CN102697888A - Traditional Chinese medicine fermented preparation for preventing and treating piglet diarrhea and preparation method thereof - Google Patents
Traditional Chinese medicine fermented preparation for preventing and treating piglet diarrhea and preparation method thereof Download PDFInfo
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- CN102697888A CN102697888A CN2012101401560A CN201210140156A CN102697888A CN 102697888 A CN102697888 A CN 102697888A CN 2012101401560 A CN2012101401560 A CN 2012101401560A CN 201210140156 A CN201210140156 A CN 201210140156A CN 102697888 A CN102697888 A CN 102697888A
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Abstract
The invention discloses a traditional Chinese medicine fermented preparation for preventing and treating piglet diarrhea and a preparation method thereof. The traditional Chinese medicine fermented preparation for preventing and treating piglet diarrhea comprises the following components in parts by weight: 10 parts of andrographis paniculata, 8 parts of punica granatum peels, 8 parts of breynia fruticosa, 8 parts of portulaca oleracea, 6 parts of sophora flavescens, 10 parts of anemone chinensis, and 10 parts of coptidis rhizome. The preparation method of the traditional Chinese medicine fermented preparation for preventing and treating piglet diarrhea comprises the following steps of: pre-treating; preparing strain fermenting liquid; and fermenting the traditional Chinese medicine. The traditional Chinese medicine fermented preparation contains traditional Chinese medicine components capable of improving intestinal environment and metabolic products of multiple probiotics; and the traditional Chinese medicine fermented preparation obviously reduces the occurrence probability of piglet diarrhea, has no toxic or side effects, no pollution, no residual and no drug resistance, and can be widely used for feeding piglets.
Description
Technical field
The present invention relates to a kind of veterinary drug, be specifically related to a kind of Chinese medicine fermentation preparation and manufacturing approach thereof of preventing and treating piglet diarrhea.
Background technology
Diarrhoea property disease takes place for annual winter-spring season sows farrowing peak period, piglet often sees, no matter piglet diarrhea is on the big pig farm of concentrating the farrowing management, still raises the medium and small pig farm under the state scattered, all is ubiquitous.Piglet diarrhea mainly occurs in two stages, i.e. in the time before and after birth back to 14 ages in days and the weaned piglet.Diarrhoea causes that piglet survival ratio is low, feed conversion rate is low, weightening finish waits slowly, has had a strong impact on the economic benefit of raising pigs.Show according to the interrelated data statistics; The sickness rate that many swinery piglet diarrhea property diseases are arranged is up to more than 50%; Case fatality rate up to 15%-20% about; The piglet death toll that is caused by diarrhoea accounts for 39.8% of the dead sum of piglet, and wherein suckling pig to wean front and back diarrhoea take place and cause the dead 20%-25% that accounts for.This shows that diarrhoea property disease is very big to the lasting generation harm of pig industry, can cause bigger economic loss.
At present, the main method of treatment piglet abdomen is to use antibiotic in a large number, it is reported, nearly all livestock and poultry cultivation family all has in breeding process adds antibiotic and hormone phenomenon in feedstuff.Yet poultry use antibiotic directly to cause bacterial drug resistance to increase in a large number, and drug resistance increases and means bacterial dissociation, when variation is accumulated to a certain degree by quantitative change back generation qualitative change, just possibly cause by this bacteriogenic disease popularity.The 2nd, can be in meat residual and health constituted a threat to.Often eat and contain antibiotic food, even trace, also can make the people untoward reaction occur.Often take in and contain antibiotic food, can make some bacterial strain produce drug resistance, Resistant strain can pass to other sensitive cellss with drug-resistance factor again, makes other xenogenesis bacterial strains become Resistant strain, thereby brings prevention and the difficulty of treating some person poultry disease.If the food of edible for a long time antibiotic remains can cause the death of some non-pathogenic bacterias in the human body, make the colony balance imbalance or cause deficiency disease such as vitamin, the infringement health causes.Particularly the abuse of chloromycetin very easily damages the hemopoietic function of human marrow, and causes the generation of aplastic anemia thus.
The typical feature of Chinese medicine fermentation pharmacy technology is exactly a biotransformation, can summarize following some: 1. the active component of medicine, active substance are able to extract, utilize to greatest extent; 2. medicine gets into the effective active component that can not directly be utilized behind the body, because of directly being utilized external being accomplished, brings into play proper efficiency rapidly; 3. preferred body probiotics itself has the function of replenishing or strengthening original medicine; 4. the Chinese medicine fermentation pharmacy is compared with original medicine and has been produced new active substance, thereby has new health care, prevention or treatment function.
Summary of the invention
The object of the present invention is to provide a kind of Chinese medicine fermentation preparation of preventing and treating piglet diarrhea; The multiple Chinese medicine ingredients of intestinal environment and the metabolite of multiple probiotic bacteria of improving contained in its inside; This Chinese medicine fermentation preparation has the generation probability of tangible reduction piglet diarrhea; And have no side effect, pollution-free, noresidue, do not produce drug resistance.
For achieving the above object, the present invention adopts following technical scheme:
A kind of Chinese medicine fermentation preparation of preventing and treating piglet diarrhea, the parts by weight of its each component consist of: 10 parts of Herba Andrographis, 8 parts of Pericarpium Granatis, 8 parts of Fruticose Breyniae Leaf and Twigs, 8 parts of Herba Portulacaes, 6 parts of Radix Sophorae Flavescentiss, 10 parts of the Radix Pulsatillaes, 10 parts of Rhizoma Coptidis.
A kind of method of making the Chinese medicine fermentation preparation of control piglet diarrhea may further comprise the steps:
The first step: pretreatment:
Choose each component material, oven dry is after 1～2 hour down respectively at 50～60 ℃, and 60 eye mesh screens are crossed in pulverizing, and are subsequent use;
Second step: preparation strain fermentating liquid:
Probiotic bacteria is inoculated in the culture medium, cultivates primary seed solution,, be inoculated into respectively and carry out amplification culture in the secondary seed fermentation tank, stop when reaching 24～36 hours cultivating when probiotic bacteria growth in the time of 20～24 hours, subsequent use;
The 3rd step: Chinese medicine fermentation:
Each component material mix homogeneously that pretreatment is good, adding accounts for 1～1.2 times water of the weight of each component material gross weight, stirs; At 100 ℃ of following steam sterilization 15～20min, after the cooling, add the weight ratio account for each component material gross weight and be total bacterium liquid measure of 10%; 1% brown sugar (Saccharum Sinensis Roxb.) 35～37 ℃ of closing anaerobic fermentations 7-10 days, whenever stirred once at a distance from 24 hours; After the fermentation ends, under 35～50 ℃ of conditions, dry, packing is up to the standards.
Described probiotic bacteria is a Clostridium butyricum.
Know-why of the present invention: microbial fermentation carries out under comparatively gentle condition such as room temperature, normal pressure, so can protect active ingredient of Chinese herbs to exempt from destruction to greatest extent; And the active component that number molecular weight is bigger in the Chinese medicine becomes small-molecule active substance after fermentation and quilt directly utilizes; This has improved the drug effect of Chinese medicine widely; The enzyme that Clostridium butyricum produces carries out structural modification to active ingredient of Chinese herbs; Generate secondary metabolite, thereby produced new active substance, thereby have new drug effect;
Adopt the Clostridium butyricum fermented tcm; Can produce like cellulase, amylase, protease etc.; Make the material of difficult absorptions such as cellulose in the Chinese medicine, lignin fully degrade, pharmaceutically active ingredient is able to discharge to greatest extent in making, and simultaneously cellulose conversion is become oligosaccharide; The growth while that promotes probiotics in the piglet gastrointestinal tract, the effective ingredient in the feedstuff was absorbed by animal more easily under the effect of the various enzymes that produced.
The present invention compares with other products has following advantage:
1, the present invention can repair the intestine of young pigs mucosa, improves the intestine of young pigs environment, effectively reduces the chance of occurrence of piglet diarrhea, promotes that piglet grows up healthy and sound, and reduces effects such as culturing input;
2, the present invention adopts Clostridium butyricum direct fermentation Chinese medicine, has protected active ingredient of Chinese herbs to exempt from destruction, produces new drug effect;
3, the present invention utilizes Chinese medicine and the probiotic bacteria new veterinary drug of development that interacts, but the advantage of nothing ratio is arranged, and it has and has no side effect, pollution-free, noresidue, do not produce security features such as drug resistance;
4, the effect of Chinese medicine fermentation preparation control piglet diarrhea of the present invention is obvious, and reliable in quality can satisfy people to the meat products requirement of safe production.
The specific embodiment
Embodiment 1:
A kind of Chinese medicine fermentation preparation of preventing and treating piglet diarrhea, the parts by weight of its each component consist of: 10 parts of Herba Andrographis, 8 parts of Pericarpium Granatis, 8 parts of Fruticose Breyniae Leaf and Twigs, 8 parts of Herba Portulacaes, 6 parts of Radix Sophorae Flavescentiss, 10 parts of the Radix Pulsatillaes, 10 parts of Rhizoma Coptidis; Probiotic bacteria: Clostridium butyricum.
A kind of method of making the Chinese medicine fermentation preparation of control piglet diarrhea may further comprise the steps:
The first step: pretreatment:
Choose each component material, oven dry is after 2 hours down respectively at 60 ℃, and 60 eye mesh screens are crossed in pulverizing, and are subsequent use;
Second step: preparation strain fermentating liquid:
Clostridium butyricum is inoculated in culture medium, and (yeast extract 3 grams, beef extract 10 grams, tryptone 10 grams, glucose 5 grams, sodium chloride 5 grams, sodium acetate trihydrate 3 grams, cysteine hydrochloride 0.5 gram, distilled water add to 1000ml; Regulate PH7.1 ± 0.1, normal pressure) in; Under 37 ℃, the condition of 200rpm, sealing, cultivate primary seed solution; Through 24h, when bacterial content reaches 5 * 10
7During the CFU/ml left and right sides, primary seed solution is inoculated into amplification culture in the secondary seed fermentation tank with inoculum concentration 5%, cultivation temperature is 37 ℃, through 36h, when bacteria growing reaches 10 to working as bacterial content
9When CFU/ml is above, stop to cultivate, subsequent use;
The 3rd step: Chinese medicine fermentation:
Each component material mix homogeneously that pretreatment is good, adding accounts for 1.2 times water of the weight of each component material gross weight, stirs; At 100 ℃ of following steam sterilization 20min, after the cooling, add the weight ratio account for each component material gross weight and be total bacterium liquid measure of 10%; 1% brown sugar (Saccharum Sinensis Roxb.) 37 ℃ of closing anaerobic fermentations 7 days, whenever stirred once at a distance from 24 hours; After the fermentation ends, under 50 ℃ of conditions, dry, packing is up to the standards.
Below through concrete test the influence of the present invention to piglet diarrhea and fertility performance is described.
Test 1:
1, the time and the place of test: tested August 27,2011 12 days to 2011 July in and accomplish on certain pig farm;
2, the selection of experimental animal and grouping: select for use from identical, age in days is identical, body weight close (about 6-7 kilogram); 120 of Du of 21 ages in days wean * big * long three way cross commodity piglet; Be divided into 5 processed group at random; Each processed group is established two repeating groups, 12 pigs of each repeating groups, public female half and half;
3, experimental period: experimental period is 14 days, piglet 21 age in days to 35 ages in days;
4, EXPERIMENTAL DESIGN and test daily ration: with reference to the nutritional need design corn bean cake type basal diet (table 1) of NRC (1998) pig.
Table 1 basal diet
In basal diet, add antibiotic 40ppm (bacitracin zinc) and constitute the 1# daily ration, the matched group of feeding examination pig; In basal diet, add the common Chinese preparation 1500ppm that the present invention prevents and treats piglet diarrhea (wherein 10 parts of Herba Andrographis, 8 parts of Pericarpium Granatis, 8 parts of Fruticose Breyniae Leaf and Twigs, 8 parts of Herba Portulacaes, 6 parts of Radix Sophorae Flavescentiss, 10 parts of the Radix Pulsatillaes, 10 parts of Rhizoma Coptidis respectively; Descend oven dry after 2 hours respectively at 60 ℃; Pulverize, cross 60 eye mesh screens and promptly get); Add Chinese medicine fermentation preparation 800ppm of the present invention, 1000ppm, 1200ppm formation 2#, 3#, 4#, 5# daily ration, respectively feeding experiment group 1, test group 2, test group 3, test group 4 examination pigs;
5, daily management: semi open model pig house, dry powder feed, free choice feeding and drinking-water; Conventional program immunity sterilization;
6, test index and method:
A, feed intake (gram/head day) they are unit with the repeating groups, the inventory of accurate recording every day, and the clear hopper of m seq calculates average daily ingestion amount;
B, daily gain (gram/head day) are unit with the repeating groups, second day early morning of same day on-test early morning and off-test, weigh on an empty stomach, by gained initial weight, last re-computation total augment weight and per day weightening finish;
C, feedstuff-meat ratio:, calculate the feedstuff-meat ratio of each repeating groups by feed intake, daily gain;
D, diarrhea rate (%): with the repeating groups is unit, and normal piglets feces water content about 60%, moisture surpass and be regarded as diarrhoea more than 90%, observe every day 3 times diarrhoea at least at any time, record diarrheal pig number of elements, and average day diarrhea rate is calculated in off-test;
E, survival rate (%): with the repeating groups is unit, is recorded as pig number of elements alive, and off-test is calculated to be motility rate.
7, the processing of data: test data adopts the spss11.5 statistical software to carry out the variance branch and analyzes and multiple comparisons.
8, result and analysis:, see table 2 to the fertility performance of ablactational baby pig and the influence of diarrhea rate:
The female different persons of shoulder marking-up represent significant difference (P＜0.05) in the notes same column, and shoulder marking-up parent phase is represented difference not remarkable (P＞0.05) with the person.
Visible by table 2, 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 4.6% and 4.1% respectively, shows the feed intake after the Chinese medicine fermentation preparation 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 can significantly improve daily gain (P＜0.05) with test group 4; The piglet feedstuff-meat ratio of test group 1 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 numeral is superior to 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 diarrhea rate between the matched group there were significant differences (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 superior to matched group (P＞0.05); Explain that Chinese medicine fermentation preparation of the present invention can improve the fertility performance of piglet and reduce diarrhea rate, and effect obviously is superior to common Chinese group (P＜0.05).
Test 2:
1, test material: sodium butyrate and product of the present invention;
2, experimental animal: provide by certain integrated culture field and to select body weight, age in days close, become the monobasic of nest assorted * last long white hybrid piglet 6 nests, be divided into three groups at random by nest;
3, test daily ration and trophic level: three groups normal feedstuff level and trophic level are identical, and testing 1 group is matched group.Test 2 groups of sodium butyrates and add in normal feedstuff 1% ratio, test 3 groups by the product of the present invention that adds 0.1% in the normal feedstuff, feedstuff is processed by sampling principle and method sampling, analyzes the conventional ingredient and the feed acidity of feedstuff then.
Feed formula and trophic level (%): corn 58, bean cake 28, whey powder 2.5, fish flour, 3, conch meal 0.2, bone meal, 1.5, Sal 0.3, mineral additive 1, multidimensional additive 10 grams; The about 3.23Mcal/kg of digestible energy, crude protein 17.8, calcium 0.81, phosphorus 0.62, pH6.84.
4, test is handled: 17 ages in days begin food calling, and the wean of 30 ages in days is weaned preceding 10 days respectively with corresponding test material food calling, and 58 ages in days finish test.The feedstuff dry feeding, feed 6 every day in early stage, 4 times/day later stages in mid-term 3 times/day.Free choice feeding, freely drink water, disease takes place in time treat, other feeding and management and diseases prevention and treatment more solito are carried out.During test, by a weighing, record beginning counterpoise, heavy eventually.Day by day each nest piglet diarrhea head number of observed and recorded as long as a hurdle has a diarrhoea promptly to write down this nest diarrhoea, and writes down a piglet number that needs treatment because of diarrhoea, writes down pig house environment temperature every day simultaneously.
5, result of the test:
Table 3 diarrhoea incidence (%)
Anti-diarrhoea effect normal piglets fecal water content about 60%, moisture is regarded as diarrhoea more than 90%.The diarrhoea index be each nest at the diarrhoea natural law of the phase of examination divided by total natural law of examination phase.Pig house environment temperature on average is 10～21.5 ℃ in the examination phase; Can find out that from table 3 wean to wean back is during two weeks, trying 2 groups of frequencies of disease development is 11.5%; Trying 3 groups of frequencies of disease development is 5.5%; Try 3 groups and try 2 groups low 6%, to initial data statistical result difference not significantly (P＞0.05), trying 1 group of frequency of disease development is 22.5%.Have a competition 2,3 groups high by 11% respectively of examinations, 17.0%, to initial data statistical result significant difference (P＜0.05).
Table 4 weightening finish and efficiency of feed utilization (kg)
Weightening finish and efficiency of feed utilization: can find out 1 group of average daily weightening finish 287 gram from table 4; 2 groups of 311 grams that daily increase weight, 3 groups is 314 grams, the initial data statistics shows difference not remarkable (P＞0.05); Test group is compared significant difference (P＜0.05) with the matched group statistics, but the food consumption rate does not have significant change.Explain that the present invention can improve the fertility performance of piglet and reduce diarrhea rate, and effect obviously is superior to matched group and sodium butyrate group (P＜0.05).
Claims (3)
1. a Chinese medicine fermentation preparation of preventing and treating piglet diarrhea is characterized in that the parts by weight of its each component consist of: 10 parts of Herba Andrographis, 8 parts of Pericarpium Granatis, 8 parts of Fruticose Breyniae Leaf and Twigs, 8 parts of Herba Portulacaes, 6 parts of Radix Sophorae Flavescentiss, 10 parts of the Radix Pulsatillaes, 10 parts of Rhizoma Coptidis.
2. a method of making the Chinese medicine fermentation preparation of the described control piglet diarrhea of claim 1 is characterized in that, may further comprise the steps:
The first step: pretreatment:
Choose each component material, oven dry is after 1～2 hour down respectively at 50～60 ℃, and 60 eye mesh screens are crossed in pulverizing, and are subsequent use;
Second step: preparation strain fermentating liquid:
Probiotic bacteria is inoculated in the culture medium, cultivates primary seed solution,, be inoculated into respectively and carry out amplification culture in the secondary seed fermentation tank, stop when reaching 24～36 hours cultivating when probiotic bacteria growth in the time of 20～24 hours, subsequent use;
The 3rd step: Chinese medicine fermentation:
Each component material mix homogeneously that pretreatment is good, adding accounts for 1～1.2 times water of the weight of each component material gross weight, stirs; At 100 ℃ of following steam sterilization 15～20min, after the cooling, add the weight ratio account for each component material gross weight and be total bacterium liquid measure of 10%; 1% brown sugar (Saccharum Sinensis Roxb.) 35～37 ℃ of closing anaerobic fermentations 7-10 days, whenever stirred once at a distance from 24 hours; After the fermentation ends, under 35～50 ℃ of conditions, dry, packing is up to the standards.
3. a kind of method of making the Chinese medicine fermentation preparation of control piglet diarrhea according to claim 2, it is characterized in that: described probiotic bacteria is a Clostridium butyricum.
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| CN103120726A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Traditional Chinese medicine composition for improving weaned pig growth performance and preparation method thereof |
| CN104171377A (en) * | 2014-07-10 | 2014-12-03 | 南宁荣港生物科技有限公司 | Traditional Chinese medicine synbiotic feed additive capable of preventing and treating diarrhea of piglest and preparation method of traditional Chinese medicine synbiotic feed additive capable of preventing and treating diarrhea of piglets |
| CN104224968A (en) * | 2014-10-14 | 2014-12-24 | 曹雨辰 | Veterinary medicine oral liquid for treating livestock and poultry diarrhea and preparation method thereof |
| CN104721443A (en) * | 2015-03-23 | 2015-06-24 | 河南省帝一方生物制药有限公司 | Traditional Chinese medicine compound preparation for preventing and treating piglet diarrhea and preparation method thereof |
| CN104758390A (en) * | 2014-12-10 | 2015-07-08 | 北京康华远景科技股份有限公司 | Botanical drug compound preparation for preventing and treating porcine diarrhea and preparation method thereof |
| CN105146216A (en) * | 2015-10-15 | 2015-12-16 | 格特生物制药（天津）有限公司 | Fermented traditional Chinese medicine preparation capable of reducing diarrhea ratio of piglets |
| CN105748742A (en) * | 2014-12-19 | 2016-07-13 | 北京大北农动物保健科技有限责任公司 | Pharmaceutical composition used for livestock and poultry dysentery and feed and preparation method thereof |
| CN106234799A (en) * | 2016-08-15 | 2016-12-21 | 吉林大学 | A kind of piglet fermentation of Chinese herbal medicine feed additive and preparation technology thereof |
| CN107509873A (en) * | 2017-09-08 | 2017-12-26 | 凤台县顺民养殖有限公司 | It is a kind of to improve the dilute black earth pig special feed of Huaihe River porkling pig tension |
| CN109258958A (en) * | 2018-12-12 | 2019-01-25 | 湖南百宜饲料科技有限公司 | A kind of feed addictive of weanling pig |
| CN110786420A (en) * | 2019-12-11 | 2020-02-14 | 重庆市畜牧科学院 | Feed additive for piglets |
| CN114053383A (en) * | 2020-12-28 | 2022-02-18 | 焦作市农科利来生物科技有限公司 | Fermented traditional Chinese medicine microecological preparation for preventing and treating piglet diarrhea and preparation method thereof |
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| CN103120726A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Traditional Chinese medicine composition for improving weaned pig growth performance and preparation method thereof |
| CN104171377A (en) * | 2014-07-10 | 2014-12-03 | 南宁荣港生物科技有限公司 | Traditional Chinese medicine synbiotic feed additive capable of preventing and treating diarrhea of piglest and preparation method of traditional Chinese medicine synbiotic feed additive capable of preventing and treating diarrhea of piglets |
| CN104224968A (en) * | 2014-10-14 | 2014-12-24 | 曹雨辰 | Veterinary medicine oral liquid for treating livestock and poultry diarrhea and preparation method thereof |
| CN104758390A (en) * | 2014-12-10 | 2015-07-08 | 北京康华远景科技股份有限公司 | Botanical drug compound preparation for preventing and treating porcine diarrhea and preparation method thereof |
| CN105748742A (en) * | 2014-12-19 | 2016-07-13 | 北京大北农动物保健科技有限责任公司 | Pharmaceutical composition used for livestock and poultry dysentery and feed and preparation method thereof |
| CN104721443A (en) * | 2015-03-23 | 2015-06-24 | 河南省帝一方生物制药有限公司 | Traditional Chinese medicine compound preparation for preventing and treating piglet diarrhea and preparation method thereof |
| CN104721443B (en) * | 2015-03-23 | 2019-01-04 | 河南省帝一方生物制药有限公司 | A kind of compound Chinese medicinal preparation and preparation method thereof for preventing and treating grice diarrhoea |
| CN105146216A (en) * | 2015-10-15 | 2015-12-16 | 格特生物制药（天津）有限公司 | Fermented traditional Chinese medicine preparation capable of reducing diarrhea ratio of piglets |
| CN106234799A (en) * | 2016-08-15 | 2016-12-21 | 吉林大学 | A kind of piglet fermentation of Chinese herbal medicine feed additive and preparation technology thereof |
| CN107509873A (en) * | 2017-09-08 | 2017-12-26 | 凤台县顺民养殖有限公司 | It is a kind of to improve the dilute black earth pig special feed of Huaihe River porkling pig tension |
| CN109258958A (en) * | 2018-12-12 | 2019-01-25 | 湖南百宜饲料科技有限公司 | A kind of feed addictive of weanling pig |
| CN110786420A (en) * | 2019-12-11 | 2020-02-14 | 重庆市畜牧科学院 | Feed additive for piglets |
| CN114053383A (en) * | 2020-12-28 | 2022-02-18 | 焦作市农科利来生物科技有限公司 | Fermented traditional Chinese medicine microecological preparation for preventing and treating piglet diarrhea and preparation method thereof |
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Application publication date: 20121003 |