CN105248877A - Micro-ecological compound preparation with immunological enhancement function - Google Patents
Micro-ecological compound preparation with immunological enhancement function Download PDFInfo
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- CN105248877A CN105248877A CN201510577909.8A CN201510577909A CN105248877A CN 105248877 A CN105248877 A CN 105248877A CN 201510577909 A CN201510577909 A CN 201510577909A CN 105248877 A CN105248877 A CN 105248877A
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Abstract
The invention discloses a micro-ecological compound preparation with an immunological enhancement function. The compound preparation is prepared from the following raw materials, by weight, 1-3 parts of lactococus lactis powder, 30-50 parts of isomaltose hypgather, 10-15 parts of yeast trace elements, 10-20 parts of NSP enzyme, 8-12 parts of radix astragali polysaccharide and 15-25 parts of fermented wheat bran/soybean meal. The fermented wheat bran/soybean meal is prepared from wheat bran and soybean meal through combined fermentation of lactobacillus plantarum and pediococcus pentosaceus. The mass ratio of wheat bran to soybean meal used for fermentation is 4:1. The yeast trace elements are a mixture of yeast chromium and yeast iron according to a mass ratio of (5-7): (5-7). The micro-ecological compound preparation can enhance organism cellular immunity, humoral immunity and mucosal immunity functions, and has obvious promotion effects on non-specific immune and specific immune response.
Description
Technical field
The present invention relates to a kind of micro-ecological compound preparation with immune enhancing function.
Background technology
Along with the development of China's poultry industry scale; abuse feeding antibiotic, content of beary metal exceed standard, the cross-infection of many cause of diseases and be with poison for a long time, to batter the phenomenons such as vaccine more and more serious; cause immunity of livestock low; immunosupress generally occurs; then cause that growth of animals or poultry is obstructed, the phenomenon of the concurrent or scabies secondary infection of vaccine immunity failure, various diseases; there is outburst and popular (showing as the variation quickening of pathogen and the enhancing of drug resistance) in communicable disease, causes huge economic loss to aquaculture.Therefore, develop a have natural, have no side effect, safe and reliable Immune-enhancing effect product is to strengthening livestock and poultry body healths level, reducing the use of clinical antibiotics, promoting that food security is significant.
Summary of the invention
For above-mentioned the deficiencies in the prior art, the object of this invention is to provide a kind of micro-ecological compound preparation with immune enhancing function.This micro-ecological compound preparation can strengthen the cellular immunity of body, humoral immunity and mucosal immunity function, all has obvious facilitation to the nospecific immunity of body and specific immune response.
For achieving the above object, the present invention adopts following technical proposals:
There is a micro-ecological compound preparation for immune enhancing function, be made up of the raw material of following weight portion:
Lactococcus bacterium powder 1-3 part, oligoisomaltose 30-50 part, yeast trace element 10-15 part, NSP enzyme 10-20 part, astragalus polyose 8-12 part, fermentation wheat bran/dregs of beans 15-25 part;
Described fermentation wheat bran/dregs of beans is formed by Lactobacillus plantarum and Pediococcus pentosaceus combined ferment wheat bran and dregs of beans, and the mass ratio for the wheat bran that ferments and dregs of beans is 4:1.
Concrete, the preparation method of described fermentation wheat bran/dregs of beans is: regulate the moisture of the wheat bran for fermenting and dregs of beans to be 38-40% (mass percent), inoculation Lactobacillus plantarum and Pediococcus pentosaceus mixed bacteria, the inoculum concentration of mixed bacteria is 1 × 10
5cfu/g, wherein, the ratio of Lactobacillus plantarum and Pediococcus pentosaceus inoculum concentration is 1:1, and normal temperature fermentation, to obtain final product.With do not ferment before with adopt conventional probiotics fermention to compare, the present invention adopts Lactobacillus plantarum and Pediococcus pentosaceus mixed culture fermentation, and the fermentation wheat bran/dregs of beans after mixed culture fermentation is rich in a large amount of beneficial bacterium and metabolite thereof, and total viable count can reach 1 × 10
9cfu/g, is rich in organic acid, and comprise lactic acid, acetic acid, propionic acid etc., pH is low to moderate 4.2, effectively can suppress the breeding of harmful bacteria, improves the palatability of compound preparation simultaneously.
The Lactobacillus plantarum used that ferments is Lactobacillus plantarum LP (Lactobacillusplantarum), be preserved in China typical culture collection center on 06 21st, 2010, its deposit number is CCTCCM2010150, is documented in another patent of invention of applicant " strain can be used for lactic acid bacteria and the application thereof of biological preservation " (CN101914475B); Pediococcus pentosaceus used of fermenting is Pediococcus pentosaceus PP (PediococcuspentosaceusPP), be preserved in China typical culture collection center on 02 22nd, 2012, its deposit number is CCTCCNO:M2012029, is documented in another patent of invention of applicant " a kind of lactic acid bacteria semisolid fermentation product and technique improving growth of animal performance " (CN102599348B).
Preferably, this has the micro-ecological compound preparation of immune enhancing function, is made up of the raw material of following weight portion:
2 parts, Lactococcus bacterium powder, oligoisomaltose 40 parts, yeast trace element 12 parts, NSP enzyme 15 parts, astragalus polyose 10 parts, fermentation wheat bran/dregs of beans 21 parts.
Described yeast trace element is by yeast chromium and yeast iron in mass ratio (5-7): (5-7) mixes; Preferably, described yeast trace element by yeast chromium and yeast iron in mass ratio 7:5 mix.In the present invention, the ratio of yeast chromium and yeast iron addition has considered the production cost of compound preparation and the two toatl proportion in the product and optimized and obtain.
Chromium content in described yeast chromium is 2000mg/kg, and in yeast iron, the content of iron is 2000mg/kg.
The active ingredient of described NSP enzyme comprises: dextranase >=3,000,000 U/g, cellulase >=70000U/g, pectase >=20000U/g, mannase >=5000U/g.
Preferably, described Lactococcus bacterium powder is Pediococcus pentosaceus (PediococcuspentosaceusPP) bacterium powder; This Pediococcus pentosaceus PP was preserved in China typical culture collection center on 02 22nd, 2012, and its deposit number is CCTCCNO:M2012029, is documented in Chinese patent application CN102599348B.
Astragalus polyose: polyoses content 50-70%, purchased from Tianjin Hua Long Medicines and Health Product Co., Ltd, authentication code is the accurate word Z20040085 of traditional Chinese medicines.
Present invention also offers the preparation method that this has the micro-ecological compound preparation of immune enhancing function, each raw material is mixed, to obtain final product.
That prepares has in the micro-ecological compound preparation of immune enhancing function, and the viable count of lactic acid bacteria is more than or equal to 1 × 10
9cfu/g.
This micro-ecological compound preparation with immune enhancing function can direct drinking water administration; Also can, with it for main ingredient, other auxiliary materials be coordinated to prepare the medicine of the various formulations such as tablet, pulvis, granule, capsule, oral agents.
In micro-ecological compound preparation of the present invention, lactic acid bacteria is the probio that a class has immune enhancing function, and it can adjust gastrointestinal bacterial flora balance, growth promoting effects, regulate the immunologic function of body, improve the resistance against diseases of body, and Lactococcus has better tolerance compared with bacillus; Oligoisomaltose can promote the growth and breeding of probio in animal body, for probio provides nutrient source; The content of trace element in body is less, and it can be combined with protein or other organic groups, can form the large biological molecules such as enzyme, hormone and vitamin; Yeast trace element is yeast and the combination of trace element, is independently absorbed and transform by yeast in growth course to various element, has advantages such as absorbing soon, have no side effect; NSP enzyme can reduce intestinal contents viscosity, improves the activity of animal endogenous digestive ferment, reduces harmful intestinal tract microorganism, and degraded NSP becomes oligosaccharides, improves the immunity function of animal; Astragalus polyose can induce body to produce interferon, viral interference copying in body, improve the immunologic function of body, promote the enhancing of hepatocuprein (SOD), activity of glutathione peroxidase, and can strengthen and stimulate the generation of lymphocyte and RE confluent monolayer cells, strengthen the phagocytic function of RE confluent monolayer cells and macrophage, and have well promotion and regulating action to body fluid, mucous membrane and cellular immunity; Fermentation wheat bran/dregs of beans is rich in a large amount of beneficial bacterial metabolic product, comprises lactein and organic acid substance, can suppress the breeding of harmful bacteria while improving local flavor.And, also there is synergy between each component in micro-ecological compound preparation of the present invention: Lactococcus, oligoisomaltose, NSP enzyme, astragalus polyose and fermentation wheat bran/dregs of beans organically combines, acting in conjunction, by increasing the quantity of beneficial bacterium, improve digestive enzyme activity simultaneously, suppress the breeding of pathogenic microorganism, improve palatability, finally reach the effect improving immunity of organisms.
Beneficial effect of the present invention:
(1) micro-ecological compound preparation of the present invention is by beneficial microbe lactic acid bacteria, prebiotics oligoisomaltose, associating yeast trace element and NSP enzyme, increase the quantity of intestinal beneficial Bacillus acidi lactici, suppress the growth and breeding of the conditioned pathogens such as Escherichia coli, promote Intestinal Mucosal Immunization, significantly can promote the growth of broiler chicken immune organ spleen, the bursa of farbricius simultaneously, in raising serum, newcastle epidemic disease antibody tires, promote the cellular immunity of body and humoral immunity, thus to the nospecific immunity of body and specific immune response, all there is obvious facilitation.
(2) micro-ecological compound preparation of the present invention can improve meat chicken production performance, improves serum immunoglobulin G (IgG) content, thus strengthens humoral immunity; Increase peripheral lymphocyte proliferation index, thus strengthen cellular immunity; Improve the content of intestinal contents secretory immunoglobulin A (sIgA), thus strengthen mucosal immunity; And then confirm that this micro-ecological compound preparation is strengthening the effect on Immune Function In Animals.
(3) the present invention strengthens Immune Function In Animals to provide a brand-new thinking, changes the single channel relying on chemical agent to strengthen immunologic function, and then provides technical foundation for the exploitation of a series of micro ecological immunopotentiator.
Accompanying drawing explanation
Fig. 1 is that in embodiment 8, various dose micro-ecological compound preparation affects comparison chart to broiler chicken index and spleen index;
Fig. 2 is that in embodiment 8, various dose micro-ecological compound preparation affects comparison chart to broiler chicken bursal index.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further illustrated, should be noted that following explanation is only to explain the present invention, not limiting its content.
Experimental technique in following embodiment, if no special instructions, is conventional method.Material used in following embodiment, reagent etc., if no special instructions, be conventional material, reagent, all can obtain from commercial channels.
Pediococcus pentosaceus PP (be preserved in China typical culture collection center on 02 22nd, 2012, its deposit number is CCTCCNO:M2012029, is documented in Chinese patent application CN102599348B) in following embodiment; Skimmed milk power (trade name: feed grade skimmed milk power; Manufacturer: Harbin Sheng Meilong commerce and trade Co., Ltd); Oligoisomaltose (trade name: oligoisomaltose; Marque: IMO-900; Manufacturer: Jinan Wei Chen Bioisystech Co., Ltd); Yeast iron (trade name: feeding level ferrum-rich saccharomyces cerevisiae; Marque: SY34; Manufacturer: Zhejiang Senyo Biotech Co., Ltd.); Yeast chromium (trade name: feeding level Cr-enriched yeast; Marque: SY33.702; Manufacturer: Zhejiang Senyo Biotech Co., Ltd.); NSP enzyme (trade name: compound NSP enzyme; Article number: 2015060802; Manufacturer: Shandong Baolai-leelai Bio-engineering Co., Ltd.); Astragalus polyose (trade name: astragalus polyose; Marque: 1254; Manufacturer: Hua Xu bio tech ltd, Shanghai); Fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) is formed by Lactobacillus plantarum LP and Pediococcus pentosaceus PP combined ferment, and fermentation condition is: regulate the moisture of the wheat bran for fermenting and dregs of beans to be 38%-40%, inoculum concentration 1 × 10
5cfu/g, Lactobacillus plantarum: Pediococcus pentosaceus is 1:1, normal temperature fermentation 72h, pH4.2.
Embodiment 1: the preparation with the micro-ecological compound preparation of immune enhancing function
Formula: 2 parts, Pediococcus pentosaceus bacterium powder, oligoisomaltose 40 parts, yeast chromium 5 parts, yeast iron 7 parts, NSP enzyme 15 parts, astragalus polyose 10 parts, fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) 21 parts.
Preparation method: Pediococcus pentosaceus PP obtains zymotic fluid through fermentation process, then adds 6% (mass percent) skimmed milk power, makes Pediococcus pentosaceus bacterium powder by freeze drying; Composite oligoisomaltose, yeast chromium, yeast iron, NSP enzyme, astragalus polyose and fermentation wheat bran/dregs of beans again, composite mode is that convention stir mixing is composite.
In the compound preparation of preparation, viable count of lactobacillus is more than or equal to 1 × 10
9cfu/g.
Embodiment 2: the preparation with the micro-ecological compound preparation of immune enhancing function
Formula: 2 parts, Pediococcus pentosaceus bacterium powder, oligoisomaltose 40 parts, yeast chromium 7 parts, yeast iron 5 parts, NSP enzyme 15 parts, astragalus polyose 10 parts, fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) 21 parts.
Preparation method is with embodiment 1.
In the compound preparation of preparation, viable count of lactobacillus is more than or equal to 1 × 10
9cfu/g.
Embodiment 3: the preparation with the micro-ecological compound preparation of immune enhancing function
Formula: 2 parts, Pediococcus pentosaceus bacterium powder, oligoisomaltose 40 parts, yeast chromium 6 parts, yeast iron 6 parts, NSP enzyme 15 parts, astragalus polyose 10 parts, fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) 21 parts.
Preparation method is with embodiment 1.
In the compound preparation of preparation, viable count of lactobacillus is more than or equal to 1 × 10
9cfu/g.
Embodiment 4: the preparation with the micro-ecological compound preparation of immune enhancing function
Formula: 1 part, Pediococcus pentosaceus bacterium powder, oligoisomaltose 50 parts, yeast chromium 5 parts, yeast iron 5 parts, NSP enzyme 20 parts, astragalus polyose 8 parts, fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) 25 parts.
Preparation method is with embodiment 1.
In the compound preparation of preparation, viable count of lactobacillus is more than or equal to 1 × 10
9cfu/g.
Embodiment 5: the preparation with the micro-ecological compound preparation of immune enhancing function
Formula: 3 parts, Pediococcus pentosaceus bacterium powder, oligoisomaltose 30 parts, yeast chromium 7 parts, yeast iron 7 parts, NSP enzyme 10 parts, astragalus polyose 12 parts, fermentation wheat bran/dregs of beans (wheat bran: dregs of beans=4:1) 15 parts.
Preparation method is with embodiment 1.
In the compound preparation of preparation, viable count of lactobacillus is more than or equal to 1 × 10
9cfu/g.
Embodiment 6: micro-ecological compound preparation is on the impact of the production performance of laying hen, Immune Organs Index and serum antibody titres to newcastle disease virus
1 test material and method
1.1 material
Micro-ecological compound preparation prepared by embodiment of the present invention 1-3.
1.2 experimental design and daily ration form
Experimental design: 240 1 blue brown laying hens in ages in days sea that body weight is even, health status is substantially identical are divided into 4 groups at random, often organize 3 repetitions, each repetition 20, namely I group: control group, basal diet of feeding; II group: feed on the basis of basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 1 preparation by 1 ‰ (mass percent) dosage drinking-water; III group: feed on the basis of basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 1 ‰ (mass percent) dosage drinking-water; IV group: feed on the basis of basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 3 preparation by 1 ‰ (mass percent) dosage drinking-water, the equal free choice feeding of each group.
1.3 feedings and managements: test chicken adopts mode of raising in cages, free choice feeding and drinking-water, illumination in house, temperature and humidity require to control in strict accordance with conventional feeding and management.Change every day before food changes water and carry out crib and water fountain virus killing sterilization.Duration of test records the health status of each group of food consumption and test chicken in detail.All chickens in 7d, 21d, 28d immunity newcastle disease vaccine, 14d immunity bursa of fabricius vaccine.
1.4 testing indexs and method:
(1) production performance is weighed to the laying hen after fasting (freely drinking water) 12h in units of repeating respectively at 14,21,28,35 ages in days, record day feed quantity, residue doses and loss doses.Calculate each stage average daily gain, average daily ingestion amount and feed-weight ratio.
(2) Immune Organs Index is weighed after randomly drawing 10 laying hen fasting (freely drinking water) 12h respectively at 14,28 and 35 ages in days from each repetition, arteria carotis sacrificed by exsanguination, spleen, the bursa of farbricius are won in aseptic dissection immediately, fresh weight is claimed, Computation immunity shoot formation after rejecting fat.
Immune Organs Index (mg/g)=immune organ weight/live body weight
(3) ND_HI antibody is tired respectively at 14,21,28,35 ages in days, randomly draws 10 chickens, wing venous blood collection, separation of serum, carry out doubling dilution from often repeat, and measures antibody titres to newcastle disease virus in serum with Hemagglutination Inhibition.Antibody titres to newcastle disease virus Log
2x represents.
2 results
2.1 micro-ecological compound preparation are on the impact of laying hen growth performance
From table 1 data: average daily gain aspect, in whole process of the test, compared with control group, micro-ecological compound preparation group (II group, III group, IV group) average daily gain is all higher than control group, feed-weight ratio is all lower than control group, all in all, the micro-ecological compound preparation that prepared by embodiment 2 significantly improves laying hen average daily gain, reduces feedstuff-meat ratio.
Table 1 micro-ecological compound preparation is on the impact of laying hen growth performance
Note: the different lowercase alphabet of colleague's data shoulder mark shows significant difference (P ﹤ 0.05), and identical lowercase alphabet differential is different not significantly (P ﹥ 0.05).Following table is same.
2.2 micro-ecological compound preparation are on the impact of laying hen Immune Organs Index
From table 2 data: during 14d, III group of index and spleen index is higher than control group; During 28d, III group of index and spleen index significantly exceeds control group 7.75%; During 35d, except II group, each test group is all higher than control group, wherein the highest with the index and spleen index of III group.All in all, the micro-ecological compound preparation that prepared by embodiment 2 is improving effect stability in laying hen index and spleen index.
Table 2 micro-ecological compound preparation is on the impact (unit: mg/g) of laying hen index and spleen index
From table 3 data: during 14d, except IV group, the bursal index of all the other each test group is all higher than control group, and III group exceeds control group 26.94%, has significant difference (P ﹤ 0.05); During 28d, III group of bursal index is higher than control group; During 35d, the bursal index of III and IV group is all higher than control group.All in all, the micro-ecological compound preparation that prepared by embodiment 2 is improving Be very effective in laying hen bursal index.
Table 3 micro-ecological compound preparation is on the impact (unit: mg/g) of laying hen bursal index
2.3 micro-ecological compound preparation are on the impact of Laying-hen Serum antibody titres to newcastle disease virus
From table 4 data: when 14d, 21d, 28d, 35d, micro-ecological compound preparation group serum antibody titres to newcastle disease virus is all significantly higher than control group, and particularly III group (drinking micro-ecological compound preparation group prepared by embodiment 2) exceeds control group 63.77%, 23.38%, 33.79%, 52.03% respectively.All in all, the micro-ecological compound preparation that prepared by embodiment 2 is improving Be very effective in serum antibody titres to newcastle disease virus.
Table 4 micro-ecological compound preparation is on the impact of Laying-hen Serum ND antibody titer
Brief summary: by finding that to interpretation of result the micro-ecological compound preparation of the embodiment of the present invention 2 preparation is to the effect of increasing significantly of laying hen Immune Organs Index (index and spleen index, bursal index) tool, to Laying-hen Serum newcastle epidemic disease antibody, there is significant humidification, show micro-ecological compound preparation Be very effective in the immunologic function improving laying hen prepared by the embodiment of the present invention 2.
Embodiment 7: the application test of micro-ecological compound preparation on broiler chicken
1 test material and method
1.1 material
Micro-ecological compound preparation prepared by the embodiment of the present invention 2; Complete feed, purchases Tai'an six and feed; Ewcastle disease (ND) LaSota IV is vaccine, purchased from Shandong animal health-care product factory; IPMI1640 culture medium, purchased from GIBCO company; Lymphocyte separation medium, purchased from Tianjin Hao ocean biological products science and technology limited Company; CCK-8 kit, purchased from Beijing Solarbio company; Concanavalin A (ConA) is Sigma Products; Hyclone: purchased from Chinese holly company; 96 porocyte culture plates are purchased from Corning company.
1.2 experimental design and daily ration form
Experimental design: 240 body weight is even, health status is substantially identical 1 age in days white meat-type chickens are divided into 4 groups at random, often organizes 3 repetitions, each repetition 20, namely I group: control group, basal diet of feeding; II group: on the basis of feeding basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 0.5 ‰ (mass percent) dosage drinking-water; III group: on the basis of feeding basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 1 ‰ (mass percent) dosage drinking-water; IV group: on the basis of feeding basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 3 ‰ (mass percent) dosage drinking-water, the equal free choice feeding of each group.
1.3 feedings and managements: test chicken adopts mode of raising in cages, free choice feeding and drinking-water, illumination in house, temperature and humidity require to control in strict accordance with conventional feeding and management.Change every day before food changes water and carry out crib and water fountain virus killing sterilization.Duration of test records the health status of each group of food consumption and test chicken in detail.All chickens in 7d, 21d, 28d immunity newcastle disease vaccine, 14d immunity bursa of fabricius vaccine.
1.4 testing indexs and method:
(1) production performance respectively at 14,21,28,35,42 ages in days by repeat in units of the broiler chicken after fasting (freely drink water) 12h is weighed, and precise and record day feed quantity, remain doses and lose doses.Calculate each stage average daily gain, average daily ingestion amount and feed-weight ratio.
(2) broiler chicken after Immune Organs Index randomly draws 5 fasting (freely drinking water) 12h respectively at 14,28 and 42 ages in days from each repetition is weighed, arteria carotis sacrificed by exsanguination, spleen, the bursa of farbricius are won in aseptic dissection immediately, fresh weight is claimed, Computation immunity shoot formation after rejecting fat.
Immune Organs Index (mg/g)=immune organ weight/live body weight
(3) ND_HI antibody is tired respectively at 14,21,28,35,42 ages in days, randomly draws 5 chickens, wing venous blood collection, separation of serum, carry out doubling dilution from often repeat, and measures antibody titres to newcastle disease virus in serum with Hemagglutination Inhibition.Antibody titres to newcastle disease virus Log
2x represents.
(4) mensuration of cecal content S-IgA (sIgA) content randomly draws 5 chickens respectively at 14d, 28d, 42d from each repeating groups, put to death, dissect, cecal content 1g is got in sterile working, adopts chicken secretory immunoglobulin A (sIgA) ELISA kit to measure the content of sIgA in intestinal juice.
(5) peripheral lymphocyte proliferation experiment randomly draws 5 chickens respectively at 14d, 28d, 42d from each repeating groups, take a blood sample under getting aseptic liquaemin anticoagulant blood-collecting pipe broiler chicken wing 1mL, lymphocyte separation medium is separated lymphocyte, add the IPMI1640 culture medium suspension cell of 10% hyclone, counted under microscope, adjustment cell concentration is 5 × 10
6.The cell suspension of every hole 100 μ L in 96 orifice plates, add the culture medium of 100 μ L containing concanavalin A in addition, or the culture medium of 100 μ L contrasts, every sample repeats 3 holes.Tissue Culture Plate is put in 37 DEG C, cultivates 60h in the incubator of 5% gas concentration lwevel.Every hole is taken out 100 μ L supernatants and is discarded, and adds the CCK-8 kit of 10 μ L, places incubator, every 2h, measures its OD
450value.
(6) mensuration of the Total albumen content is respectively at 14,21,28,35,42 ages in days, randomly draws 5 chickens, wing venous blood collection, separation of serum from often repeat, and adopts total protein detection reagent box to detect.
(7) mensuration of gut flora randomly draws 5 chickens respectively at 14,28,42 ages in days from often repeat, slaughter through venesection after weighing, aseptic separation caecum stage casing, gets content 1g, adopts slat chain conveyor counting method to count enteron aisle lactobacillus and coliform count respectively.Microflora represents with the logarithm of total number of bacterial colonies contained by every gram of intestinal contents [(lgCFU)/g].
(8) serum immunoglobulin IgG content detection randomly draws 6 chickens respectively at 14d, 28d, 42d in each repeating groups, wing venous blood collection, separation of serum, adopts Chicken immunoglobulin ELISA kit to measure the content of IgG in serum.
2 results
2.1 micro-ecological compound preparation are on the impact of meat chicken production performance
From table 5 data: average daily gain aspect, in whole process of the test, compared with control group, each compared with other group of III and IV group of average daily gain has rising trend, but difference is not remarkable; Show that micro-ecological compound preparation prepared by the embodiment of the present invention 2 can improve broiler chicken daily gain according to 1 ‰ and 3 ‰ dosage interpolations.
Table 5 micro-ecological compound preparation is on the impact of broiler chicken average daily gain
2.2 micro-ecological compound preparation are on the impact of broiler chicken immune organ
As shown in Table 6: during 14 age in days, II, III, IV group of index and spleen index exceeds control group 17.46%, 17.46%, 20.63% respectively, but difference is not all significantly (P>0.05); During 28 age in days, test group index and spleen index is all higher than control group, and wherein III group exceeds control group 30.67%, and IV group significantly exceeds control group 65.33% (P<0.05); During 42 age in days, II, III, IV group of index and spleen index exceeds control group 45.04%, 48.65%, 54.05% respectively, has significant difference (P<0.05).Result of the test shows, the growth of 1 ‰ and 3 ‰ dosage to broiler chicken immune organ spleen of micro-ecological compound preparation prepared by the embodiment of the present invention 2 has significant facilitation.
Table 6 micro-ecological compound preparation is on the impact (unit: mg/g) of broiler chicken index and spleen index
As shown in Table 7: during 14 age in days, II, III, IV group of bursal index is all higher than control group, and particularly IV group of broiler chicken bursal index is the highest, exceeds control group 31.25%, has significant difference (P<0.05); During 28 age in days, test group bursal index is all higher than control group, and particularly IV group of broiler chicken bursal index exceeds control group 42.24%, has significant difference (P<0.05); During 42 age in days, except II group, each test group bursal index higher than control group, but without conspicuousness (P>0.05).Result of the test shows that the growth of 1 ‰ and 3 ‰ dosage to the broiler chicken immune organ bursa of farbricius of micro-ecological compound preparation prepared by the embodiment of the present invention 2 has significant facilitation.
Table 7 micro-ecological compound preparation is on the impact (unit: mg/g) of broiler chicken bursal index
2.3 micro-ecological compound preparation are on the impact of serum of broilers ND antibody titer
As shown in Table 8: 14,21,28,35,42 ages in days, in each test group serum, antibody titres to newcastle disease virus is all higher than control group, during 21 age in days, test III group, IV group serum antibody titres to newcastle disease virus and exceed control group 38.02%, 42.59% respectively, there is significant difference (P<0.05); During 28 age in days, test IV group of serum antibody titres to newcastle disease virus and exceed control group 46.77%; Result of the test shows that 1 ‰ and 3 ‰ dosage of micro-ecological compound preparation prepared by the embodiment of the present invention 2 can significantly improve tiring of newcastle epidemic disease antibody in serum.
Table 8 micro-ecological compound preparation is on the impact (Log of serum N D antibody titer
2x)
2.4 micro-ecological compound preparation are on the impact of broiler chicken cecal content sIgA
As shown in Table 9: during 14 age in days, test group IV cecal content sIgA content is higher than control group; During 28 age in days, test group cecal content sIgA content is all higher than control group, but difference is not significantly (P>0.05); During 42 age in days, each test group cecal content sIgA content, all higher than control group, is wherein tested IV group of cecal content sIgA content the highest, is exceeded control group 21.50%, significant difference (P<0.05).Result of the test shows the content of the raising cecal content sIgA of 3 ‰ dosage energy conspicuousnesses of micro-ecological compound preparation prepared by the embodiment of the present invention 2.
Table 9 micro-ecological compound preparation is on the impact (pg/mL) of cecal content sIgA
2.5 micro-ecological compound preparation are on the impact of broiler chicken peripheral lymphocyte proliferation index
As shown in Table 10: 14,28,42 age in days time, high dose group (testing IV group) peripheral lymphocyte proliferation index exceeds control group 7.54%, 43%, 7.84% respectively, has significant difference (P<0.05); Low dose group (testing II group) human peripheral blood lymphopoiesis index does not affect.Result of the test shows that 3 ‰ dosage of micro-ecological compound preparation prepared by the embodiment of the present invention 2 can promote the proliferation index of PBLC significantly, thus promotes cellular immunity.
The impact of table 10 micro-ecological compound preparation human peripheral blood lymphopoiesis index
2.6 micro-ecological compound preparation are on the impact of serum of broilers total protein content
As shown in Table 11: along with the increase of chicken age in days, the Total albumen content also increases.When 14d, 21d, 1 ‰ and 3 ‰ dosage group the Total albumen content of micro-ecological compound preparation prepared by the embodiment of the present invention 2 are higher than control group, but there was no significant difference, illustrate that the interpolation of micro-ecological compound preparation prepared by the embodiment of the present invention 2 improves the utilization rate of protein and amino acids, strengthen the anabolic action of body internal protein, be conducive to promoting growing and reducing the food consumption of per gain weight of broiler chicken.
Table 11 micro-ecological compound preparation is on the impact of serum of broilers total protein content
2.7 micro-ecological compound preparation are on the impact of serum of broilers immunoglobulin G content
As shown in Table 12: during 14d, the content of test III, IV group of serum of broilers IgG exceeds control group 20.32%, 19.28%, has the difference (P ﹤ 0.05) of conspicuousness; During 28d, the content testing IV group of serum of broilers IgG exceeds control group 30.35%, has the difference (P ﹤ 0.05) of conspicuousness; During 42d, the content of test III, IV group of serum of broilers IgG exceeds control group 24.36%, 24.47%, has the difference (P ﹤ 0.05) of conspicuousness.All in all, micro-ecological compound preparation 1 ‰ and 3 ‰ dosage can significantly improve the content of serum of broilers IgG.
Table 12 micro-ecological compound preparation is on the impact (pg/mL) of serum of broilers IgG content
2.8 micro-ecological compound preparation are on the impact of broiler chicken cecum flora
As shown in Table 13: during 14d, the caecum lactobacillus number of test group is all higher than control group, and wherein IV group of caecum Bacillus acidi lactici number is the highest, exceeds control group 7.02%, but there are no significant difference (P ﹥ 0.05); During 28d, in III group and IV group of caecum, Bacillus acidi lactici number is higher than control group, but there are no significant difference (P ﹥ 0.05), during 42d, in III group and IV group of caecum, Bacillus acidi lactici number exceeds control group 7.01% and 7.60%, significant difference (P ﹤ 0.05) respectively.
As shown in Table 14: during 14d, in test group caecum, coliform count is all lower than control group, and wherein in III group and IV group of caecum, coliform count, respectively lower than control group 7.89% and 9.25%, has significant difference (P ﹤ 0.05); During 28d, in test group caecum, the equal conspicuousness of coliform count is lower than control group (P ﹤ 0.05), and wherein in IV group of caecum, coliform count is minimum, lower than control group 13.10%; During 42d, in test group caecum, coliform count is all lower than control group, and wherein in IV group of caecum, coliform count, lower than control group 8.19%, has significant difference (P ﹤ 0.05).All in all, the reduction caecum coliform count of micro-ecological compound preparation 1 ‰ and 3 ‰ dosage energy conspicuousness, increases the quantity of caecum Bacillus acidi lactici.
Table 13 micro-ecological compound preparation is on the impact (lgcfu/g) of broiler chicken caecum Bacillus acidi lactici number
Table 14 micro-ecological compound preparation is on the impact (lgcfu/g) of broiler chicken caecum coliform count
Brief summary
(1) micro-ecological compound preparation can improve the growth performance of broiler chicken, and outstanding behaviours is significantly increasing the average daily gain of broiler chicken.Increase the Total albumen content, improve the utilization rate of protein and amino acids, strengthen the anabolic action of body internal protein, be conducive to promoting growing and reducing the food consumption of per gain weight of broiler chicken, optimum addition is 1 ‰ and 3 ‰ (mass percents).
(2) micro-ecological compound preparation significantly can promote the growth of broiler chicken immune organ (spleen and the bursa of farbricius), to the effect of increasing significantly of serum of broilers ND antibody titer tool, high dose group significantly can promote peripheral lymphocyte proliferation, the secretion of remarkable promotion cecal content sIgA, show that micro-ecological compound preparation can the enhancing Humoral Immunity in Broiler of conspicuousness, cellular immunity and mucosal immunity function, thus strengthening the immunologic function of broiler chicken, optimum addition is 1 ‰ and 3 ‰ (mass percents).
(3) probio of adding in micro-ecological compound preparation, in the increase enteron aisle of energy conspicuousness, the quantity of lactic acid bacteria, reduces colibacillary quantity, thus the health of protection enteron aisle.
Embodiment 8: the protectiveness test of micro-ecological compound preparation optimal dose prevention broiler chicken bacillus coli
1 test material and method
1.1 material
Micro-ecological compound preparation prepared by the embodiment of the present invention 2; Complete feed, purchases Tai'an six and feed; Ewcastle disease (ND) LaSota IV is vaccine, purchased from Shandong animal health-care product factory.Pathogenic bacteria chicken colibacillosis O1, O2, O78, O26 are the bacterial strain that Shandong Agricultural University is so kind as to give.
1.2 experimental design and daily ration form
Experimental design: 240 body weight is even, health status is substantially identical 1 age in days white meat-type chickens are divided into 4 groups at random, often organizes 3 repetitions, each repetition 20.Namely I group: control group, basal diet of feeding; II group: model group, basal diet of feeding; III group: on the basis of feeding basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 1 ‰ (mass percent) dosage drinking-water; IV group: on the basis of feeding basal diet, add the micro-ecological compound preparation of the embodiment of the present invention 2 preparation by 3 ‰ (mass percent) dosage drinking-water, the equal free choice feeding of each group.
1.3 feedings and managements: test chicken adopts mode of raising in cages, free choice feeding and drinking-water, illumination in house, temperature and humidity require to control in strict accordance with conventional feeding and management.Change every day before food changes water and carry out crib and water fountain virus killing sterilization.Duration of test records the health status of each group of food consumption and test chicken in detail.All chickens in 9d, 22d, 29d immunity newcastle disease vaccine, 15d immunity bursa of fabricius vaccine.During 31 age in days, challenge test is carried out to II, III, IV group of chicken, the gavage inoculation 1mL Escherichia coli bacteria suspension (mixed liquor 1.0 × 10 of O1, O2, O78, O26
9cfu/mL).
1.4 testing indexs and method:
(1) production performance respectively at 17,24,31,38 ages in days by repeat in units of the broiler chicken after fasting (freely drink water) 12h is weighed, and precise and record day feed quantity, remain doses and lose doses.Calculate each stage average daily gain, average daily ingestion amount and feed-weight ratio.
(2) broiler chicken after Immune Organs Index randomly draws 10 fasting (freely drinking water) 12h respectively at 17,31 and 38 ages in days from each repetition is weighed, arteria carotis sacrificed by exsanguination, spleen, the bursa of farbricius are won in aseptic dissection immediately, fresh weight is claimed, Computation immunity shoot formation after rejecting fat.
Immune Organs Index (mg/g)=immune organ weight/live body weight
(3) ND_HI antibody is tired respectively at 17,24,31,38 ages in days, randomly draws 10 chickens, wing venous blood collection, separation of serum, carry out doubling dilution from often repeat, and measures antibody titres to newcastle disease virus in serum with Hemagglutination Inhibition.Antibody titres to newcastle disease virus Log
2x represents.
(4) mensuration of cecal content S-IgA content randomly draws 10 chickens respectively at 17,24,31,38 ages in days from each repeating groups, put to death, dissect, cecal content 1g is got in sterile working, adopts chicken secretory immunoglobulin A (sIgA) ELISA kit to measure the content of sIgA in intestinal juice.
(5) mensuration of the Total albumen content is respectively at 17,24,31,38 ages in days, randomly draws 10 chickens, wing venous blood collection, separation of serum from often repeat, and adopts total protein detection reagent box to detect.
(6) mensuration of gut flora randomly draws 10 chickens respectively at 17,31,38 ages in days from often repeat, slaughter through venesection after weighing, aseptic separation caecum stage casing, gets content 1g, adopts slat chain conveyor counting method to count enteron aisle lactobacillus and coliform count respectively.Microflora represents with the logarithm of total number of bacterial colonies contained by every gram of intestinal contents [(lgCFU)/g].
2 results
2.1 micro-ecological compound preparation are on the impact of meat chicken production performance
As shown in Table 15: the initial chicken of each group broiler chicken is heavy basically identical, and during 17d, III, IV group exceeds control group 3.33%, 3.95% respectively, but there was no significant difference (P>0.05); During 31d, III, IV group of average chicken heavily respectively conspicuousness exceeds control group 16.08%, 19.79%; The average chicken weight average of each test group of 38d is higher than control group.Comprehensively it seems that micro-ecological compound preparation 3 ‰ dosage group has and significantly improve the heavy effect of the average chicken of broiler chicken.
The impact that table 15 micro-ecological compound preparation is heavy on the average chicken of broiler chicken
From table 16 data: in whole process of the test compared with control group, each compound group (III group, IV group) average daily gain conspicuousness raises, and feed-weight ratio conspicuousness reduces.All in all micro-ecological compound preparation has the effect improving broiler chicken average daily gain, reduce feed-weight ratio, and optimum addition is 3 ‰ (mass percents).
Table 16 micro-ecological compound preparation is on the impact of growth of meat chicken performance
2.2 micro-ecological compound preparation are on the impact of broiler chicken Immune Organs Index
From table 17 data: during 31d, micro-ecological compound preparation III, IV group of index and spleen index is higher than control group, and particularly III group exceeds control group 13.55%, has significant difference (P ﹤ 0.05); 38d, each compound group index and spleen index, all higher than control group, is particularly tested IV group and is exceeded control group 16.54%, has significant difference (P ﹤ 0.05); All in all, micro-ecological compound preparation can improve the index and spleen index of broiler chicken, and optimum addition is 3 ‰ (mass percents).
Table 17 micro-ecological compound preparation is on the impact (unit: mg/g) of broiler chicken index and spleen index
From table eighteen data: during 17d, micro-ecological compound preparation III, IV group of broiler chicken bursal index exceeds control group 44.13%, 51.72% respectively, there is significant difference (P ﹤ 0.01), during 38d, the bursal index that micro-ecological compound preparation is III, IV group exceeds control group 10.64%, 12.77%, but there was no significant difference (P ﹥ 0.05), all in all micro-ecological compound preparation can significantly improve broiler chicken bursal index, and with more remarkable effect during 17d.
Table 18 micro-ecological compound preparation is on the impact (unit: mg/g) of broiler chicken bursal index
2.3 micro-ecological compound preparation are on the impact of serum of broilers ND antibody titer
As shown in Table 19: 17d, 24d, 31d, 38d, in micro-ecological compound preparation III, IV group of serum antibody titres to newcastle disease virus all higher than control group; During 24d, micro-ecological compound preparation III, IV group of serum antibody titres to newcastle disease virus exceeds control group 75.19%, 78.94% respectively, has significant difference (P ﹤ 0.05); During 38d, micro-ecological compound preparation III, IV group of serum antibody titres to newcastle disease virus exceeds control group 49.55%, 110.45% respectively; All in all, micro-ecological compound preparation can significantly improve serum N D antibody titer, and optimum addition is 3 ‰ (mass percents).
Table 19 micro-ecological compound preparation is on the impact (Log of serum N D antibody titer
2x)
2.4 micro-ecological compound preparation are on the impact of broiler chicken cecal content sIgA content
As shown in Table 20: during 17d, micro-ecological compound preparation III, IV group of cecal content sIgA content exceeds control group 48.23%, 62.50% respectively, there is significant difference (P ﹤ 0.05), during 31d, micro-ecological compound preparation III, IV group of cecal content sIgA content exceeds control group 42.37%, 55.93% respectively, has significant difference (P ﹤ 0.05); During 38d, micro-ecological compound preparation III, IV group of cecal content sIgA content conspicuousness is higher than control group and model group (P ﹤ 0.05); All in all, micro-ecological compound preparation can significantly improve cecal content sIgA content, and optimum addition is 3 ‰ (mass percents).
Table 20 micro-ecological compound preparation is on the impact (pg/mL) of cecal content sIgA content
2.5 micro-ecological compound preparation are on the impact of serum of broilers total protein
As shown in Table 21: before 31d, along with the increase of chicken age in days, the Total albumen content also increases, compared with control group, when 17d, 24d, 31d, 38d, micro-ecological compound preparation III, IV group significantly increases the Total albumen content, illustrate that the interpolation of micro-ecological compound preparation improves the utilization rate of protein and amino acids, the anabolic action of body internal protein is strengthened, be conducive to promoting growing and reducing the food consumption of per gain weight of broiler chicken, optimum addition is 3 ‰ (mass percents).
Table 21 micro-ecological compound preparation is on the impact of serum of broilers total protein
2.6 micro-ecological compound preparation are on the impact of broiler chicken cecum flora
From table 22 data, in 17d, 31d, 38d III, IV group of caecum, the quantity of Bacillus acidi lactici is all higher than control group.17d tests IV group and exceeds control group 8.56%, has significant difference (P ﹤ 0.05); During 31d, III and IV group exceeds a control group 1-2 order of magnitude, has significant difference (P ﹤ 0.05); During 38d, compared with control group, in model group caecum, the quantity of lactic acid bacteria is significantly lower than control group (P ﹤ 0.05), and the quantity of Bacillus acidi lactici is higher than control group in compound group (III group, IV group) caecum, particularly IV group exceeds control group 18.57% (P ﹤ 0.05).All in all micro-ecological compound preparation can improve the quantity of lactobacillus in broiler chicken caecum, and prevention broiler chicken is from colibacillary infection.
Table 22 micro-ecological compound preparation is on the impact (lgcfu/g) of broiler chicken caecum lactobacillus number
From table 23 data, in the caecum of 17d, 31d, 38d each compound group (III group, IV group), colibacillary quantity is all lower than control group; During 31d IV group lower than control group 20.80%, there is significant difference (P ﹤ 0.05); During 38d, compared with control group, in model group caecum, colibacillary quantity conspicuousness raises, and in the caecum of IV group Escherichia coli conspicuousness lower than control group (P ﹤ 0.05).All in all, micro-ecological compound preparation significantly can reduce the colibacillary quantity of broiler chicken caecum, and optimum addition is 3 ‰ (mass percents).
Table 23 micro-ecological compound preparation is on the impact (lgcfu/g) of broiler chicken caecum coliform count
Comprehensive broiler chicken indices measurement result shows: 1. micro-ecological compound preparation can improve the growth performance of broiler chicken, and show the average daily gain of significantly increase broiler chicken, reduce feedstuff-meat ratio aspect, optimum addition is 3 ‰ (mass percents); 2. micro-ecological compound preparation, increases the Total albumen content, improves the utilization rate of protein and amino acids, strengthen the anabolic action of body internal protein, is conducive to promoting growing and reducing the food consumption of per gain weight of broiler chicken; 3. micro-ecological compound preparation significantly can promote the growth of broiler chicken immune organ spleen, the bursa of farbricius, significant raising serum of broilers ND antibody titer, the secretion of remarkable promotion cecal content sIgA, significantly improve serum IgG content, show that micro-ecological compound preparation can the enhancing Humoral Immunity in Broiler of conspicuousness, cellular immunity and mucosal immunity function, thus strengthening the immunologic function of broiler chicken, optimum addition is 3 ‰ (mass percents); The probio of 4. adding in micro-ecological compound preparation, in the increase E. coli Infected Broiler enteron aisle of energy conspicuousness, the quantity of Bacillus acidi lactici, reduces colibacillary quantity, thus the health of protection enteron aisle.
Claims (10)
1. there is a micro-ecological compound preparation for immune enhancing function, it is characterized in that, be made up of the raw material of following weight portion:
Lactococcus bacterium powder 1-3 part, oligoisomaltose 30-50 part, yeast trace element 10-15 part, NSP enzyme 10-20 part, astragalus polyose 8-12 part, fermentation wheat bran/dregs of beans 15-25 part;
Described fermentation wheat bran/dregs of beans is formed by Lactobacillus plantarum and Pediococcus pentosaceus combined ferment wheat bran and dregs of beans.
2. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 1, it is characterized in that, be made up of the raw material of following weight portion:
2 parts, Lactococcus bacterium powder, oligoisomaltose 40 parts, yeast trace element 12 parts, NSP enzyme 15 parts, astragalus polyose 10 parts, fermentation wheat bran/dregs of beans 21 parts.
3. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 1 or 2, it is characterized in that, the preparation method of described fermentation wheat bran/dregs of beans is: regulate the moisture of the wheat bran for fermenting and dregs of beans to be 38-40%, inoculation Lactobacillus plantarum and Pediococcus pentosaceus mixed bacteria, fermentation, to obtain final product.
4. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 1 or 2, it is characterized in that, described yeast trace element by yeast chromium and yeast iron in mass ratio (5-7): (5-7) mixes.
5. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 4, it is characterized in that, described yeast trace element by yeast chromium and yeast iron in mass ratio 7:5 mix.
6. have the micro-ecological compound preparation of immune enhancing function as claimed in claim 4, it is characterized in that, the chromium content in described yeast chromium is 2000mg/kg, and in yeast iron, the content of iron is 2000mg/kg.
7. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 1, it is characterized in that, the active ingredient of described NSP enzyme comprises: dextranase >=3,000,000 U/g, cellulase >=70000U/g, pectase >=20000U/g, mannase >=5000U/g.
8. have the micro-ecological compound preparation of immune enhancing function as claimed in claim 1, it is characterized in that, described Lactococcus bacterium powder is Pediococcus pentosaceus bacterium powder.
9. there is the micro-ecological compound preparation of immune enhancing function as claimed in claim 8, it is characterized in that, the bacterial strain of described Pediococcus pentosaceus to be deposit number be CCTCCNO:M2012029.
10. the micro-ecological compound preparation described in any one of claim 1 to 9 is as the application had in the feed addictive of immune enhancing function.
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