CN106566791A - Porcine compound synbiotics micro-ecological preparation and preparation method thereof - Google Patents
Porcine compound synbiotics micro-ecological preparation and preparation method thereof Download PDFInfo
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- CN106566791A CN106566791A CN201610993492.8A CN201610993492A CN106566791A CN 106566791 A CN106566791 A CN 106566791A CN 201610993492 A CN201610993492 A CN 201610993492A CN 106566791 A CN106566791 A CN 106566791A
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- probiotic bacteria
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- 150000001875 compounds Chemical class 0.000 title claims abstract description 51
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 235000019722 synbiotics Nutrition 0.000 title abstract 2
- 235000018291 probiotics Nutrition 0.000 claims abstract description 84
- 239000006041 probiotic Substances 0.000 claims abstract description 83
- 230000000529 probiotic effect Effects 0.000 claims abstract description 78
- 241001465754 Metazoa Species 0.000 claims abstract description 42
- 229920001542 oligosaccharide Polymers 0.000 claims abstract description 41
- 150000002482 oligosaccharides Chemical class 0.000 claims abstract description 40
- 235000013406 prebiotics Nutrition 0.000 claims abstract description 30
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 15
- 241000186660 Lactobacillus Species 0.000 claims abstract description 12
- 210000001035 gastrointestinal tract Anatomy 0.000 claims abstract description 12
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000008901 benefit Effects 0.000 claims abstract description 10
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 claims abstract description 9
- 241000282887 Suidae Species 0.000 claims abstract description 7
- DLRVVLDZNNYCBX-RTPHMHGBSA-N isomaltose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-RTPHMHGBSA-N 0.000 claims abstract description 7
- 239000004310 lactic acid Substances 0.000 claims abstract description 6
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 6
- 239000001530 fumaric acid Substances 0.000 claims abstract description 5
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims abstract description 5
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- 239000000843 powder Substances 0.000 claims description 3
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
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- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 abstract 1
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- ZGBSOTLWHZQNLH-UHFFFAOYSA-N [Mg].S(O)(O)(=O)=O Chemical compound [Mg].S(O)(O)(=O)=O ZGBSOTLWHZQNLH-UHFFFAOYSA-N 0.000 description 1
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- 235000010439 isomalt Nutrition 0.000 description 1
- HPIGCVXMBGOWTF-UHFFFAOYSA-N isomaltol Natural products CC(=O)C=1OC=CC=1O HPIGCVXMBGOWTF-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
-
- C—CHEMISTRY; METALLURGY
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- 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
-
- 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
- A23V2400/51—Bifidobacterium
Abstract
The invention discloses a porcine compound synbiotics micro-ecological preparation and a preparation method thereof. The micro-ecological preparation consists of probiotics and prebiotics, wherein the probiotics and the prebiotics are mixed at a ratio of 1 to (1-5); the probiotics include lactobacillus, bifidobacterium, bacillus and saccharomycetes; the prebiotics include oligosaccharide and an acidifying agent; the oligosaccharide includes mannose-oligosaccharide, fructooligosaccharide and isomaltooligosaccharide; and the acidifying agent is is a compound acidifying agent which is composed of lactic acid, citric acid and fumaric acid. The micro-ecological preparation provided by the invention can overcome weaknesses of probiotic products which are independently used for breeding commercial pigs and the like, and the micro-ecological preparation is applicable to commercial pigs of various types; the micro-ecological preparation is capable of enhancing advantages and avoiding disadvantages, and promoting the planting of exogenous probiotics in animal intestinal tracts; in addition, various probiotics can be selectively stimulated to rapidly grow or reproduce in pig bodies, so that the purpose of promoting the health of the commercial pigs is achieved, and the production performance of the commercial pigs is also greatly improved; and the micro-ecological preparation can relieve the influence of conventional animal edible additives to the quality of pork; therefore, the micro-ecological preparation is more economic and applicable.
Description
Technical field
The invention belongs to microbial ecological agent field, more particularly to a boar compound symphysis unit microbial ecological agent and its system
Preparation Method.
Background technology
The appearance of microbial ecological agent is that, due to the improvement of people's living standards, people gradually recognize that antibiotic adds for a long time
The health of the mankind can be worked the mischief by food chain in feedstuff.At present, life of China's symphysis unit microbial ecological agent in animal
Applied research in product is also less, and in farm animals edible additive, the service condition of microbial ecological agent is also less, people couple
Microbial ecological agent is suitable for addition and relatively obscures, that is, obscure, cause the effect of growth of animal often low
In Expected Results.The research application of microbial ecological agent overcomes a variety of drawbacks that life-time service antibiotic is caused to aquaculture, right
Development green feed additive has broad application prospects, and oneself is classified as 21st century emphasis promotional technique item by the Ministry of Agriculture
Mesh.This means that the research to microbial ecological agent, popularization meet the general requirement of China's sustainable development.In document " Kang Feng, Lu
Win bright, Zhang Haifeng, An Ligang, Hu Jianwu. functional oligosaccharide and its mechanism of action. Laboratory Animal Science .2011 (28). " in, refer to
The functional oligosaccharide gone out in microbial ecological agent is a kind of safe, new green environment protection additive for having no toxic side effect, Neng Gougai
Microecological environment in kind animal intestinal, promotes profitable strain growth, suppresses the breeding of pathogen, improve animal immunizing power, from
And promote animal health growth.Document " Zhao Yanhong. the physiological function of oligosaccharide and the application in pig production. cultural technique is turned round and look at
Ask .2012. " in, author points out oligosaccharide as the excellent succedaneum of antibiotic, in pig production, in its premunition of raising,
There is in terms of reducing feedstuff-meat ratio, and weightening preferably effect.
The defect of prior art is that the stability of bacterial strain in microbial ecological agent is not ensured well, is thrown necessary to bacterium
The form of viable bacteria reaches gut metabolism by stomach, and colonization is trained in intestinal and grown, for this antibacterial to general spore form
It is not easily achieved.Furthermore microbial ecological agent does not targetedly add object, and the domestic application to microbial ecological agent is not enough
Fully, not enough, the not a height of solution abuse of antibiotics of the degree that received by raiser causes increasing pathogenic bacterium to popularization degree
Develop immunity to drugs, drug residue, reduce pork product quality and due to food chain accumulation human body health is brought it is tight
Ghost image rings, and produces the microbial ecological agent of substitute antibiotics.
The content of the invention
It is an object of the invention to provide a boar compound symphysis unit microbial ecological agent and preparation method thereof, it is intended to solve
Certainly abuse of antibiotics causes increasing pathogenic bacterium to develop immunity to drugs, drug residue, reduce pork product quality and due to
The problem having a strong impact on that food chain accumulation is brought to human body health.
The present invention is achieved in that the compound symphysis unit microbial ecological agent of pig, and the pig is micro- with compound symphysis unit
Ecological agent is made up of probiotic bacteria and prebioticses；
According to probiotic bacteria described in mass ratio：Prebioticses=1:1～5 ratio mixing.
Further, pig probiotic bacteria described in compound symphysis unit microbial ecological agent is by lactobacilluss, bacillus bifiduss, spore bar
Bacterium and yeast composition (reference culture is purchased from Beijing North Na Chuanlian Bioteknologisk Institut).
Further, the prebioticses are made up of oligosaccharide and acidulant；
The oligosaccharide is MOS, fruit oligose, oligomeric isomaltose, and acidulant is by lactic acid, citric acid, Fumaric acid
The compound acidulant of composition.
Further, the probiotic bacteria is 0.02-15g/kg, and probiotic bacteria nutrient substance is 0.05-80g/kg, and oligosaccharide is
0.01-90g/kg。
Further, the addition of oligosaccharide be MOS 0.25-2g/kg, fruit oligose 0.01-3g/kg, oligomeric Isomalt
Sugared 0.01-3g/kg；The compound acidulant that addition is made up of lactic acid, citric acid, Fumaric acid, compound acidulant account for Tiny ecosystem system
The 0.1%-0.5% of agent ratio.
Another object of the present invention is to a kind of preparation method of pig with compound symphysis unit microbial ecological agent is provided,
The pig preparation method of compound symphysis unit microbial ecological agent filters out Intestinum Sus domestica road benefit first respectively in different environments
Raw bacterium, and the function and biological characteristicses to probiotic bacteria be compared, and probiotic bacteria is studied under simulation pig intestinal environment
Viable count changes, and filters out optimal bacterial strain；Then all kinds of oligosaccharide are measured, select optimal oligosaccharide consumption, most
Probiotic bacteria and oligosaccharide are mixed and made into into compound symphysis unit microbial ecological agent afterwards；Specifically include following steps：
Step one, the soil from the plant of pig, the trough for feeding pig, gastrointestinal region separate newborn bar
Bacterium, bacillus bifiduss, bacillus cereuss and yeast probiotic bacteria, by isolated probiotic bacteria carry out biological characteristicses, function and
The identification of safety, and carry out external probiotic effects evaluation, preliminary screening probiotic bacteria；
Step 2, using pig enteric microorganism and carbohydrate metabolism, the relation of vitamin, filter out raising pig obtain from daily ration
The high-quality probiotic bacteria of energy, the acquisition and storage that improve energy, raising pig using high-quality probiotic bacteria in Intestinum Sus domestica road is from daily ration
The middle ability for obtaining energy, further according to microbial technique index screening quality pigs beneficial bacteria of intestinal tract；
Step 3, the pig beneficial bacteria of intestinal tract in the pig gastrointestinal fluid of manual simulation after culture preliminary screening, determine all kinds of
The viable count of probiotic bacteria, reproductive order of generation are further screened, and screen optimal bacterial strain；
Step 4：Species, the functional characteristic of all kinds of prebioticses according to probiotic bacteria and the utilization journey with corresponding probiotic bacteria
Degree is selected by all kinds of oligosaccharide and the prebioticses for constituting, and selects all kinds of oligosaccharide optimum additions；
Step 5, the prebioticses that probiotic bacteria and step 4 are selected are filtered out to step 3, be mixed and made into compound symphysis
First microbial ecological agent.
Further, in step 5, probiotic bacteria nutrient substance each component is by weight：Peptone 1-100g/kg, Carnis Bovis seu Bubali cream
2-8g/kg, sucrose 1-50g/kg, yeast powder 1-20g/kg, potassium iodate 1-3g/kg, sodium dihydrogen phosphate 0.5-2.5g/kg, sulphuric acid
Magnesium 0.1-1g/kg, compound vitamin 0.01-10g/kg, aminoacids complex 0.01-10g/kg, mammal NaCl concentration is
0.85-0.9%, amphibian animal NaCl concentration are 0.67-0.7%.
Pig compound symphysis unit microbial ecological agent and preparation method thereof that the present invention is provided, solves raising market pig list
Solely using the weakness of probiotic products, and it is suitable for feeding for all kinds of market pigs, maximizes favourable factors and minimizes unfavourable ones, promote exogenous probiotic bacteria
Field planting in Intestinum Sus domestica road, and growth and breeding of the various probiotic bacterias of selective stimulating in pig body, so as to reach promotion
The purpose of animal health, reduces impact of the animal edible additive to meat quality, more economical to be suitable for；The pig is with compound
Symphysis unit microbial ecological agent can adjust pig intestinal microbial balance, maintain intestinal health, stimulate the growth of profitable strain, competition
Property suppress harmful bacteria, be finally reached promote pig growth, improve pig immunity purpose；The strain of acquisition directly can be used
Microbial ecological agent is prepared in fermentation, pollution-free, noresidue is the green animal edible additive of standard at this stage, meets people
Health standards to food, meet the food safety and prescription for increasingly improving；This boar is with compound symphysis unit Tiny ecosystem
Preparation is played an important role in animal husbandry, pig not only can be promoted to grow, moreover it is possible to improve the conversion ratio of feedstuff, and meet
The standard of Food Quality and Safety.
The present invention substitutes traditional pig edible additive to find, for solving that livestock faces because antibiotic is indiscriminate
With safety and the problem of quality of the Carnis Sus domestica for causing, a kind of compound symphysis unit microbial ecological agent is proposed.It is intended to make up benefit
Raw bacterium or prebioticses are added directly into the not enough and impact to meat quality in feedstuff, it is possible to increase the Carnis Sus domestica quality of production,
And farming industry animal tradition edible additive is updated with facilitation.The present invention relates to one kind is beneficial to pig life
The invention of compound symphysis unit microbial ecological agent that is long, improving meat quality amount.Symphysis unit microbial ecological agent as one kind it is new,
The pig edible additive of green, Main Function are the intestinal microbial populations for optimizing pig, adjust pig intestinal microbial balance, maintain its intestinal
Road health, stimulates the growth of profitable strain, competitive inhibition harmful bacteria to be finally reached the life for promoting pig using oligosaccharide probiotics
It is long, resistance of the animal body to cause of disease material is improved, body immune system is adjusted, so as to improve the immunity of animal, is reached
Improve the quality purpose of animal flesh；This compound symphysis unit microbial ecological agent is played an important role in animal husbandry, in food
There is guide effect in product industry, growth of animal not only can be promoted, moreover it is possible to improve the conversion ratio of feedstuff, increase measurement techniques for quality detection of meat and carry
The safe mass of high food.After present invention simulation Intestinum Sus domestica road environmental screening goes out probioticss, selected properly according to all kinds of probiotic bacterias
Prebioticses, the such as oligosaccharide such as xylooligosaccharide, MOS, fruit oligose synthesizes a kind of compound symphysis unit microbial ecological agent, for me
State's animal husbandry solves new animal edible additive to provide new selection, and solves the medicine brought by traditional antibiotic
Thing is remained, is developed immunity to drugs, animal intestinal micro-ecology balance is destroyed and reduces the food safetys such as meat productss quality and quality
Problem.
Description of the drawings
Fig. 1 is preparation method flow chart of the pig provided in an embodiment of the present invention with compound symphysis unit microbial ecological agent.
Specific embodiment
For the content of the invention, feature and effect of the present invention can be further appreciated that, following examples are hereby enumerated, and coordinates accompanying drawing
Describe in detail as follows：
The symphysis unit microbial ecological agent of the present invention, first, various probiotic bacterias of screening are filtered out through test of many times
The optimal species of condition, so combines the good characteristic of various probiotic bacterias, complements each other and play a multiplier effect, prebiotic
Characteristic is more significantly；Secondly, add the prebioticses based on oligosaccharide, solve oligosaccharide be difficult to it is digested utilize ask
Topic, appropriate oligosaccharide can promote the intestinal beneficial bacterium propagation of pig, and act on to body health；Finally, according to various benefits
The difference of prebioticses needed for raw bacterium, configures more preferably ratio, brings the compound prebioticses of more economic benefit and the suitability
Microbial ecological agent.
Increasing pathogenic bacterium are caused to develop immunity to drugs in order to solve abuse of antibiotics, drug residue reduces animal and produces
Product quality and to having a strong impact on that human body health is brought, produce the animal edible additive of substitute antibiotics.The addition
Agent effect is good, the green additive of drug residue free.The present invention is solved to find the new animal edible additive of substituted type
The vital task that certainly livestock faces provides a kind of compound symphysis unit microbial ecological agent.It is intended to make up probiotic bacteria or benefit
The impact directly as the not enough of additive and to animal of raw unit, and it is suitable for various domestic animals, it is possible to increase the quality of production,
Develop new symphysis unit microbial ecological agent and the replacement is updated to farming industry.
Below in conjunction with the accompanying drawings the present invention is elaborated.
With compound symphysis unit microbial ecological agent, the pig is with compound symphysis unit Tiny ecosystem for pig provided in an embodiment of the present invention
Preparation component by weight is pig beneficial bacteria of intestinal tract, oligosaccharide；Beneficial bacteria of intestinal tract by weight component by lactobacilluss, bacillus bifiduss,
Bacillus cereuss and yeast composition.
Another object of the present invention is the preparation method for providing the compound symphysis unit microbial ecological agent of a boar,
Filter out pig beneficial bacteria of intestinal tract, and the function and biological characteristicses to probiotic bacteria first respectively in different environments
It is compared, and the viable count change of probiotic bacteria is studied under simulation pig intestinal environment, filters out optimal bacterial strain；Then will
All kinds of oligosaccharide are measured, and select optimal oligosaccharide consumption, probiotic bacteria and oligosaccharide are mixed and made into compound conjunction finally
Raw unit's microbial ecological agent.
As shown in Figure 1：The pig is specifically included with the preparation method of compound symphysis unit microbial ecological agent：
S101:Soil from the plant of pig, the trough for feeding pig, gastrointestinal region separate lactobacilluss,
Isolated probiotic bacteria is carried out biological characteristicses, function and safety by bacillus bifiduss, bacillus cereuss and yeast probiotic bacteria
The identification of property, and external probiotic effects evaluation is carried out, preliminary screening probiotic bacteria；
S102:Using pig enteric microorganism and carbohydrate metabolism, the relation of vitamin, filter out with the acquisition that can improve energy
The high-quality probiotic bacteria of the ability of energy is obtained with storage, raising pig from daily ration, using high-quality probiotic bacteria carrying in Intestinum Sus domestica road
The acquisition and storage of high-energy, raising pig obtain the ability of energy from daily ration, further according to microbial technique index screening high-quality
Pig beneficial bacteria of intestinal tract；
S103:The pig beneficial bacteria of intestinal tract after preliminary screening is cultivated in the pig gastrointestinal fluid of manual simulation, all kinds of benefits are determined
The viable count of raw bacterium, reproductive order of generation are further screened, and screen optimal bacterial strain；
S104:Species, the functional characteristic of all kinds of prebioticses and the producing level with corresponding probiotic bacteria according to probiotic bacteria
Select, by all kinds of oligosaccharide and the prebioticses for constituting, to select all kinds of oligosaccharide optimum additions；
S105:The prebioticses that probiotic bacteria is selected with S104 are filtered out to S103, the compound symphysis unit of pig is mixed and made into
Microbial ecological agent.
Further, in the optimal bacterial strain of S103 screenings preferably：1 saccharomycete, 1 bacillus subtilis, 2 plants of lactobacilluss
With 2 plants of bacillus bifiduss.
Further, in S104, all kinds of oligosaccharide are：MOS, fruit oligose, oligomeric isomaltose；
The preferred MOS 0.25-2g/kg of all kinds of oligosaccharide optimum additions, fruit oligose 0.02-2g/kg, oligomeric different wheat
Bud sugar 0.5-3g/kg.
And the product being eventually fabricated is added in pig feed materials is tested.
General survey：Lactobacilluss are Gram-positive sporeless bacterium, and cellular morphology is in chain is shaft-like or club shape；Bifid bar
Bacterium is gram-positive sporeless bacterium, and varying length, bacillus bifiduss have V-shape or a Y-shaped, arrangement in single, double, short chain,
X, Y, V or palisade；Bacillus cereuss breeding is quick, breeds 100,000 times within four hours, vitality by force without wet condition can low temperature resistant -60 DEG C,
+ 280 DEG C of high temperature resistant, resistance to strong acid, resistance to highly basic, antibiotic and sterilizing, resistance to hyperoxia (thermophilic oxygen breeding), lower oxygen concentration resistance (anaerobism breeding).Volume ratio
The four times greater number of general nosophyte numerator, take up space advantage, suppresses the growth and breeding of harmful bacteria.Yeast is unicellular, in circle
Shape, elliptical or oval shape inside have nucleus, vacuole and granule material.Some species can produce ascospore or ballistopore, winter
Spore, basidiospore etc..Pseudohypha or fungal filament are formed under some species certain conditions.It is general that oval is presented, can be had
Oxygen is breathed, and can also carry out anaerobic respiration, and colonial morphology is similar to antibacterial but larger thicker, is creamy white or red, and surface is wet
Profit, it is sticky, easily provoked.5 grams of soil in vineyard are taken, is added in 100ml sterilized water, after being sufficiently stirred for, is used aseptic straw
Take 1ml to be linked in 9ml lactic acid potato dextrose broths, 24h is cultivated in 28-30 DEG C of incubator, it is seen that culture fluid becomes
It is muddy.
Lactobacilluss, bacillus bifiduss, bacillus cereuss and yeast probiotic bacteria and the prebioticses selected are combined,
Adding is beneficial to the nutrient substance for being conducive to growth of probiotics, makes compound symphysis unit microbial ecological agent.
Nutrient substance：As pig belongs to mammal, dilution normal saline used slightly has area with mammalss, Amphibian
Not, General N aCl concentration is 0.7%, peptone 1-100g/kg, Carnis Bovis seu Bubali cream 2-8g/kg, sucrose 1-50g/kg, yeast powder 1-
20g/kg, potassium iodate 1-3g/kg, sodium dihydrogen phosphate 0.5-2.5g/kg, magnesium sulfate 0.1-1g/kg, compound vitamin 0.01-
10g/kg, aminoacids complex 0.01-10g/kg.
The present invention solve brought by traditional antibiotic drug residue, develop immunity to drugs, animal intestinal micro-ecology
Balance is destroyed and is reduced the food safetys such as meat productss quality and quality problems；
The compound symphysis unit microbial ecological agent is avoided that the weakness that probiotic products are used alone, and is suitable for various
Domestic animal, maximizes favourable factors and minimizes unfavourable ones, and promotes field planting of the exogenous probiotic bacteria in animal intestinal, and the life of the various probiotic bacterias of selective stimulating
Long and breeding, so as to reach the purpose for promoting animal health, reduces impact of the animal edible additive to animal itself, more
It is economic and practical；
The compound symphysis unit microbial ecological agent can adjust animal intestinal microbial balance, maintain intestinal health, stimulate
The growth of profitable strain, competitive inhibition harmful bacteria are finally reached the growth for promoting animal, improve the mesh of the immunity of animal
's；The strain of acquisition can directly apply to fermentation and prepare microbial ecological agent, and pollution-free, noresidue is the green of standard at this stage
Color animal edible additive, meets health standards of the people to food, meets the food safety and prescription for increasingly improving；This
Plant compound symphysis unit microbial ecological agent to play an important role in animal husbandry, growth of animal not only can be promoted, moreover it is possible to carry
The conversion ratio of high feedstuff, and meet the standard of Food Quality and Safety.
With reference to specific embodiment, the present invention is further described.
Embodiment 1：Soil preliminary screening budding fungus probiotic bacteria from pig plant.
The first step：Collection containing yeast pedotheque.The soil on Adult Pig cultivation ground in large-scale plant that raises is selected to adopt
Collection destination, randomly select 3-5 zonule, each region takes 20-40g initial soils, be divided into 2 be assembled into it is sterilized self-styled
In bag, take back rapidly laboratory and be placed in 4 DEG C of refrigerators and save backup；
Second step：Saccharomycetic separation.Aseptically, 2 groups of pedotheques are added separately in enrichment culture liquid,
Bacterium colony is formed, then strain is inoculated into into bacteria suspension made by physiological saline solution respectively, then be diluted to not with 10 sequence diminishing methods
Same Concentraton gradient, different solubility gradient sample plate streaks is seeded in culture medium, and each dilution factor does 3 and puts down
OK, thus made by flat board be put into couveuse in cultivate 24-72h；
3rd step：It is saccharomycetic to isolate and purify.Aseptically, in 2 groups of picking, single purebred bacterial strain is identified,
Probiotic bacteria inoculating loop after discriminating is inoculated in physiological saline solution, bacteria suspension is made, is inoculated on flat board, trained
Observe after supporting, circulate operation, until obtaining pure bacterium colony, inclined plane inoculating is saved backup；
4th step：Saccharomycetic discriminating.Yeast after isolating and purifying is carried out into morphologic observation, physio-biochemical characteristics are determined
With 16S rDNA sequencings etc. method, respectively different bacterium colonies is identified, the purpose for knowing that to which class distinguishes name is reached.
Embodiment 2
From preliminary screening bacillus subtilises in pig feed grain.
The first step：The collection of sample in trough.It is sample collecting mesh with selecting each large-scale slaughterhouse Adult Pig cultivation
Ground, randomly choose a few place's troughs, sampling 20-30g in everywhere is respectively charged in sterilized valve bag, takes back laboratory
It is placed in 4 DEG C of refrigerators and saves backup；
Second step：The enrichment of bacillus subtilises, initial gross separation.Aseptically, 2 groups of trough samples are added respectively
Enter in enrichment culture liquid, form bacterium colony, then strain is inoculated in physiological saline solution respectively makes bacteria suspension, then use 10 sequences
Row diminishing method is diluted to different Concentraton gradient, and different solubility gradient sample plate streaks is seeded in culture medium,
Each dilution factor do 3 it is parallel, thus made by flat board be put into couveuse in cultivate 24-72h；
3rd step：Bacillus subtilises isolate and purify.Aseptically, in 2 groups of picking, single purebred bacterial strain enters
Row identification, the bacillus subtilises after discriminating is inoculated in physiological saline solution with inoculating loop, bacteria suspension is made, then is connect
Plant on flat board, observe after culture, circulate operation, until obtaining the bacterium colony that purebred bacillus subtilises are formed, inclined plane inoculating is protected
Deposit standby；
4th step：The discriminating of bacillus subtilises.Bacillus subtilises after isolating and purifying are carried out into morphologic observation, physiology
Biochemical characteristic determine and 16S rDNA sequencings etc. method, respectively different bacterium colonies is identified, is reached and class is known to which
Distinguish the purpose of name.
Example 3
Preliminary screening lactobacilluss, bacillus bifiduss probiotic bacteria from animal intestinal.
The first step：Collection containing probiotic bacteria sample.Select each large-scale slaughterhouse to be sample collecting destination, cut at random
Different health now kill the stomach intestinal tissues of pig, are divided into 2 and are assembled in sterilized valve bag, and quickly take back laboratory and are placed in 4
Save backup in DEG C refrigerator；
Second step：The enrichment of probiotic bacteria, screening.Aseptically, 2 groups of Gaster Sus domestica intestinal tissue sample inner surfacies are distinguished
The enrichment culture in corresponding culture fluid, forms bacterium colony, then strain is inoculated in bacteria suspension made by addition physiological saline solution,
Different Concentraton gradient are diluted to 10 sequence diminishing methods again, different solubility gradient sample plate streaks is seeded to into phase
Answer in culture medium, each dilution factor do 3 it is parallel, thus made by flat board be put into couveuse in cultivate 24-72h；
3rd step：Probiotic bacteria is isolated and purified.Aseptically, in 2 groups of picking, single purebred bacterial strain is identified,
Probiotic bacteria inoculating loop after discriminating is inoculated in physiological saline solution, bacteria suspension is made, then is connect with plate streak
Plant on flat board, observe after culture, circulate operation, until obtaining pure bacterium colony, inclined plane inoculating is saved backup；
4th step：The discriminating of probiotic bacteria.Lactobacilluss after isolating and purifying are carried out into morphologic observation, physio-biochemical characteristics are determined
With 16S rDNA sequencings etc. method, respectively different bacterium colonies is identified, the purpose for knowing that to which class distinguishes name is reached.
Embodiment 4
The further screening of probiotics strain.
The first step：The probiotic bacteria of preliminary screening is carried out into function test measure respectively, for example, the probiotic bacteria of new culture is connect
Plant in the culture fluid of different temperatures, determine the tolerance to temperature；Probiotic bacteria is cultivated 3-5 days in the environment of different pH, so
The degree of probiotic bacteria acid and alkali-resistance change is judged afterwards；
Second step：The probiotic bacteria after preliminary screening is cultivated in manual simulation pig gastrointestinal microecological environment, to all kinds of
The survival rate of probiotic bacteria, reproductive order of generation are measured, and filter out the preferable probiotic bacteria of upgrowth situation；
3rd step：The above is every to determine the probiotic bacteria that qualified probiotic bacteria is final screening, finally give 1 saccharomycete, 1
Bacillus subtilis, 2 plants of lactobacilluss and 2 plants of bacillus bifiduss.
Embodiment 5
The preparation of symphysis unit microbial ecological agent.
The first step：Species, the functional characteristic of all kinds of prebioticses according to probiotic bacteria and the utilization journey with corresponding probiotic bacteria
Degree selects the prebioticses that are made up of oligosaccharide, and selectable oligosaccharide is MOS, fruit oligose, oligomeric isomaltose.
Second step：The addition of all kinds of oligosaccharide is tested, the addition of all kinds of oligosaccharide is MOS 0.25-2g/kg,
Fruit oligose 0.02-2g/kg, oligomeric isomaltose 0.5-3g/kg.
3rd step：4 kinds of probiotic bacterias and the prebioticses selected are combined, add be beneficial to be conducive to it is prebiotic
The nutrient substance of bacteria growing, makes compound symphysis unit microbial ecological agent；
4th step：The product being eventually fabricated is tested in being added to pig feed materials, the gain in weight, feedstuff according to experiment pig
Consumption and feedstuff-meat ratio etc. carry out the production performance situation of judgment experiment pig.
The above is only, to presently preferred embodiments of the present invention, not to make any pro forma restriction to the present invention,
Every technical spirit according to the present invention is belonged to any simple modification made for any of the above embodiments, equivalent variations and modification
In the range of technical solution of the present invention.
Claims (7)
1. a boar is with compound symphysis unit microbial ecological agent, it is characterised in that the pig is with compound symphysis unit Tiny ecosystem system
Agent is made up of probiotic bacteria and prebioticses；
According to probiotic bacteria described in mass ratio：Prebioticses=1:1～5 ratio mixing.
2. pig as claimed in claim 1 is with compound symphysis unit microbial ecological agent, it is characterised in that the compound symphysis of the pig
Described in first microbial ecological agent, probiotic bacteria is made up of lactobacilluss, bacillus bifiduss, bacillus cereuss and yeast.
3. pig as claimed in claim 1 is with compound symphysis unit microbial ecological agent, it is characterised in that the prebioticses are by oligomeric
Sugar and acidulant are constituted；
The oligosaccharide is MOS, fruit oligose, oligomeric isomaltose, and acidulant is made up of lactic acid, citric acid, Fumaric acid
Compound acidulant.
4. the pig as described in claim 1 or 3 is with compound symphysis unit microbial ecological agent, it is characterised in that the probiotic bacteria is
0.02-15g/kg, probiotic bacteria nutrient substance are 0.05-80g/kg, and oligosaccharide is 0.01-90g/kg.
5. pig as claimed in claim 3 is with compound symphysis unit microbial ecological agent, it is characterised in that the addition of oligosaccharide is
MOS 0.25-2g/kg, fruit oligose 0.01-3g/kg, oligomeric isomaltose 0.01-3g/kg；Addition by lactic acid, citric acid,
The compound acidulant of Fumaric acid composition, compound acidulant account for the 0.1%-0.5% of microbial ecological agent ratio.
6. preparation method of a kind of pig as claimed in claim 1 with compound symphysis unit microbial ecological agent, it is characterised in that described
Pig filters out pig beneficial bacteria of intestinal tract first respectively in different environments with the preparation method of compound symphysis unit microbial ecological agent,
And the function and biological characteristicses to probiotic bacteria is compared, and the viable count of probiotic bacteria is studied under simulation pig intestinal environment
Change, filters out optimal bacterial strain；Then all kinds of oligosaccharide are measured, select optimal oligosaccharide consumption, finally by benefit
Raw bacterium and oligosaccharide are mixed and made into compound symphysis unit microbial ecological agent；Specifically include following steps：
Step one, the soil from the plant of pig, feed the trough of pig, gastrointestinal region and separate lactobacilluss, double
Isolated probiotic bacteria is carried out the mirror of biological characteristicses, function and safety by discrimination bacillus, bacillus cereuss and yeast
It is fixed, and carry out external probiotic effects evaluation, preliminary screening probiotic bacteria；
Step 2, using pig enteric microorganism and carbohydrate metabolism, the relation of vitamin, filter out raising pig energy is obtained from daily ration
High-quality probiotic bacteria, using high-quality probiotic bacteria in Intestinum Sus domestica road raising energy acquisition and storage, improve pig obtain from daily ration
The ability of energy is taken, further according to microbial technique index screening quality pigs beneficial bacteria of intestinal tract；
Step 3, the pig beneficial bacteria of intestinal tract in the pig gastrointestinal fluid of manual simulation after culture preliminary screening, determine all kinds of prebiotic
The viable count of bacterium, reproductive order of generation are further screened, and screen optimal bacterial strain；
Step 4：Species, the functional characteristic of all kinds of prebioticses according to probiotic bacteria and the producing level choosing with corresponding probiotic bacteria
Select by all kinds of oligosaccharide and the prebioticses for constituting, select all kinds of oligosaccharide optimum additions；
Step 5, the prebioticses that probiotic bacteria and step 4 are selected are filtered out to step 3, be mixed and made into compound symphysis unit micro-
Ecological agent.
7. the preparation method of compound symphysis unit microbial ecological agent as claimed in claim 6, it is characterised in that benefit in step 5
Raw bacterium nutrient substance each component is by weight：Peptone 1-100g/kg, Carnis Bovis seu Bubali cream 2-8g/kg, sucrose 1-50g/kg, yeast
Powder 1-20g/kg, potassium iodate 1-3g/kg, sodium dihydrogen phosphate 0.5-2.5g/kg, magnesium sulfate 0.1-1g/kg, compound vitamin
0.01-10g/kg, aminoacids complex 0.01-10g/kg, mammal NaCl concentration be 0.85-0.9%, amphibian animal
NaCl concentration is 0.67-0.7%.
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Application publication date: 20170419 |