CN103555641B - A kind of mycoplasma hyopneumoniae culture medium and preparation method thereof - Google Patents
A kind of mycoplasma hyopneumoniae culture medium and preparation method thereof Download PDFInfo
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Abstract
A kind of mycoplasma hyopneumoniae culture medium and preparation method thereof, belongs to veterinary biologics technical field.Containing PPLO 20 ~ 50g, yeast powder 2 ~ 10g, glucose 3 ~ 15g, 1% phenol red 0.5 ~ 4ml, combination of amino acids 2 ~ 10g, herbal mixture polysaccharide 15 ~ 30g and porcine blood serum 50 ~ 100ml in its every 1000ml phosphoric acid buffer.Mycoplasma hyopneumoniae culture medium of the present invention, can improve mycoplasma hyopneumoniae tolerance when cultivating mycoplasma hyopneumoniae, reduces mycoplasma hyopneumoniae apoptosis rate; Mycoplasma hyopneumoniae culture medium of the present invention, can improve mycoplasma hyopneumoniae breeding bacteria, cultivates bacterial concentration and reaches 10
10-11/ ml, improves more than 5-10 times than ordinary culture medium; Mycoplasma hyopneumoniae culture medium of the present invention, can suppress other bacterial growths when cultivating mycoplasma hyopneumoniae simultaneously, reduces the Pollution risk of cultivating mycoplasma hyopneumoniae and cultivating.
Description
Technical field
The invention belongs to veterinary biologics technical field, be specifically related to a kind of mycoplasma hyopneumoniae culture medium and preparation method thereof.
Background technology
Porcine mycoplasmal pneumonia is a kind of very common respiratory tract disease in pig farm, popular in worldwide, be commonly called as swine enzootic pneumonia, caused by mycoplasma hyopneumoniae (Mhp), this cause of disease mainly resides in tracheae in pig body, on segmental bronchus and bronchiolar cilium, physics defense mechanism-defence the mucous membrane of havoc porcine respiratory, add the susceptibility of swinery to other respiratory pathogenic agent, as swine influenza virus, pig breeding and respiratory syndrome virus, II type pig circular ring virus, bordetella branchiseptica, actinobacillus pleuropneumoniae and haemophilus parasuis etc., so usually cause the multiplicity of infection of multiple cause of disease, cause porcine respiratory syndrome (PRDC) or other diseases, mycoplasma pneumoniae can reduce pig growth rate 12.7% and reduce the price of deed 13.8%, the development of serious harm pig industry.
Mycoplasma hyopneumoniae is facultative anaerobe, and growth demand is very harsh, and be separated quite difficulty, it is higher than general mycoplasma to nutritional requirement.Although mycoplasma hyopneumoniae can utilize the material needed for the breeding of nutrition synthesis own growth in substratum, but due to its genomic molecular weight, the quantity of information of carrying is few, biosynthesis ability is limited, therefore need from materials such as external picked-up cholesterol, amino acid, lipid acid, Nucleic acid precurser and VITAMIN, this just impels the medium component more complicated of mycoplasma, must add some special nutrition materials.
Because mycoplasma hyopneumoniae is higher than general mycoplasma to nutritional requirement, medium component more complicated, incubation time is long, and culture concentration is lower, have impact on people widely and studies further it, also counteracts that i (mycoplasma hyopneumoniae) vaccine exploitation simultaneously and produces.Many scholars devise multiple substratum to for its growth at present, although they differ from one another, generally speaking all do not solve the problem of cultivating difficulty.So also will make further research in the cultivation of mycoplasma hyopneumoniae.
Polysaccharide is one of active ingredient of Chinese herbs, and large quantity research shows, polysaccharide and saccharide complex are not only in vivo as energy resource and constituent material, the more important thing is that it is present in all membrane structures, participates in the various activities of cell in biological phenomena.Polysaccharose substance is the important component part of all Living organisms, has scavenging free radicals, improves the ability of activities of antioxidant enzymes and anti-lipid peroxidation.In addition, herbal polysaccharide has special provide protection to unicellular lower eukaryote.Herbal polysaccharide is rich in hydroxyl, can combine, build multidimensional network space hydrogen bond structure with low isoreactivity living organisms, forms protective membrane, realize the provide protection to active bio on active bio live body surface.
The present invention is based on above-mentioned technical background, propose a kind of mycoplasma culture medium, specifically a kind of substratum being specially adapted to mycoplasma hyopneumoniae, improves mycoplasma hyopneumoniae bacterial concentration.
Summary of the invention
For prior art Problems existing, the object of the invention is to design the technical scheme that a kind of mycoplasma hyopneumoniae culture medium and preparation method thereof is provided, by this culture medium culturing mycoplasma hyopneumoniae, mycoplasma hyopneumoniae bacterial concentration can be improved, improve mycoplasma hyopneumoniae Yield of Antigen.
Described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that the component containing following weight proportion in every 1000ml phosphoric acid buffer:
PPLO 20～50g
Yeast powder 2 ~ 10g
Glucose 3 ~ 15g
1% phenol red 0.5 ~ 4ml
Combination of amino acids 2 ~ 10g
Herbal mixture polysaccharide 15 ~ 30g
Porcine blood serum 50 ~ 100ml;
Containing L-arginine, glycine, tyrosine, Cys and Serine in described combination of amino acids;
Described herbal mixture polysaccharide extracts and obtains from following Chinese medicine composition: the coptis, Ginkgo Leaf, Radix Achyranthis Bidentatae, Radix Codonopsis, Poria cocos, matrimony vine, Radix Glycyrrhizae and the red sage root.
Described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that the component containing following weight proportion in every 1000ml phosphoric acid buffer:
PPLO 30～40g
Yeast powder 4 ~ 8g
Glucose 5 ~ 10g
1% phenol red 1 ~ 3ml
Combination of amino acids 4 ~ 8g
Herbal mixture polysaccharide 20 ~ 25g
Porcine blood serum 60 ~ 80ml.
Described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that described combination of amino acids contains L-arginine 10 ~ 30%, glycine 5 ~ 15%, tyrosine 10 ~ 30%, Cys 20 ~ 40% and Serine 10 ~ 30%.
Described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that described herbal mixture polysaccharide extracts from following Chinese medicine composition and obtains: the coptis 5 ~ 15 parts, Ginkgo Leaf 10 ~ 30 parts, Radix Achyranthis Bidentatae 10 ~ 20 parts, Radix Codonopsis 10 ~ 30 parts, 5 ~ 50 parts, Poria cocos, matrimony vine 10 ~ 30 parts, 10 ~ 20 parts, Radix Glycyrrhizae and the red sage root 5 ~ 15 parts.
Described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that described herbal mixture polysaccharide is obtained by following steps:
A, take each Chinese medicine material, chopping, clean after spend the night by cold water soak, then add the purified water of raw material weight 15 times, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out alcohol precipitation, precipitation is rough herbal polysaccharide;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add gac 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal mixture polysaccharide.
The preparation method of described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that comprising following processing step:
1) phosphate buffer solution is prepared;
2) from the coptis 5 ~ 15 parts, Ginkgo Leaf 10 ~ 30 parts, Radix Achyranthis Bidentatae 10 ~ 20 parts, Radix Codonopsis 10 ~ 30 parts, 5 ~ 50 parts, Poria cocos, matrimony vine 10 ~ 30 parts, 10 ~ 20 parts, Radix Glycyrrhizae and the red sage root 5 ~ 15 parts, extraction obtains herbal mixture polysaccharide;
3) PPLO 20 ~ 50g, yeast powder 2 ~ 10g, glucose 3 ~ 15g, combination of amino acids 2 ~ 10g, herbal mixture polysaccharide 15 ~ 30g is taken, be dissolved in the obtained 1000mL phosphate buffer solution of step 1), add 1% phenol red 0.5 ~ 4ml fully to mix, regulate between pH to 7.4 ~ 7.8 with acid or alkali, by 0.22 μm of filter membrane under aseptic condition, rearmounted 4 DEG C of filtration sterilization saves backup, and adds health pig serum 50 ~ 100ml, obtain mycoplasma hyopneumoniae culture medium before using.
The preparation method of described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that described step 2) in herbal mixture polysaccharide by following steps obtain:
A, take each Chinese medicine material, chopping, clean after spend the night by cold water soak, then add the purified water of raw material weight 15 times, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out alcohol precipitation, precipitation is rough herbal polysaccharide;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add gac 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal mixture polysaccharide.
The preparation method of described a kind of mycoplasma hyopneumoniae culture medium, is characterized in that the combination of amino acids described in described step 3) contains L-arginine 10 ~ 30%, glycine 5 ~ 15%, tyrosine 10 ~ 30%, Cys 20 ~ 40% and Serine 10 ~ 30%.
The preparation method of described a kind of mycoplasma hyopneumoniae culture medium, it is characterized in that in described step 3), taking PPLO 30 ~ 40g, yeast powder 7 ~ 8g, glucose 5 ~ 10g, combination of amino acids 4 ~ 8g, herbal mixture polysaccharide 20 ~ 25g, be dissolved in the obtained 1000mL phosphate buffer solution of step 1), add 1% phenol red 1 ~ 3ml fully to mix, regulate between pH to 7.4 ~ 7.8 with acid or alkali, by 0.22 μm of filter membrane under aseptic condition, rearmounted 4 DEG C of filtration sterilization saves backup, and adds health pig serum 60 ~ 80ml before using.
In the present invention, PPLO is that PPLO substratum is buied by Sen Beijia (Nanjing) bio tech ltd.
Compared with prior art, the present invention has following beneficial effect:
(1) mycoplasma hyopneumoniae culture medium prepared by the present invention, can improve mycoplasma hyopneumoniae tolerance when cultivating mycoplasma hyopneumoniae, reduces mycoplasma hyopneumoniae apoptosis rate.
(2) mycoplasma hyopneumoniae culture medium prepared by the present invention, can improve mycoplasma hyopneumoniae breeding bacteria, cultivates bacterial concentration and reaches 10
10-11/ ml(ccu counting process), improve more than 5-10 times than ordinary culture medium.
(3) mycoplasma hyopneumoniae culture medium prepared by the present invention, can suppress other bacterial growths when cultivating mycoplasma hyopneumoniae simultaneously, reduces the Pollution risk of cultivating mycoplasma hyopneumoniae and cultivating.
Embodiment
In order to make the present invention easier to understand, below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.
Embodiment 1
A kind of mycoplasma hyopneumoniae culture medium, the component containing following weight proportion in this substratum every 1000ml phosphoric acid buffer:
PPLO 20g
Yeast powder 6g
Glucose 8g
1% phenol red 4ml
Combination of amino acids 4g
Herbal mixture polysaccharide 15g
Porcine blood serum 50ml
A preparation method for mycoplasma hyopneumoniae culture medium, the steps include:
(1) preparation of phosphate buffer solution: accurately take sodium-chlor 8g, Repone K 0.2g, Sodium phosphate dibasic 1.15g, potassium primary phosphate 0.2g, calcium chloride 0.1g, the magnesium chloride 0.1g containing 6 crystal water is dissolved in 1000ml distilled water, by 0.22 μm of filter membrane under aseptic condition, filtration sterilization is for subsequent use.
(2) preparation of herbal mixture polysaccharide
Each Chinese medicine material is taken by the coptis 15 parts, Ginkgo Leaf 30 parts, Radix Achyranthis Bidentatae 10 parts, Radix Codonopsis 10 parts, 20 parts, Poria cocos, matrimony vine 10 parts, 10 parts, Radix Glycyrrhizae and the red sage root 15 parts, chopping, clean after spend the night by cold water soak, add the purified water of raw material weight 15 times again, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out alcohol precipitation, precipitation is rough herbal polysaccharide;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add gac 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal mixture polysaccharide.
(3) preparation of mycoplasma hyopneumoniae culture medium: take PPLO 20g, yeast powder 6g, glucose 8g, 1% phenol red 4ml, combination of amino acids 4g(L-arginine 0.8g, glycine 0.4g, tyrosine 1.0g, Cys 1.2g and Serine 0.6g in proportion), herbal mixture polysaccharide 15g, be dissolved in the obtained phosphate buffer solution of step (1), add 1% phenol red 4mL fully to mix, regulate between pH to 7.4 ~ 7.8 with acid or alkali, by 0.22 μm of filter membrane under aseptic condition, rearmounted 4 DEG C of filtration sterilization saves backup, and adds health pig serum 50ml before using.
Embodiment 2
A kind of mycoplasma hyopneumoniae culture medium, the component containing following weight proportion in this substratum every 1000ml phosphoric acid buffer:
PPLO 35g
Yeast powder 2g
Glucose 15g
1% phenol red 0.5ml
Combination of amino acids 10g
Herbal mixture polysaccharide 30g
Porcine blood serum 80ml.
A preparation method for mycoplasma hyopneumoniae culture medium, its step is with embodiment 1.Wherein herbal mixture polysaccharide extracts and obtains from the coptis 10 parts, Ginkgo Leaf 20 parts, Radix Achyranthis Bidentatae 15 parts, Radix Codonopsis 20 parts, 50 parts, Poria cocos, matrimony vine 20 parts, 15 parts, Radix Glycyrrhizae and the red sage root 5 parts.
Embodiment 3
A kind of mycoplasma hyopneumoniae culture medium, the component containing following weight proportion in this substratum every 1000ml phosphoric acid buffer:
PPLO 50g
Yeast powder 10g
Glucose 3g
1% phenol red 2.5ml
Combination of amino acids 2g
Herbal mixture polysaccharide 25g
Porcine blood serum 100ml.
A preparation method for mycoplasma hyopneumoniae culture medium, its step is with embodiment 1.Wherein herbal mixture polysaccharide extracts and obtains from the coptis 5 parts, Ginkgo Leaf 10 parts, Radix Achyranthis Bidentatae 20 parts, Radix Codonopsis 30 parts, 5 parts, Poria cocos, matrimony vine 30 parts, 20 parts, Radix Glycyrrhizae and the red sage root 10 parts.
Substratum prepared by comparative example 1 the present invention and conventional substratum comparison test
1 Mhp bacterial strain and culture condition
Mycoplasma hyopneumoniae J strain purchased from ATCC is inoculated respectively the mycoplasma hyopneumoniae culture medium of embodiment 1 and 2 kinds of mycoplasma hyopneumoniae culture mediums of prior art, in 5%(V/V) ratio Pigs Inoculated mycoplasma pneumoniae in each substratum, 37 DEG C of cultivations, when substratum its colour changed into yellow, when pH is down to 6.4-6.8, aseptic taking-up culture.
2 live bacterial counts (CCU method)
Each strain culture gets 14 sterile test tube, often pipe dress 4.5ml is containing mycoplasma hyopneumoniae culture medium, the well-grown culture of 0.5ml is added at the 1st pipe, after mixing, draw 0.5ml and add the 2nd by all means, so carry out 10 times of serial dilutions to manage to last, establish the mycoplasma hyopneumoniae culture medium not adding bacterium liquid as negative control simultaneously.Developmental tube does three repetitions.Test tube is placed static gas wave refrigerator in 37 DEG C of constant incubators after serial dilution, every day observes once, the colour-change of main detection substratum and opacity, the Continuous Observation time is 14, and the tubule extent of dilution that colour-change finally occurs is the CCU titre of this culture.
3 results
3.1 mycoplasma hyopneumoniae Z strains inoculate 3 kinds of substratum 3 secondary growth experiment CCU measurement results in table 1.
3 kinds of substratum 3 secondary growth experiment CCU measurement results are inoculated in table 1 mycoplasma hyopneumoniae Z strain
Mycoplasma hyopneumoniae culture medium prepared by the present invention, can improve mycoplasma hyopneumoniae breeding bacteria, cultivates bacterial concentration and reaches 10
10-11/ ml(ccu counting process), improve more than 5-10 times than ordinary culture medium.
Embodiment 2 carries out the application identical with comparative example 1 with the mycoplasma hyopneumoniae culture medium in 3, and it finally also can reach the technique effect identical with comparative example 1.
Claims (2)
1. a preparation method for mycoplasma hyopneumoniae culture medium, is characterized in that comprising following processing step:
1) phosphate buffer solution is prepared;
2) from the coptis 5 ~ 15 parts, Ginkgo Leaf 10 ~ 30 parts, Radix Achyranthis Bidentatae 10 ~ 20 parts, Radix Codonopsis 10 ~ 30 parts, 5 ~ 50 parts, Poria cocos, matrimony vine 10 ~ 30 parts, 10 ~ 20 parts, Radix Glycyrrhizae and the red sage root 5 ~ 15 parts, extraction obtains herbal mixture polysaccharide;
3) PPLO 20 ~ 50g, yeast powder 2 ~ 10g, glucose 3 ~ 15g, combination of amino acids 2 ~ 10g, herbal mixture polysaccharide 15 ~ 30g is taken, be dissolved in the obtained 1000mL phosphate buffer solution of step 1), add 1% phenol red 0.5 ~ 4ml fully to mix, regulate between pH to 7.4 ~ 7.8 with acid or alkali, by 0.22 μm of filter membrane under aseptic condition, rearmounted 4 DEG C of filtration sterilization saves backup, and adds health pig serum 50 ~ 100ml, obtain mycoplasma hyopneumoniae culture medium before using;
Described step 2) in herbal mixture polysaccharide by following steps obtain:
A, take each Chinese medicine material, chopping, clean after spend the night by cold water soak, then add the purified water of raw material weight 15 times, water-bath to 90 DEG C, and remain on 90 DEG C, boil 2 hours, boil in process every 10 minutes and stir once;
B, discard a Chinese medicine slag, collect liquid, after room temperature cooling through 10000rpm centrifugal 10 minutes, discard precipitation, collect supernatant;
C, supernatant in b is carried out alcohol precipitation, precipitation is rough herbal polysaccharide;
D, with rough herbal polysaccharide in sterilizing purified water washing c, dissolves with 42 DEG C of sterilizing purified water of 20 times amount after washing, after dissolving, add gac 4 DEG C of attach overnight in 0.5% ratio, after absorption through 10000rpm centrifugal 10 minutes, discard precipitation, collection supernatant;
E, by supernatant in d after 0.22um membrane filtration is degerming, carry out 10 times and concentrate, obtained herbal polysaccharide concentrated solution;
F, herbal polysaccharide concentrated solution in e is carried out vacuum lyophilization namely obtain herbal mixture polysaccharide;
In described step 3), combination of amino acids contains L-arginine 10 ~ 30%, glycine 5 ~ 15%, tyrosine 10 ~ 30%, Cys 20 ~ 40% and Serine 10 ~ 30%.
2. the preparation method of a kind of mycoplasma hyopneumoniae culture medium as claimed in claim 1, it is characterized in that in described step 3), taking PPLO 30 ~ 40g, yeast powder 7 ~ 8g, glucose 5 ~ 10g, combination of amino acids 4 ~ 8g, herbal mixture polysaccharide 20 ~ 25g, be dissolved in the obtained 1000mL phosphate buffer solution of step 1), add 1% phenol red 1 ~ 3ml fully to mix, regulate between pH to 7.4 ~ 7.8 with acid or alkali, by 0.22 μm of filter membrane under aseptic condition, rearmounted 4 DEG C of filtration sterilization saves backup, and adds health pig serum 60 ~ 80ml before using.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560835A (en) * | 2015-01-27 | 2015-04-29 | 新疆天康畜牧生物技术股份有限公司 | Culture medium for culturing mycoplasma hyopneumoniae and preparation method thereof |
| CN105462840A (en) * | 2015-12-10 | 2016-04-06 | 浙江卫信生物药业有限公司 | Method for preparing living freeze-dried body of mycoplasma orale |
| CN105779362A (en) * | 2016-05-16 | 2016-07-20 | 南京大爻网络科技有限公司 | Culture medium for low-serum culture of mycoplasma hyopneumoniae and preparation method and application thereof |
| CN106591177B (en) * | 2016-11-29 | 2020-04-21 | 中国农业科学院兰州兽医研究所 | Low-serum efficient culture medium for mycoplasma capricolum goat pneumonia subspecies and preparation method thereof |
| CN106434502A (en) * | 2016-12-23 | 2017-02-22 | 瑞普（保定）生物药业有限公司 | Swine mycoplasma hyopneumoniae culture medium and preparation method and application thereof |
| CN109010814B (en) * | 2018-08-31 | 2021-11-16 | 武汉科前生物股份有限公司 | Production method of haemophilus parasuis and mycoplasma hyopneumoniae bivalent inactivated vaccine |
| CN109294955A (en) * | 2018-10-30 | 2019-02-01 | 南京大爻网络科技有限公司 | A kind of mycoplasma hyopneumoniae Pseudonocardia and preparation method thereof |
| CN109468248A (en) * | 2018-12-14 | 2019-03-15 | 南京天邦生物科技有限公司 | A kind of porcine mycoplasmal pneumonia antigen high-efficient culture method |
| CN110607260A (en) * | 2019-09-23 | 2019-12-24 | 山东甲骨文生物科技有限公司 | Culture medium and method for low-serum culture of mycoplasma hyopneumoniae |
| CN110804563A (en) * | 2019-11-13 | 2020-02-18 | 山东滨州沃华生物工程有限公司 | Culture medium for low-serum culture of mycoplasma hyopneumoniae |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824119A (en) * | 2005-12-30 | 2006-08-30 | 广东工业大学 | Preparation method of compound Chinese medicinal polysaccharide and its use |
| CN1928076A (en) * | 2006-09-28 | 2007-03-14 | 中国兽医药品监察所 | Live vaccine for mycoplasma pneumonia of swine and production method thereof |
| CN102154167A (en) * | 2011-01-05 | 2011-08-17 | 北京大北农科技集团股份有限公司 | Mycoplasma hyopneumoniae culture medium and preparation method thereof |
| CN103060220A (en) * | 2012-08-31 | 2013-04-24 | 南京天邦生物科技有限公司 | Low-serum culture medium for efficiently culturing mycoplasma hyopneumoniae and preparation method thereof |
-
2013
- 2013-11-19 CN CN201310579845.6A patent/CN103555641B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824119A (en) * | 2005-12-30 | 2006-08-30 | 广东工业大学 | Preparation method of compound Chinese medicinal polysaccharide and its use |
| CN1928076A (en) * | 2006-09-28 | 2007-03-14 | 中国兽医药品监察所 | Live vaccine for mycoplasma pneumonia of swine and production method thereof |
| CN102154167A (en) * | 2011-01-05 | 2011-08-17 | 北京大北农科技集团股份有限公司 | Mycoplasma hyopneumoniae culture medium and preparation method thereof |
| CN103060220A (en) * | 2012-08-31 | 2013-04-24 | 南京天邦生物科技有限公司 | Low-serum culture medium for efficiently culturing mycoplasma hyopneumoniae and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| "中药多糖对鼠李糖乳杆菌性质的影响";赵玉萍 等;《食品科技》;20050831(第8期);摘要 * |
| "中药多糖的提取、分离纯化及其含量测定方法概述";郑晓艳 等;《福建分析测试》;20130831;第22卷(第4期);第58页右栏附图和第59页左栏第1.1.1节 * |
| "中药多糖研究进展";王彦军;《医学理论与实践》;20090331;第22卷(第3期);第279页左栏第1段和第280页右栏第3段 * |
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