CN1724647A - Preparation method of parachicken blood phili bacillus liquid culture medium - Google Patents
Preparation method of parachicken blood phili bacillus liquid culture medium Download PDFInfo
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- CN1724647A CN1724647A CN 200510027329 CN200510027329A CN1724647A CN 1724647 A CN1724647 A CN 1724647A CN 200510027329 CN200510027329 CN 200510027329 CN 200510027329 A CN200510027329 A CN 200510027329A CN 1724647 A CN1724647 A CN 1724647A
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Abstract
The invention relates to a fit chicken hemophilus liquid cultivating medium that belongs to bioengineering field. It includes the following steps: washing chicken pectoral muscle and grinding, adding distilled water, boiling, filtering to remove meat residue and adjusting chicken soup capacity; under the condition of asepsis, getting animal blood and freezing to form degradation blood; adding peptone, yeastrel, dextrose, sodium chloride, into chicken soup, after solving, sterilizing in high pressure sterilizer; after cooling, adding degradation blood into the soup, the fit chicken hemophilus liquid cultivating medium would be made. The invention improves the fungus containing quantity of the cultivating medium and lowers cost.
Description
Technical field
That the present invention relates to is a kind of preparation method of biological medium, specifically, is a kind of preparation method of parachicken blood phili bacillus liquid culture medium.Be used for bioengineering field.
Background technology
At present, the substratum that the breeding para bacillus fowl blood phili is used generally uses the solid chocolate culture-medium, also have the heart and brain of use to soak soup, chicken embryo or cell cultures, but the biomass of preparation is few, and general bacteria containing amount is 10
8CFU/ml, (CFU is a colony-forming unit) could prepare vaccine, the cost height after needing to concentrate.Para bacillus fowl blood phili CVCC258 and CVCC269 are that " Chinese bacterial classification catalogue " compiling bacterial classification (English edition, in October, 1992, China Machine Press published, China Committee for Culture Collection of Microorganisms writes), China veterinary microorganism preservation center is teaching both at home and abroad as the subordinate assurance of China Committee for Culture Collection of Microorganisms throughout the year, scientific research, production unit provides the microorganism in " Chinese bacterial classification catalogue ", the microorganism of listing " Chinese bacterial classification catalogue " in is with prolonged preservation, externally supply always, (CVCC: Chinese veterinary microorganism preservation center, this center are production to the microbial strains that the public can buy from Chinese veterinary microorganism preservation center over more than 20 year, scientific research, teaching provides strain more than 60,000 all kinds of veterinary microorganism bacterial classifications).Para bacillus fowl blood phili CVCC258 and CVCC269 are the pathogenies of infectious rhinitis of fowl.CVCC258 separated serotype C type from morbidity the chicken in 1984; CVCC269 separates the serotype A type from the morbidity chicken.
Find through literature search prior art, Chinese patent publication number CN1297041Y, open day is May 30 calendar year 2001, denomination of invention is: a kind of substratum that utilizes the chicken embryo to make bacterium such as infectious coryza of chicken, the substratum of bacteriums such as this patent utilization chicken embryo making infectious coryza of chicken utilizes chick embryo extract to make bacteria culture medium.Get suitable instar chicken embryo, boil after smashing to pieces, remove residue, add other nutrient substances in the supernatant liquor in proportion.Its weak point is: para bacillus fowl blood phili bacteria containing amount after it is cultivated is few, the cost height.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of preparation method of parachicken blood phili bacillus liquid culture medium is provided, make it improve preparation antigen bacteria containing amount, reduce cost, can provide high-quality antigen for clinical preparation Heamophilus paragallinarum vaccine.
The present invention is achieved by the following technical solutions, and concrete steps of the present invention are as follows:
1) pigeon breast muscle 100 gram is cleaned, and rubs, and adds 800 ml distilled waters, boils 10 minutes, filters and removes the meat slag, and chicken soup is adjusted capacity to 900 milliliter.
2) under aseptic condition, get 100 milliliters of animal blood (sheep or chicken), put into-40 ℃ of refrigerator freeze thawing 3 times cracking blood.
3) in 900 milliliters of chicken soup, add peptone 5 grams, yeast extract 2 grams, glucose 5 grams, sodium-chlor 10 grams, dissolving back adjust pH is 7.4, sterilizes 20 minutes for 121 ℃ in autoclave, cooling back adds 100 milliliters of cracking blood, parachicken blood phili bacillus liquid culture medium.
Para bacillus fowl blood phili is the bacterium that a kind of enzyme system is incomplete, need the somatomedin in the blood to grow, and this bacterium needs the V factor.The V factor is Reduced nicotinamide-adenine dinucleotide and Triphosphopyridine nucleotide, reduced, works the hydrogen effect of passing in the bacterial respiratory process.Animal blood can discharge the V factor after cracking, contain the various nutrient substances of thalli growth simultaneously in the serum, and para bacillus fowl blood phili can fully be grown in this substratum.
The present invention adopts preparation chicken soup, adds nutrient substances such as peptone, yeast extract paste, glucose, sodium-chlor in chicken soup, after the germ-free animal blood lysate is handled, add in the chicken soup after the sterilization, parachicken blood phili bacillus liquid culture medium.The parachicken blood phili bacillus liquid culture medium of preparation is suitable for the growth of para bacillus fowl blood phili, and para bacillus fowl blood phili was cultivated 36 hours for 37 ℃ in this substratum, and biomass reaches as high as 10
11CFU/ml is for clinical preparation Heamophilus paragallinarum vaccine provides high-quality antigen.
Embodiment
Embodiment 1
Get pigeon breast muscle 100 gram, clean and rub, add 800 ml distilled waters, boiled 10 minutes, filter and remove the meat slag, chicken soup is adjusted capacity to 900 milliliter.Under aseptic condition, get 100 milliliters of sheep blood, put into-40 ℃ of refrigerator freeze thawing 3 times cracking blood.In 900 milliliters of chicken soup, add peptone 5 grams, yeast extract 2 grams, glucose 5 grams, sodium-chlor 10 grams, dissolving back adjust pH is 7.4, sterilizes 20 minutes for 121 ℃ in autoclave, cooling back adds 100 milliliters of germ-free animal cracking blood, parachicken blood phili bacillus liquid culture medium.
Embodiment 2
Get pigeon breast muscle 100 gram, clean and rub, add 800 ml distilled waters, boiled 10 minutes, filter and remove the meat slag, chicken soup is adjusted capacity to 900 milliliter.Under aseptic condition, get 100 milliliters of chicken blood, put into-40 ℃ of refrigerator freeze thawing 3 times cracking blood.Following preparation method is with embodiment 1.
The parachicken blood phili bacillus liquid culture medium of above embodiment 1,2 preparations is suitable for the growth of para bacillus fowl blood phili, and para bacillus fowl blood phili was cultivated 36 hours for 37 ℃ in this substratum, and biomass is up to 10
11CFU/ml is for clinical preparation Heamophilus paragallinarum vaccine provides high-quality antigen.
Claims (4)
1, a kind of preparation method of parachicken blood phili bacillus liquid culture medium is characterized in that, concrete steps are as follows:
1) pigeon breast muscle is cleaned, rubbed, add distilled water, boil, filter and remove the meat slag, adjust chicken soup capacity;
2) under aseptic condition, get animal blood, put into the refrigerator freeze thawing cracking blood;
3) in chicken soup, add peptone, yeast extract, glucose, sodium-chlor after the dissolving, is sterilized in autoclave, cooling back adds cracking blood, parachicken blood phili bacillus liquid culture medium.
2, the preparation method of parachicken blood phili bacillus liquid culture medium according to claim 1 is characterized in that, in the described step 1), pigeon breast muscle 100 grams add 800 milliliters of distilled water, boil 10 minutes, and chicken soup capacity is adjusted to 900 milliliters.
3, the preparation method of parachicken blood phili bacillus liquid culture medium according to claim 1 is characterized in that, described step 2) in, get 100 milliliters of sheep or chicken blood, put into-40 ℃ of refrigerator freeze thawing 3 times.
4, the preparation method of parachicken blood phili bacillus liquid culture medium according to claim 1, it is characterized in that, in the described step 3), in 900 milliliters of chicken soup, add peptone 5 grams, yeast extract 2 grams, glucose 5 grams, sodium-chlor 10 grams, dissolving back adjust pH is 7.4, sterilizes 20 minutes for 121 ℃ in autoclave, cooling back adds 100 milliliters of cracking blood, parachicken blood phili bacillus liquid culture medium.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200510027329 CN1724647A (en) | 2005-06-30 | 2005-06-30 | Preparation method of parachicken blood phili bacillus liquid culture medium |
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| CN 200510027329 CN1724647A (en) | 2005-06-30 | 2005-06-30 | Preparation method of parachicken blood phili bacillus liquid culture medium |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948781A (en) * | 2010-09-01 | 2011-01-19 | 四川农业大学实验动物工程技术中心 | Culture medium for production of riemerella anatipestifer vaccine and preparation method thereof |
| CN106190909A (en) * | 2016-07-21 | 2016-12-07 | 山东滨州沃华生物工程有限公司 | Haemophilus paragallinarum Type B strain fermentation medium, its preparation method and application thereof |
-
2005
- 2005-06-30 CN CN 200510027329 patent/CN1724647A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948781A (en) * | 2010-09-01 | 2011-01-19 | 四川农业大学实验动物工程技术中心 | Culture medium for production of riemerella anatipestifer vaccine and preparation method thereof |
| CN106190909A (en) * | 2016-07-21 | 2016-12-07 | 山东滨州沃华生物工程有限公司 | Haemophilus paragallinarum Type B strain fermentation medium, its preparation method and application thereof |
| CN106190909B (en) * | 2016-07-21 | 2019-08-27 | 山东滨州沃华生物工程有限公司 | Haemophilus paragallinarum Type B strain fermentation medium, preparation method and its application |
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