CN103535185B - A kind of liquid separation method of China pilose spore bacterial classification - Google Patents
A kind of liquid separation method of China pilose spore bacterial classification Download PDFInfo
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- CN103535185B CN103535185B CN201310455421.9A CN201310455421A CN103535185B CN 103535185 B CN103535185 B CN 103535185B CN 201310455421 A CN201310455421 A CN 201310455421A CN 103535185 B CN103535185 B CN 103535185B
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- bacterial classification
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- china pilose
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Abstract
A liquid separation method for China pilose spore bacterial classification, the present invention relates to the separation method of a kind of microorganism, is mainly used in the liquid separation method of China pilose spore bacterial classification; Be characterized in comprising the steps: step one: fresh natural cordyceps is removed outside bacterium cover, brush away Chinese caterpillar fungus surface impurity, carry out disinfection with thimerosal, aqua sterilisa cleaning 1 ~ 4 time, excision Chinese caterpillar fungus face tissue, stripping and slicing, size is at 1 ~ 4mm
3, for subsequent use; Step 2: birds, beasts and eggs are shelled, bacterial filter, insert the culture vessel after sterilizing, nutrient solution accounts for 1/10 ~ 9/10 of culture vessel volume, access 1 ~ 5 piece of Chinese caterpillar fungus block cut, at 0 ~ 18 DEG C, rotating speed 60 ~ 180r/min shaking table is cultivated 1 ~ 15 day, namely covers with in mycelia and filtered liquid on the wall of culture vessel and forms bacterium ball; Carry out bacterium inspection, be separated the China pilose spore bacterial classification obtained, solid or liquid fermenting production can be directly used in; Effect is: 1, material easily obtains, and cost is low.2, the spawn culture time is short, can be directly used in production without the need to purifying, rejuvenation.3, growth is vigorous, and obtain bacterial classification many, substratum is simple.
Description
Technical field
The present invention relates to the separation method of a kind of microorganism, particularly a kind of liquid separation method of China pilose spore bacterial classification.
Background technology
Cordyceps sinensis be Hepialus (HepialusarmoricamusOberthur) the larva infection of Chinese of lepidopteran bat Genus Hepialus by after hair spore fungi, in vivo amount reproduction formed entomogenous fungi complex body.The method of Cordyceps fungus imperfect stage strain separating disclosed in Chinese patent CN102283022A.Above-mentioned patent is separated the method for China pilose spore imperfect stage bacterial classification from infection of Chinese the body fluid of the bat moth larvae live body of hair spore; Its shortcoming is that the bat moth larvae difficulty that collection China pilose spore infects is large, and the time is long, and the bacterial classification of separation must by being used for scale operation after domestication, rejuvenation.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of difference novel method that China pilose spore bacterial classification is in the past separated is provided.
Technical solution of the present invention realizes by the following method, is characterized in comprising the steps:
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, carry out disinfection with thimerosal, aqua sterilisa cleaning 1 ~ 4 time, excision Chinese caterpillar fungus face tissue, stripping and slicing, size is at 1 ~ 4mm
3, for subsequent use;
Step 2: birds, beasts and eggs are shelled, with bacterial filter, insert the culture vessel after sterilizing, nutrient solution accounts for 1/10 ~ 9/10 of culture vessel volume, access 1 ~ 5 piece of Chinese caterpillar fungus block cut, at 0 ~ 18 DEG C, rotating speed 60 ~ 180r/min shaking table is cultivated 1 ~ 15 day, namely covers with in mycelia and filtered liquid on the wall of culture vessel and forms bacterium ball; Carry out bacterium inspection, be separated the China pilose spore bacterial classification obtained and can be directly used in solid or liquid fermenting production.
Another feature of the present invention is the thimerosal described in step one is javelle water.
Another feature of the present invention is the thimerosal described in step one is mercury chloride.
Another feature of the present invention is the thimerosal described in step one is ethanol.
Another feature of the present invention is the thimerosal described in step one is hydrogen peroxide.
Another feature of the present invention is the birds, beasts and eggs described in step 2 is egg.
Another feature of the present invention is the birds, beasts and eggs described in step 2 is duck's egg.
Another feature of the present invention is the birds, beasts and eggs described in step 2 is quail egg.
Another feature of the present invention is the birds, beasts and eggs described in step 2 is goose egg.
Another feature of the present invention is the birds, beasts and eggs described in step 2 is Ostrich egg.
Effect of the present invention is: 1, material easily obtains, and cost is low.2, the spawn culture time is short, can be directly used in production without the need to purifying, rejuvenation.3, growth is vigorous, and obtain bacterial classification many, substratum is simple.
Embodiment
Embodiment 1
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, with 5% hypochlorite disinfectant 15 seconds, aqua sterilisa cleaned 4 times, and then excise Chinese caterpillar fungus face tissue, stripping and slicing, size is at 4mm
3, for subsequent use;
Step 2: shelled by egg, with bacterial filter, insert the culture vessel after sterilizing, filtered liquid accounts for 3/10 of culture vessel volume, accesses 2 pieces of 4mm
3the Chinese caterpillar fungus block of size, 4 DEG C, cultivate 4 days in the shaking table of rotating speed 180r/min, formed in the filtered liquid of culture vessel on bacterium ball and wall of container and cover with mycelia; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
Embodiment 2
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, sterilize 7 seconds with 2 ‰ mercury chloride, aqua sterilisa cleans 3 times, and then excise Chinese caterpillar fungus face tissue, stripping and slicing, size is at 3mm
3, for subsequent use;
Step 2: shelled by duck's egg, with bacterial filter, insert the culture vessel after sterilizing, filtered liquid accounts for 5/10 of culture vessel volume, accesses 2 pieces of 3mm
3the Chinese caterpillar fungus block of size, 18 DEG C, cultivate 4 days in the shaking table of rotating speed 180r/min, formed in the filtered liquid of culture vessel on bacterium ball and wall of container and cover with mycelia; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
Embodiment 3
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, 75% ethanol disinfection 11 seconds, aqua sterilisa cleans 2 times, and then excise Chinese caterpillar fungus face tissue, stripping and slicing, size is at 1mm
3, for subsequent use;
Step 2: shelled by goose egg, with bacterial filter, insert the culture vessel after sterilizing, filtered liquid accounts for 7/10 of culture vessel volume, accesses 4 pieces of 1mm
3the Cordyceps block of size, 8 DEG C, cultivate 10 days in the shaking table of rotating speed 80r/min, formed in the filtered liquid of culture vessel on bacterium ball and wall of container and cover with mycelia; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
Embodiment 4
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, sterilize 10 seconds with 3% hydrogen peroxide, aqua sterilisa cleans 1 time, and then excise Chinese caterpillar fungus face tissue, stripping and slicing, size is at 3mm
3, for subsequent use;
Step 2: shelled by Ostrich egg, with bacterial filter, insert the culture vessel after sterilizing, filtered liquid accounts for 9/10 of culture vessel volume, accesses 5 pieces of 3mm
3the Chinese caterpillar fungus block of size, 0 DEG C, cultivate 15 days in the shaking table of rotating speed 180r/min, formed in the filtered liquid of culture vessel on bacterium ball and wall of container and cover with mycelia; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
Embodiment 5
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Chinese caterpillar fungus surface impurity, with 75% ethanol disinfection 15 seconds, aqua sterilisa cleaned 2 times, and then excise Chinese caterpillar fungus face tissue, stripping and slicing, size is at 2mm
3, for subsequent use;
Step 2: shelled by quail egg, with bacterial filter, insert the culture vessel after sterilizing, filtered liquid accounts for 1/10 of culture vessel volume, accesses 3 pieces of 2mm
3the Chinese caterpillar fungus block of size, 4 DEG C, cultivate 7 days in the shaking table of rotating speed 100r/min, formed in the filtered liquid of culture vessel on bacterium ball and wall of container and cover with mycelia; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
Claims (10)
1. a liquid separation method for China pilose spore bacterial classification, is characterized in that comprising the steps:
Step one: fresh natural cordyceps is removed outside bacterium cover, brushes away Cordyceps sinensis surface impurity, carry out disinfection with thimerosal, aqua sterilisa cleaning 1 ~ 4 time, excision Cordyceps sinensis face tissue, stripping and slicing, size is at 1 ~ 4mm
3, for subsequent use;
Step 2: birds, beasts and eggs are shelled, with bacterial filter, insert the culture vessel after sterilizing, nutrient solution accounts for 1/10 ~ 9/10 of culture vessel volume, access 1 ~ 5 piece of Cordyceps sinensis block cut, at 0 ~ 18 DEG C, rotating speed 60 ~ 180r/min shaking table is cultivated 1 ~ 15 day, namely covers with in mycelia and filtered liquid on the wall of culture vessel and forms bacterium ball; Namely China pilose spore bacterial classification is obtained after bacterium inspection.
2. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that thimerosal described in step one is javelle water.
3. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the thimerosal described in step one is mercury chloride.
4. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the thimerosal described in step one is ethanol.
5. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the thimerosal described in step one is hydrogen peroxide.
6. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the birds, beasts and eggs described in step 2 are egg.
7. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the birds, beasts and eggs described in step 2 are duck's egg.
8. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the birds, beasts and eggs described in step 2 are quail egg.
9. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the birds, beasts and eggs described in step 2 are goose egg.
10. the liquid separation method of a kind of China pilose spore bacterial classification as claimed in claim 1, is characterized in that the birds, beasts and eggs described in step 2 are Ostrich egg.
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| CN112342145A (en) * | 2020-11-04 | 2021-02-09 | 青海久实虫草生物科技有限公司 | Rejuvenation method of hirsutella hepiali Chen et Shen fungi |
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| CN85101971A (en) * | 1985-04-01 | 1987-03-04 | 青海省畜牧兽医科学院 | China Cordyceps sinensis fungi and artificial culture method thereof |
| CN1348988A (en) * | 2000-10-17 | 2002-05-15 | 刑桂兰 | Artificial cultivation of cordyceps |
| CN1935978A (en) * | 2006-10-16 | 2007-03-28 | 曾树生 | Method for culturing Chinese caterpillar domestic fungus from fresh egg |
| CN101165163A (en) * | 2007-10-17 | 2008-04-23 | 杨国栋 | Method for preparing fowl ovum like culture medium and application thereof |
| CN101407765A (en) * | 2007-10-10 | 2009-04-15 | 涂峰 | Method for producing Chinese piliferous mycelium powder |
| CN101423799A (en) * | 2008-11-29 | 2009-05-06 | 涂峰 | Method for producing Chinese piliferous mycelium powder |
| CN101530042A (en) * | 2009-04-16 | 2009-09-16 | 吕树山 | Fermented processing technique of hirsutslla sinensis |
| CN101695257A (en) * | 2009-10-30 | 2010-04-21 | 上海芝草生物技术有限公司 | Solid fermentation method for cordyceps sinensis |
| CN102220249A (en) * | 2011-06-13 | 2011-10-19 | 杭州柯氏生物科技有限公司 | Method for producing Hirsutella sinensis |
| CN102283022A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for isolating Cordyceps sinensis Sacc. asexual-stage spawn |
-
2013
- 2013-09-30 CN CN201310455421.9A patent/CN103535185B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85101971A (en) * | 1985-04-01 | 1987-03-04 | 青海省畜牧兽医科学院 | China Cordyceps sinensis fungi and artificial culture method thereof |
| CN1348988A (en) * | 2000-10-17 | 2002-05-15 | 刑桂兰 | Artificial cultivation of cordyceps |
| CN1935978A (en) * | 2006-10-16 | 2007-03-28 | 曾树生 | Method for culturing Chinese caterpillar domestic fungus from fresh egg |
| CN101407765A (en) * | 2007-10-10 | 2009-04-15 | 涂峰 | Method for producing Chinese piliferous mycelium powder |
| CN101165163A (en) * | 2007-10-17 | 2008-04-23 | 杨国栋 | Method for preparing fowl ovum like culture medium and application thereof |
| CN101423799A (en) * | 2008-11-29 | 2009-05-06 | 涂峰 | Method for producing Chinese piliferous mycelium powder |
| CN101530042A (en) * | 2009-04-16 | 2009-09-16 | 吕树山 | Fermented processing technique of hirsutslla sinensis |
| CN101695257A (en) * | 2009-10-30 | 2010-04-21 | 上海芝草生物技术有限公司 | Solid fermentation method for cordyceps sinensis |
| CN102220249A (en) * | 2011-06-13 | 2011-10-19 | 杭州柯氏生物科技有限公司 | Method for producing Hirsutella sinensis |
| CN102283022A (en) * | 2011-07-22 | 2011-12-21 | 重庆市中药研究院 | Method for isolating Cordyceps sinensis Sacc. asexual-stage spawn |
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Effective date of registration: 20170927 Address after: 810003 Qinghai Xining biological science and Technology Industrial Park, No. four, No. 26, No. 4 building, Qinghai long Cordyceps biological science and Technology Co., Ltd. Patentee after: Qinghai Jiushi worm grass biotechnology Co., Ltd. Address before: 810003 Qinghai Xining economic and Technological Development Zone biological science and Technology Industrial Park, 4 road 26 Qinghai long Cordyceps biological science and Technology Co., Ltd. Patentee before: Tu Feng |