CN1903058A - Ternary active microbiological prepns. for animals and birds - Google Patents
Ternary active microbiological prepns. for animals and birds Download PDFInfo
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- CN1903058A CN1903058A CNA2005100319253A CN200510031925A CN1903058A CN 1903058 A CN1903058 A CN 1903058A CN A2005100319253 A CNA2005100319253 A CN A2005100319253A CN 200510031925 A CN200510031925 A CN 200510031925A CN 1903058 A CN1903058 A CN 1903058A
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- bacillus
- lactobacillus plantarum
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
A three-element active microecologic food of domestic animals and fowls for improving immune function contains living bacteria component (94-98 Wt%) prepared from vegetative lactobacillus liquid, yeast liquid and bacillus liquid, oligose (0.1-1) and yolk immuno globulin (1-5). Its features that said living bacteria are in sleep state, resulting in high stability.
Description
Technical field
The present invention relates to livestock and poultry and use feed addictive, be meant that further the livestock and poultry that are used as described additive use ternary active microbiological preparations.
Background technology
Antibiotic for world's Developing of Animal Industry has been made huge contribution, yet has also brought great hidden danger in the application of livestock and poultry breeding industry: the first, cause autogenous infection and suprainfection; The second, the livestock and poultry immunity of organisms is descended, cell produces drug resistance, and transmits by plasmid, can be transferred to human body, threatens human body health; The 3rd, chemical residual can pollute meat, egg, milk, reduces the quality of animal products.Therefore, world developed country began to completely forbid the application of antibiotic in feed in 1989.
In order to overcome antibiotic various drawback, countries in the world successively carried out have no side effect, the research of the green feed additive of noresidue, and in its research and development process, first-selected research be probiotics.Probiotics is a kind ofly to contain a large amount of beneficial bacteriums and metabolite and positive growth factor, and the active bacteria formulation of keeping the intestinal microecology balance, improving the body health level is arranged.Immunoglobulin (Ig) is the anti-infective material that animal body produces, and in recent years, foreign study proves, adds polyclonal antibody in daily ration, can prevent the Escherichia coli diarrheal disease of piglet and calf.U.S. food and drug administration listed Yolk immunoglobulin (Igy) in 1996 " It is generally accepted safe material " category, Germany in nineteen ninety-five approval use it for feed addictive.
This novel biological product of probiotics, though with the growing animal husbandry of its unique function effect, but also exist many remaining further to study a question: first, specific aim is not strong, not as antibiotic can adopt corresponding product at certain intestines problem like that, specific aim is treated very doughtily; The second, the probiotics The stability of strain is not high, and is low as the ability of the extraneous poor environments of opposing such as the heat resistance of live bacteria agent, acid resistance; The 3rd, the shelf life of probiotics product is not long, and along with the increase of storage period, number of viable progressively reduces.
From external exploitation and result of use, the more single microbial inoculum action effect of composite bacteria preparation is better, and composite bacteria preparation generally all has synergy.In order to improve the validity of probiotics, people are just putting forth effort on the development and the exploitation of probiotics assistant.Test as to oligosaccharide and the compound interpolation of active bacteria formulation obtains certain effect; The also synergy between R and D live bacteria agent and other preparations simultaneously.Also pass through technique for gene engineering abroad at present, some useful foreign genes are imported in the bacterial classification of active bacteria formulations, enlarge its biology specific function, thereby reach the purpose of the multiple function of a kind of preparation in research.
Summary of the invention
The technical problem to be solved in the present invention is, deficiency at the prior art existence, propose a kind of livestock and poultry and use ternary active microbiological preparations, it organically combines active bacteria formulation, oligosaccharide, antibody globulin, with the non-specific immune function of the specific immune function of giving full play to antibody globulin and live bacteria agent, oligosaccharide, to have improved the specific aim of probiotics; Also can adopt the cryogenic vacuum Freeze Drying Technique, live bacteria agent is made dry powder, make viable bacteria be in resting state, to improve the stability of probiotics; Also can adopt microcapsule embedded technology, the embedding of ternary active preparation is got up, to prolong the shelf life of probiotics.
Technical solution of the present invention is that described livestock and poultry are with the weight portion composition of ternary active microbiological preparations:
Active bacteria formulation 94%-98%;
Oligosaccharide 0.1%-1%;
Yolk immunoglobulin 1%-5%;
Wherein, described active bacteria formulation by Lactobacillus plantarum liquid, yeast liquid, bacillus liquid by 1: the weight ratio of 0.9-1.1: 0.9-1.1 is formed, and contains Lactobacillus plantarum 〉=8 * 10 in every gram Lactobacillus plantarum liquid
8Individual, contain saccharomycete 〉=7 * 10 in every gram yeast liquid
8Individual, contain bacillus 〉=4 * 10 in every gram bacillus liquid
8Individual.
Below the present invention made further specify.
Described oligosaccharide can be a kind of in xylo-oligosaccharide, FOS, the oligomeric sweet dew, also can be other oligosaccharide.
The present invention can above-mentioned ternary active microbiological preparations effective ingredient be core, be the wall material with the homogeneous mixture of following weight parts composition:
Converted starch 35%-45%;
Arabic gum 10%-20%;
Maltodextrin 20%-30%;
Corn syrup 15%-25%;
With above-mentioned core and wall material by 1: after the weight ratio of 0.9-1.1 is mixed homogeneous, make microcapsules state product through conventional cold nebulization technology.
Among the present invention, described active bacteria formulation can adopt the culture medium of following parts by weight of component:
Main body carbon source 5%-9%; Glucose 1%-3%;
Peptone 0.4-0.6%; Vegetable oil 0.2-0.4%;
K
2HPO
4 0.1-0.3％； CaCo
3 1％-3％；
MgSO
4 0.05-0.07％； MnSO
4 0.02-0.03％；
Water 84%-90%;
Wherein, described main body carbon source is a kind of in mealy potato, soy meal, the corn flour, and implants corresponding bacterial classification, and the bacterial classification content in every gram bacterium liquid is: Lactobacillus plantarum 〉=8 * 10
8Individual, saccharomycete 〉=7 * 10
8Individual, bacillus 〉=4 * 10
8Individual.
Among the present invention, pig can be adopted mealy potato with ternary active microbiological preparations main body carbon source, and chicken can be adopted soy meal with ternary active microbiological preparations main body carbon source, and ruminant ternary active microbiological preparations main body carbon source can adopt corn flour.
The present invention has carried out processing parameters such as single strain culturing temperature, incubation time, cultivation initial p H value, oxygen demand, culture medium composition and has studied to described Lactobacillus plantarum (PC0586-2), bacillus subtilis (BL0067), candida utili bacterium (2.120), and the correction of technological parameter is carried out in their compound cultivation.The result shows: the time that three kinds of bacterium reach the output peak is 48-54 hour, saccharomycetic fermentation optimum temperature is 32 ℃, bacillus is 38 ℃, and lactic acid bacteria is 36 ℃, and optimum temperature has nothing in common with each other, therefore, during compound cultivation, carry out 32 ℃ of phase I temperature in two stages, 31 ℃ of second stage temperature, the time half and half (28 hours).The initial cultivation pH value of three kinds of bacterium is respectively lactic acid bacteria 5.6, saccharomycete 5.8, bacillus 6.0.Therefore, pH value will be controlled at about 6.0 during composite fermentation.The oxygen demand of three kinds of bacterium sweats has nothing in common with each other, and lactic acid bacteria is the amphimicrobe based on anaerobic respiration, and saccharomycete and bacillus are the bacterial classifications of aerobic respiration.Therefore, compound cultivation is carried out in two steps, and the phase I (preceding 28 hours) ventilates and stopped 2 hours in 1 hour when cultivating, and the second stage ventilation stopped 1 hour in 2 hours.
Among the present invention, described Lactobacillus plantarum, saccharomycete, bacillus can be adopted reference culture, also can obtain by following strain separating screening:
Get animal and bird intestines or pedotheque, dilute the back and separate steps such as cultivation, bacterium colony collection, purifying subculture separation cultivation, pilot production cultivation, qualitative and quantitative analysis by enrichment domestication cultivation, selection cultivation, dull and stereotyped (line), from 126 parts of chitling, chicken intestines, bovine rumen sample, in nearly 20,000 strain bacterial strains, separate Lactobacillus plantarum (being numbered PC0586 temporarily) the lactic acid production 90.7g/L of a plant height lactic acid producing, higher by 16% than reference culture (Chinese Academy of Sciences microorganism fungus kind preservation center provides, and lactic acid production is 78g/L) lactic acid production.From 27 parts of pedotheques, in nearly 4000 strain bacterial strains, separate the very strong bacillus subtilis variant of a strain bacteriostasis, be numbered BL0067 temporarily.Compare with reference culture (Chinese Academy of Sciences microorganism fungus kind provides in preserving), bacteriostasis improves 31%.
The present invention also can adopt ultraviolet that the above-mentioned Lactobacillus plantarum PC0586 that brings out is carried out mutagenic treatment, and treatment dosage is that survival rate is below 1%.Ultraviolet wavelength is about 260nm, and uviol lamp power is 15W, and fixed distance is in sample 30nm eminence.Mutation time 0-72h.Part material, select the Lactobacillus plantarum of a plant height lactic acid producing surplus 2000, lactic acid production has reached 130g/L, is 86%-90% to the conversion ratio of sugar, and shows genetic stability preferably.The tentative PC0586-1 by name of this bacterial strain.
The also available genetic engineering of the present invention is carried out genetic improvement to described bacterial classification.Wherein, turn: the excellent bacillus (AS1.536) of high yield lysine is provided by Institute of Microorganism, Academia Sinica bacterial classification preservation center, the restriction endonuclease of various DNA, the key gene in the lysine synthetic pathway: dihydrodipicolinic acid synthase's gene (dapA) is provided by Hunan Academy of Agricultural Sciences biological study chamber; Acceptor: the Lactobacillus plantarum PC0586-1 that can adopt mutation breeding to select.Carrier: Escherichia coli.The technique for gene engineering that utilizes recombinant DNA is with the key gene in excellent bacillus (AS1.563) lysine synthetic pathway-dihydrodipicolinic acid synthase's gene (dapA), adopt the random integration method, with Escherichia coli as carrier, successfully imported aimed strain Lactobacillus plantarum PC0586-1, turn out a strain gene engineering bacterial strain, be numbered PC0586-2 temporarily.The result shows, exogenous plasmid conversion ratio height can be preserved in the object bacteria midium or long term, and high level expression.Make the Lactobacillus plantarum of no protein decomposition enzyme can synthetic lysine, and output reaches 27g/L, be applied in the production, played both high lactic acid producing, can produce the double action of lysine again.
Probiotics is a kind of novel biological agent of developing over past ten years, also all is the active bacteria formulation of unit mostly both at home and abroad at present, and typical case's representative is " the former dew of EM " of Japan's stream ball university research exploitation, American-European and China's wide popularization and application in Japan.At this, the present invention is compared as follows with every technical indicator of " the former dew of EM ":
1. homology is relatively:
One of bacterial classification in the product of the present invention source is made the ternary active preparation and is made an addition in the feed for deriving from animal and bird intestines, returns animal and bird intestines again after getting food, because good homology is arranged, has improved the field planting ability of microorganism in fowl is raiseeed enteron aisle.And be that other probiotics bacterial classifications of representative mostly derive from soil with " the former dew of EM ", the field planting ability in animal and bird intestines is lower than product of the present invention.
2. compare targetedly:
Table 1: ternary active preparation (the present invention) compares targetedly with the former dew of EM
| Compare index | The ternary active preparation | The former dew of EM |
| Bacterial classification quantity antibody globulin Oilgosaccharkdes nonspecific immunity function specific immune function scope of application specific aim | 3 have pig, chicken, ruminant strong | 80 do not have not to have and to have or not all animals, all plants poor |
From above-mentioned comparative result as can be seen, " the former dew of EM " contains 80 bacterial classifications, can be used for animal feed, also can make the bio-feritlizer of plant, and its universal is good, specific aim is poor; And contain antibody ball egg in the ternary active preparation at Escherichia coli and Salmonella, and have special immunologic function, with strong points.
3. heat endurance relatively
Ternary active preparation of the present invention and the former dew of EM were taked 70 ℃ of high-temperature process 10 minutes, and the result shows that the viable bacteria survival rate in the ternary active preparation is doubled many up to 88.7% than the former dew of EM (41.5%).
Table 2: ternary active preparation (the present invention) claims qualitative comparison with the heat of the former dew of EM
| Compare index | The ternary active preparation | The former dew of EM |
| Viable count before the heat treatment (individual/g) viable count after the heat treatment is (individual/g) viable bacteria survival rate (%) | 4.98×10 8 4.22×10 8 88.7 | 5.11×10 8 2.12×10 8 41.5 |
4. absolute acid stability relatively
After livestock and poultry were got food, probiotics was killed by a large amount of hydrochloric acid in gastric juice, pepsin, trypsase, chymotrypsin in the stomach of animal or is digested, and the number of viable that can reach enteron aisle can reduce in a large number.The present invention has carried out the SGF test of ternary active preparation and the former dew of EM.
SGF is formed: hydrochloric acid in gastric juice 10%, pepsin 1%, trypsase 0.1%, chymotrypsin 0.05%, pH value 4.0-5.0, time 1-8h.
Table 3: ternary active preparation (the present invention) compares with the former dew hydrochloric acid in gastric juice stability of EM
| Processing time (h) | 0 | 1 | 2 | 3 | 4 | 3 | 6 | 7 | 8 |
| Ternary active preparation viable bacteria survival rate (%) EM former dew viable bacteria survival rate (%) | 100 100 | 98.2 82.4 | 96.7 77.4 | 82.1 66.0 | 73.4 50.9 | 57.0 30.1 | 40.1 11.7 | 37.6 0 | 32 0 |
As can be seen from Table 3, the anti-ability of supporting hydrochloric acid in gastric juice of ternary active preparation, be higher than the former dew of EM far away, after SGF is handled 6 hours, viable bacteria rate in the ternary active preparation also has 41.0%, improves 4 times than the former dew of EM, handles after 7 hours, do not had viable bacteria in the former dew of EM, and the ternary active preparation also has 37.6% viable bacteria survival rate.
5. the comparison of bacteriostasis
Table 4: the comparison of the former dew bacteriostasis of ternary active preparation and EM
| The indication bacterial classification | Antibacterial circle diameter (mm) | |
| The ternary active preparation | The former dew of EM | |
| Escherichia coli lactobacillus acidophilus bacillus subtilis Salmonella bifidobacterium adolescentis | 27.3 - 20.2 31.8 - | 10.6 - 11.6 13.7 - |
As can be seen from Table 4, the former dew of ternary active preparation and EM all shows inhibitory action to the harmful bacterium in the enteron aisle, and beneficial bacterium is not showed inhibitory action.From bacteriostasis relatively, the ternary active preparation is 2.5 times of the former dew of EM (antibacterial circle diameter 10.6) to colibacillary inhibition ability (antibacterial circle diameter 27.3mm).
6. the comparison of shelf life
Table 5: the comparison of the former dew shelf life of ternary active preparation and EM
| Item compared | Before the storage | January | March | May |
| Ternary active preparation viable bacteria survival rate (%) EM former dew viable bacteria survival rate (%) | 100 100 | 97.2 93.0 | 94.5 78.77 | 89.9 61.1 |
From the result of table 5 as can be seen, the ternary active preparation through 6 months shelf life after, the viable bacteria survival rate has still reached 89.8%, and after the former dew of EM stored through 6 months, the viable bacteria survival rate had only 61.1%.Ternary active preparation shelf storage capability has improved 45%.
As known from the above, the present invention uses ternary active microbiological preparations for livestock and poultry, it organically combines active bacteria formulation, oligosaccharide, antibody globulin, give full play to the non-specific immune function of the specific immune function of antibody globulin and live bacteria agent, oligosaccharide, improved the specific aim of probiotics; Also adopt the cryogenic vacuum Freeze Drying Technique, live bacteria agent has been made dry powder, made viable bacteria be in resting state, improved the stability of probiotics; Also adopt microcapsule embedded technology, the embedding of ternary active preparation has been got up, prolonged the shelf life of probiotics.
The specific embodiment
Embodiment 1: the weight portion of ternary active microbiological preparations is formed and is: the active bacteria formulation 98% that the main body carbon source is cultivated with mealy potato, xylo-oligosaccharide 1%, Yolk immunoglobulin (320 read/g) 1%; Wherein, described active bacteria formulation is made up of by 1: 1: 1 weight ratio Lactobacillus plantarum liquid, yeast liquid, bacillus liquid, and contains Lactobacillus plantarum 〉=8 * 10 in every gram Lactobacillus plantarum liquid
8Individual, contain saccharomycete 〉=7 * 10 in every gram yeast liquid
8Individual, contain bacillus 〉=4 * 10 in every gram bacillus liquid
8Individual; Effective ingredient with above-mentioned ternary active microbiological preparations is a core, is the wall material with the homogeneous mixture of following weight parts composition: converted starch 45%, Arabic gum 20%, maltodextrin 20%, corn syrup 15%;
Above-mentioned core with after the wall material mixes homogeneous by 1: 1 weight ratio, is made microcapsules state product through conventional low temperature (5 ℃) spray art.Product is used:
Place: Taoyuan county, Hunan Province farms producing good poultry and animal strains
Object: 28 age in days two-way cross (long * big) weanling pig
Time: 28 days (on May 28th, 1 2005 on May 1st, 2005)
Daily ration is formed: Yueyang nine ancient cooking vessel SH112 type sucking pig concentrate feeds
Grouping and raising: 12 of test group, 15 of control groups, every day, feeding was 2 times.
Application result:
Table 6: the effect of ternary active preparation
| Compare index | Test group (ternary active preparation) | Control group (not using) | Comparative result |
| The average daily ingestion amount of average daily gain (g/ head) (g/ day. head) feedstuff-meat ratio diarrhea rate (%) survival rate (%) weightening finish 1kg body weight cost (unit) weightening finish 1kg body weight net income (unit) | 495.71 796.07 1.61∶1 4.2 100 13.46 2.54 | 298.93 564.4 1.83∶1 14.3 83％ 15.19 0.81 | +196.78 +249.67 -0.22 -11.1 +17.0 -1.73 +1.73 |
As can be seen from Table 6, test group is than the control group weightening finish of Duoing every day 196.78 grams, and comparison is according to having improved 65%, feedstuff-meat ratio has reduced by 0.22, and feed conversion rate has improved 12.02%, and the diarrhea rate comparison is according to having reduced by 3 times, survival rate has improved 20%, every increase by one kg body weight, and net income increases by 1.73 yuan.
Embodiment 2: the weight portion of ternary active microbiological preparations is formed and is: the weight portion of the active bacteria formulation that the main body carbon source is cultivated with soy meal is formed and is: active bacteria formulation 94%, FOS 1%, Yolk immunoglobulin (320 read/g) 5%; Wherein, described active bacteria formulation is made up of by 1: 0.9: 1 weight ratio Lactobacillus plantarum liquid, yeast liquid, bacillus liquid, and contains Lactobacillus plantarum 〉=8 * 10 in every gram Lactobacillus plantarum liquid
8Individual, contain saccharomycete 〉=7 * 10 in every gram yeast liquid
8Individual, contain bacillus 〉=4 * 10 in every gram bacillus liquid
8Individual.Effective ingredient with above-mentioned ternary active microbiological preparations is a core, is the wall material with the homogeneous mixture of following weight parts composition: converted starch 35%, Arabic gum 10%, maltodextrin 30%, corn syrup 25%; Above-mentioned core with after the wall material mixes homogeneous by 1: 0.9 weight ratio, is made microcapsules state product through conventional low temperature (0--10 ℃) spray art.
Embodiment 3: the weight portion composition of ternary active microbiological preparations is: the main body carbon source with the weight portion composition of the active bacteria formulation that corn flour is cultivated is: active bacteria formulation 96%, oligomeric sweet dew 0.1%, Yolk immunoglobulin 3.9%; Wherein, described active bacteria formulation is made up of by 1: 1: 0.9 weight ratio Lactobacillus plantarum liquid, yeast liquid, bacillus liquid, and contains Lactobacillus plantarum 〉=8 * 10 in every gram Lactobacillus plantarum liquid
8Individual, contain saccharomycete 〉=7 * 10 in every gram yeast liquid
8Individual, contain bacillus 〉=4 * 10 in every gram bacillus liquid
8Individual.Effective ingredient with above-mentioned ternary active microbiological preparations is a core, is the wall material with the homogeneous mixture of following weight parts composition: converted starch 40%, Arabic gum 15%, maltodextrin 25%, corn syrup 20%; Above-mentioned core with after the wall material mixes homogeneous by 1: 1.1 weight ratio, is made microcapsules state product through conventional low temperature (20 ℃) spray art.
Claims (3)
1, a kind of livestock and poultry are used ternary active microbiological preparations, it is characterized in that, its weight portion composition is:
Active bacteria formulation 94%-98%;
Oligosaccharide 0.1%-1%;
Yolk immunoglobulin 1%-5%;
Wherein, described active bacteria formulation by Lactobacillus plantarum liquid, yeast liquid, bacillus liquid by 1: the weight ratio of 0.9-1.1: 0.9-1.1 is formed, and contains Lactobacillus plantarum 〉=8 * 10 in every gram Lactobacillus plantarum liquid
8Individual, contain saccharomycete 〉=7 * 10 in every gram yeast liquid
8Individual, contain bacillus 〉=4 * 10 in every gram bacillus liquid
8Individual.
2, livestock and poultry according to claim 1 are used ternary active microbiological preparations, it is characterized in that, and be core with described composition, be the wall material with the homogeneous mixture of following weight parts composition:
Converted starch 35%-45%;
Arabic gum 10%-20%;
Maltodextrin 20%-30%;
Corn syrup 15%-25%;
With above-mentioned core and wall material by 1: after the weight ratio of 0.9-1.1 is mixed homogeneous, make microcapsules state product through conventional cold nebulization technology.
3, livestock and poultry according to claim 1 are used ternary active microbiological preparations, it is characterized in that, described oligosaccharide is a kind of in xylo-oligosaccharide, FOS, the oligomeric sweet dew.
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| CN106578350A (en) * | 2016-11-15 | 2017-04-26 | 漳州傲农牧业科技有限公司 | Enterobactin preparation used for animals, and preparation method thereof |
| CN107136302A (en) * | 2017-04-21 | 2017-09-08 | 大连三仪动物药品有限公司 | It can increase weight and reduce the biological compound preparation of aquatic bird raising and preparation method of feedstuff-meat ratio |
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