CN106381271A - Making method for selenium-rich Cordyceps militaris - Google Patents
Making method for selenium-rich Cordyceps militaris Download PDFInfo
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention relates to a making method for selenium-rich Cordyceps militaris. The making method comprises the following steps: (1) making of a solid medium; (2) making of a mother seed; (3) making of a liquid spawn; (4) inoculation; (5) spawn running and coloring; (6) growth; (7) harvesting; and (8) drying. The making method provided by the invention can increase the selenium content of Cordyceps militaris, enables Cordyceps militaris to be good in growth, easy to survive and high in yield and nutrient content, can be easily promoted and is beneficial for increasing the income of farmers breeding Cordyceps militaris.
Description
Technical field
The invention belongs to cultural technique field, it is related to a kind of manufacture method of Cordyceps militaris (L.) Link. and in particular to a kind of selenium-rich pupa worm
The manufacture method of grass.
Background technology
Cordyceps militaris (L.) Link. also known as Cordyceps militaris (L.) Link., biotaxy is belonged to together with Chinese Cordyceps funguses Ascomycetes,
Cordyceps.Cordycepin is one kind 3 '-deoxyadenosine separated from the culture fluid of Cordyceps militaris in nineteen fifty, and it has
There is significant pharmacological action, such as bacteriostasis, antitumor action, antiviral effect, and strengthen immunologic function, preventing and treating heart and brain
Angiopathy etc..Modern scientific research shows, cordycepin is the important active component of Cordyceps militaris (L.) Link., and in Cordyceps militaris (L.) Link., cordycepin content is
The decades of times of Cordyceps, is the main source obtaining cordycepin.Cordyceps militaris (L.) Link. has resisting fatigue, antitumor, anoxia enduring, enhancing are exempted from
The effects such as the condensation of epidemic disease power, sexual function improving and suppression human blood platelets., as the succedaneum of wild Chinese caterpillar fungus, market is empty for Cordyceps militaris (L.) Link.
Between big.Cordyceps militaris (L.) Link. artificial culture small investment, high efficiency, with short production cycle, all can cultivate throughout the year.
But, the Cordyceps militaris (L.) Link. Se content obtained by the artificial cultivation method of existing Cordyceps militaris (L.) Link. is few, and selenium to be human body necessary
Elements of limit quantity, there is anticancer, cardioprotection, treating liver necrosis, preventing and treating myopia and cataract, removing toxic substances, enhance immunity, delay to decline
Always with multiple pharmacological effect such as enhancing reproductive functions, thus the exploitation of selenium-rich funguses should cause the attention of people, cultivates selenium-rich
Cordyceps militaris (L.) Link. then can improve nutrition and the medical value of Cordyceps militaris (L.) Link..
But, the artificial cultivation method presence of existing selenium-enriched cordceps militaris yields poorly, of poor quality, high cost, nutritive value
Low multiple shortcoming.
Therefore, it is badly in need of finding a kind of manufacture method of new selenium-enriched cordceps militaris.
Content of the invention
It is an object of the invention to overcoming shortcoming present in prior art, provide a kind of making side of selenium-enriched cordceps militaris
Method, this manufacture method can improve the selenium content of Cordyceps militaris (L.) Link., and Cordyceps militaris (L.) Link. is grown, easily survival, yield is high, nutrient content
Height, beneficial to the income promoting and being favorably improved raiser.
To achieve these goals, the present invention provides following technical scheme：A kind of manufacture method of selenium-enriched cordceps militaris, it is special
Levy and be, comprise the following steps：
(1) solid culture matrix manufacturing
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, take
Filtrate；Add 20 grams of agar toward in described filtrate and heated and stirred to described agar melts completely, add 1 gram of yeast extract, Fructus Citri Limoniae
1g and 20 grams of goose egg are received in acid, continue heated and stirred to melting completely, and adjusting PH afterwards is to obtain solid medium after 7-8, then will
Described solid medium be dispensed in test tube cool down standby；
(2) parent species make
In sterile board, rice-shaped block is taken to put into from the cordyceps militaris fruit body after sterilization peeling described invisible spectro
On the basal plane of solid medium, it is stoppered the opening of described test tube with silica gel plug, it is prosperous that lucifuge cryopreservation selected mycelia after 18 days
That contains carries out tube, carries out that lucifuge low temperature is standby to deposit use in the test tube after tube；
(3) liquid spawn makes
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, take
Filtrate；Described filtrate adds 2 grams of potassium dihydrogen phosphate, citric acid receive 1 gram, 3 grams of peptone, 30 grams of glucose, vitamin B1
2 grams and 20 grams of goose egg, continue heated and stirred to melting completely, adjust PH afterwards and obtain culture medium, then by described training for after 7-8
Foster base loads in triangular flask, and the parent species that described step (2) is made to be placed into lucifuge in the culture medium in described triangular flask quiet
Put 24 hours, after going up shaking table culture afterwards 5 days, select as clear as crystal, the uniform liquid spawn of fungus ball；
(4) inoculate
In aseptic interior, described liquid spawn is injected directly into sterilized or after high temperature sterilize Bombyxmori Linnaeuss pupal cell
Interior；
(5) send out bacterium annesl
Postvaccinal mulberry silkworm chrysalis are put between growth 15 days afterwards, in the case that mycelia is plentiful, carries out illumination annesl, turn
The color time is 5-7 days, after in crocus, initially enters growth stage；
(6) grow
Carry out the growth of 40-50 days after annesl；
(7) harvest
Treat that turtleback decorative pattern in cordyceps militaris fruit body apical head, and occur illustrating during the ascospore of powdery
Through maturation, harvested in time；
(8) dry
Cordyceps militaris (L.) Link. after harvesting is put in drying oven and is dried to moisture for 5～10% with 50 DEG C of temperature, cool
Packed in time afterwards, prevent from getting damp.
Further, wherein, in described step (2), will be sub for the Cordyceps militaris (L.) Link. after peeling of sterilizing with aseptic inoculation knife
Entity vertical profile into two, takes interior tissue 1-2 cubic millimeter to put into described invisible spectro solid medium from middle sterile region
Basal plane on, be then placed within lucifuge cryopreservation 18 days in constant incubator, wherein, first 3 days temperature controls at 25 DEG C,
At 24 DEG C, after the week, temperature is down to 22 DEG C to 4-7 days temperature controls, and after fortnight, temperature is down to 16 DEG C；And, wherein,
Carry out that lucifuge low temperature is standby to deposit use in the test tube after tube with 4 DEG C of temperature.
Further, wherein, the shake speed of the shaking table in described step (3) is 130r/min.
Yet further, wherein, in described step (4), described mulberry silkworm chrysalis are placed in glass preserving jar, and toward each
Liquid spawn described in injection 7-8ml in described mulberry silkworm chrysalis.
Again further, wherein, in described step (5), interior in described growth, relative air humidity is 65%,
And the temperature of first 7 days is 15-18 DEG C, the temperature of latter 8 days is 20-22 DEG C；During annesl, day temperature is made to be 20- within first 3 days
21 DEG C, make within latter 2-4 days day temperature be 22-23 DEG C, and make nocturnal temperature be 8-10 DEG C during whole annesl, meanwhile,
During annesl, the relative air humidity of first 3 days is 65%, and the relative air humidity of latter 2-4 days is 85%, and whole colour-change period
Between intensity of illumination reach 200-250 lux, light application time is daily more than 12 hours and has the dark period of 6 hours daily.
Finally, wherein, in the growth period of described step (6), first 20 days temperature controls 22-23 DEG C, air relatively wet
Degree controls and controls in 18 hours/day in 80-90%, light application time；20 days afterwards, and temperature control is in 23 DEG C, relative air humidity
Control and control in 20 hours/day in 85-90%, light application time.
With existing Cordyceps militaris (L.) Link. manufacture method, the selenium-enriched cordceps militaris manufacture method of the present invention has following Advantageous effect
Really：
1st, the riched Cordyceps Militaris obtained by the manufacture method of the selenium-enriched cordceps militaris of the present invention contain selenium so that its nutritive value and medicine
Higher with being worth；
2nd, the manufacture method low cost of the selenium-enriched cordceps militaris of the present invention, be easy to utilization and extention.
3rd, the manufacture method of the selenium-enriched cordceps militaris of the present invention makes the yield height of Cordyceps militaris (L.) Link., survival rate is high, be easy to cultivation, has
Beneficial to the income improving raiser.
Specific embodiment
With reference to embodiment, the present invention is further described, and the content of embodiment is not as to protection scope of the present invention
Restriction.
The manufacture method of the selenium-enriched cordceps militaris of the present invention comprises the following steps：
First, solid culture matrix manufacturing
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, directly
To can not filtering filtrate, take filtrate afterwards.Then, add 20 grams of agar toward in described filtrate and heated and stirred is to described agar
Melt completely, add 1 gram of yeast extract, citric acid receives 1g and 20 grams of goose egg, continue heated and stirred to melting completely.Adjust afterwards
Its PH obtains solid medium for after 7-8.Finally, described solid medium is dispensed in test tube cool down standby.Normal conditions
Under, chilling temperature is 4 DEG C.
2nd, parent species make
In sterile board, rice-shaped block is taken to put into from the cordyceps militaris fruit body after sterilization peeling described invisible spectro
On the basal plane of solid medium, it is stoppered the opening of described test tube, lucifuge cryopreservation 18 days with silica gel plug.Afterwards, select bacterium
Silk is vigorous to carry out tube, carries out that lucifuge low temperature is standby to deposit use in the test tube after tube.
Wherein, specifically, with aseptic inoculation knife by the cordyceps militaris fruit body vertical profile after peeling of sterilizing into two, therefrom
Between sterile region take interior tissue 1-2 cubic millimeter to put on the basal plane of described invisible spectro solid medium, use silica gel plug
It is stoppered the opening of described test tube.Then, described test tube is placed on lucifuge cryopreservation 18 days in constant incubator.Wherein, front 3
At 25 DEG C, at 24 DEG C, after the week, temperature is down to 22 DEG C to the 4-7 days temperature controls to its temperature control, temperature after fortnight
It is down to 16 DEG C.So, through the culture of 18 days, it is vigorous that most of strain can become mycelia.Afterwards, select vigorous the carrying out of mycelia
Tube, it is, strain vigorous for mycelia is transferred in other test tube；What mycelia was not vigorous gives up to fall.And, wherein, exist
Carry out with 4 DEG C of temperature in test tube after tube that lucifuge low temperature is standby to deposit use.
3rd, liquid spawn makes
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, directly
To can not filtering filtrate position, take filtrate afterwards.2 grams of potassium dihydrogen phosphate, citric acid is added to receive 1 gram, albumen in described filtrate
3 grams of peptone, 30 grams of glucose, 2 grams of vitamin B1 and 20 grams of goose egg, continue heated and stirred to melting completely.Adjusting its PH afterwards is
Culture medium is obtained after 7-8.Then described culture medium is loaded in triangular flask, and the parent species making in above-mentioned steps are placed into institute
State in the culture medium in triangular flask lucifuge and stand 24 hours, after going up shaking table culture afterwards 5 days, select as clear as crystal, fungus ball uniform
Liquid spawn.
Wherein, the shake speed of described shaking table is 130r/min.So, by shake and the culture of five days, can obtain
The uniform liquid spawn of as clear as crystal, fungus ball.
4th, inoculate
In aseptic interior, described liquid spawn is injected directly into sterilized or after high temperature sterilize Bombyxmori Linnaeuss pupal cell
Interior.
Specifically, described mulberry silkworm chrysalis are placed in glass preserving jar, and described glass preserving jar carries out high temperature at 130 DEG C
Sterilizing.Meanwhile, in the present invention, it is preferred to, toward liquid spawn described in injection 7-8ml in mulberry silkworm chrysalis each described, in order to bacterium
The growth planted.
5th, send out bacterium annesl
Postvaccinal mulberry silkworm chrysalis are put between growth 15 days afterwards, in the case that mycelia is plentiful, carries out illumination annesl, turn
The color time is 5-7 days, after in crocus, initially enters growth stage.
Wherein, interior in described growth, relative air humidity is 65%, and the temperature of first 7 days is 15-18 DEG C, latter 8 days
Temperature is 20-22 DEG C.So, through 15 days so that the mycelia of strain is very plentiful.
Meanwhile, in the present invention, during annesl, make day temperature be 20-21 DEG C within first 3 days, make daytime within latter 2-4 days
Temperature is 22-23 DEG C, and makes nocturnal temperature be 8-10 DEG C during whole annesl, meanwhile, during annesl, the sky of first 3 days
Gas relative humidity is 65%, and the relative air humidity of latter 2-4 days is 85%, and during whole annesl, intensity of illumination reaches 200-
250 luxs, light application time is daily more than 12 hours and has the dark period of 6 hours daily.So, through the annesl of 5-7 days
Afterwards, it is in crocus, just can initially enter growth stage afterwards.
6th, grow
Carry out the growth of 40-50 days after annesl.
In the present invention, in described growth period, first 20 days temperature controls 22-23 DEG C, relative air humidity controls
80-90%, light application time control in 18 hours/day；20 days are afterwards, temperature control 23 DEG C, relative air humidity controls in 85-
90%th, light application time controls in 20 hours/day.Through the growth of 40-50 days, Cordyceps militaris (L.) Link. just can become ripe.
7th, harvest
Through the growth of 40-50 days, treat that turtleback decorative pattern in cordyceps militaris fruit body apical head, and powdery occurs
Ascospore when explanation ripe, harvested in time.
8th, dry
Cordyceps militaris (L.) Link. after harvesting is put in drying oven and is dried to moisture for 5～10% with 50 DEG C of temperature, cool
Packed in time afterwards, prevent from getting damp.
Riched Cordyceps Militaris obtained by the manufacture method of the selenium-enriched cordceps militaris of the present invention contain selenium so that its nutritive value and medicinal
It is worth higher；Meanwhile, its low cost, be easy to utilization and extention；And, it makes the yield height of Cordyceps militaris (L.) Link., survival rate height, is easy to support
Grow, be conducive to improving the income of raiser, deeply welcome by raiser.
The above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not to the present invention
Embodiment restriction.For those of ordinary skill in the field, can also make on the basis of the above description
The change of other multi-forms or variation.Here all of embodiment cannot be exhaustive.Every skill belonging to the present invention
Obvious change that art scheme is extended out or change the row still in protection scope of the present invention.
Claims (6)
1. a kind of manufacture method of selenium-enriched cordceps militaris is it is characterised in that comprise the following steps：
(1) solid culture matrix manufacturing
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, take filter
Liquid；Add 20 grams of agar toward in described filtrate and heated and stirred to described agar melts completely, add 1 gram of yeast extract, citric acid
Receive 1g and 20 grams of goose egg, continue heated and stirred to melting completely, adjusting PH afterwards is to obtain solid medium after 7-8, then by institute
State solid medium be dispensed in test tube cool down standby；
(2) parent species make
In sterile board, rice-shaped block is taken to put into described invisible spectro solid from the cordyceps militaris fruit body after sterilization peeling
On the basal plane of culture medium, it is stoppered the opening of described test tube with silica gel plug, it is vigorous that lucifuge cryopreservation selected mycelia after 18 days
Carry out tube, carry out that lucifuge low temperature is standby to deposit use in the test tube after tube；
(3) liquid spawn makes
200 grams of Rhizoma Solani tuber osis are cleaned peeling section, add water 1000ml, boil not rotten to shortcake, then through multilamellar filtered through gauze, take filter
Liquid；Described filtrate adds 2 grams of potassium dihydrogen phosphate, citric acid receive 1 gram, 3 grams of peptone, 30 grams of glucose, vitamin B1 2
Gram and 20 grams of goose egg, continue heated and stirred to melting completely, afterwards adjust PH be 7-8 after obtain culture medium, then by described training
Foster base loads in triangular flask, and the parent species that described step (2) is made to be placed into lucifuge in the culture medium in described triangular flask quiet
Put 24 hours, after going up shaking table culture afterwards 5 days, select as clear as crystal, the uniform liquid spawn of fungus ball；
(4) inoculate
Aseptic indoor described liquid spawn is injected directly into sterilized or in the mulberry silkworm chrysalis body after high temperature sterilize；
(5) send out bacterium annesl
Postvaccinal mulberry silkworm chrysalis are put between growth 15 days afterwards, in the case that mycelia is plentiful, carries out illumination annesl, during annesl
Between be 5-7 days, after in crocus, initially enter growth stage；
(6) grow
Carry out the growth of 40-50 days after annesl；
(7) harvest
Treat that turtleback decorative pattern in cordyceps militaris fruit body apical head, and explanation during the ascospore of powdery occur to have become
Ripe, harvested in time；
(8) dry
Cordyceps militaris (L.) Link. after harvesting is put in drying oven and is dried to moisture for 5～10% with 50 DEG C of temperature, cool afterwards and
Shi Jinhang packs, and prevents from getting damp.
2. the manufacture method of selenium-enriched cordceps militaris according to claim 1, is characterized in that, in described step (2), with aseptic
Vaccinating lancet by the cordyceps militaris fruit body vertical profile after peeling of sterilizing into two, takes interior tissue 1-2 to stand from middle sterile region
Square millimeter is put on the basal plane of described invisible spectro solid medium, is then placed within lucifuge cryopreservation in constant incubator
18 days, wherein, at 25 DEG C, at 24 DEG C, after the week, temperature was down to 22 DEG C to the 4-7 days temperature controls to first 3 days temperature controls, and two
After individual week, temperature is down to 16 DEG C；And, wherein, carrying out standby the depositing of lucifuge low temperature with 4 DEG C of temperature in the test tube after tube makes
With.
3. the manufacture method of selenium-enriched cordceps militaris according to claim 2, is characterized in that, the shaking table in described step (3)
Shake speed is 130r/min.
4. the manufacture method of selenium-enriched cordceps militaris according to claim 3, is characterized in that, in described step (4), described Mulberry
Pupa bombycises are placed in glass preserving jar, and liquid spawn described in the injection 7-8ml toward in mulberry silkworm chrysalis each described.
5. the manufacture method of selenium-enriched cordceps militaris according to claim 4, is characterized in that, in described step (5), described
Growth is interior, and relative air humidity is 65%, and the temperature of first 7 days is 15-18 DEG C, and the temperature of latter 8 days is 20-22 DEG C；Turning
During color, make day temperature be 20-21 DEG C within first 3 days, make within latter 2-4 days day temperature be 22-23 DEG C, and whole colour-change period
Between make nocturnal temperature be 8-10 DEG C, meanwhile, during annesl, the relative air humidity of first 3 days is 65%, the sky of latter 2-4 days
Gas relative humidity is 85%, and during whole annesl, intensity of illumination reaches 200-250 lux, and light application time is daily 12 hours
There is the dark period of 6 hours above and daily.
6. the manufacture method of selenium-enriched cordceps militaris according to claim 5, is characterized in that, in the trophophase of described step (6)
Between, first 20 days temperature controls 22-23 DEG C, relative air humidity controls and controls in 18 hours/day in 80-90%, light application time；
20 days are afterwards, temperature control 23 DEG C, relative air humidity controls and controls in 20 hours/day in 85-90%, light application time.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610670960.8A CN106381271A (en) | 2016-08-16 | 2016-08-16 | Making method for selenium-rich Cordyceps militaris |
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| CN201610670960.8A CN106381271A (en) | 2016-08-16 | 2016-08-16 | Making method for selenium-rich Cordyceps militaris |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107155639A (en) * | 2017-07-13 | 2017-09-15 | 山东省蚕业研究所 | A kind of cultural method for the Cordceps militaris for being rich in 1 DNJ |
| CN107251757A (en) * | 2017-06-12 | 2017-10-17 | 刘进军 | A kind of eutrophy element Cordyceps militaris chewable tablets and preparation method and application |
| CN107333568A (en) * | 2017-09-19 | 2017-11-10 | 张家港市藏联生物研究所有限公司 | A kind of more batches of culture techniques of Cordyceps militaris constant temperature |
| CN107373002A (en) * | 2017-08-25 | 2017-11-24 | 周保福 | A kind of selenium-enriched cordceps militaris fish oil comfit |
| CN107502554A (en) * | 2017-09-19 | 2017-12-22 | 张家港市藏联生物研究所有限公司 | A kind of Cordyceps militaris spawn high yield cultural method |
| CN107529410A (en) * | 2017-08-31 | 2018-01-02 | 周保福 | A kind of Cordyceps militaris fish oil comfit |
| CN108450934A (en) * | 2017-02-17 | 2018-08-28 | 北京神州航天食品技术研究院 | A kind of selenium-enriched cordceps militaris piece |
| CN109007823A (en) * | 2018-08-09 | 2018-12-18 | 张斌 | Preparation method of selenium-rich cordyceps militaris powder |
| CN110447455A (en) * | 2019-08-12 | 2019-11-15 | 江门市山海堂虫草有限公司 | A kind of preparation method of Cordyceps militaris Lysimachia sikokiana |
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| CN1861782A (en) * | 2006-05-26 | 2006-11-15 | 李宏 | Artificial cultivating method of enriched selenium Cordyceps militaris and application of fruiting body |
| CN101050426A (en) * | 2007-03-28 | 2007-10-10 | 邹玉华 | Method for breeding north caterpillar fungus rich in selenium |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108450934A (en) * | 2017-02-17 | 2018-08-28 | 北京神州航天食品技术研究院 | A kind of selenium-enriched cordceps militaris piece |
| CN107251757A (en) * | 2017-06-12 | 2017-10-17 | 刘进军 | A kind of eutrophy element Cordyceps militaris chewable tablets and preparation method and application |
| CN107155639A (en) * | 2017-07-13 | 2017-09-15 | 山东省蚕业研究所 | A kind of cultural method for the Cordceps militaris for being rich in 1 DNJ |
| CN107373002A (en) * | 2017-08-25 | 2017-11-24 | 周保福 | A kind of selenium-enriched cordceps militaris fish oil comfit |
| CN107529410A (en) * | 2017-08-31 | 2018-01-02 | 周保福 | A kind of Cordyceps militaris fish oil comfit |
| CN107333568A (en) * | 2017-09-19 | 2017-11-10 | 张家港市藏联生物研究所有限公司 | A kind of more batches of culture techniques of Cordyceps militaris constant temperature |
| CN107502554A (en) * | 2017-09-19 | 2017-12-22 | 张家港市藏联生物研究所有限公司 | A kind of Cordyceps militaris spawn high yield cultural method |
| CN109007823A (en) * | 2018-08-09 | 2018-12-18 | 张斌 | Preparation method of selenium-rich cordyceps militaris powder |
| CN110447455A (en) * | 2019-08-12 | 2019-11-15 | 江门市山海堂虫草有限公司 | A kind of preparation method of Cordyceps militaris Lysimachia sikokiana |
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Application publication date: 20170208 |