical and immunological indexes of weaned piglets
As can be seen from Table 4, the addition of the fermented feed to the daily ration can increase the contents of total protein, globulin, IgG and IgM in the serum of the weaned piglets, and reduce the contents of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase with significant difference (P < 0.05). The total protein in serum consists of albumin and globulin, the level of which can reflect the strength of protein anabolism of an organism, and the globulin is secreted after B cells are converted into plasma cells and can reflect the resistance of the organism; glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase are 2 indexes for detecting liver functions, and the 2 enzyme activities are increased to indicate that liver cell functions are damaged; the immune globulin is the main immune molecule for mediating the humoral immunity of animals and also is an important index for reflecting the immune function of organisms, and the increase of the content of IgG and IgM in serum indicates that the immune function of the organisms is enhanced. Therefore, the composite lactobacillus fermented feed is added into the feed for the weaned pigs according to a certain proportion to feed the weaned pigs, so that the liver function of the weaned pigs can be protected, the immunity of the weaned pigs is improved, and the animal health is promoted.
Claims (10)
1. The compound microbial inoculum for solid-state fermentation of the feed for weaned piglets is characterized by comprising pediococcus acidilactici ZPA017 with the preservation number of CGMCC No.9040 and lactobacillus paracasei ZLP019 with the preservation number of CGMCC No. 11532.
2. The complex microbial inoculant according to claim 1, wherein the complex microbial inoculant is freeze-dried powder, and the effective viable bacteria content of the pediococcus acidilactici ZPA017 and the lactobacillus paracasei ZLP019 are both 2.0 x 1010CFU/g and above.
3. The preparation method of the solid-state fermented weaned piglet feed is characterized by comprising the following steps:
(1) preparing fermented weaned pig feed raw materials: preparing raw materials for fermentation according to parts by weight: 0.2 part of pediococcus acidilactici ZPA017 freeze-dried preparation, 0.2 part of lactobacillus paracasei ZLP019 freeze-dried preparation, 5 parts of glucose, 1000 parts of water and 1000 parts of weaned pig compound feed; wherein, the preservation number of the pediococcus acidilactici ZPA017 is as follows: CGMCC No.9040, the preservation number of lactobacillus paracasei ZLP019 is as follows: CGMCC No. 11532;
(2) according to the weight ratio, the pediococcus acidilactici preparation, the lactobacillus paracasei preparation and the glucose are put into water, and after being fully dissolved, the weaned pig compound feed is added and stirred;
(3) putting the uniformly mixed feed into a clean plastic barrel, compacting, and sealing by a cover;
(4) putting the barrel into a constant-temperature, sealed and clean culture device, and culturing at a constant temperature of 34 ℃ for 24h to obtain the fermented feed.
4. The method of claim 3, wherein the milk is prepared byThe viable bacteria content of the lyophilized preparation of Pediococcus acidilactici ZPA017 is 2.0 × 1010CFU/g, the viable bacteria content of the lactobacillus paracasei ZLP019 freeze-drying preparation is 2.0 multiplied by 1010CFU/g。
5. The production method according to claim 3, wherein in the step (4), the incubation is carried out at 34 ℃ for 24 hours.
6. Use of the fermented feed obtained by the production method according to any one of claims 3 to 5 as an additive in feed.
7. The use according to claim 6, wherein the fermented feed is used in pig feed.
8. The use according to claim 6, wherein the fermented feed is used in the feed of weaned piglets.
9. The use according to claim 6, wherein the total number of viable bacteria in the fermented feed is 3.0 x 109CFU/g and above.
10. The use of claim 6, wherein the use is the addition of the fermented feed to a basal ration in an amount of 1% of the basal ration.
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