CN102232998A - Micro-ecological traditional Chinese medicine preparation for enhancing immunity of livestock and poultry, and preparation method thereof - Google Patents
Micro-ecological traditional Chinese medicine preparation for enhancing immunity of livestock and poultry, and preparation method thereof Download PDFInfo
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Abstract
The invention provides a micro-ecological traditional Chinese medicine preparation for enhancing the immunity of livestock and poultry, and a preparation method thereof. The invention is characterized in that traditional Chinese medicines are combined with the modern microbial fermentation technology to develop a novel feed additive, i.e. traditional Chinese medicines such as astragalus root, eucommia, licorice, bupleurum and the like are directly fermented by two or more of sporeformer, microzyme and/or lactobacillus to obtain the product. The micro-ecological traditional Chinese medicine preparation provided by the invention has the efficacies of enhancing the body immunity of livestock and poultry, maintaining the gastrointestinal microbial flora balance of animals, preventing and treating alimentary and respiratory diseases of livestock and poultry, promoting healthy and quick growth of the animals, and improving the qualities of meat, egg and milk; the micro-ecological traditional Chinese medicine preparation has no toxic side effect, no pollution and no residue, can not generate drug resistance, and is suitable for use at each growth stage of the livestock and poultry; and the effect is stable and reliable and can meet the requirements of people for safety in production of livestock and poultry products. The micro-ecological traditional Chinese medicine preparation for enhancing the immunity of livestock and poultry can be increased in dose to be used as a veterinary medicine for application.
Description
Technical field
The invention belongs to feed additive field, relate to a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock and preparation method thereof, relate more specifically to the composition and method of making the sames of Chinese medicine behind probiotics fermention such as the Radix Astragali, the Cortex Eucommiae, Radix Glycyrrhizae, Rhizoma Zingiberis Recens.
Background technology
In the breeding process of poultry, diseases such as digestive tract, respiratory tract often appear, and this has seriously influenced the growth of poultry, even can cause poultry death, causes very big loss for numerous raisers.The use of feeding antibiotic has improved the Production of Livestock and Poultry level greatly, and it promotes growth mechanism and effect to become common recognition, makes animal husbandry scale and intensification become possibility simultaneously.Antibiotic has been made significant contribution to aquaculture in the use history in more than 50 year in the past of China.Supply falls short of demand, urban and rural residents protein product source is deficient from the past animal products, and drug on the market to domestic market meat, fowl, egg, milk now, and realized outlet in batches.Yet the counter productive of its initiation has become the focus of concern.Fastbacteria growth and drug resistant gene that life-time service antibiotic or abuse of antibiotics cause shift, may cause the decline of humans and animals immunity of organisms, the increase of secondary dosage, drug residue, dual and a series of problems such as autogenous infection, sensitization, health is constituted a threat to.The animal derived food potential safety hazard that these factors cause enjoys consumer to pay close attention to.
Drug residue has become one of major reason that influences the export trade, and there is safety problem in the exported product major part of China, directly influence foreign exchange earning.On August 1st, 1996, European Union forbids also being contained in states such as Japan and Korea S., Russia subsequently from China's import broiler and aquatic products; Only January in August, 2000 to calendar year 2001, U.S. FDA has just been detained 634 batches of Chinese imported foods, and the overwhelming majority is because safety issue; On April 16th, 2002, the Chinese frozen meat that total value is 4,210,000 dollars is committed to the flames at Dutch Rui Dankeluodeshi, and its reason is to detect wherein to contain chloromycetin.Such example happens occasionally in recent years, has beaten alarm bell all for the aquaculture of China at every turn.
Meanwhile, along with the progress of development and national economy and society, the people's living standard improves constantly, the progressively enhancing of self health consciousness, people more and more pay attention to diet and healthy relation, to meat, egg, milk consumption demand forward high-quality, nutrition, secure side to development.
Summary of the invention
The objective of the invention is to overcome the deficiency of above-mentioned technology, a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock and preparation method thereof is provided, Chinese medicine ingredients and multiple beneficial bacterium and the metabolite thereof with raising immunity contained in its inside.This Chinese medicine microbial ecological agent has significant immunoregulation effect, and has no side effect, pollution-free, noresidue, do not produce drug resistance.
Another object of the present invention provides the preparation method of the Chinese medicine microbial ecological agent of this enhancing immunity of livestock.
The present invention also provides a kind of feedstuff that comprises the Chinese medicine microbial ecological agent of this enhancing immunity of livestock.
To achieve these goals, the technical solution adopted in the present invention is as follows:
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that the method that adopts Chinese medicine to pass through probiotics fermention obtains.
Described a kind of preparation method that strengthens the Chinese medicine microbial ecological agent of immunity of livestock may further comprise the steps:
(1) Chinese crude drug pretreatment
Choose Chinese crude drug, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross 80～120 mesh sieves, standby;
(2) preparation strain fermentating liquid
In the optimum medium that is inoculated in respectively in sporeformer, yeast, the lactobacillus separately, cultivate primary seed solution under suitable condition, carry out amplification culture in the secondary seed fermentation tank when each bacteria growing is transferred to respectively during to exponential phase, stop when reaching exponential phase cultivating, standby;
(3) Chinese medicine fermentation
With the mix homogeneously according to a certain ratio of the Chinese crude drug after pulverizing, add water to water content between 50～70%, after soaking 12～24h, regulate between the pH value 6.0～7.5, at 121 ℃ of following steam sterilization 15～20min, after the cooling, add the total strain liquid that accounts for Chinese crude drug quality 8%～20%, 28～37 ℃ fermented 24～72 hours, stirred once every 8h; After the fermentation ends,, promptly get the Chinese medicine microbial ecological agent 35～50 ℃ of oven dry down.
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that described Chinese medicine comprises 1～10 part of the Radix Astragali, 1～10 part of the Cortex Eucommiae, 1～10 part in Radix Glycyrrhizae, 1～10 part of Radix Bupleuri by mass fraction.
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that described probiotic bacteria comprises two or more in sporeformer, yeast and/or the lactobacillus.
The preparation method of described Chinese medicine microbial ecological agent is characterized in that total strain liquid is by in sporeformer secondary seed solution, yeast secondary seed solution and/or the lactobacillus secondary seed solution two or more is mixed and made into seed mixture liquid by equal proportion.
The Chinese medicine microbial ecological agent of described enhancing immunity of livestock is characterized in that described Chinese medicine also comprises one or more in 1～5 part of 1～5 part of Herba Ecliptae, 1～10 part of Fructus Ligustri Lucidi, 1～5 part of Radix Salviae Miltiorrhizae, 1～10 part of Fructus Lycii, 1～5 part of Semen Coicis, 1～5 part of Radix Scrophulariae, 1～5 part of Radix Saposhnikoviae, 1～5 part in Rhizoma Zingiberis Recens, 1～5 part of Radix Rehmanniae, 1～3 part of Fructus Schisandrae Chinensis, 1～10 part of Herba Taraxaci, 1～10 part of Rhizoma Dioscoreae and the Semen Allii Tuberosi.
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that described sporeformer comprises Bacillus licheniformis, bacillus subtilis and/or bacillus cereus.
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that described yeast comprises Candida utilis and/or saccharomyces cerevisiae.
Described a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that described lactobacillus comprises enterococcus faecalis, lactobacillus lactis, bacillus acidophilus, lactobacillus casei, lactobacillus lactis and/or Lactobacillus bulgaricus.
Described a kind of feedstuff that strengthens the Chinese medicine microbial ecological agent of immunity of livestock is characterized in that, comprises the constituent of following mass ratio: the Chinese medicine microbial ecological agent 0.02～0.2% of described enhancing immunity of livestock and adjuvant 99.98～99.8%.
The invention provides a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock and preparation method thereof, have the following advantages:
1, Chinese medicine microbial ecological agent of the present invention can strengthen the immunocompetence of animal body, keep animal intestines and stomach microorganism species balance, effectively prevent and treat livestock and poultry alimentary tract and respiratory tract disease, promote livestock birds health, growth apace, the meat, eggs and milk quality better reduces effects such as culturing input;
2, the present invention's Chinese medicine microbial ecological agent of strengthening immunity of livestock each growth stage of being adapted at livestock and poultry animal is used;
3, the present invention adopts probiotic bacteria direct fermentation Chinese medicine, has protected active ingredient of Chinese herbs to exempt from destruction, produces new drug effect;
Microbial fermentation is to carry out under comparatively gentle condition such as room temperature, normal pressure, and active component exempts from destruction in the Chinese medicine so can protect to greatest extent.And the bigger active component of the number molecular weight in the Chinese medicine becomes small-molecule active substance and directly utilized after fermentation, and this has improved the drug effect of Chinese medicine greatly; The enzyme of microorganisms carries out structural modification to active ingredient of Chinese herbs, generates secondary metabolite, thereby has produced new active substance, thereby have new drug effect;
4, the present invention adopts multiple probiotic mixed fermentation Chinese medicine, can produce as cellulase, amylase, protease etc., make the material of difficult absorptions such as cellulose in the Chinese medicine, lignin fully to degrade, pharmaceutically active ingredient is discharged to greatest extent in making, simultaneously cellulose conversion can be become oligosaccharide, promote the growth of probiotics in the animal gastrointestinal tract; Under the effect of the various enzymes that produced, the effective ingredient in the feedstuff is easier to be absorbed by animal simultaneously;
5, the present invention utilizes the novel fodder additive of the interaction development of Chinese medicine and probiotic bacteria, and incomparable advantage is arranged, and it has and has no side effect, pollution-free, noresidue, do not produce security features such as drug resistance;
6, it is reliable that the present invention strengthens the effect stability of immunity of livestock Chinese medicine microbial ecological agent, can satisfy people to meat, egg, milk requirement of safe production;
7, the present invention strengthens immunity of livestock Chinese medicine microbial ecological agent and can pass through the raising using dosage, uses as veterinary drug.
, it should be understood that described embodiment only for the present invention is described, rather than limit the scope of the invention by any way more specific description the present invention by following examples.
The specific embodiment
Embodiment 1
Chinese medicine: 10 parts of the Radixs Astragali, 6 parts of the Cortexs Eucommiae, 6 parts in Radix Glycyrrhizae, 3 parts of Radix Bupleuri, 5 parts of Herba Ecliptaes.
Fermentation technology:
Bacillus licheniformis, bacillus subtilis are inoculated into respectively in the common nutrient broth medium, and at 37 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, primary seed solution is inoculated into amplification culture in the secondary seed fermentation tank that nutrient broth is housed with inoculum concentration 5%, cultivation temperature is 28～35 ℃, and rotating speed is 200rpm, when sporeformer content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Saccharomyces cerevisiae is inoculated in the malt extract medium, and 28 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, primary seed solution is inoculated into amplification culture in the secondary seed fermentation tank that nutrient broth is housed with inoculum concentration 6%, cultivation temperature is 28～35 ℃, and rotating speed is 200rpm, when yeast content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Choose above-mentioned each Chinese crude drug, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross the 80-120 mesh sieve, standby;
Take by weighing Chinese medicinal powder after the pulverizing by above-mentioned prescription, add water to water content at 50-60%, behind the mix homogeneously, at room temperature soak 24h, regulate about pH7.0, at 121 ℃ of following steam sterilization 15～20min, after the cooling, add the Bacillus licheniformis bacterium liquid account for Chinese crude drug quality 5%, 5% bacillus subtilis bacterium liquid and 5% saccharomyces cerevisiae bacterium liquid respectively,, stir once every 8h at 28～35 ℃ of fermentation 36h; When total viable count reaches 1.0 * 10
8～9Stop fermentation during CFU/ml, 45 ℃ of following vacuum dryings make the Chinese medicine microbial ecological agent.
Embodiment 2
Chinese medicine: 8 parts of the Radixs Astragali, 6 parts of the Cortexs Eucommiae, 10 parts in Radix Glycyrrhizae, 3 parts of Radix Bupleuri, 4 parts of Herba Ecliptaes, 5 parts of Fructus Ligustri Lucidi, 3 parts of Radix Salviae Miltiorrhizaes, 10 parts of Fructus Lyciis, 6 parts of Semen Coicis, 3 parts of Radix Scrophulariaes, 2 parts of Radix Saposhnikoviaes, 6 parts of Rhizoma Dioscoreaes, 5 parts of Semen Allii Tuberosi, 3 parts of Radix Rehmanniae, 6 parts of Herba Taraxacis.
Fermentation technology:
Bacillus licheniformis is inoculated in the common nutrient broth medium, and at 37 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, amplification culture in the secondary seed fermentation tank that primary seed solution is inoculated into inoculum concentration 3%, cultivation temperature is 30～32 ℃, rotating speed is 200rpm, when bacterial content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Candida utilis is inoculated in the malt extract medium, and 28 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, amplification culture in the secondary seed fermentation tank that primary seed solution is inoculated into inoculum concentration 8%, cultivation temperature is 30～32 ℃, rotating speed is 200rpm, when yeast content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Lactobacillus lactis and Lactobacillus bulgaricus are inoculated into respectively in the MRS culture medium, and 37 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, primary seed solution is inoculated into amplification culture in the secondary seed fermentation tank that nutrient broth is housed with inoculum concentration 5%, cultivation temperature is 30～32 ℃, and rotating speed is 200rpm, when bacterial content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Choose above-mentioned each Chinese crude drug, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross the 80-120 mesh sieve, standby;
Take by weighing Chinese medicinal powder after the pulverizing by above-mentioned prescription, add water to water content at 60-70%, behind the mix homogeneously, at room temperature soak 24h, regulate PH6.5～6.9, at 121 ℃ of following steam sterilization 20min, after the cooling, add the Bacillus licheniformis bacterium liquid account for Chinese crude drug quality 2.5%, 2.5% Candida utilis bacterium liquid, 2.5% lactobacillus lactis bacterium liquid and 2.5% Lactobacillus bulgaricus bacterium liquid respectively,, stir once every 8h at 30～32 ℃ of fermentation 48h; When total viable count reaches 1.0 * 10
8～9Stop fermentation during CFU/ml, 40 ℃ of following vacuum dryings make the Chinese medicine microbial ecological agent.
Embodiment 3
Chinese medicine: 5 parts of the Radixs Astragali, 8 parts of the Cortexs Eucommiae, 10 parts in Radix Glycyrrhizae, 2 parts of Radix Bupleuri, 2 parts in Rhizoma Zingiberis Recens, 4 parts of Herba Ecliptaes, 5 parts of Fructus Ligustri Lucidi, 3 parts of Radix Salviae Miltiorrhizaes, 9 parts of Fructus Lyciis, 5 parts of Semen Coicis.
Fermentation technology:
Bacillus licheniformis is inoculated in the common nutrient broth medium, and at 37 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, amplification culture in the secondary seed fermentation tank that primary seed solution is inoculated into inoculum concentration 3%, cultivation temperature is 30 ± 2 ℃, rotating speed is 200rpm, when bacterial content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Candida utilis and saccharomyces cerevisiae are inoculated in the malt extract medium, and 28 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, amplification culture in the secondary seed fermentation tank that primary seed solution is inoculated into inoculum concentration 8%, cultivation temperature is 30 ± 2 ℃, rotating speed is 200rpm, when yeast reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Enterococcus faecalis, lactobacillus casei and Lactobacillus bulgaricus are inoculated into respectively in the MRS culture medium, and 37 ℃, the 200rpm shaking table is cultivated, and the production primary seed solution is when bacterial content reaches 10
7-8During the CFU/ml left and right sides, primary seed solution is inoculated into amplification culture in the secondary seed fermentation tank that nutrient broth is housed with inoculum concentration 5%, cultivation temperature is 30 ± 2 ℃, and rotating speed is 200rpm, when bacterial content reaches 10
9During the CFU/ml left and right sides, stop fermentation, standby;
Choose above-mentioned each Chinese crude drug, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross the 80-120 mesh sieve, standby;
Take by weighing Chinese medicinal powder after the pulverizing by above-mentioned prescription, add water to water content 60%, behind the mix homogeneously, at room temperature soak 24h, regulate pH6.5～6.9, at 121 ℃ of following steam sterilization 20min, after the cooling, add the lactobacillus casei bacterium liquid of the Bacillus licheniformis bacterium liquid account for Chinese crude drug quality 2.5%, 2.5% Candida utilis bacterium liquid, 2.5% saccharomyces cerevisiae liquid, 2.5% enterococcus faecalis bacterium liquid and 2.5% and 2.5% Lactobacillus bulgaricus bacterium liquid respectively, at 30 ± 2 ℃ of fermentation 40h, stir once every 8h; When total viable count reaches 1.0 * 10
8～9Stop fermentation during CFU/ml, 50 ℃ of following vacuum dryings make the Chinese medicine microbial ecological agent.
Embodiment 4
Feedstuff comprises the constituent of following mass percent: the Chinese medicine microbial ecological agent 0.05% and the adjuvant 99.92% that strengthen immunity of livestock.The addition that promptly strengthens the Chinese medicine microbial ecological agent of immunity of livestock in 1 ton of feedstuff is 500g.Feedstuff can specifically comprise the constituent of following mass fraction: by 0.05 part of the Chinese medicine microbial ecological agent of the prepared enhancing immunity of livestock of the embodiment of the invention 2,55 parts of Semen Maydis powder, 1.1 parts of wheat-middlings, 30 parts of bean cake, 9 parts of cottonseed meal, 1 part in corn bean white lead, 0.6 part of oils and fats, 0.9 part of hydrogen calcium, 1.1 parts of stone powders, 0.3 part of Sal, 0.3 part of lysine, 0.3 part of methionine, 0.05 part of choline chloride, 0.02 part of compound cellulose pre-mixing agent, 0.28 part of compound microelement pre-mixing agent.
Embodiment 5
A kind of feedstuff (piglet is used) that contains the Chinese medicine microbial ecological agent that strengthens immunity of livestock comprises the constituent of following mass percent: the Chinese medicine microbial ecological agent 0.12% and the adjuvant 99.92% that strengthen immunity of livestock.The addition that promptly strengthens the Chinese medicine microbial ecological agent of immunity of livestock in 1 ton of feedstuff is 1200g.Feedstuff can specifically comprise the constituent of following mass fraction: by 0.12 part of the Chinese medicine microbial ecological agent of the prepared enhancing immunity of livestock of the embodiment of the invention 3,59 parts of corns, 16 parts of puffing Semen sojae atricolor, 13 parts of dehulled soybean meal, 4 parts of imported fish meals, 3.88 parts of glucoses, 4 parts of 4% piglet premixes.
Embodiment 6
A kind of feedstuff (nursing sow is used) that contains the Chinese medicine microbial ecological agent that strengthens immunity of livestock comprises the constituent of following mass percent: the Chinese medicine microbial ecological agent 0.1% and the adjuvant 99.9% that strengthen immunity of livestock.The addition that promptly strengthens the Chinese medicine microbial ecological agent of immunity of livestock in 1 ton of feedstuff is 1000g.Feedstuff can specifically comprise the constituent of following mass fraction: by 0.1 part of the Chinese medicine microbial ecological agent of the prepared enhancing immunity of livestock of the embodiment of the invention 1,61.9 parts of corns, 22 parts of soybean meal, 10 parts in Testa Tritici, 1 part of imported fish meal, 1 part in Oleum Glycines, 4 parts of 4% nursing sow premix material.
Below by concrete test example the beneficial effect of the Chinese medicine microbial ecological agent of enhancing immunity of livestock of the present invention is described.
Test example 1
The Chinese medicine microbial ecological agent of the enhancing immunity of livestock that the routine method of having studied embodiment 2 of this test prepares is to the influence of fertility performance and the premunition of ablactational baby pig.
1 materials and methods
1.1 the time and the place of test
Tested on May 26,12 days to 2010 May in 2010 and finish on Nan Gezhuan pig farm, Beijing.
1.2 the selection of experimental animal and grouping
Select for use the source identical, age in days is identical, body weight close (about 6-7 kilogram), 120 of Du of 21 ages in days that are in a good state of health wean * big * long three way cross commodity piglet are divided into 5 processed group at random, and each processed group is established 2 repeating groups, 12 pigs of each repeating groups, male and female half and half.
1.3 experimental period
Experimental period is 14 days, piglet 21 age in days to 35 ages in days.
1.4 EXPERIMENTAL DESIGN and test daily ration
Nutritional need design corn-bean cake type basal diet (table 1) with reference to NRC (1998) pig.In basal diet, add antibiotic and constitute the 1# daily ration, the matched group of feeding examination pig, (wherein the common Chinese preparation is: 8 parts of the Radixs Astragali, 6 parts of the Cortexs Eucommiae, 10 parts in Radix Glycyrrhizae, 3 parts of Radix Bupleuri, 4 parts of Herba Ecliptaes, 5 parts of Fructus Ligustri Lucidi, 3 parts of Radix Salviae Miltiorrhizaes, 10 parts of Fructus Lyciis, 6 parts of Semen Coicis, 3 parts of Radix Scrophulariaes, 2 parts of Radix Saposhnikoviaes, 6 parts of Rhizoma Dioscoreaes, 5 parts of Semen Allii Tuberosi, 3 parts of Radix Rehmanniae, 6 parts of Herba Taraxacis to add common Chinese preparation 1500ppm in basal diet respectively, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross the 80-120 mesh sieve promptly.), Chinese medicine microbial ecological agent 400ppm, 600ppm, 800ppm constitute 2#, 3#, 4#, 5# daily ration, feeding experiment group 1, test group 2, test group 3 and test group 4 examination pigs respectively, the basal diet prescription sees Table 1.
Table 1 basal diet
| Raw material | Proportioning | Nutritive index | Trophic level |
| The one-level corn | 59.00 | Crude protein (%) | 19.20 |
| Expanded full-fat bean | 16.00 | Pig digestible energy (Mcal/kg) | 3380 |
| Dehulled soybean meal (46%) | 13.00 | Calcium (%) | 0.80 |
| Imported fish meal (62%) | 4.00 | Available phosphorus (%) | 0.45 |
| Glucose | 4.00 | Lysine (%) | 1.25 |
| 4% premix material | 4.00 | ||
| Add up to | 100.00 |
1.5 daily management
The semi open model pig house, online flat foster, dry powder feed, free choice feeding and drinking-water; Conventional program immunity sterilization.
1.6 testing index and method
1.6.1 feed intake (gram/head day): with the repeating groups is unit, the inventory of accurate recording every day, and the clear hopper of m seq calculates average daily ingestion amount.
1.6.2 daily gain (gram/head day): with the repeating groups is unit, second day early morning of same day on-test early morning and off-test, weighs on an empty stomach, by gained initial weight, last re-computation total augment weight and average daily gain.
1.6.3 feedstuff-meat ratio:, calculate the feedstuff-meat ratio of each repeating groups by feed intake, daily gain.
1.6.4 diarrhea rate (%): with the repeating groups is unit, and every day, at least 3 observation diarrhoea at any time write down diarrheal pig number of elements, off-test calculating diarrhea rate of average day.
1.6.5 survival rate (%): with the repeating groups is unit, the pig number of elements that record survives, and off-test is calculated to be motility rate.
1.6.6 lymphocyte transformation test: pig is only during 28 ages in days, and each processed group blood sampling is 4 before the morning feeding, adopts aseptic vena cava anterior blood sampling, injects the biochemical pipe of vacuum EDTA anticoagulant tube and vacuum respectively and makes anticoagulation and serum.Adopt mtt assay to measure the peripheral blood lymphocyte conversion ratio.
1.6.7 antibody titer test: all test piglets are waited dosage swine fever cell vaccine immunity inoculation at 25 ages in days, each processed group blood sampling in 10 days is 4 after immunity inoculation, with the blood sampling of the biochemical pipe of vacuum vena cava anterior, the separation of serum packing,-80 ℃ frozen, with the swine fever ELISA antibody kit detection hog cholera antibody of American I DEXX company.
The hog cholera antibody level is represented with the blocking-up rate: blocking-up rate/%=(OD
Neg-OD
Test)/OD
Neg* 100%
1.7 the processing of data
Test data adopts SPSS 11.5 statistical softwares to carry out variance analysis and multiple comparisons.
2 results and analysis
2.1 influence to ablactational baby pig fertility performance and diarrhea rate
The influence of table 2 pair ablactational baby pig fertility performance and diarrhea rate
| Handle | Average daily ingestion amount (g) | Average daily gain (g) | Feedstuff-meat ratio | Diarrhea rate (%) | Survival rate (%) |
| Contrast | 356.5±41.0 a | 221.5±39.2 a | 1.61±0.15 a | 5.56±1.15 a | 95.8 ab |
| Test group 1 | 352.9±47.8 a | 205.2±20.1 c | 1.72±0.16 b | 9.59±2.77 b | 91.6 b |
| Test group 2 | 359.3±22.6 a | 217.8±21.8 a | 1.65±0.23 a | 7.34±1.97a b | 95.8 ab |
| Test group 3 | 373.7±22.6 a | 239.6±34.2 b | 1.56±0.17 a | 6.73±2.17 a | 100 a |
| Test group 4 | 372.7±22.6 a | 234.4±38.1 b | 1.59±0.19 a | 6.34±1.85 a | 100 a |
Annotate: the female different persons of shoulder marking-up represent significant difference (P＜0.05) in the same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
By table 2 as seen, the average daily ingestion amount of piglet between each group of test does not have significant difference (P＞0.05).But numerical analysis is compared with matched group, and the piglet feed intake of test group 3 and test group 4 improves 4.8% and 4.5%, shows the feed intake after the Chinese medicine microbial ecological agent can improve weaned piglet to a certain extent; The piglet daily gain of test group 1 significantly is lower than matched group (P＜0.05), compares with matched group, and test group 3 and test group 4 can significantly improve piglet daily gain (P＜0.05); Test group 1 piglet feedstuff-meat ratio is significantly higher than matched group (P＜0.05), but feedstuff-meat ratio does not have significant difference (P＞0.05) between test group 3 and test group 4 and matched group, and numerical value is better than matched group.The piglet diarrhea rate of test group 3 and test group 4 significantly is lower than test group 1 (P＜0.05), and the piglet diarrhea rate between the matched group does not have significant difference (P＞0.05); The piglet survival ratio of test group 3 and test group 4 is significantly higher than test group 1 (P＜0.05), is better than matched group (P＞0.05); Illustrate that Chinese medicine microbial ecological agent of the present invention can improve the fertility performance of ablactational baby pig and reduce diarrhea rate, and effect obviously is better than common Chinese group (P＜0.05).
2.2 influence to the ablactational baby pig premunition
The influence of table 3 pair ablactational baby pig premunition
Annotate: the female different persons of shoulder marking-up represent significant difference (P＜0.05) in the same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
By table 3 as seen, during 28 ages in days, test group 3, test group 4 periphery blood T lymphocyte conversion ratios and peripheral blood B lymhocyte transformation rate are significantly higher than matched group and test group 1 (P＜0.05), test group 2 and test group 1 lymhocyte transformation rate increase than matched group, but difference not significantly (P＞0.05).Test group 1, test group 2, test group 3, test group 4 antibody horizontals all are higher than matched group, and test group 3, test group 4 antibody horizontals are than test group 2 height; Illustrate that interpolation common Chinese preparation and Chinese medicine microbial ecological agent all can improve the serum antibody level, and Chinese medicine microbial ecological agent group is better than the common Chinese preparation on raising serum antibody level.
Test example 2
This test example has been studied by the influence to broiler chicken fertility performance and immune indexes of the Chinese medicine microbial ecological agent of the enhancing immunity of livestock of the method preparation of embodiment 3.
Test method: choose 1500 of Luo Si 308 broiler chicken of 1 age in days body condition health, be divided into 5 processing (being designated as 1#, 2#, 3#, 4#, 5#) at random, each handles 4 repetitions, and each repeats 75 chickens; 49 days experimental periods, divide 3 stages, 0～21 age in days stage, 22～42 age in days stages, 43～49 age in days stages.Recommendation feeding standard according to this kind, nutritional need in conjunction with NRC (1994) broiler chicken, make three stage metabolizable energy concentration and be respectively 2.88Mcal/kg, 2.99Mcal/kg, 3.05Mcal/kg, crude protein content is 20.37%, 18.97%, 17.44% basal diet, and basal diet sees Table 4.Basal diet is made as matched group, be designated as the 1# daily ration, in basal diet, add common Chinese preparation 1000ppm, Chinese medicine microbial ecological agent 300ppm, 500ppm, 800ppm respectively, constitute 4 kinds of test daily rations (being called 2#, 3#, 4#, 5# daily ration successively), give respectively and raise the chicken of above 5 processed group.Three-layer stereo is raised in cages, dry powder feed, and free choice feeding and drinking-water, the routine immunization sterilization, normal condition is with the chicken sterilization weekly once, if meet strong wind weather, then wind is after sterilization at once.Adopt the hot-blast stove heating between brood time, all mean temperatures are respectively 32.7 ℃, 28.5 ℃, 27.6 ℃, 27.3 ℃, 25.6 ℃, 25.5 ℃, 25.8 ℃ in the house in 7 weeks.After the off-test, detect body weight, feed intake, feedstuff-meat ratio, death rate and newcastle epidemic disease antibody titre.When broiler chicken 21 ages in days and 42 ages in days, from each repeating groups, extract 2 respectively and butcher, peel off and weighing thymus, method fabricius bursa and spleen fresh weight, calculate the immune organ index with the approaching test chicken of average weight.The result is shown in table 5, table 6 and table 7.
Table 4 basal diet prescription
The influence of table 5 pair broiler chicken fertility performance and diarrhea rate
Annotate: the female different persons of shoulder marking-up represent significant difference (P＜0.05) in the same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
Result of the test shows: each test group is than matched group chicken feed intake decrease (P＞0.05); 3#, 4# and 5# test group improve 2.0%, 1.8%, 3.7% respectively than the matched group daily gain, and wherein 5# test group daily gain significantly improves (P＜0.05) than matched group.2#, 3#, 4#, 5# test group feedstuff-meat ratio reduce by 3.9%, 6.0%, 7.5%, 8.0% respectively than matched group, wherein significantly reduce (P＜0.05) with 4#, 5# test group feedstuff-meat ratio than matched group; 2#, 3#, 4#, 5# test group can significantly reduce the diarrhea rate (P＜0.05) of broiler chicken than matched group, and 4#, 5# test group survival rate significantly improve than matched group.In the whole breeding cycle, wherein the Chinese medicine microbial ecological agent group of 500ppm and 800ppm show aborning more outstanding, the efficiency of feed utilization height, it is great to increase day by day, feedstuff-meat ratio is low.
The influence of table 6 pair broiler chicken newcastle epidemic disease antibody level
Annotate: the female different persons of shoulder marking-up represent significant difference (P＜0.05) in the same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
Table 6 shows: after each immunity, the newcastle epidemic disease antibody titre of 4# processed group and 5# processed group broiler chicken illustrates that all than significantly improve (P＜0.05) of matched group and common Chinese preparation group Chinese medicine microbial ecological agent of the present invention can improve the antibody titer of broiler chicken.
The table 7 pair exponential influence of broiler chicken immune organ
Annotate: the female different persons of shoulder marking-up represent significant difference (P＜0.05) in the same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
Result of the test shows, during 21 ages in days, 2#, 3#, 4#, 5# test group are than matched group thymus, fabricius bursa and index and spleen index all increase (P＞0.05).During 42 ages in days, 4#, 5# processed group thymus index and fabricius bursa index significantly improve (P＜0.05) than matched group, be tending towards improving index and spleen index (P＞0.05), 2#, 3# processed group all increase than matched group thymus, fabricius bursa and index and spleen index, but compare difference not remarkable (P＞0.05) with matched group, wherein, 4#, 5# processed group fabricius bursa index are significantly higher than 2#, 3# processed group (P＜0.05).
Conclusion: Chinese medicine by fermentation after, newcastle epidemic disease antibody level, immune organ index are higher than the unfermentable Chinese medicine preparation of simple pulverizing, fertility performance to improve effect also very obvious.
Claims (10)
1. a Chinese medicine microbial ecological agent that strengthens immunity of livestock is characterized in that the method that adopts Chinese medicine to pass through probiotics fermention obtains.
2. according to right 1 described a kind of preparation method that strengthens the Chinese medicine microbial ecological agent of immunity of livestock, may further comprise the steps:
(1) Chinese crude drug pretreatment
Choose Chinese crude drug, behind 50～60 ℃ of following oven dry 2～3h, pulverize, cross 80～120 mesh sieves, standby;
(2) preparation strain fermentating liquid
In the optimum medium that is inoculated in respectively in sporeformer, yeast, the lactobacillus separately, cultivate primary seed solution under suitable condition, carry out amplification culture in the secondary seed fermentation tank when each bacteria growing is transferred to respectively during to exponential phase, stop when reaching exponential phase cultivating, standby;
(3) Chinese medicine fermentation
With the mix homogeneously according to a certain ratio of the Chinese crude drug after pulverizing, add water to water content between 50～70%, after soaking 12～24h, regulate between the pH value 6.0～7.5, at 121 ℃ of following steam sterilization 15～20min, after the cooling, add the total strain liquid that accounts for Chinese crude drug quality 8%～20%, 28～37 ℃ fermented 24～72 hours, stirred once every 8h; After the fermentation ends,, promptly get the Chinese medicine microbial ecological agent 35～50 ℃ of oven dry down.
3. a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock according to claim 1 is characterized in that described Chinese medicine comprises by mass fraction:
1～10 part of the Radix Astragali, 1～10 part of the Cortex Eucommiae, 1～10 part in Radix Glycyrrhizae, 1～10 part of Radix Bupleuri.
4. a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock according to claim 1 is characterized in that described probiotic bacteria comprises two or more in sporeformer, yeast and/or the lactobacillus.
5. the preparation method of Chinese medicine microbial ecological agent according to claim 2 is characterized in that total strain liquid is by in sporeformer secondary seed solution, yeast secondary seed solution and/or the lactobacillus secondary seed solution two or more is mixed and made into seed mixture liquid by equal proportion.
6. the Chinese medicine microbial ecological agent of enhancing immunity of livestock according to claim 3 is characterized in that described Chinese medicine also comprises one or more in 1～5 part of 1～5 part of Herba Ecliptae, 1～10 part of Fructus Ligustri Lucidi, 1～5 part of Radix Salviae Miltiorrhizae, 1～10 part of Fructus Lycii, 1～5 part of Semen Coicis, 1～5 part of Radix Scrophulariae, 1～5 part of Radix Saposhnikoviae, 1～5 part in Rhizoma Zingiberis Recens, 1～5 part of Radix Rehmanniae, 1～3 part of Fructus Schisandrae Chinensis, 1～10 part of Herba Taraxaci, 1～10 part of Rhizoma Dioscoreae and the Semen Allii Tuberosi.
7. a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock according to claim 4 is characterized in that described sporeformer comprises Bacillus licheniformis, bacillus subtilis and/or bacillus cereus.
8. a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock according to claim 4 is characterized in that described yeast comprises Candida utilis and/or saccharomyces cerevisiae.
9. a kind of Chinese medicine microbial ecological agent that strengthens immunity of livestock according to claim 4 is characterized in that described lactobacillus comprises enterococcus faecalis, lactobacillus lactis, bacillus acidophilus, lactobacillus casei, lactobacillus lactis and/or Lactobacillus bulgaricus.
10. feedstuff that comprises the Chinese medicine microbial ecological agent of each described enhancing immunity of livestock of claim 1～9, it is characterized in that, comprise the constituent of following mass ratio: the Chinese medicine microbial ecological agent 0.02～0.2% of described enhancing immunity of livestock and adjuvant 99.98～99.8%.
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