CN108165513A - A kind of composite bacteria preparation and prepare the method for compound bacteria-fermented feed using it - Google Patents
A kind of composite bacteria preparation and prepare the method for compound bacteria-fermented feed using it Download PDFInfo
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- CN108165513A CN108165513A CN201810115421.7A CN201810115421A CN108165513A CN 108165513 A CN108165513 A CN 108165513A CN 201810115421 A CN201810115421 A CN 201810115421A CN 108165513 A CN108165513 A CN 108165513A
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- composite bacteria
- cervisiae
- bacillus subtilis
- bacterium solution
- lactobacillus salivarius
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- 241000894006 Bacteria Species 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims abstract description 29
- 239000002131 composite material Substances 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 9
- 150000001875 compounds Chemical class 0.000 title claims description 21
- 241000282887 Suidae Species 0.000 claims abstract description 25
- 241000186869 Lactobacillus salivarius Species 0.000 claims abstract description 21
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 20
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 20
- 238000000855 fermentation Methods 0.000 claims abstract description 17
- 230000004151 fermentation Effects 0.000 claims abstract description 17
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 14
- 239000000463 material Substances 0.000 claims abstract description 7
- 239000001963 growth medium Substances 0.000 claims abstract 2
- 239000000203 mixture Substances 0.000 claims abstract 2
- 239000006041 probiotic Substances 0.000 claims description 8
- 235000018291 probiotics Nutrition 0.000 claims description 8
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
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- 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
- A23V2400/181—Salivarius
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Abstract
The present invention relates to a kind of composite bacteria preparations, are made of Lactobacillus salivarius, bacillus subtilis, S. cervisiae, and the mass ratio of three kinds of bacterium is 2：2：1.Preparation method：Lactobacillus salivarius, bacillus subtilis, S. cervisiae are inoculated in fermentation culture, quiescent culture after culture medium activates respectively, when cell concentration reaches exponential phase 1 × 109~2.94 × 109During cfu/ml, fermentation is terminated, three kinds of bacterium solutions are respectively obtained, by 3 kinds of bacterium solutions with mass ratio 2：2：1 ratio mix to get.The composite bacteria preparation is further used to make growing and fattening pigs fermentation material by the present invention, the fermented feed can be effectively improved fattening meat quality, reduce the feed cost in breeding process, promote the relative abundance of beneficial bacterium in growing and fattening pigs intestinal mucosa and content, inhibit the proliferation of harmful bacteria by competition mechanism, to adjust intestinal microecology, ensure health, so as to reduce the mortality in feeding process, production cost is saved.
Description
Technical field
The present invention relates to a kind of composite bacteria preparations, belong to feed additive field.
Background technology
At present, the non-nutritive feed addictive used in husbandry sector mainly includes feeding antibiotic, expelling parasite health care
Agent, feed preservative and other additives.Since requirement of the global every country to food and Environmental security is increasingly harsh, animal
The nonreactive of feed has become trend；Probiotics has nontoxic, pollution-free, noresidue etc. as a kind of novel feed addictive
While the characteristics of small to environmental pressure, the immunity of animal can be improved, improves the production performance of animal, thus, probiotic quilt
It is considered a kind of additive agent for feeding in aquaculture with wide application prospect.
It is found in practical feeding process, the probiotic product directly used is asked there are shelf life is short, effect is unstable etc.
Topic.Therefore, to avoid such issues that, commonly uses probiotic fermented feed in production at present.Fermented feed is a kind of by benefit
The novel fodder formed after raw bacterium fermented feed, the probiotic tunning in this kind of feed can be provided directly or indirectly for host
Nutriment promotes host growth development；And probiotics therein can prevent harmful bacteria from being colonized in alimentary canal, ensure host
Alimentary canal microecological balance；And the anti-nutritional factors in feed can be also reduced by fermentation, so as to improve nutrient profit in feed
With rate, feed waste is reduced.
At present, fermented feed has become a new development direction in feed industry, and culturist is hoped by above-mentioned fermentation
The advantage of feed solves the problems, such as that intestinal health present in Pig Industry industry and meat are bad.However, due to Vehicles Collected from Market
The background information of the fermented feed production bacterial strain of upper sale is unclear, and culture environment, mechanism of action and the metabolite of bacterial strain are not
It is bright, the problem of fermented feed product quality is irregular and using effect is unstable is as a result directly resulted in, it is final to current hair
Being widely applied for ferment feed is brought a negative impact.Therefore, it is necessary to obtain production hair using means such as separation, screening and optimizations
The excellent species of ferment feed by compound compatibility, solve existing for the most fermented feed sold on Vehicles Collected from Market on source
The problem of quality is bad and functional characteristic is unknown.
Invention content
It is an object of the invention to solve the deficiencies in the prior art, a kind of composite bacteria preparation is provided, growing and fattening pigs can be improved
Meat and intestinal microecology health.
Another object of the present invention also resides in the preparation method for providing the composite bacteria preparation.
To solve above-mentioned purpose, the technical solution adopted by the present invention is as follows：
A kind of composite bacteria preparation is made of Lactobacillus salivarius, bacillus subtilis, S. cervisiae；
Further, in the composite bacteria preparation, the total quantity of probiotics is 1 × 109~2.94 × 109cfu/g；
Further, in the composite bacteria preparation, Lactobacillus salivarius, bacillus subtilis, S. cervisiae mass ratio be
2：2：1.
The Lactobacillus salivarius is Lactobacillus salivarius (Lactobacillus salivarius), the withered grass gemma of NO.L79
Bacillus subtilis (Bacillus subtilis) that bacillus is NO.B1121, the wine brewing ferment that S. cervisiae is NO.S1145
Female bacterium (Saccharomyces cerevisiae).
The present invention also provides the preparation methods of above-mentioned composite bacteria preparation, include the following steps：
(1) take Lactobacillus salivarius, bacillus subtilis, S. cervisiae be inoculated in respectively MRS culture mediums, BPY culture mediums,
After being activated in YPD culture mediums, activation bacterium solution is taken to be inoculated in fermentation culture respectively, 37 DEG C of cultures, when cell concentration reaches
Exponential phase 1 × 109~2.94 × 109During cfu/ml, fermentation is terminated, respectively obtains Lactobacillus salivarius bacterium solution, bacillus subtilis
Bacterium bacterium solution and S. cervisiae bacterium solution；
(2) by Lactobacillus salivarius bacterium solution, bacillus subtilis bacterium solution and S. cervisiae bacterium solution with mass ratio 2：2：1 ratio
Example mixing to get.
The method that compound bacteria-fermented feed is prepared using above-mentioned composite bacteria preparation：By composite bacteria preparation with 0.1~0.5%
Additive amount is added in growing and fattening pigs adequate diet, is adjusted feed humidity as 30~50%, 37 DEG C of sealing and fermenting 48h, is fermented
Material, then by fermentation material and adequate diet with 1：4 mass ratio is mixed to get compound bacteria-fermented feed.
Advantageous effect：The composite bacteria preparation of the present invention can be used as the additive of fermented feed for fattening pigs, not influence to educate
In the case of hog growth performance, health status and carcass quality, feed cost is effectively reduced, and improves growing and fattening pigs longissimus dorsi muscle
Quality and intestinal health, and additive amount is low.
Specific embodiment
According to following embodiments, the present invention may be better understood.It is however, as it will be easily appreciated by one skilled in the art that real
It applies the described content of example and is merely to illustrate the present invention, without sheet described in detail in claims should will not be limited
Invention.
Embodiment 1
Prepare composite bacteria preparation：
(1) by three plants of bacterium [NO:The Lactobacillus salivarius (Lactobacillus salivarius) of L79, bacillus subtilis
For NO:The bacillus subtilis (Bacillus subtilis) of B1121, S. cervisiae NO:The S. cervisiae of S1145
(Saccharomyces cerevisiae)] it is inoculated in respectively in MRS culture mediums, BPY culture mediums, YPD culture mediums after activation, then
It is inoculated in fermentation culture, 37 DEG C of constant temperature incubations, Lactobacillus salivarius is stood, and S. cervisiae and bacillus subtilis are placed in 37
DEG C shaking table 150r/min culture, makes cell concentration reach exponential phase 1 × 109cfu/ml；Respectively obtain Lactobacillus salivarius
Bacterium solution, bacillus subtilis bacterium solution and S. cervisiae bacterium solution.
By taking Lactobacillus salivarius as an example, scribing line culture is carried out in MRS solid mediums, 37 DEG C are cultivated for 24 hours, picking colony knot
Crystalline substance carries out microscopy；It is more than that 2,000,000,000/milliliter can be used to detect bacterial concentration；The MRS culture mediums：Glucose 2.0%, citric acid
Sodium 0.2%, sodium acetate 0.5%, dipotassium hydrogen phosphate 0.5%, manganese sulfate 0.02%, magnesium sulfate 0.05%, peptone 1.0%, ox
Meat extract 1.0%, yeast extract 0.5%, Tween 80 0.1%；BPY culture mediums：Beef extract 0.05%, peptone 1.0%, yeast extract
0.05%th, glucose 0.05%, sodium chloride 0.05%；YPD culture mediums：Yeast extract 1.0%, peptone 2.0%, glucose
2.0%；Solid medium adds 1.5% agar powder.Medium sterilization condition is 0.10-0.15MPa, 121 DEG C of sterilizing 20min.
(2) by Lactobacillus salivarius bacterium solution, bacillus subtilis bacterium solution and S. cervisiae bacterium solution with mass ratio 2：2：1 ratio
Example mixing is to get composite bacteria preparation.
Prepare compound bacteria-fermented feed：Above-mentioned composite bacteria preparation is added to growing and fattening pigs full price day with 0.2% additive amount
In grain (corn 67%, wheat bran 10%, dregs of beans 18%, premix 5%), it is 30-50% to adjust feed humidity, and 37 DEG C are sealed by fermentation
48h obtains fermentation material, then by fermentation material and adequate diet with 1：4 mass ratio is mixed to get fermented feed.
Application test：
Test method：The Ternary Pig three way cross growing and fattening pigs that 24 original body mass are 66.5 ± 2.0kg are chosen, are randomly divided into
2 groups of test group and control group, every group of 4 repetitions (circle) each repeat 3 pigs.Wherein 1 gained of test group feeding embodiment is compound
Bacterium fermented feed, control group fed basal diet.The nutritional ingredient of two kinds of Diet Formulas is satisfied by the battalion of NRC (2012) growing and fattening pigs
The demand of supporting.Feeding period is 28 days, and feeding daily twice, ensures to be freely eaten and drinking-water, periodically sterilizes, and keeps colony house ventilation and clear
It is clean；Feeding experiment terminates the same day, the Slaughter weight of every pig will be weighed after test pig taboo feeding 12h, prohibits and ensure freely to drink during raising
Water；Test the growth performance index of growing and fattening pigs；Then, it is each to repeat to extract weight close to 1 pig (4/place of average weight
Reason), it is butchered according to old process, carcass trait index is measured after inspection and quarantine, and bulk longissimus dorsi muscle meat sample is taken to preserve
It in 4 DEG C of ice chests, is measured for meat, dissection abdomen collects colonic mucosa and content, liquid nitrogen cryopreservation.
1. growth performance index
To repeat (circle) as unit, record the end weight after the original body mass and feeding period that often enclose pig and examination foster period is every
The total feed feed intake of pig is enclosed, average daily gain (average daily weight gain, ADG) is calculated according to these data,
Average daily gain (average daily feed intake, ADFI) and feed-weight ratio (F/G).
Average daily gain=often enclosing pig always whole opisthosoma weight-often total original body mass/3 pig/28 day of circle pig；
Average daily gain=often encloses pig total feed feed intake/3 pig/28 day；
Feed-weight ratio=average daily gain/average daily gain.
Test result is shown in Table 1：
Influence of the 1 compound bacteria-fermented feed of table to growing-finishing pigs performance
Note：All indexs are (n=4) for statistical analysis as unit of colony house in table.
As shown in Table 1：Compared with the growing and fattening pigs for conventional base daily ration of searching for food, the fattening for 20% compound bacteria-fermented feed of searching for food
Pig has higher end weight, average daily gain, while has relatively low feed-weight ratio and feed cost, but difference not significantly (P>
0.05)。
2. carcass trait index
Slaughter weight, carcass weight, dressing percentage, average backfat be thick and the measure reference of eye muscle area《NY/T 825-2004 are thin
Meat type hog on hook property determination technical specification》.Test result is shown in Table 2：
Influence of the 2 compound bacteria-fermented feed of table to growing and fattening pigs carcass trait
Note：All indexs are (n=4) for statistical analysis as unit of single pig in table.
Carcass trait is the important indicator for weighing growing and fattening pigs Meat Performance, is directly related to the economy profit of pig production person
Benefit.As shown in Table 2, compared with the growing and fattening pigs for conventional base daily ration of searching for food, the trunk for 20% compound bacteria-fermented feed growing and fattening pigs of searching for food
Weight, dressing percentage are higher, but difference not significantly (P>0.05).Meanwhile compared to conventional base daily ration, compound bacteria-fermented feed
The thickness of backfat of growing and fattening pigs is reduced, increases eye muscle area, but influences not significantly (P>0.05).
3. meat index
Longissimus dorsi muscle pH45min、pH24h, brightness value L *, red scale value a*, yellow value degree b*, drip loss, cooking loss and shearing
The measure reference of force value《The edible quality method for objectively evaluating of NY/T 2793-2015 meat》；NPPC yellowish pinks and marble grain scoring
It is scored using U.S. version NPPC colorimetric cards.Test result is shown in Table 3：
Influence of the 3 compound bacteria-fermented feed of table to growing and fattening pigs longissimus dorsi muscle meat
Note：All indexs are (n=4) for statistical analysis as unit of single pig in table.
Edible quality refers to the attribute for embodying meat quality quality, is to determine the most important one side of meat commodity value, reflection
The consumption performance and potential value of meat, the higher meat of edible quality are easy to be esthetically acceptable to the consumers, the market price often compared with
Height, table 3 are compound bacteria-fermented feeds to edible qualities indexs such as growing and fattening pigs longissimus dorsi muscle pH, color, shearing force and water-retaining propertys
It influences.As can be seen from Table 3, compared with conventional daily ration, compound bacteria-fermented feed significantly improves pH45minValue, red scale value a* and
NPPC Color Scores (P<0.05), meanwhile, the growing and fattening pigs for compound bacteria-fermented feed of searching for food, longissimus dorsi muscle meat has relatively low cut
Shear value (P<0.05).Compound bacteria-fermented feed is to pH24hValue, yellowish pink L and b* values, NPPC marble grains scoring, drip loss and
Cooking loss does not make significant difference (P>0.05).
4. Vfa Concentration in colonic contents
Using gas chromatography, (Japan, GC-14B gas chromatographs, color column type number are Agilent＆W GC Columns；
30m × 0.32mm × 0.25 μm, 110 DEG C of column temperature, gasify 180 DEG C of room temperature, detects 180 DEG C of room temperature) measure volatile fat
Acid concentration.It the results are shown in Table 4：
Influence of the 4 compound bacteria-fermented feed of table to growing and fattening pigs colonic contents volatile fatty acid
Note：All indexs are (n=4) for statistical analysis as unit of single pig in table.
As can be seen from Table 4, compared with the growing and fattening pigs for conventional base daily ration of searching for food, 20% compound bacteria-fermented feed of feeding is educated
The only horizontal significantly raising (P of butyric acid in hog colonic contents<0.05), acetic acid, propionic acid, isobutyric acid, valeric acid, isovaleric acid level
Without significant changes (P>0.05).
5. colonic contents Bacterial community is analyzed
Colonic contents grind sample with bead mill method machinery, spare with phenol and chloroform/isoamyl alcohol extracting total DNA afterwards.Sequencing
Work is assisted to complete by Tian Yi health sciences research institute (Zhenjiang) Co., Ltd.It the results are shown in Table 5：
5 growing and fattening pigs faecal flora of table belongs to horizontal relative abundance
Note：All indexs are (n=4) for statistical analysis as unit of single pig in table.
It detects 116 Pseudomonas altogether in content, 35 superiority bacteria spps is selected for standard screen with relative abundance 0.5%,
In addition to non-classified Pseudomonas, Subdoligranulum Pseudomonas and Wei Si Bordetella relative abundance significantly increase in fermented feed group
(P<0.05).Subdoligranulum Pseudomonas and Wei Si Bordetella are beneficial bacteriums in enteron aisle, and abundance raising is shown, the fermentation
Feed can be effectively improved fattening pig intestinal health.
The foregoing is merely the examples of the application, are not limited to the application, for those skilled in the art,
The application can have various modifications and variations.All any modifications within spirit herein and principle, made equally are replaced
It changes, improve, should be included within the protection domain of the application.
Claims (5)
1. a kind of composite bacteria preparation, which is characterized in that be made of Lactobacillus salivarius, bacillus subtilis, S. cervisiae.
2. composite bacteria preparation as described in claim 1, which is characterized in that in the composite bacteria preparation, the total quantity 1 of probiotics
× 109~2.94 × 109cfu/g；
3. composite bacteria preparation as claimed in claim 1 or 2, which is characterized in that in the composite bacteria preparation, Lactobacillus salivarius,
Bacillus subtilis, S. cervisiae mass ratio be 2：2：1.
4. the preparation method of the composite bacteria preparation described in claims 1 or 2 or 3, which is characterized in that include the following steps：
(1) Lactobacillus salivarius, bacillus subtilis, S. cervisiae is taken to be inoculated in MRS culture mediums, BPY culture mediums, YPD respectively
After being activated in culture medium, activation bacterium solution is taken to be inoculated in fermentation culture respectively, 37 DEG C of cultures, when cell concentration reaches logarithm
During 1 × 109~2.94 × 109cfu/ml of growth period, fermentation is terminated, respectively obtains Lactobacillus salivarius bacterium solution, bacillus subtilis
Bacterium solution and S. cervisiae bacterium solution；
(2) by Lactobacillus salivarius bacterium solution, bacillus subtilis bacterium solution and S. cervisiae bacterium solution with mass ratio 2：2：1 ratio is mixed
Close to get.
5. the method for compound bacteria-fermented feed is prepared using the composite bacteria preparation described in claims 1 or 22 or 3, which is characterized in that
Composite bacteria preparation is added to 0.1~0.5% additive amount in growing and fattening pigs adequate diet, adjust feed humidity for 30~
50%, 37 DEG C of sealing and fermenting 48h obtain fermentation material, then by fermentation material and adequate diet with 1：4 mass ratio mix to get
Compound bacteria-fermented feed.
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Application publication date: 20180615 |