CN107494936A - A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof - Google Patents
A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof Download PDFInfo
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Abstract
A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof, including the dandelion of parts by weights meter, the Radix Astragali, cordate houttuynia, humic acid fermentation raw material, bean cake powder, corn flour, water are prepared into culture medium；Then culture medium is sterilized with steam, culture medium cooled down afterwards；Microorganism mixed fermentation liquid is inoculated on culture medium, advanced act charitably aerobe fermentation and anaerobic fermentation, anaerobic fermentation terminate the Chinese medicine humic acid degraded, and Chinese medicine humic acid product is made through low temperature drying, is mixed afterwards with yeast cell wall, obtains Chinese medicine humic acid feed.Abundant raw material of the present invention, cost is low, simple production process, easy to operate；There is antibacterial, anti-inflammatory, be not easy to make pathogenic bacteria produce drug resistance and Immune-enhancing effect, there is the characteristics of safe efficient；Have no drug resistance, noresidue, toxic side effect it is small, the advantage of uniqueness is shown in terms of livestock and poultry cultivation；Mastitis for milk cows can effectively be prevented and treated.
Description
Technical field
The present invention relates to field of feed, more particularly to a kind of Chinese medicine humic acid feed for treating mastitis for milk cows and its
Preparation method.
Background technology
Mastitis for milk cows is the inflammation of mammary glandular cell and mammary gland interstitial.Cause the pathogen of breast inflammation, about 85% is hammer
Bacterium, micrococcus scarlatinae etc., 5% is staphylococcus.Other have a tubercle bacillus, Brucella, corynebacterium pyogenes, Escherichia coli,
Mycoplasma and mould etc..Economic loss caused by mastitis for milk cows is very big, is that the output of milk is reduced first, next to that sick breast can not
Drink, or even give people to infect disease, people's health is damaged.If mammitis can not treat in time, often make mastatrophy, milk
Juice can not be sold, or even completely lose galactogogue action.Therefore the work of preventing and treating mammitis is performed.At present, more using antibiotic
Mastitis for milk cows is treated, therapeutic effect is undesirable, and drug tolerant bacteria can also be given people by milk-born infection, residual in milk cow in addition
The antibiotic stayed can also induce human body and hypersensitivity occurs, and therefore, the non-antibiotic therapy of mastitis for milk cows gradually causes people
Concern.
The content of the invention
To solve the above problems, the invention provides a kind of Chinese medicine humic acid feed for treating mastitis for milk cows and its preparation
Method.Preparation method is：
Step 1：Prepare culture medium
Radix Astragali 30-60 parts of parts by weights meter, dandelion 30-50 parts, cordate houttuynia 10-40 parts, sodium humate 10-30 parts are passed through
40-80 mesh sieve, the above-mentioned substance after sieving and bean cake powder 6-10 parts, corn flour 6-10 parts, the stirring of water 120-180 parts is mixed
Conjunction is uniformly prepared into culture medium；
Step 2：Prepare Chinese medicine humic acid product
Then culture medium is sterilized with steam, 100-135 DEG C of sterilizing 15-30 minute, culture medium is cooled to afterwards
28 DEG C of -35 DEG C of cultivation temperatures；Microorganism mixed fermentation liquid is inoculated on culture medium, inoculative proportion 0.9-1.2:0.05-
0.2, advanced aerobe fermentation of acting charitably, fermentation condition is mixing speed 160-200r/min, starts ventilation ratio and is adjusted to 1：0.4-0.7,
Pressure is filled during culture will be maintained at 0.03-0.06pa, and tune throughput is 15m3/h and mixing speed is 200r/min, makes dissolved oxygen
Relative value 20%-40%, fermentation time 24-48 hours, 32-37 DEG C of fermentation temperature；Then anaerobic fermentation is carried out, ferment bar
Part is standing for fermentation, fermentation time 48-72 hours, 32 DEG C -37 DEG C of fermentation temperature, 0.5-3 minutes is stirred per 5-7 hours；
Anaerobic fermentation terminates the Chinese medicine humic acid fermentation product degraded, through low temperature（100 DEG C of ＜）Dry, moisture is not higher than 15%, is made
Chinese medicine humic acid product.
Step 3：By Chinese medicine humic acid product 50-90 parts obtained by step 2 and yeast cell wall 10-40 parts by weight
Match and produce.
Microorganism mixed fermentation liquid is sent out by bacillus subtilis fermentation liquor, the lichen bacillus ferments liquid, Lactobacillus plantarum
Zymotic fluid, by weight 40-60%: 40-60%: 10-15% ratio mix.
Microorganism mixed fermentation liquid, its production technology are as follows：
A：The production of bacillus subtilis fermentation liquor：
1) solid medium is：Glucose 1-3%, peptone 1-3%, yeast extract 0.3-0.6%, agar 1-4%,
Distilled water 90% -97%, regulation medium pH is 6.5- 7.0,110-120 DEG C of sterilizing 16-20 minute；Condition of culture：
It is put into 25-35 DEG C of constant incubator and cultivates 22-26h；
2) above-mentioned gained material is subjected to Shaking culture, culture medium is：Glucose 1-3%, peptone 0.5-2%, yeast
Cream 0.3-0.8%, water 94-99%, regulation Shake flask medium pH are 6.5-7.0,110-120 DEG C of sterilizing 16-20 minutes
；Condition of culture：Shaking flask is accessed in above-mentioned gained strain, is put into 25-35 DEG C of constant-temperature table, 180-230r/ min vibrations
Cultivate 22-26h；
3) above-mentioned gained material progress seeding tank, fermentation tank are expanded numerous：Culture medium is：Dregs of beans 0.5-1.3%, corn 0.2-
0.8%th, sucrose 1-3%, peptone 0.5-1.2%, yeast extract 0.3-0.8%, potassium dihydrogen phosphate 0.05-0.8%, ammonium sulfate
0.1-0.3%, sodium chloride 0.02-0.05%, defoamer 0.02-0.07%, water 90-98%, culture medium starting pH=
7.0-7.5,115-130 DEG C of sterilizing 20-40 minutes；Condition of culture：Shaking flask is to seeding tank inoculum concentration and seeding tank to hair
Fermentation tank is inoculated with by 3-7%, and the seed tank culture time is 22-26h, and the fermentation tank culture time is 30-40h；Viable count is not low
In 3.0 × 109cfu/ml；
B：The production of the lichen bacillus ferments liquid：
1) solid medium is：Peptone 8-12%, beef extract 1-5%, NaCl 3-7%, agar 12-18%, distilled water
58% -76%, regulation medium pH 7.0-7.5,105-125 DEG C of sterilizing 15-20 minute；Condition of culture：It is put into 30-35
22-24h is cultivated in DEG C constant incubator；
2) above-mentioned gained material is subjected to shake-flask seed culture：Culture medium is peptone 8-12%, beef extract 1-5%, NaCl
3-7%, agar 12-18%, distilled water 58% -76%, regulation medium pH 7.0-7.5,105-125 DEG C of 15-20 points of sterilizing
Clock；Condition of culture：It is put into 30-35 DEG C of constant incubator and cultivates 22-24h；Condition of culture：By above-mentioned gained strain
Shaking flask is accessed, is put into 30-35 DEG C of constant-temperature table, 180-220r/min shaken cultivations 22-26h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 2-7%, molasses 0.5-1.5%, peptone
0.3-0.8%, yeast extract 0.1-0.5%, potassium dihydrogen phosphate 0.05-0.2%, ammonium sulfate 0.05-0.5%, sodium chloride 0-
0.08%th, soya-bean oil 0.05-0.25%, defoamer 0-0.1%, water 89-97%, medium pH=6.5-7.5,115- is adjusted
125 DEG C of sterilizing 25-35 minutes；Condition of culture：Shaking flask is connect to seeding tank and seeding tank to fermentation tank inoculum concentration by 3-7%
Kind, the seed tank culture time is 22-26h, and the fermentation tank culture time is 32-40h；Viable count is not less than 3.0 × 109 cfu/
ml；
C：The production of Lactobacillus plantarum zymotic fluid：
1) solid medium is：Glucose 0.5-1.5%, peptone 0.5-1.5%, yeast extract 0.1-0.8%, beef
Cream 0.1-0.8%, potassium dihydrogen phosphate 0.05-0.15%, agar 1-3%, water 93%-98%, regulation Medium's PH Value to 6.5-
7.0th, 110-120 DEG C of sterilizing 15-25 minute；Condition of culture：It is put into Anaerobic culturel in 30-35 DEG C of constant incubator
22-24h ；
2) shake-flask seed culture：Culture medium is：Glucose 0.5-1.5%, peptone 0.5-1.5%, yeast extract
0.1-0.8%, beef extract 0.1-0.8%, potassium dihydrogen phosphate 0.05-0.15%, agar 1-3%, water 93%-98%, regulation training
Base pH value is supported to 6.5-7.0,110-120 DEG C of sterilizing 15-25 minutes；Condition of culture：Above-mentioned gained strain is accessed into shaking flask
Anaerobic culturel 22-26h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 0.5-1.5%, corn 0.2-0.7%, sucrose
0.5-1.5%, peptone 1-2%, yeast extract 0.2-0.7%, potassium dihydrogen phosphate 0.1-0.5%, ammonium sulfate 0.1-0.5%, second
Sour sodium 0.3-0.8%, trisodium citrate 0.1-0.5%, Tween 80 0.05-0.2%, magnesium sulfate 0.01-0.05%, soya-bean oil
0.1-0.3%, defoamer 0.03-0.07%, water 93%-97%, starting pH=6.5-7.0,110-125 DEG C of sterilizing 25-40
Minute；Condition of culture：Shaking flask is inoculated with to seeding tank and seeding tank to fermentation tank inoculum concentration by 3-7%, seed tank culture
Time is 22-24h, and the fermentation tank culture time is 30-40h, and viable count is not less than 2.0 × 109cfu/ml；
Chinese medicine humic acid product includes：Culture medium and microorganism mixed fermentation liquid, its ratio are 0.9-1.2:0.05-0.2；It is described
Culture medium include：By Radix Astragali 30-60 parts of parts by weights meter, dandelion 30-50 parts, cordate houttuynia 10-40 parts, sodium humate
10-30 parts, bean cake powder 6-10 parts, corn flour 6-10 parts, water 120-180 parts.
The advantage of the invention is that：
1. abundant raw material, cost is low, simple production process, easy to operate；
2. using cordate houttuynia, dandelion, the Radix Astragali and humic acid as raw material, there is antibacterial, anti-inflammatory, be not easy to make pathogenic bacteria produce drug resistance
With the effect of Immune-enhancing effect, there is the characteristics of safe efficient；
3. probiotics has the culture medium of oneself, a variety of probiotics organically combine with Chinese medicine；
4. the present invention after animal use, has no drug resistance, noresidue, toxic side effect are small, uniqueness is shown in terms of livestock and poultry cultivation
Advantage.
Embodiment：
Embodiment 1
A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof, its preparation method is：
Step 1：Prepare culture medium
, will by 60 mesh sieve by 50 parts of the Radix Astragali of parts by weights meter, 40 parts of dandelion, 25 parts of cordate houttuynia, 20 parts of sodium humate
8 parts of above-mentioned substance and bean cake powder after sieving, 8 parts of corn flour, 150 parts of water, which is uniformly mixed, is prepared into culture medium；
Step 2：Prepare Chinese medicine humic acid product
Then culture medium is sterilized with steam, 120 DEG C sterilize 20 minutes, culture medium are carried out being cooled to 30 DEG C of cultures afterwards
Temperature；Microorganism mixed fermentation liquid is inoculated on culture medium, inoculative proportion 1:0.1, described microorganism mixed fermentation liquid
By bacillus subtilis fermentation liquor, the lichen bacillus ferments liquid, Lactobacillus plantarum zymotic fluid, by weight 50%: 40%:
10% ratio mixes.
Microorganism mixed fermentation liquid, its production technology are as follows：
A：The production of bacillus subtilis fermentation liquor：
1) solid medium is：Glucose 3%, peptone 1%, yeast extract 0.5%, agar 3.5%, distilled water
93%, regulation medium pH is 7.0,110 DEG C and sterilized 20 minutes；Condition of culture：It is put into 30 DEG C of constant incubators and trains
Support 24h；
2) above-mentioned gained material is subjected to Shaking culture, culture medium is：Glucose 2%, peptone 0.5%, yeast extract
0.5%, water 97%, regulation Shake flask medium pH is 7.0,110 DEG C and sterilized 20 minutes；Condition of culture：In above-mentioned gained
Strain accesses shaking flask, is put into 30 DEG C of constant-temperature tables, 200r/ min shaken cultivations 24h；
3) above-mentioned gained material progress seeding tank, fermentation tank are expanded numerous：Culture medium is：Dregs of beans 1%, corn 0.5%, sucrose
2%th, peptone 0.5%, yeast extract 0.5%, potassium dihydrogen phosphate 0.3%, ammonium sulfate 0.1%, sodium chloride 0.05%, defoaming
Agent 0.05%, water 95%, culture medium starting pH=7.0,120 DEG C sterilize 30 minutes；Condition of culture：Shaking flask to kind
Sub- tank inoculum concentration and seeding tank are inoculated with to fermentation tank by 5%, and the seed tank culture time is 24h, and the fermentation tank culture time is 35h
；Viable count is 5.0 × 109cfu/ml；
B：The production of the lichen bacillus ferments liquid：
1) solid medium is：Peptone 10%, beef extract 5%, NaCl 5%, agar 15%, distilled water 65%, regulation
7.0,110 DEG C of medium pH sterilizes 18 minutes；Condition of culture：It is put into 35 DEG C of constant incubators and cultivates 22h；
2) above-mentioned gained material is subjected to shake-flask seed culture：Culture medium is peptone 10%, beef extract 5%, NaCl 5%,
Agar 15%, distilled water 65%, 7.0,110 DEG C of regulation medium pH sterilize 18 minutes；Condition of culture：It is put into 35 DEG C of constant temperature
22h is cultivated in incubator；Condition of culture：Above-mentioned gained strain is accessed into shaking flask, is put into 35 DEG C of constant-temperature tables, 200r/
Min shaken cultivations 24h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 5%, molasses 0.5%, peptone 0.7%,
Yeast extract 0.3%, potassium dihydrogen phosphate 0.05%, ammonium sulfate 0.15%, sodium chloride 0.05%, soya-bean oil 0.15%, defoamer
0.1%, water 93%, regulation medium pH=7.0,120 DEG C sterilize 30 minutes；Condition of culture：Shaking flask is to seeding tank and kind
Sub- tank is to fermentation tank inoculum concentration by 5% inoculation, and the seed tank culture time is 24h, and the fermentation tank culture time is 35h；Viable count
For 4.0 × 109cfu/ml；
C：The production of Lactobacillus plantarum zymotic fluid：
1) solid medium is：Glucose 1%, peptone 1%, yeast extract 0.4%, beef extract 0.5%, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.1%, agar 2%, water 95%, regulation Medium's PH Value sterilize 25 minutes to 7.0,110 DEG C；Condition of culture：
It is put into Anaerobic culturel 24h in 30 DEG C of constant incubators；
2) shake-flask seed culture：Culture medium is：Glucose 1%, peptone 1%, yeast extract 0.4%, beef extract
0.5%th, potassium dihydrogen phosphate 0.1%, agar 2%, water 95%, regulation Medium's PH Value sterilize 25 minutes to 7.0,110 DEG C；
Condition of culture：By above-mentioned gained strain access shaking flask Anaerobic culturel 24h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 1%, corn 0.5%, sucrose 1.5%, albumen
Peptone 1.5%, yeast extract 0.5%, potassium dihydrogen phosphate 0.5%, ammonium sulfate 0.35%, sodium acetate 0.6%, trisodium citrate
0.2%th, Tween 80 0.1%, magnesium sulfate 0.02%, soya-bean oil 0.2%, defoamer 0.03%, water 93%, starting pH=
7.0th, 110 DEG C sterilize 40 minutes；Condition of culture：Shaking flask connects to seeding tank and seeding tank to fermentation tank inoculum concentration by 5%
Kind, the seed tank culture time is 24h, and the fermentation tank culture time is 35h, and viable count is 3 × 109cfu/ml；
Advanced aerobe fermentation of acting charitably, fermentation condition are mixing speed 180r/min, start ventilation ratio and are adjusted to 1：0.5, during culture
0.05pa will be maintained at by filling pressure, and tune throughput is 15m3/h and mixing speed is 200r/min, makes dissolved oxygen relative value 30%, is sent out
36 hours ferment time, 35 DEG C of fermentation temperature；Then anaerobic fermentation is carried out, fermentation condition is standing for fermentation, and fermentation time 55 is small
When, 35 DEG C of fermentation temperature, stir 1 minute within every 6 hours；Anaerobic fermentation terminates the Chinese medicine humic acid fermentation product degraded,
Through 85 DEG C of dryings of low temperature, moisture 10%, Chinese medicine humic acid product is made.
Step 3：It is by weight proportioning for 30 parts with yeast cell wall by 70 parts of Chinese medicine humic acid product obtained by step 2
.
Embodiment 2
A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof, its preparation method is：
Step 1：Prepare culture medium
, will by 50 mesh sieve by 40 parts of the Radix Astragali of parts by weights meter, 35 parts of dandelion, 10 parts of cordate houttuynia, 15 parts of sodium humate
6 parts of above-mentioned substance and bean cake powder after sieving, 7 parts of corn flour, 130 parts of water, which is uniformly mixed, is prepared into culture medium；
Step 2：Prepare Chinese medicine humic acid product
Then culture medium is sterilized with steam, 130 DEG C sterilize 15 minutes, culture medium are carried out being cooled to 33 DEG C of cultures afterwards
Temperature；Microorganism mixed fermentation liquid is inoculated on culture medium, inoculative proportion 1.2:0.1, described microorganism mixed fermentation
Liquid is by bacillus subtilis fermentation liquor, the lichen bacillus ferments liquid, Lactobacillus plantarum zymotic fluid, by weight 40%: 45%:
15% ratio mixes.
Microorganism mixed fermentation liquid, its production technology are as follows：
A：The production of bacillus subtilis fermentation liquor：
1) solid medium is：Glucose 1%, peptone 3%, yeast extract 0.3%, agar 3.7%, distilled water
92%, regulation medium pH is 6.5,115 DEG C and sterilized 18 minutes；Condition of culture：It is put into 25 DEG C of constant incubators and trains
Support 26h；
2) above-mentioned gained material is subjected to Shaking culture, culture medium is：Glucose 3%, peptone 1.2%, yeast extract
0.8%, water 95%, regulation Shake flask medium pH is 6.5,115 DEG C and sterilized 18 minutes；Condition of culture：In above-mentioned gained
Strain accesses shaking flask, is put into 35 DEG C of constant-temperature tables, 180r/ min shaken cultivations 22h；
3) above-mentioned gained material progress seeding tank, fermentation tank are expanded numerous：Culture medium is：Dregs of beans 1.2%, corn 0.7%, sugarcane
Sugar 3%, peptone 1%, yeast extract 0.3%, potassium dihydrogen phosphate 0.6%, ammonium sulfate 0.15%, sodium chloride 0.03%, disappear
Infusion 0.02%, water 93%, culture medium starting pH=7.5,130 DEG C sterilize 20 minutes；Condition of culture：Shaking flask arrives
To fermentation tank by 7% inoculation, the seed tank culture time is 22h for seeding tank inoculum concentration and seeding tank, and the fermentation tank culture time is
30h ；Viable count is 4.0 × 109cfu/ml；
B：The production of the lichen bacillus ferments liquid：
1) solid medium is：Peptone 8%, beef extract 3%, NaCl 7%, agar 12%, distilled water 70%, regulation
7.5,125 DEG C of medium pH sterilizes 15 minutes；Condition of culture：It is put into 30 DEG C of constant incubators and cultivates 24h；
2) above-mentioned gained material is subjected to shake-flask seed culture：Culture medium is peptone 8%, beef extract 3%, NaCl 7%,
Agar 12%, distilled water 70%, 7.5,125 DEG C of regulation medium pH sterilize 15 minutes；Condition of culture：It is put into 30 DEG C of constant temperature
24h is cultivated in incubator；Condition of culture：Above-mentioned gained strain is accessed into shaking flask, is put into 30 DEG C of constant-temperature tables, 220r/
Min shaken cultivations 26h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 7%, molasses 1.3%, peptone 0.35%,
Yeast extract 0.4%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.5%, sodium chloride 0.07%, soya-bean oil 0.23%, defoamer
0.05%, water 90%, regulation medium pH=7.5,115 DEG C sterilize 35 minutes；Condition of culture：Shaking flask to seeding tank and
Seeding tank is to fermentation tank inoculum concentration by 7% inoculation, and the seed tank culture time is 22h, and the fermentation tank culture time is 40h；Viable bacteria
Number is 5.0 × 109cfu/ml；
C：The production of Lactobacillus plantarum zymotic fluid：
1) solid medium is：Glucose 0.5%, peptone 1.1%, yeast extract 0.6%, beef extract 0.75%, phosphorus
Acid dihydride potassium 0.05%, agar 3%, water 94%, regulation Medium's PH Value sterilize 15 minutes to 6.5,120 DEG C；Cultivate bar
Part：It is put into Anaerobic culturel 22h in 35 DEG C of constant incubators；
2) shake-flask seed culture：Culture medium is：Glucose 0.5%, peptone 1.1%, yeast extract 0.6%, beef
Cream 0.75%, potassium dihydrogen phosphate 0.05%, agar 3%, water 94%, regulation Medium's PH Value sterilize 15 points to 6.5,120 DEG C
Clock；Condition of culture：By above-mentioned gained strain access shaking flask Anaerobic culturel 22h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 0.5%, corn 0.2%, sucrose 0.8%, egg
White peptone 1.8%, yeast extract 0.7%, potassium dihydrogen phosphate 0.35%, ammonium sulfate 0.4%, sodium acetate 0.3%, trisodium citrate
0.5%th, Tween 80 0.05%, magnesium sulfate 0.05%, soya-bean oil 0.3%, defoamer 0.05%, water 94%, starting pH=
6.5th, 125 DEG C sterilize 25 minutes；Condition of culture：Shaking flask connects to seeding tank and seeding tank to fermentation tank inoculum concentration by 7%
Kind, the seed tank culture time is 22h, and the fermentation tank culture time is 40h, and viable count is 5.0 × 109cfu/ml；
Advanced aerobe fermentation of acting charitably, fermentation condition are mixing speed 200r/min, start ventilation ratio and are adjusted to 1：0.7, during culture
0.06pa will be maintained at by filling pressure, and tune throughput is 15m3/h and mixing speed is 200r/min, makes dissolved oxygen relative value 25%, is sent out
45 hours ferment time, 32 DEG C of fermentation temperature；Then anaerobic fermentation is carried out, fermentation condition is standing for fermentation, and fermentation time 60 is small
When, 32 DEG C of fermentation temperature, stir 2 minutes within every 7 hours；Anaerobic fermentation terminates the Chinese medicine humic acid fermentation product degraded,
Through 60 DEG C of dryings of low temperature, moisture 11%, Chinese medicine humic acid product is made.
Step 3：It is by weight proportioning for 20 parts with yeast cell wall by 80 parts of Chinese medicine humic acid product obtained by step 2
.
Embodiment 3
A kind of Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof, its preparation method is：
Step 1：Prepare culture medium
, will by 80 mesh sieve by 35 parts of the Radix Astragali of parts by weights meter, 50 parts of dandelion, 40 parts of cordate houttuynia, 30 parts of sodium humate
10 parts of above-mentioned substance and bean cake powder after sieving, 10 parts of corn flour, 180 parts of water, which is uniformly mixed, is prepared into culture medium；
Step 2：Prepare Chinese medicine humic acid product
Then culture medium is sterilized with steam, 100 DEG C sterilize 30 minutes, culture medium are carried out being cooled to 28 DEG C of cultures afterwards
Temperature；Microorganism mixed fermentation liquid is inoculated on culture medium, inoculative proportion 0.9:0.05, described microorganism mixed fermentation
Liquid is by bacillus subtilis fermentation liquor, the lichen bacillus ferments liquid, Lactobacillus plantarum zymotic fluid, by weight 45%: 43%:
12% ratio mixes.
Microorganism mixed fermentation liquid, its production technology are as follows：
A：The production of bacillus subtilis fermentation liquor：
1) solid medium is：Glucose 1%, peptone 1%, yeast extract 0.2%, agar 1.8%, distilled water
96%, regulation medium pH is 7.0,120 DEG C and sterilized 16 minutes；Condition of culture：It is put into 25 DEG C of constant incubators and trains
Support 26h；
2) above-mentioned gained material is subjected to Shaking culture, culture medium is：Glucose 1%, peptone 1.7%, yeast extract
0.3%, water 97%, regulation Shake flask medium pH is 7.0,120 DEG C and sterilized 16 minutes；Condition of culture：In above-mentioned gained
Strain accesses shaking flask, is put into 25 DEG C of constant-temperature tables, 230r/ min shaken cultivations 26h；
3) above-mentioned gained material progress seeding tank, fermentation tank are expanded numerous：Culture medium is：Dregs of beans 0.5%, corn 0.2%, sugarcane
Sugar 1%, peptone 1%, yeast extract 0.8%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.3%, sodium chloride 0.04%, defoaming
Agent 0.06%, water 96%, culture medium starting pH=7.0,115 DEG C sterilize 40 minutes；Condition of culture：Shaking flask to kind
Sub- tank inoculum concentration and seeding tank are inoculated with to fermentation tank by 3%, and the seed tank culture time is 26h, and the fermentation tank culture time is 40h
；Viable count is 5.0 × 109cfu/ml；
B：The production of the lichen bacillus ferments liquid：
1) solid medium is：Peptone 12%, beef extract 1%, NaCl 3%, agar 18%, distilled water 76%, regulation
7.0,105 DEG C of medium pH sterilizes 20 minutes；Condition of culture：It is put into 33 DEG C of constant incubators and cultivates 23h；
2) above-mentioned gained material is subjected to shake-flask seed culture：Culture medium is peptone 12%, beef extract 1%, NaCl3%,
Agar 18, distilled water 76%, 7.0,105 DEG C of 20 clocks of sterilizing of regulation medium pH；Condition of culture：It is put into 33 DEG C of constant temperature trainings
Support in case and cultivate 23h；Condition of culture：Above-mentioned gained strain is accessed into shaking flask, is put into 33 DEG C of constant-temperature tables, 200r/min
Shaken cultivation 22h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 2%, molasses 1%, peptone 0.4%, ferment
Female cream 0.1%, potassium dihydrogen phosphate 0.2%, ammonium sulfate 0.3%, sodium chloride 0.05%, soya-bean oil 0.05%, defoamer
0.1%, water 95.8%, regulation medium pH=6.5,125 DEG C sterilize 25 minutes；Condition of culture：Shaking flask to seeding tank and
Seeding tank is to fermentation tank inoculum concentration by 3% inoculation, and the seed tank culture time is 26h, and the fermentation tank culture time is 32h；Viable bacteria
Number is 5.0 × 109cfu/ml；
C：The production of Lactobacillus plantarum zymotic fluid：
1) solid medium is：Glucose 1.24%, peptone 0.9%, yeast extract 0.7%, beef extract 0.55%, phosphorus
Acid dihydride potassium 0.11%, agar 2.5%, water 94%, regulation Medium's PH Value sterilize 20 minutes to 7.0,115 DEG C；Culture
Condition：It is put into Anaerobic culturel 23h in 33 DEG C of constant incubators；
2) shake-flask seed culture：Culture medium is：Glucose 1.24%, peptone 0.9%, yeast extract 0.7%, beef
Cream 0.55%, potassium dihydrogen phosphate 0.11%, agar 2.5%, water 94%, regulation Medium's PH Value to 7.0,115 DEG C of sterilizings 20
Minute；Condition of culture：By above-mentioned gained strain access shaking flask Anaerobic culturel 26h；
3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 0.8%, corn 0.6%, sucrose 1.04%, egg
White peptone 1.6%, yeast extract 0.6%, potassium dihydrogen phosphate 0.45%, ammonium sulfate 0.45%, sodium acetate 0.7%, trisodium citrate
0.4%th, Tween 80 0.12%, magnesium sulfate 0.03%, soya-bean oil 0.15%, defoamer 0.06%, water 93%, starting pH=
6.6th, 116 DEG C sterilize 30 minutes；Condition of culture：Shaking flask connects to seeding tank and seeding tank to fermentation tank inoculum concentration by 6%
Kind, the seed tank culture time is 23h, and the fermentation tank culture time is 35h, and viable count is 4.0 × 109cfu/ml；
Advanced aerobe fermentation of acting charitably, fermentation condition are mixing speed 160r/min, start ventilation ratio and are adjusted to 1：0.4, during culture
0.03pa will be maintained at by filling pressure, and tune throughput is 15m3/h and mixing speed is 200r/min, makes dissolved oxygen relative value 40%, is sent out
24 hours ferment time, 37 DEG C of fermentation temperature；Then anaerobic fermentation is carried out, fermentation condition is standing for fermentation, and fermentation time 72 is small
When, 37 DEG C of fermentation temperature, stir 0.5 minute within every 5 hours；Anaerobic fermentation terminates the Chinese medicine humic acid fermentation degraded
Product, through 50 DEG C of dryings of low temperature, moisture 8%, Chinese medicine humic acid product is made.
Step 3：It is by weight proportioning for 10 parts with yeast cell wall by 90 parts of Chinese medicine humic acid product obtained by step 2
.
Clinical trial result is as follows：
Embodiment one：
Using 60, Inner Mongol Liangcheng County pasture milking cows as subjects, two groups are randomly divided into, A groups and B groups, every group
30；The full mixing TMR daily rations of A groups feeding, B groups increase 150g Chinese medicine humic acid feeds on the basis of A groups, and a point early, evening raises twice
Hello, other rearing conditions are constant.Before experiment, feeding 7d, feeding 15d are respectively fallen ill with CTM methods two groups of mammitises of measure and body cell
Situation；Result of the test shows：In A groups 15d, recessive mastitis body cell is totally in rising 9.8%, the clinical mammitis incidence of disease
For 3.3%；After B groups 7d, after overall somatic number declines 8%, 15d, overall somatic number declines 22%, clinical mammitis morbidity
Rate is 0%.
Embodiment two：
Using 16, the Fengning pasture clinical mammitis milk cows of He Sitan as subjects, two groups are randomly divided into, A groups 8
With B groups 8；Every beef injection cephalo parenteral solution 60ml, B group additionally adds Chinese medicine humic acid feed 200g to A groups daily daily, point
Early, evening secondary feeding, other rearing conditions are constant.Experiment before and feeding 10d respectively with CTM methods determine two groups of mammitis cure rates and
Body cell situation；Result of the test shows：In A groups 10d, clinical mammitis cure rate is 50%；After B groups 10d, clinical mammitis
Cure rate is 50%, and body cell also significantly reduces in remaining milk cow milk.Conclusion：Chinese medicine humic acid feed is imitated with antibiosis extract for treating
Fruit is suitable.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc., it should be included in the scope of the protection.
Claims (4)
- A kind of 1. Chinese medicine humic acid feed for treating mastitis for milk cows and preparation method thereof, it is characterised in that：Preparation method is：Step 1：Prepare culture mediumRadix Astragali 30-60 parts of parts by weights meter, dandelion 30-50 parts, cordate houttuynia 10-40 parts, sodium humate 10-30 parts are passed through 40-80 mesh sieve, the above-mentioned substance after sieving and bean cake powder 6-10 parts, corn flour 6-10 parts, the stirring of water 120-180 parts is mixed Conjunction is uniformly prepared into culture medium；Step 2：Prepare Chinese medicine humic acid productThen culture medium is sterilized with steam, 100-135 DEG C of sterilizing 15-30 minute, culture medium is cooled to afterwards 28 DEG C of -35 DEG C of cultivation temperatures；Microorganism mixed fermentation liquid is inoculated on culture medium, inoculative proportion 0.9-1.2:0.05- 0.2, advanced aerobe fermentation of acting charitably, fermentation condition is mixing speed 160-200r/min, starts ventilation ratio and is adjusted to 1：0.4-0.7, Pressure is filled during culture will be maintained at 0.03-0.06pa, and tune throughput is 15m3/h and mixing speed is 200r/min, makes dissolved oxygen Relative value 20%-40%, fermentation time 24-48 hours, 32-37 DEG C of fermentation temperature；Then anaerobic fermentation is carried out, ferment bar Part is standing for fermentation, fermentation time 48-72 hours, 32 DEG C -37 DEG C of fermentation temperature, 0.5-3 minutes is stirred per 5-7 hours； Anaerobic fermentation terminates the Chinese medicine humic acid fermentation product degraded, through low temperature（100 DEG C of ＜）Dry, moisture is not higher than 15%, is made Chinese medicine humic acid product；Step 3：By Chinese medicine humic acid product 50-90 parts obtained by step 2 and yeast cell wall 10-40 parts by weight proportioning Produce.
- 2. a kind of Chinese medicine humic acid feed for treating mastitis for milk cows according to claim 1 and preparation method thereof, it is special Sign is：The microorganism mixed fermentation liquid is by bacillus subtilis fermentation liquor, the lichen bacillus ferments liquid, Lactobacillus plantarum Zymotic fluid, by weight 40-60%: 40-60%: 10-15% ratio mix.
- A kind of 3. microbial degradation humic acid feed according to claim 1 or 2 and preparation method thereof, it is characterised in that： Described microorganism mixed fermentation liquid, its production technology are as follows：A：The production of bacillus subtilis fermentation liquor：1) solid medium is：Glucose 1-3%, peptone 1-3%, yeast extract 0.3-0.6%, agar 1-4%, Distilled water 90% -97%, regulation medium pH is 6.5- 7.0,110-120 DEG C of sterilizing 16-20 minute；Condition of culture： It is put into 25-35 DEG C of constant incubator and cultivates 22-26h；2) above-mentioned gained material is subjected to Shaking culture, culture medium is：Glucose 1-3%, peptone 0.5-2%, yeast Cream 0.3-0.8%, water 94-99%, regulation Shake flask medium pH are 6.5-7.0,110-120 DEG C of sterilizing 16-20 minutes ；Condition of culture：Shaking flask is accessed in above-mentioned gained strain, is put into 25-35 DEG C of constant-temperature table, 180-230r/ min vibrations Cultivate 22-26h；3) above-mentioned gained material progress seeding tank, fermentation tank are expanded numerous：Culture medium is：Dregs of beans 0.5-1.3%, corn 0.2- 0.8%th, sucrose 1-3%, peptone 0.5-1.2%, yeast extract 0.3-0.8%, potassium dihydrogen phosphate 0.05-0.8%, ammonium sulfate 0.1-0.3%, sodium chloride 0.02-0.05%, defoamer 0.02-0.07%, water 90-98%, culture medium starting pH= 7.0-7.5,115-130 DEG C of sterilizing 20-40 minutes；Condition of culture：Shaking flask is to seeding tank inoculum concentration and seeding tank to hair Fermentation tank is inoculated with by 3-7%, and the seed tank culture time is 22-26h, and the fermentation tank culture time is 30-40h；Viable count is not low In 3.0 × 109cfu/ml；B：The production of the lichen bacillus ferments liquid：1) solid medium is：Peptone 8-12%, beef extract 1-5%, NaCl 3-7%, agar 12-18%, distilled water 58% -76%, regulation medium pH 7.0-7.5,105-125 DEG C of sterilizing 15-20 minute；Condition of culture：It is put into 30-35 22-24h is cultivated in DEG C constant incubator；2) above-mentioned gained material is subjected to shake-flask seed culture：Culture medium is peptone 8-12%, beef extract 1-5%, NaCl 3-7%, agar 12-18%, distilled water 58% -76%, regulation medium pH 7.0-7.5,105-125 DEG C of 15-20 points of sterilizing Clock；Condition of culture：It is put into 30-35 DEG C of constant incubator and cultivates 22-24h；Condition of culture：By above-mentioned gained strain Shaking flask is accessed, is put into 30-35 DEG C of constant-temperature table, 180-220r/min shaken cultivations 22-26h；3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 2-7%, molasses 0.5-1.5%, peptone 0.3-0.8%, yeast extract 0.1-0.5%, potassium dihydrogen phosphate 0.05-0.2%, ammonium sulfate 0.05-0.5%, sodium chloride 0- 0.08%th, soya-bean oil 0.05-0.25%, defoamer 0-0.1%, water 89-97%, medium pH=6.5-7.5,115- is adjusted 125 DEG C of sterilizing 25-35 minutes；Condition of culture：Shaking flask is connect to seeding tank and seeding tank to fermentation tank inoculum concentration by 3-7% Kind, the seed tank culture time is 22-26h, and the fermentation tank culture time is 32-40h；Viable count is not less than 3.0 × 109 cfu/ ml；C：The production of Lactobacillus plantarum zymotic fluid：1) solid medium is：Glucose 0.5-1.5%, peptone 0.5-1.5%, yeast extract 0.1-0.8%, beef Cream 0.1-0.8%, potassium dihydrogen phosphate 0.05-0.15%, agar 1-3%, water 93%-98%, regulation Medium's PH Value to 6.5- 7.0th, 110-120 DEG C of sterilizing 15-25 minute；Condition of culture：It is put into Anaerobic culturel in 30-35 DEG C of constant incubator 22-24h ；2) shake-flask seed culture：Culture medium is：Glucose 0.5-1.5%, peptone 0.5-1.5%, yeast extract 0.1-0.8%, beef extract 0.1-0.8%, potassium dihydrogen phosphate 0.05-0.15%, agar 1-3%, water 93%-98%, regulation training Base pH value is supported to 6.5-7.0,110-120 DEG C of sterilizing 15-25 minutes；Condition of culture：Above-mentioned gained strain is accessed into shaking flask Anaerobic culturel 22-26h；3) seeding tank, fermentation tank expand numerous culture：Culture medium is：Dregs of beans 0.5-1.5%, corn 0.2-0.7%, sucrose 0.5-1.5%, peptone 1-2%, yeast extract 0.2-0.7%, potassium dihydrogen phosphate 0.1-0.5%, ammonium sulfate 0.1-0.5%, second Sour sodium 0.3-0.8%, trisodium citrate 0.1-0.5%, Tween 80 0.05-0.2%, magnesium sulfate 0.01-0.05%, soya-bean oil 0.1-0.3%, defoamer 0.03-0.07%, water 93%-97%, starting pH=6.5-7.0,110-125 DEG C of sterilizing 25-40 Minute；Condition of culture：Shaking flask is inoculated with to seeding tank and seeding tank to fermentation tank inoculum concentration by 3-7%, seed tank culture Time is 22-24h, and the fermentation tank culture time is 30-40h, and viable count is not less than 2.0 × 109 cfu/ml。
- 4. a kind of Chinese medicine humic acid feed for treating mastitis for milk cows according to claim 1 and preparation method thereof, it is special Sign is：Described Chinese medicine humic acid product includes：Culture medium and microorganism mixed fermentation liquid, its ratio are 0.9-1.2: 0.05-0.2；Described culture medium includes：By Radix Astragali 30-60 parts of parts by weights meter, dandelion 30-50 parts, cordate houttuynia 10-40 Part, sodium humate 10-30 parts, bean cake powder 6-10 parts, corn flour 6-10 parts, water 120-180 parts.
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