CN107173522A - A kind of fermented feed containing probiotics and its preparation method and application - Google Patents
A kind of fermented feed containing probiotics and its preparation method and application Download PDFInfo
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- 239000006041 probiotic Substances 0.000 title claims abstract description 70
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 70
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- 230000037213 diet Effects 0.000 claims abstract description 21
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- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 17
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 14
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 12
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 12
- 238000000855 fermentation Methods 0.000 claims description 23
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- 239000002609 medium Substances 0.000 claims description 10
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 8
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- 239000001963 growth medium Substances 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
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- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 6
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- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 claims description 5
- KVCWSJZUKMSPLM-UHFFFAOYSA-N O.O[PH2]=O Chemical class O.O[PH2]=O KVCWSJZUKMSPLM-UHFFFAOYSA-N 0.000 claims description 5
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 claims description 5
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- 229920002261 Corn starch Polymers 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
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- 239000005913 Maltodextrin Substances 0.000 claims description 3
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
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- 229910052749 magnesium Inorganic materials 0.000 claims 1
- 239000001632 sodium acetate Substances 0.000 claims 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical class S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims 1
- 206010012735 Diarrhoea Diseases 0.000 abstract description 12
- 230000001143 conditioned effect Effects 0.000 abstract description 12
- 244000052769 pathogen Species 0.000 abstract description 12
- 230000001717 pathogenic effect Effects 0.000 abstract description 12
- 241000588724 Escherichia coli Species 0.000 abstract description 9
- 241000607142 Salmonella Species 0.000 abstract description 9
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 8
- 241001465754 Metazoa Species 0.000 abstract description 6
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- 235000014590 basal diet Nutrition 0.000 description 3
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- 238000009360 aquaculture Methods 0.000 description 1
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- 238000009395 breeding Methods 0.000 description 1
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- 230000018109 developmental process Effects 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/02—Breeding vertebrates
-
- 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
- A23V2400/169—Plantarum
Abstract
The invention belongs to animal and fowl fodder field, it is related to a kind of fermented feed containing probiotics and its preparation method and application.Specific method is, after bacillus subtilis, VREF, 4 kinds of probiotics of Lactobacillus plantarum and S. cervisiae are individually fermented, according to 1 ~ 3：1~3：1~3：1 ~ 3 ratio is mixed and made into mixing probiotics fermention liquid.Probiotics fermention liquid will be mixed with complete feed according to 3.5 ~ 5.0：After 10 ratio mixing is mixed thoroughly, the h of sealed fermenting 24 30, is made fermented feed under the conditions of 25 40 DEG C.The total viable count of fermented feed is up to 1.21 × 1010 CFU/g, and pH is minimum to be down to 4.2.The fermented feed made can be added to feeding weanling pig and growing and fattening pigs in daily ration directly or with 5 10% ratio.Compared with common complete diet pellet, using fermented feed of the present invention, the feedstuff-meat ratio and diarrhea rate of pig are significantly reduced；The quantity of the conditioned pathogen such as Escherichia coli, salmonella and staphylococcus aureus declines to a great extent in excrement.
Description
Technical field：
The invention belongs to animal and fowl fodder field, it is related to a kind of fermented feed containing probiotics and its preparation method and application.
Background technology：
The continuous lifting that fast development and people with China's livestock and poultry breeding industry are required for quality of the life, animal products
Quality attracts attention more and more, and the quality of feed directly decides the quality of animal products.In recent years, American-European flourishing state
Family widelys popularize application of the fermented feed in cultivation, and China also has part plant to start recognition and acceptance to use fermented feed
Instead of conventional feed.
In brief, fermented feed is product of the conventional feed after probiotics fermention.Compared to conventional feed, fermentation is raised
Material has obvious advantage：Probiotics can utilize the macromoleculars such as protein, fat and the polysaccharide in feed, be degraded into more
The small-molecule substance absorbed beneficial to animal, improves the utilization rate of nutriment；The bacteriocin that fermentation of bacillus is produced can
Suppress the growth of harmful bacteria；Lactobacillus-fermented produces organic acid in addition to having bacteriostasis, moreover it is possible to improve the palatability of feed；
A large amount of protein and vitamin are produced during saccharomycetes to make fermentation, more sufficient nutrition is provided to animal.Due to fermented feed
Have the advantages that above conventional feed does not possess, have huge application prospect in following aquaculture.
The content of the invention：
（One）Goal of the invention：
It is an object of the invention to provide a kind of fermented feed containing probiotics and its preparation method and application, the fermented feed of preparation is agreeable to the taste
Property it is good, be favorably improved the feed intake and efficiency of feed utilization of pig, strengthen the resistance against diseases of pig, promote growth and the health of pig.
（Two）Technical scheme：
The purpose of the present invention is realized by following scheme：
1. the invention provides a kind of fermented feed containing probiotics, by mixing probiotics viable bacteria and its metabolite and antibiotic-free
Ferment complete feed is constituted, and water content is 35%-50%, and the viable count of fermented feed reaches as high as 1.21 × 1010 CFU/g, pH are
4.2-4.8；Above-mentioned mixing probiotics includes bacillus subtilis WEI-2（CGMCC No.7744）, VREF WEI-10
（CGMCC No.7746）, Lactobacillus plantarum WEI-70（CGMCC No.11747）With saccharomyces cerevisiae WJ-113.
Wherein bacillus subtilis WEI-2（CGMCC No.7744）It is documented in the special of Application No. 201310719942.0
In sharp application documents；VREF WEI-10（CGMCC No.7746）It is documented in the patent of Application No. 201310719899.8
In application documents；Forage plant breast bacterium Lactobacillus plantarum Lactobacillus plantarum are sent on November 26th, 2015
China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, numbering is CGMCC No. 11747, preservation
Location：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of Chinese Academy of Sciences postcode:100101 phones：86-10-
64807596；Saccharomyces cerevisiae is to be separated according to method disclosed in Application No. 200510010940.X patent application document
Screening is obtained, and is named as saccharomyces cerevisiae WJ-113.
2. the invention provides a kind of preparation method of fermented feed containing probiotics, preparation method includes：Mix probiotics hair
The preparation of zymotic fluid and the preparation of fermented feed containing probiotics, are carried out according to the following steps respectively：
（1）The culture of probiotics.
The method of fermented and cultured is distinguished using each bacterial strain：
Bacillus subtilis WEI-2 CGMCC No.7744：LB culture mediums, 30-40 DEG C, 160-180 rpm cultivate 12-18
h.LB culture medium prescriptions are as follows：Tryptone 10 g/L, Yeast extract 5 g/L, sodium chloride 10 g/L, pH7.2,121
DEG C sterilizing 20 min.
VREF WEI-10 CGMCC No.7746：VREF fermentation medium, 35-42 DEG C, quiescent culture 8-
12 h.VREF fermentation medium is as follows：Sucrose 14-21 g/L, peptone 14-21 g/L, Triammonium citrate 1-3 g/L,
Sodium acetate trihydrate 3-7 g/L, bitter salt 0.1-0.4 g/L, Manganous sulfate monohydrate 0.02-0.07 g/L, three hydrations
8.0,121 DEG C of 20 min of sterilizing of dipotassium hydrogen phosphate 1-3 g/L, Tween-80 1-3 g/L, pH.
Lactobacillus plantarum WEI-70（CGMCC No.11747）：Lactobacillus plantarum fermentation medium, stands by 30-40 DEG C
Cultivate 16-20 h.Lactobacillus plantarum fermentative medium formula is as follows：Maltodextrin 15-25 g/L, corn starch 30-40 g/L,
Sodium acetate trihydrate 2-5 g/L, three hypophosphite monohydrate hydrogen dipotassium 2-4 g/L, ammonium citrate 1-4 g/L, bitter salt 0.2-1
G/L, Manganous sulfate monohydrate 0.05-0.2 g/L, 6.5,121 DEG C of 20 min of sterilizing of Tween 80 0.5-2 g/L, pH.
S. cervisiae WJ-113：YPD culture mediums, 28-32 DEG C, 190-220 rpm, culture 18-20 h.YPD is cultivated
Based formulas is as follows：The g/L of peptone 20, the g/L of dusty yeast 10, glucose 20 g/L, natural pH., 115 DEG C of 20 min of sterilizing.
（2）The preparation of mixed fermentation liquid.
By cultivate respectively four kinds of bacterium solutions according to（1-3）：（1-3）：（1-3）：（1-3）Ratio be sufficiently mixed, be made mixed
Close probiotics fermention liquid.
（3）The preparation of fermented feed containing probiotics.
The complete diet pellet of antibiotic-free is put into clean container, adds mixing probiotics fermention liquid, the ratio of the two is
Bacterium solution：Complete diet pellet=（3.5-5.0）：10, final water content is 35%-45%, is stirred and evenly mixed.Lid lid, sealing.Environment during fermentation
Temperature is 25-40 DEG C, and fermentation time is 24-30 h.The viable count of complete diet pellet after fermentation reaches as high as 1.21 × 1010 CFU/
G, water content is 35%-50%, and pH is 4.2-4.8, there is strong alcohol fragrance.
（4）The application of fermented feed containing probiotics.
The fermented feed made can be used for Direct-fed or be added in daily ration with 5-10% ratio to feed weanling pig
And growing and fattening pigs.The palatability of fermented feed is good, nutritious, and the feed intake increase of pig after feeding, feedstuff-meat ratio is remarkably decreased, enteron aisle
Flora is significantly improved, and diarrhea rate reduction, the general level of the health is improved.
（Three）Meaning：
A kind of fermented feed containing probiotics that the present invention is provided are made of complete diet pellets by 4 kinds of probiotic mixed fermentations, effective viable bacteria
Number is high, and palatability is good, it is adaptable to the pig of each growth phase, especially suitable weanling pig and growing and fattening pigs, can significantly reduce material
Meat ratio, reduces diarrhea rate, improves gut flora.The zymotechnique that the present invention is provided, the cycle is short, low to device dependence, is applicable
In the pig farm of various scales, beneficial to popularization.
Embodiment:
Following instance is that, in order to preferably explain the present invention, but the present invention is not restricted to the scope described in following instance.
Experiment case study one, for fermented feed probiotics culture：
4 kinds of probiotics in the present invention for fermented feed are specific as follows using the method cultivated respectively：
Bacillus subtilis WEI-2 CGMCC No.7744：LB is seeded to from the bacillus subtilis inclined-plane of fresh activation
Culture medium, 37 DEG C, 180 rpm cultivate 16 h, viable count is up to 9.8 × 108CFU/mL.LB culture medium prescriptions are as follows：
The g/L of Tryptone 10 g/L, Yeast extract 5, g/L, pH7.2,121 DEG C of 20 min of sterilizing of sodium chloride 10.
VREF WEI-10 CGMCC No.7746：Dung intestines ball is seeded to from the VREF inclined-plane of fresh activation
Bacteria fermentation culture medium, 40 DEG C, the h of quiescent culture 10, viable count is up to 2.31 × 109CFU/mL.VREF fermentation medium
It is as follows：Sucrose 14-21 g/L, peptone 14-21 g/L, Triammonium citrate 1-3 g/L, sodium acetate trihydrate 3-7 g/L, seven water
Close magnesium sulfate 0.1-0.4 g/L, Manganous sulfate monohydrate 0.02-0.07 g/L, three hypophosphite monohydrate hydrogen dipotassium 1-3 g/L, Tween-80
8.0,121 DEG C of 20 min of sterilizing of 1-3 g/L, pH.
Lactobacillus plantarum WEI-70 CGMCC No.11747：It is seeded to from the Lactobacillus plantarum inclined-plane of fresh activation
Lactobacillus plantarum fermentation medium, 37 DEG C, the h of quiescent culture 18, viable count is up to 2.12 × 109CFU/mL.Plant breast bar
Bacteria fermentation culture medium formula is as follows：Maltodextrin 15-25 g/L, corn starch 30-40 g/L, sodium acetate trihydrate 2-5 g/L,
Three hypophosphite monohydrate hydrogen dipotassium 2-4 g/L, ammonium citrate 1-4 g/L, bitter salt 0.2-1 g/L, Manganous sulfate monohydrate
0.05-0.2 g/L, 6.5,121 DEG C of 20 min of sterilizing of Tween 80 0.5-2 g/L, pH.
S. cervisiae WJ-113：It is seeded to YPD culture mediums from the saccharomyces cerevisiae inclined-plane of fresh activation, 30 DEG C,
200 rpm, cultivate 20 h, viable count is up to 3.60 × 108CFU/mL.YPD culture medium prescriptions are as follows：The g/L of peptone 20,
The g/L of dusty yeast 10, glucose 20 g/L, natural pH, 115 DEG C of 20 min of sterilizing.
The preparation of experiment case study two, fermented feed containing probiotics：
By cultured bacillus subtilis WEI-2 CGMCC No.7744, VREF WEI-10 CGMCC No.7746, plant
Thing lactobacillus WEI-70 CGMCC No.11747 and S. cervisiae WJ-113 nutrient solution press 1：1：1：1 ratio is well mixed,
Mixing probiotics fermention liquid is made.
By cultured bacillus subtilis WEI-2 CGMCC No.7744, VREF WEI-10 CGMCC
No.7746, Lactobacillus plantarum WEI-70 CGMCC No.11747 and S. cervisiae WJ-113 nutrient solution press 2：1：2：1 ratio
Example is well mixed, and mixing probiotics fermention liquid is made.
By cultured bacillus subtilis WEI-2 CGMCC No.7744, VREF WEI-10 CGMCC
No.7746, Lactobacillus plantarum WEI-70 CGMCC No.11747 and S. cervisiae WJ-113 nutrient solution press 3：2：2：3 ratio
Example is well mixed, and mixing probiotics fermention liquid is made.
By cultured bacillus subtilis WEI-2 CGMCC No.7744, VREF WEI-10 CGMCC
No.7746, Lactobacillus plantarum WEI-70 CGMCC No.11747 and S. cervisiae WJ-113 nutrient solution press 1：2：3：2 ratio
Example is well mixed, and mixing probiotics fermention liquid is made.
The complete feed of a certain amount of antibiotic-free is weighed, is fitted into clean container, mixing probiotics fermention liquid is added,
The mass ratio of complete feed and zymotic fluid is 10：3.5 ~ 5.0, it is thoroughly mixed, plastic cloth sealing, 37 DEG C of 24 h of fermentation.
The water content that fermented feed is detected after fermentation ends is 39.2%；Total number of bacteria is 1.03 × 1010 CFU/g, pH are 4.4.
The influence of experiment case study three, fermented feed containing probiotics to Intestinal Microflora of Weanling Piglets and growth performance.
The weanling pig of close 54 first 28 ages in days of body weight is chosen, 2 groups are randomly divided into, every group of 3 repetitions, each repeatedly 6
Head pig, male and female half and half.First group is control group（C）, feed the weanling pig complete diet pellet of antibiotic-free；Second group is that probiotics sends out
Ferment feed group（FF）, feed the nonreactive weanling pig complete diet pellet through 4 kinds of probiotics fermentions.
After the complete diet pellet of weanling pig antibiotic-free is raised seven days in advance, different daily rations are fed respectively by above-mentioned packet and are started just
Formula is tested.Experiment periods 1 totally month, whole free choice feeding drinking-water.Experiment starts and terminated to gather fecal specimens respectively, detects excrement
In Escherichia coli, the clump count of salmonella and staphylococcus aureus；Counting statistics feedstuff-meat ratio and diarrhea rate.
Colony counting uses the method that selective medium is coated on after gradient dilution, and E. coli selectable nutrient chemical is
Maconkey agar, salmonella selective medium is SS agar, and staphylococcus aureus selective medium is high salt sweet dew
Alcohol agar.
Administering transgenic result shows that the feedstuff-meat ratio and diarrhea rate of fermented feed containing probiotics group are substantially less than control group（Table
1）.In the detection of enteron aisle opportunistic flora, the enteron aisle conditioned pathogen of control group is substantially unchanged before and after feeding；Feeding
Conditioned pathogen in the swine excrement of fermented feed containing probiotics before feeding than have dropped 15 times（Table 2）.
Influence of the fermented feed containing probiotics of table 1. to Growth Performance of Weaning Piglets
Influence of the fermented feed containing probiotics of table 2. to weanling pig enteron aisle conditioned pathogen
The influence of experiment case study four, fermented feed containing probiotics to growing and fattening pigs gut flora and growth performance.
The growing and fattening pigs of 54 close kg of average weight 63.10 ± 0.13 of body weight are chosen, 2 groups, every group 3 are randomly divided into
Repeat, each repeat 6 pigs, male and female half and half.First group is control group（C）, feed the complete diet pellet of antibiotic-free；Second group is benefit
Raw bacterium fermented feed group（FF）, feed the nonreactive complete diet pellet through 4 kinds of probiotics fermentions.
After the complete diet pellet of finishing pigs antibiotic-free is raised seven days in advance, different daily rations are fed respectively by above-mentioned packet and are started formally
Experiment.Experiment periods 1 totally month, whole free choice feeding drinking-water.Experiment starts and terminated to gather fecal specimens respectively, in detection excrement
Escherichia coli, the clump count of salmonella and staphylococcus aureus；Counting statistics feedstuff-meat ratio and diarrhea rate.
Colony counting uses the method that selective medium is coated on after gradient dilution, and E. coli selectable nutrient chemical is
Maconkey agar, salmonella selective medium is SS agar, and staphylococcus aureus selective medium is high salt sweet dew
Alcohol agar.
Administering transgenic result shows that the feedstuff-meat ratio of fermented feed containing probiotics group is substantially less than control group, and probiotics fermention is raised
The diarrhea rate of material group is substantially less than control group（Table 3）.In the detection of enteron aisle opportunistic flora, the enteron aisle condition of control group
Pathogenic bacteria are substantially unchanged before and after feeding；Before the conditioned pathogen in the swine excrement of fermented feed containing probiotics is fed than feeding
It has dropped 18 times（Table 4）.
Influence of the fermented feed containing probiotics of table 3. to growing-finishing pigs performance
Influence of the fermented feed containing probiotics of table 4. to growing and fattening pigs enteron aisle conditioned pathogen
Experiment case study five, fermented feed containing probiotics mixed by 10% addition with basal diet after to Intestinal Microflora of Weanling Piglets and
The influence of growth performance.
The weanling pig of close 54 first 28 ages in days of body weight is chosen, 2 groups are randomly divided into, every group of 3 repetitions, each repeatedly 6
Head pig, male and female half and half.First group is control group（C）, feed the weanling pig complete diet pellet of antibiotic-free；Second group is that probiotics sends out
Ferment feed group（FF）, by the nonreactive weanling pig complete diet pellet of 4 kinds of probiotics fermentions according to 10% addition and the base of weanling pig
Weanling pig is fed after the mixing of plinth daily ration.
After the complete diet pellet of weanling pig antibiotic-free is raised seven days in advance, different daily rations are fed respectively by above-mentioned packet and are started just
Formula is tested.Experiment periods 1 totally month, whole free choice feeding drinking-water.Experiment starts and terminated to gather fecal specimens respectively, detects excrement
In Escherichia coli, the clump count of salmonella and staphylococcus aureus；Counting statistics feedstuff-meat ratio and diarrhea rate.
Colony counting uses the method that selective medium is coated on after gradient dilution, and E. coli selectable nutrient chemical is
Maconkey agar, salmonella selective medium is SS agar, and staphylococcus aureus selective medium is high salt sweet dew
Alcohol agar.
Administering transgenic result shows that the feedstuff-meat ratio and diarrhea rate of fermented feed containing probiotics group are substantially less than control group（Table
5）.In the detection of enteron aisle opportunistic flora, the enteron aisle conditioned pathogen of control group is substantially unchanged before and after feeding；Feeding
Conditioned pathogen in the swine excrement of fermented feed containing probiotics before feeding than have dropped 10 times（Table 6）.
The fermented feed containing probiotics of table 5.（10% addition）Influence to Growth Performance of Weaning Piglets
The fermented feed containing probiotics of table 6.（10% addition）Influence to weanling pig enteron aisle conditioned pathogen
Experiment case study six, fermented feed containing probiotics mixed by 10% addition with basal diet after to growing and fattening pigs gut flora and life
The influence of long performance.
The growing and fattening pigs of 54 close kg of average weight 63.10 ± 0.13 of body weight are chosen, 2 groups, every group 3 are randomly divided into
Repeat, each repeat 6 pigs, male and female half and half.First group is control group（C）, feed the complete diet pellet of antibiotic-free；Second group is benefit
Raw bacterium fermented feed group（FF）, addition and growing and fattening pigs by the nonreactive growing and fattening pigs complete diet pellet of 4 kinds of probiotics fermentions according to 10%
Growing and fattening pigs are fed after basal diet mixing.
After the complete diet pellet of finishing pigs antibiotic-free is raised seven days in advance, different daily rations are fed respectively by above-mentioned packet and are started formally
Experiment.Experiment periods 1 totally month, whole free choice feeding drinking-water.Experiment starts and terminated to gather fecal specimens respectively, in detection excrement
Escherichia coli, the clump count of salmonella and staphylococcus aureus；Counting statistics feedstuff-meat ratio and diarrhea rate.
Colony counting uses the method that selective medium is coated on after gradient dilution, and E. coli selectable nutrient chemical is
Maconkey agar, salmonella selective medium is SS agar, and staphylococcus aureus selective medium is high salt sweet dew
Alcohol agar.
Administering transgenic result shows that the feedstuff-meat ratio of fermented feed containing probiotics group is substantially less than control group, two groups of diarrhea rate
Without significant difference（Table 7）.In the detection of enteron aisle opportunistic flora, the enteron aisle conditioned pathogen of control group is in the front and rear base of feeding
This is unchanged；The conditioned pathogen in the swine excrement of fermented feed containing probiotics is fed than have dropped 19 times before feeding（Table 8）.
The fermented feed containing probiotics of table 7.（10% addition）Influence to growing-finishing pigs performance
| C groups | FF groups | |
| Feedstuff-meat ratio | 3.52±0.03a | 3.23±0.01b |
| Diarrhea rate（%） | 2.10±0.02a | 2.09±0.03a |
The fermented feed containing probiotics of table 8.（10% addition）Influence to growing and fattening pigs enteron aisle conditioned pathogen
Claims (6)
1. a kind of fermented feed containing probiotics, it is characterised in that by mixing probiotics viable bacteria and its metabolite and antibiotic-free
Ferment complete feed is constituted, and the viable count of complete diet pellet reaches as high as 1.21 × 1010CFU/g, water content is 35%-50%, and pH is
4.2-4.8。
2. fermented feed containing probiotics according to claim 1, it is characterised in that described mixing probiotics includes withered grass bud
Spore bacillus WEI-2 CGMCC No.7744, VREF WEI-10 CGMCC No.7746, Lactobacillus plantarum WEI-70 CGMCC
No.11747 and saccharomyces cerevisiae WJ-113.
3. a kind of preparation method of fermented feed containing probiotics, it is characterised in that preparation method includes：Mix probiotics fermention liquid
Prepare the preparation with fermented feed containing probiotics.
4. preparation method according to claim 3, it is characterised in that the preparation method of described mixing probiotics fermention liquid
For：
1）Bacillus subtilis WEI-2 CGMCC No.7744：LB culture mediums, 30-40 DEG C, 160-180 rpm cultivate 12-18
h；LB culture medium prescriptions are as follows：Tryptone 10 g/L, Yeast extract 5 g/L, sodium chloride 10 g/L, pH7.2,121
DEG C sterilizing 20 min；
2）VREF WEI-10 CGMCC No.7746：VREF fermentation medium, 35-42 DEG C, quiescent culture 8-12
h；VREF fermentation medium is as follows：Sucrose 14-21 g/L, peptone 14-21 g/L, Triammonium citrate 1-3 g/L, three water
Close sodium acetate 3-7 g/L, bitter salt 0.1-0.4 g/L, Manganous sulfate monohydrate 0.02-0.07 g/L, three hypophosphite monohydrates
8.0,121 DEG C of 20 min of sterilizing of hydrogen dipotassium 1-3 g/L, Tween-80 1-3 g/L, pH；
3）Lactobacillus plantarum Lactobacillus plantarum WEI-70 CGMCC No.11747：Lactobacillus plantarum fermentation medium, 30-40
DEG C, quiescent culture 16-20 h；Lactobacillus plantarum fermentative medium formula is as follows：Maltodextrin 15-25 g/L, corn starch 30-
40 g/L, sodium acetate trihydrate 2-5 g/L, three hypophosphite monohydrate hydrogen dipotassium 2-4 g/L, ammonium citrate 1-4 g/L, seven hydrated sulfuric acids
Magnesium 0.2-1 g/L, Manganous sulfate monohydrate 0.05-0.2 g/L, 6.5,121 DEG C of sterilizings 20 of Tween 80 0.5-2 g/L, pH
min；
4）S. cervisiae WJ-113：YPD culture mediums, 28-32 DEG C, 190-220 rpm, culture 18-20 h；YPD culture mediums are matched somebody with somebody
Side is as follows：The g/L of peptone 20, the g/L of dusty yeast 10, glucose 20 g/L, natural pH, 115 DEG C of 20 min of sterilizing； 5）It will divide
The four kinds of bacterium solutions of bacillus subtilis, VREF, Lactobacillus plantarum Lactobacillus plantarum, S. cervisiae do not cultivated are according to 1 ~ 3：
1~3：1~3：1 ~ 3 ratio is sufficiently mixed, and mixing probiotics fermention liquid is made.
5. preparation method according to claim 3, it is characterised in that the preparation process of fermented feed containing probiotics is as follows：Will
Complete diet pellet is put into clean container, adds mixing probiotics fermention liquid, and the mass ratio of the two is bacterium solution：Complete diet pellet=3.5 ~
5.0：10, final water content is 35%-50%, is stirred and evenly mixed, lid lid, sealing, and environment temperature is 25-40 DEG C during fermentation, fermentation
Time is 24-30 h；The viable count of complete diet pellet after fermentation reaches as high as 1.21 × 1010 CFU/g, pH are 4.2-4.8.
6. a kind of application of fermented feed containing probiotics, it is characterized in that, the fermented feed is new nonreactive green feed, is made
Fermented feed Direct-fed or feeding weanling pig and growing and fattening pigs in daily ration can be added to 5-10% ratio.
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