CN107319103A - Promote feed addictive of growth of animals or poultry and intestinal growth and its preparation method and application - Google Patents

Promote feed addictive of growth of animals or poultry and intestinal growth and its preparation method and application Download PDF

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CN107319103A
CN107319103A CN201710648118.9A CN201710648118A CN107319103A CN 107319103 A CN107319103 A CN 107319103A CN 201710648118 A CN201710648118 A CN 201710648118A CN 107319103 A CN107319103 A CN 107319103A
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growth
animals
poultry
feed addictive
feed
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杨建平
赵志军
史素荣
李新锋
黄好强
刘健
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Henan University of Animal Husbandry and Economy
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Henan University of Animal Husbandry and Economy
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/174Vitamins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry

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  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
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  • Fodder In General (AREA)
  • Feed For Specific Animals (AREA)

Abstract

The invention belongs to field of feed additive technology, disclose feed addictive of a kind of promotion growth of animals or poultry and intestinal growth and its preparation method and application, it is intended to improve animal and bird intestines Bacterial community, solves the harmful effect that antibiotic is caused to growth of animals or poultry, promotes livestock birds health growth.The feed addictive is 500 by mass ratio:(1~3)Cabbage slurries and solid-state bacillus subtilis be made, into every 100g cabbage slurries add 0.2~0.6g solid-state bacillus subtilises, mix thoroughly;By the feed addictive to add 10~40g in per kilogram feed, the application of additive for farm animal feed is used as.Extensively, fully, digestive utilization ratio is high for nutrition for raw material sources of the present invention;Used as additive for farm animal feed, the palatability of feed can be improved, moreover it is possible to adjust the microecological balance of animal intestinal tract, promote growth of animals or poultry and intestinal growth, reduce aquaculture cost, laid the first stone to improve recycling of the vegetables in Animal husbandry production.

Description

Promote feed addictive of growth of animals or poultry and intestinal growth and its preparation method and application
Technical field
The present invention relates to field of feed additive technology, and in particular to be the feed for promoting growth of animals or poultry and intestinal growth Additive and its preparation method and application.
Background technology
In recent years, there is abuse in order to promote growth of animal and keep livestock birds health and excessively use in many breeding enterprises The phenomenon of antibiotic and chemicals, as a result causes livestock products medicament residue too high, and healthy to consumer is caused not Good influence, so, seek effective, safety, green, natural feed addictive substitute antibiotics product, become livestock and poultry cultivation The inexorable trend of industry development.
As the potential replacer of antibiotic, probiotics embodies many superiority, for example, can overcome abuse of antibiotics A variety of drawbacks such as caused animal autogenous infection or the drug-fast generation of suprainfection, animal and immunologic function decline, and tool Have notable disease-resistant, growth promotion, improve efficiency of feed utilization, it is nontoxic, have no adverse reaction and the features such as noresidue, be a kind of natural life Long accelerator.
Bacillus subtilis is the Ministry of Agriculture《Catalogue of feed additive varieties》Listed strain, is additive for microbe feedstuff Conventional strain.Bacillus subtilis does not have toxicity to humans and animals, and the protease of high activity can be produced in growth course, is formed sediment Powder enzyme and cellulase, can promote livestock and poultry to the digestive utilization ratio of feed;Antibacterial metabolin can be produced, suppresses enteropathogenic The growth of bacterium;The bacterium is strict aerobic bacteria, into enteron aisle after by it is biological take oxygen by force act as enteron aisle lactic acid bacteria class and bifid Bacillus class anaerobe provides anaerobic environment to promote the growth of beneficial bacterium, regulation intestinal microecology balance.
Meanwhile, plant extracts as feed addictive so that application effect is good, low-residual, be not likely to produce the resistance to the action of a drug and nothing Pollute and turn into a kind of preferable feed addictive, be also one of focus of present feed supplement research.
Single bacillus subtilis or plant extracts is limited as the action effect of feed addictive, it is difficult to meet The requirement of complete substitute antibiotics product.Therefore, the natural material source for coordinating synergistic effect can be played in the urgent need to finding Complex additive.
The content of the invention
The invention provides feed addictive of a kind of promotion growth of animals or poultry and intestinal growth and its preparation method and application, By additive addition in feed, growth, intestinal growth and the improvement intestinal microflora of livestock and poultry can be promoted, growth-promoting is reduced The use of long antibiotic, is conducive to the feed-weight ratio of food security and reduction livestock and poultry.
In order to realize foregoing invention purpose, the technical solution adopted by the present invention is as follows,
The feed addictive of a kind of promotion growth of animals or poultry of design and intestinal growth, is 500 by mass ratio:The knot of (1~3) Cabbage slurries and solid-state bacillus subtilis are made.
It is preferred that, the solid-state bacillus subtilis number of viable is 0.5~2 × 109CFU/g。
Present invention also offers the preparation method of a kind of promotion growth of animals or poultry and the feed addictive of intestinal growth, including such as Lower step:
Fresh cabbage blade is ground into after slurry, cabbage slurries are obtained, to every 100g cabbage slurries Middle addition 0.2~0.6g solid-state bacillus subtilises, mix thoroughly.
It is preferred that, the solid-state bacillus subtilis is made by following methods:
(1) test tube strains culture:Under gnotobasis, withered grass bud is inoculated with the slant tube culture medium of beef extract protein Spore bacillus, 20h is cultivated under the conditions of 37 DEG C;
(2) culture is expanded:By liquid beef-protein medium after sterilizing 20min under the conditions of 0.1MPa, 121 DEG C, The test tube strains that aseptic inoculation step (1) is obtained, expand culture 24h on 37 DEG C, 120r/min constant-temperature table;
(3) fermentation tank culture:By fermentation tank culture medium under the conditions of 0.1MPa, 121 DEG C, after sterilizing 25min, cooling and fermentation Tank culture medium is to temperature to less than 40 DEG C, and inoculation step (2) cultivates gained strain, in speed of agitator 120r/min, ventilation Under conditions of 0.8vvm, 36~48h of fermented and cultured, wherein, inoculum concentration is the 5% of fermentation tank culture medium quality;
(4) solid substrate fermentation culture:Solid medium is loaded in high-pressure sterilizing pot, in 0.1MPa, 121 DEG C of conditions Under, after sterilizing 30min, solid medium is cooled to less than 40 DEG C, by weight the 10% of inoculation step (3) gained strain, stirred Mix uniform, be fitted into aerated koji making pond, under conditions of 30 DEG C of ventilation temperature, humidity 80%~90%, every 10~15h stirrings Once, 36~48h of fermented and cultured, obtains solid-state bacillus subtilis;
(5) the solid state bacterial crushed after being dried for obtaining step (4), crosses 0.2mm sieve apertures, and viable bacteria content during microscopy ≥2×109CFU/g, miscellaneous bacteria content < 1%.
It is further preferred that in step (1), the bacillus subtilis derives from Institute of Microorganism, Academia Sinica DSMZ.
It is further preferred that in step (3), the formula of the fermentation tank culture medium is 1% bean cake powder, 1% starch, 0.01% potassium dihydrogen phosphate, 0.01% magnesium sulfate.
It is further preferred that in step (4), the formula of the solid medium is wheat bran 430kg, bean cake powder 50kg, Flour 20kg, potassium dihydrogen phosphate 100g, magnesium sulfate 100g, 400~450kg of water.
It is further preferred that in step (5), during the solid state bacterial aeration-drying that step (4) is obtained:5h before drying, Feng Wen≤50 DEG C are controlled, 80 DEG C are then progressively warming up to, dried to Shui Fen≤12%.
Present invention also offers the feed addition for promoting growth of animals or poultry and intestinal growth described in a kind of above-mentioned technical proposal Prepared by agent or the method recorded by above-mentioned technical proposal promotes the feed addictive of growth of animals or poultry and intestinal growth as livestock and poultry The application of feed addictive.
It is preferred that, the addition of the feed addictive for promoting growth of animals or poultry and intestinal growth is to add in per kilogram feed Plus 10~40g.
Cabbage:Be the plant of Cruciferae, Brassica genus, be the mutation of wild cabbage, also known as cabbage, cabbage, cabbage, Cabbage etc..The features such as wild cabbage has cold-resistant, disease-resistant, strong adaptability, easily stores high resistance to fortune, yield, quality better, it is general all over China All over cultivation.The rich in nutrition content of cabbage, per hectogram edible part 1.2~2.4g containing protein, fat is about 0.3g, carbon Hydrate is about 3.5g.It has been investigated that, cabbage is also containing abundant vitamin, and vitamin has well to ulcer Therapeutic action, can accelerate the healing of ulcer, to improving livestock and poultry production performance and promoting animal and bird intestines healthy development to have well Effect.
Bacillus:B family vitamin and vitamin C vitamin K can be produced during in growth and breeding2, it is animal Vitamin is provided;Bacillus can produce the materials such as lactic acid, propionic acid, acetic acid in stomach and enteron aisle portion drops the pH in enteron aisle It is low;Volatile fatty acid is produced in lower intestine, suppresses harmful microbe growth, and help animal particularly brood battalion Support digesting and assimilating for the digestion of material, especially protein;Bacillus can secrete substantial amounts of ectoenzyme, participate in the drop of feed Solution, digestion, improve efficiency of feed utilization;The trophosome of bacillus can secrete cellulase, protease, amylase of complexity etc. Extracellular enzyme;Bacillus, which enters, can consume substantial amounts of oxygen after enteron aisle, maintain enteron aisle anaerobic environment, suppress the bacteria growing that causes a disease, The effect of balance, stable Bacillus acidi lactici, maintains intestinal microecology balance.
Compared with prior art, advantageous effects of the invention are embodied in:
1) raw material sources of the present invention extensively, it is rich in nutrition content, with low cost, be conducive to the feed addictive Widely used in livestock and poultry cultivation;Also, cabbage slurries are added in the additive, are conducive to substituting or reducing antibiotic Use in livestock and poultry, it is to avoid or residual of the antibiotic in livestock products is reduced, meet the food-safe demand of consumer.
2) the cabbage slurries in feed addictive of the present invention contain vitamin, and vitamin is that important biology is living Property material, be conducive to the reparation of damaged intestine, promote animal and bird intestines healthy development, increase the abilities of digestive and absorption of enteron aisle, improve Livestock and poultry daily gain.
3) probiotics bacillus subtilis is used, intestinal growth can be stimulated, the immunologic function of livestock and poultry is improved;It can press down The growth of partial enteral harmful bacteria processed, promotes the growth and breeding of beneficial bacterium, reduces the generation of ammonia and other corrupt substances, hinders intestines Absorption of the road to harmful substance and waste, improves the production performance of livestock and poultry, and reduces the discharge of pernicious gas, reduces plant pair The harm of environment.
4) present invention realizes the combination of cabbage and probiotics functional characteristics, the two mutual cooperation, increases Effect is notable, all significant to animal and bird intestines healthy development, improvement intestinal microflora and promotion growth of animals or poultry.
Brief description of the drawings
Fig. 1 is 21 Day-old Broiler Chickens duodenum intestinal growths section electron micrograph;
Fig. 2 is that 21 Day-old Broiler Chickens test groups and control group cecum microorganisms belong to flora comparison between community structures figure in level;
Fig. 3 is the LDA Distribution value block diagrams for testing III group of significant difference species between control group;
Fig. 4 is the flora cladogram for testing III group of significant difference species between control group.
Wherein, D represents control group, Y1Represent I group of experiment, Y2Represent II group of experiment, Y3Represent III group of experiment.
Embodiment
Technical scheme is described in detail with reference to specific embodiment.It should be appreciated that described herein specific Embodiment only to explain the present invention, is not intended to limit the present invention.In following embodiments involved test method or point Analysis method, is conventional method unless otherwise instructed, and raw materials used and reagent is commercially available unless otherwise instructed.
Embodiment 1:A kind of feed addictive for promoting growth of animals or poultry and intestinal growth, by 100g cabbage slurries and 0.2g solid-state bacillus subtilises, which are mixed with, to be formed.
Embodiment 2:A kind of feed addictive for promoting growth of animals or poultry and intestinal growth, by 100g cabbage slurries and 0.4g solid-state bacillus subtilises, which are mixed with, to be formed.
Embodiment 3:A kind of feed addictive for promoting growth of animals or poultry and intestinal growth, by 100g cabbage slurries and 0.6g solid-state bacillus subtilises, which are mixed with, to be formed.
Promote the preparation method of the feed addictive of growth of animals or poultry and intestinal growth as follows described in above-described embodiment 1-3:
(1) cabbage slurries are prepared
Fresh cabbage blade is taken, slurry is smashed into, cabbage slurries are obtained, storage is standby;
(2) solid-state bacillus subtilis is prepared
A. slant tube Spawn incubation:Under aseptic technique, Institute of Microorganism, Academia Sinica's strain will be derived from Collection, Bacillus subtilis subspecies, AS1.2163 bacillus subtilis are lined in slant tube culture medium, are connect Kind, 20h is cultivated at 37 DEG C, wherein, slant tube culture medium is beef extract protein free medium;
B. triangular flask expands culture:Load 100mL liquid beef-protein mediums in 250mL triangular flasks, in 0.1MPa, sterilize under the conditions of 121 DEG C 20min, in gnotobasis in inoculation step a gained slant tube strain, then will connect The triangular flask planted is placed on constant-temperature table, and in 37 DEG C, 24h is cultivated under the conditions of 120r/min, obtains bacillus subtilis liquid Triangular flask bacterial;
C. fermentation tank culture:It is less than 40 DEG C of fermentation tank culture medium in temperature, using inoculum concentration as fermentation tank culture medium quality 5% inoculation b step obtained by bacillus subtilis liquid triangular flask bacterial, 37 DEG C of temperature in tank, speed of agitator 120r/ Under conditions of min, ventilation 0.8vvm, 36~48h of fermented and cultured obtains liquid Bacillus subtilis strain, wherein, fermentation The formula of tank culture medium is 1% bean cake powder, and 1% starch, 0.01% potassium dihydrogen phosphate, 0.01% magnesium sulfate puts the culture medium Enter in 50L air blow tank with machinery agitation, under the conditions of 0.1MPa, 121 DEG C, real tank sterilizing 25min;
D. solid substrate fermentation culture:It is less than in temperature in 40 DEG C of fermentation tank solid medium, c steps is inoculated with by weight Liquid Bacillus subtilis strain 10% in rapid, mixing turns uniform, is fitted into aerated koji making pond, in 30 DEG C of ventilation temperature, Under the conditions of humidity 80%~90%, once, 36~48h of fermented and cultured obtains solid state bacterial to every 10~15h stirrings.The solid-state is trained Foster base, which is pressed, is formulated wheat bran 430kg, bean cake powder 50kg, flour 20kg, potassium dihydrogen phosphate 100g, magnesium sulfate 100g, and water 400~ 450kg is formulated, and the solid state rheology is based under the conditions of 0.1MPa, 121 DEG C, and sterilize 30min;
E. solid state bacterial step d obtained is dried under ventilated environment, wherein, the control of 5h wind-warm syndrome is not more than before drying 50 DEG C, 80 DEG C are then progressively warming up to, dries to moisture and is less than or equal to 12%;
F. step e is dried into obtained solid state bacterial to crush, by 0.2mm sieve apertures, and carries out viable count inspection, wherein, Viable bacteria content >=2 × 109CFU/g, miscellaneous bacteria content < 1%.
(3) according to the addition of embodiment 1 to 3, step is added in the cabbage slurries prepared to every 100g steps (1) (2) solid-state bacillus subtilis made from, mixes thoroughly.
According to tests below method, the feed addictive prepared for above-described embodiment 1 to 3 promote poultry growth and Following performance test is carried out in terms of intestinal growth.
1. experimental group is set
Experiment uses single factor test completely randomized design, in Zhengzhou City Zhongmou County Central Asia chicken farm, from 800 health, work 1 close Day-old Broiler Chickens of bold and vigorous, body weight, are randomly divided into 4 groups, every group of 5 repetitions each repeat 40 chickens, and group feeding situation is shown in Table 1.
The group feeding situation of the experiment broiler chicken of table 1
Note:The scale of feeding all same of every group of feeding.
Wherein, basal diet is prepared from by the raw material of following parts by weight:56 parts of corn, 22 parts of dregs of beans, 5 parts of Cottonseed Meal, 3 parts of corn protein powder, 3 parts of vegetable oil, 7 parts of wheat-middlings, 0.3 part of salt, 0.8 part of medical stone, 1.3 parts of stone flour, calcium monohydrogen phosphate 1.5 Part, 1 part of premix, the constituent mass proportioning of the premix are as follows:Lysine 14~16%, methionine 3~4% is combined dimension life Element 3%, cupric sulfate pentahydrate 0.7%, ferrous sulfate monohydrate 4.8%, monohydrate zinc sulphate 3.1%, manganese sulfate monohydrate 4.2%, purity For 1% sodium selenite 1.8%, purity is 1% KI 1.1%, and litter shell powder 25%, surplus is medical stone powder.
2. growth performance index determining method
Experimental period is 42 days, and the state of mind of daily observation broiler chicken, behavior, weigh and record often for 8 points every night during experiment The feed intake of individual repeating groups broiler chicken, daily material consumption;
(1) weigh and record the weight of each age in days of repeating groups broiler chicken 21 on an empty stomach, calculate total increasing of four groups of experiment broiler chicken Weight, average weight, average daily gain, wherein, average daily gain=total augment weight/(experiment number of days × experiment number of elements);
(2) recorded according to feed intake, calculate total feed consumption amount (total feed intake), the average feed consumption rate of every broiler chicken and (be averaged Feed intake) and feed-weight ratio, wherein, feed-weight ratio=total feed consumption amount/total augment weight, average daily gain=total feed intake/(experiment day Number × experiment number of elements).
(3) 21 Day-old Broiler Chickens growth performances are determined according to the method described above, and test result is shown in Table 2.
The age in days production performance result of the test of 2 broiler chicken of table 21
Note:The different letters of data shoulder mark of going together represent significant difference (P < 0.05), and shoulder mark same letter represents that difference does not show Write (P > 0.05).
(4) interpretation of result
As shown in Table 2, compared with control group, experiment II group, test III group of 21 Day-old Broiler Chickens feed intake and body weight it is bright Aobvious raising, daily gain also substantially increase, and significant difference (P<0.05);Test II group, test the material weight of III group of 21 Day-old Broiler Chickens The trend increased than having compared with control group, but the not notable (P of difference>0.05).Thus illustrate, made by embodiment 2, embodiment 3 The feed addictive obtained can promote the conspicuousness of meat chicken production performance to improve.
3. Intestine of Broiler development experiment
(1) test method
In 21 age in days of broiler chicken, in above-mentioned 4 groups of experiment broiler chicken, every group randomly selects 10 and butchers, in Henan animal husbandry economy Institute's physiology dissecting room, each 3cm of position intestinal tube such as clip duodenum, jejunum and ileum, is removed with normal saline flushing Content, is put into 10% formalin fixed preservation, the measure for Intestinal morphology index.
Go bail for keep it is broken-hearted, be dehydrated step by step with ethanol, dimethylbenzene is transparent, embedding machine embedding is cut into 4 μm of section, used H and E is dyed, gummy mounting.
First 40 × and 100 × optical microphotograph Microscopic observation, then use electron microscope observation instead and analyze and gather figure Picture, is shown in Fig. 1;
The survey of histotomy height of naps, Crypt depth and smooth muscle thickness is carried out with the softwares of Motic Images 2000 Amount, measurement result is shown in Table 3.
The age in days Broiler chicks intestinal growth result of the test of table 3 21
Note:The different letters of data shoulder mark of going together represent significant difference (P < 0.05), and shoulder mark same letter represents that difference does not show Write (P > 0.05).
(2) interpretation of result
Analysis chart 1 and table 3 are understood, compared with control group, II group of experiment and experiment III group of Duodenal villi height, 12 Duodenum 12 smooth muscle thickness, jejunum villi are highly significantly improved, significant difference (P<0.05);Test I group and II group of jejunum of experiment is put down Sliding flesh thickness is significantly improved compared with control group, significant difference (P<0.05);Compared with control group, test group ileum fine hair is high Degree, Crypt depth are significantly improved, significant difference (P<0.05).
Therefore, the feed addictive that prepared by above-mentioned three kinds of embodiments can be effectively facilitated Intestine of Broiler development, especially in fact The feed addictive of the offer of example 3 is provided.
4. broiler chicken cecal content microorganism species structure detection
(1) test method
Select 21 Day-old Broiler Chickens that body weight is close in 10 groups from control group and in testing III group respectively to slaughter, cecal ligation Two ends, are transferred to superclean bench after cutting, take out cecal content sample, and liquid nitrogen flash freezer delivers to Beijing Nuo Hezhi sources company, The cecum microorganisms flora structure of community in cecum flora diversity detection, category level is carried out, result of the test is referring to table 4 and Fig. 2.
The age in days control group of table 4 21 belongs to cecum microorganisms flora structure of community in level with test group broiler chicken
Pseudomonas type Control group content (%) Test group content (%)
Bacteroides Bacteroides 0.47 34.17
Clostridium leptum belongs to Faecalibacterium 1.17 3.97
Lachnoclostridium 13.8 12.23
Rare Mycosphaerella Subdoligranulum 7.23 2.0
Blautia 2.27 2.63
Clostridiaceae Ruminococcus Ruminococcaceae_UCG-014 6.17 5.97
Eisenbergiella 6.67 4.47
Escherichia Escherichia Shigella 0.5 2.87
Eubacterium coprostanoligenes group 4.87 1.6
Fusobacterium clostruidium Anaerotruncus 5.1 2.17
Other Pseudomonas 51.75 27.92
(2) interpretation of result
From table 4 and Fig. 2, in category level, the of a relatively high Pseudomonas of content is in control group cecum microorganisms group: Lachnoclostridium (13.8%), rare Mycosphaerella (7.23%), Clostridiaceae Ruminococcus (6.17%), Eisenbergiella (6.67%), Eubacterium coprostanoligenes group (4.87%), fusobacterium shuttle Shape bacillus (5.1%);The of a relatively high Pseudomonas of test group content is:Bacteroides (34.17%), Lachnoclostr Idium (12.23%), Clostridiaceae Ruminococcus (5.97%), Eisenbergiella (4.47%), Clostridium leptum category (3.97%), Escherichia Shigella (2.87%).Also, it is Bacteroides in test group, tender compared to control group Fusobacterium, Escherichia proportion are significantly improved.
Domestic and international numerous studies show that bacteroid is helping host to decompose polysaccharide raising nutrient utilization, accelerating intestinal mucosa Vascularization and developing immune system, maintain to play the role of in terms of intestinal microecology balance it is very important, especially To the utilization aspect of polysaccharide.
Clostridium leptum category is one of strain of a small number of energy synthetic butyric acid salt.Butyrate is intestinal epithelial tissue cytothesis and repaiied Multiple Major Nutrient material, energetic supersession and development to intestinal epithelial cell play very important effect, can suppress enteron aisle Interior pathogen or the growth of spoilage organisms, promote development and the propagation of probiotics (such as Bifidobacterium, lactic acid bacteria), the generation of these bacterium Thank active healthy closely related with host, butyrate is in protection colon cell, prevention colonic diseases such as inflammatory enteritis, colon cancer In terms of also tool have certain effect.
Therefore, the result of the test of table 4 shows, the feed addictive containing embodiment 3 can influence enteric microorganism to belong to water Flat upper structure of community, hence it is evident that Bacteroides in enteron aisle, Clostridium leptum category are improved in the ratio shared by intestinal microflora, to dimension Shield animal and bird intestines health plays an important role.
5. the III group of difference species analysis of control group and experiment
Using LEfSe analysis methods, the cecum microorganisms to testing III group and the Day-old Broiler Chickens of control group 21 in above-mentioned steps 4 Flora group carries out group difference species analysis, and draws test group and the notable species analysis chart of control group group difference, referring to Fig. 3 and Fig. 4.
LEfSe is a kind of software for being used to find higher-dimension biological label and disclose genome signature.LEfSe analyses can The Biomarker with significant difference, the i.e. significant species of group difference are found between group and group.
LDA Distribution values block diagram and cladogram are drawn using LEfSe statistical result.Wherein, LDA Distribution values column The species that LDA Score are more than setting value (being defaulted as 4) are illustrated in figure, i.e., there is significant difference between group Biomarker, illustrates the significant species of abundance difference in different groups, and the influence of the length representative difference species of block diagram is big It is small.
In cladogram, the circle radiated from the inside to the outside is represented by door to the category level for belonging to (or kind).Not A classification under the level, small circle diameter and relative abundance size are represented with each small circle in category level It is proportional.Colour principle:Species without significant difference are uniformly colored as yellow, and difference species Biomarker, which is followed, group to be carried out Color, red node represents the microbe groups played an important role in red group, and green node is represented in green group The microbe groups played an important role.
Analyzed from Fig. 3 and Fig. 4, in cecum microorganisms flora group group difference species, control group plays an important role Microbe groups be cud Cordycepps (Ruminococcaceae), clostridium guiding principle (Clostridiales), clostridium mesh (Clostridia);It is Bacteroides (Bateroidaceae), bacteroid mesh to test III group of microbe groups played an important role (Bateroidales), Bacteroidetes (Bateroidia), Bacteroides (Bacteroides), enterobacteriaceae And enterobacteria mesh (Enterobacteriales) (Enterobacteriaceae).Further demonstrate what is provided by embodiment 3 Feed addictive can improve bacteroid ratio and richness shared in intestinal microflora group, to improving livestock and poultry intestines Road microorganism species structure of community plays an important role.
In summary, the present invention will contain the wild cabbage slurries of bacillus subtilis as feed addictive, to promoting enteron aisle There is positive meaning in terms of development, growth of animals or poultry and improvement intestinal microflora.

Claims (10)

1. a kind of feed addictive for promoting growth of animals or poultry and intestinal growth, it is characterised in that by mass ratio be 500:(1~3) Cabbage slurries and solid-state bacillus subtilis be made.
2. the feed addictive of promotion growth of animals or poultry according to claim 1 and intestinal growth, it is characterised in that described solid State bacillus subtilis number of viable is 0.5~2 × 109 CFU/g。
3. promoting the preparation method of the feed addictive of growth of animals or poultry and intestinal growth as claimed in claim 1, its feature exists In, fresh cabbage blade is crushed and broken into after slurry, addition 0.2~0.6g solid-state withered grass in every 100g cabbage slurries Bacillus, stirs.
4. the preparation method of the feed addictive of promotion growth of animals or poultry according to claim 3 and intestinal growth, its feature It is, the solid-state bacillus subtilis is made by following methods:
(1)Test tube strains culture:Under gnotobasis, bacillus subtilis is inoculated with slant tube culture medium, in 37 DEG C of conditions Lower culture 20h;
(2)Expand culture:Aseptic inoculation step on liquid culture medium after the 20min that sterilized with conventional method(1)Obtained examination Pipe strain, expands culture 24h on 37 DEG C, 120r/min constant-temperature table;
(3)Fermentation tank culture:Fermentation tank culture medium is cooled to less than 40 DEG C, inoculation step after conventional method sterilizing 25min (2)Culture gained strain, under conditions of speed of agitator 120r/min, ventilation 0.8vvm, 36~48h of fermented and cultured, wherein, Inoculum concentration is the 5% of fermentation tank culture medium quality;
(4)Solid substrate fermentation culture:By solid medium after conventional sterilant 30min but to less than 40 DEG C, connect by weight Plant step(3)The 10% of gained strain, stirs, and is fitted into aerated koji making pond, in 30 DEG C of ventilation temperature, humidity 80%~90% Under conditions of, once, 36~48h of fermented and cultured obtains solid-state bacillus subtilis to every 10~15h stirrings;
(5)By step(4)Obtained solid state bacterial crushed after being dried, crosses 0.2mm sieve apertures, and viable bacteria content >=2 during microscopy ×109CFU/g, miscellaneous bacteria content < 1%.
5. the preparation method of the feed addictive of promotion growth of animals or poultry according to claim 4 and intestinal growth, its feature It is, in step(1)In, the bacillus subtilis derives from DSMZ of Institute of Microorganism, Academia Sinica, withered Careless subtilis subspecies, AS1.2163.
6. the preparation method of the feed addictive of promotion growth of animals or poultry according to claim 4 and intestinal growth, its feature It is, in step(3)In, the formula of the fermentation tank culture medium is 1% bean cake powder, 1% starch, 0.01% potassium dihydrogen phosphate, 0.01% magnesium sulfate.
7. the preparation method of the feed addictive of promotion growth of animals or poultry according to claim 4 and intestinal growth, its feature It is, in step(4)In, the formula of the solid medium is wheat bran 430kg, bean cake powder 50kg, flour 20kg, biphosphate Potassium 100g, magnesium sulfate 100g, 400~450kg of water.
8. the preparation method of the feed addictive of promotion growth of animals or poultry according to claim 4 and intestinal growth, its feature It is, in step(5)In, by step(4)During obtained solid state bacterial aeration-drying:5h before drying, control Feng Wen≤50 DEG C, so 80 DEG C are progressively warming up to afterwards, are dried to Shui Fen≤12%.
9. promote the feed addictive of growth of animals or poultry and intestinal growth as claimed in claim 1 or prepared by claim 3 Promote the feed addictive of growth of animals or poultry and intestinal growth as the application of additive for farm animal feed.
10. application according to claim 9, it is characterised in that the addition of the feed addictive is per kilogram feed 10~40g of middle addition.
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