MD4329C1 - Process for cultivation of yeast strain Saccharomyces cerevisiae CNMN-Y-20 - Google Patents
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- MD4329C1 MD4329C1 MDA20130081A MD20130081A MD4329C1 MD 4329 C1 MD4329 C1 MD 4329C1 MD A20130081 A MDA20130081 A MD A20130081A MD 20130081 A MD20130081 A MD 20130081A MD 4329 C1 MD4329 C1 MD 4329C1
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 35
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 11
- 235000013405 beer Nutrition 0.000 claims abstract description 8
- 238000003756 stirring Methods 0.000 claims abstract description 5
- 235000015097 nutrients Nutrition 0.000 claims abstract description 4
- 230000001678 irradiating effect Effects 0.000 claims 1
- 230000002906 microbiologic effect Effects 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 2
- 229920002498 Beta-glucan Polymers 0.000 description 15
- 239000002028 Biomass Substances 0.000 description 8
- 239000002609 medium Substances 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 description 4
- 239000003651 drinking water Substances 0.000 description 4
- 235000020188 drinking water Nutrition 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001503 Glucan Polymers 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000013528 artificial neural network Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Invenţia se referă la biotehnologii microbiene, în special la un procedeu de cultivare a tulpinii de levuri Saccharomyces cerevisiae CNMN-Y-20, şi poate fi utilizată în industria microbiologică, alimentară şi farmaceutică. The invention relates to microbial biotechnologies, in particular to a method of cultivating the yeast strain Saccharomyces cerevisiae CNMN-Y-20, and can be used in the microbiological, food and pharmaceutical industries.
Este cunoscut un procedeu de cultivare submersă a tulpinilor de levuri pe medii care conţin precursori ai biosintezei β-glucanilor (monozaharide, extracte din şroturi etc.) [1]. Dezavantajul procedeului constă în conţinutul mic de β-glucani acumulat în biomasa celulară. There is a known process of submerged cultivation of yeast strains on media containing precursors of β-glucan biosynthesis (monosaccharides, meal extracts, etc.) [1]. The disadvantage of the process consists in the small content of β-glucans accumulated in the cellular biomass.
Soluţia cea mai apropiată este procedeul de cultivare a tulpinii de levuri Saccharomyces cerevisiae CNMN-Y-20 ce include etapele: prepararea materialului semincer pe must de bere, durata de cultivare de 3 zile la temperatura de 20° C, inocularea germenilor în mediul de fermentare steril YPD sau Rieder. Durata cultivării în profunzime, pe agitator (200 r.p.m), la temperatura de 20 sau 25° C este de 120 ore [2]. The closest solution is the cultivation process of the yeast strain Saccharomyces cerevisiae CNMN-Y-20 which includes the steps: preparation of the seed material on beer wort, cultivation period of 3 days at a temperature of 20° C, inoculation of the germs in the fermentation medium sterile YPD or Rieder. The duration of cultivation in depth, on a shaker (200 r.p.m.), at a temperature of 20 or 25° C is 120 hours [2].
Cultivarea în mediile de fermentare şi condiţiile indicate asigură acumularea a 15,38…22,5% β-glucani în biomasa uscată a levurii. Cultivation in the indicated fermentation media and conditions ensure the accumulation of 15.38...22.5% β-glucans in the dry yeast biomass.
Dezavantajul acestui prototip constă în aceea că nu se realizează pe deplin potenţialul biosintetic al tulpinii şi conţinutul de β-glucani nu atinge valoarea maximă. The disadvantage of this prototype is that the biosynthetic potential of the strain is not fully realized and the β-glucan content does not reach the maximum value.
Problema pe care o rezolvă invenţia este de a elabora un procedeu de cultivare a tulpinii de levuri Saccharomyces cerevisiae CNMN-Y-20, care să asigure sporirea capacităţii de biosinteză a β-glucanilor. The problem that the invention solves is to develop a method of cultivating the yeast strain Saccharomyces cerevisiae CNMN-Y-20, which ensures the increase of the biosynthesis capacity of β-glucans.
Esenţa invenţiei constă în aceea că se cultivă levurile pe must de bere timp de 24 ore, la temperatura de 25°C, apoi se iradiază cu unde milimetrice cu frecvenţa de 53,3 GHz, emise în regim continuu timp de 20 min, după care cultura de levuri, cu concentraţia de 2x106 celule/ml, se transferă pe mediul de fermentare steril YPD sau Rieder în cantitate de 5%vol. şi se cultivă în profunzime în condiţii de agitare continuă (200 r.p.m.), la temperatura de 25°C, timp de 120 ore. The essence of the invention is that the yeasts are cultivated on beer wort for 24 hours, at a temperature of 25°C, then irradiated with millimeter waves with a frequency of 53.3 GHz, emitted continuously for 20 min, after which the yeast culture, with a concentration of 2x106 cells/ml, is transferred to the sterile YPD or Rieder fermentation medium in an amount of 5%vol. and cultivated in depth under conditions of continuous stirring (200 r.p.m.), at a temperature of 25°C, for 120 hours.
Efectul de activare a biosintezei β-glucanilor la acţiunea undelor milimetrice cu frecvenţa menţionată se asociază cu modificările oscilaţiilor de frecvenţă ale biomembranei celulare a levurii. The effect of activating the biosynthesis of β-glucans under the action of millimeter waves with the mentioned frequency is associated with changes in the frequency oscillations of the yeast cell biomembrane.
Conţinutul de β-glucani în biomasa levurii Saccharomyces cerevisiae CNMN-Y-20, la cultivarea submersă în condiţii proxime şi conform procedeului elaborat este prezentat în tabel. The content of β-glucans in the biomass of the yeast Saccharomyces cerevisiae CNMN-Y-20, during submerged cultivation under close conditions and according to the developed procedure, is presented in the table.
Rezultatul tehnic al invenţiei constă în sporirea conţinutului de β-glucani cu 17,5...25,7% faţă de cea mai apropiată soluţie. The technical result of the invention consists in increasing the content of β-glucans by 17.5...25.7% compared to the closest solution.
Exemple de realizare a invenţiei Examples of realization of the invention
Exemplul I Example I
Cultura de levuri Saccharomyces cerevisiae CNMN-Y-20, crescută submers pe must de bere timp de 24 ore, la temperatura de 25° C este supusă acţiunii undelor milimetrice cu frecvenţa de 53,3 GHz, emise în regim continuu, timp de 10 min. După iradiere, materialul semincer (2x106 celule ml-1), în concentraţie de 5% vol., se transferă în mediul de fermentare steril YPD cu următoarea compoziţie, g L-1: peptonă - 20,0; glucoză - 20,0; extract de drojdie - 10 ml; apă potabilă - 1000 ml. Cultivarea se efectuează în condiţii de agitare continuă (200 r.p.m.), în baloane Erlenmayer cu capacitatea de 1,0 L, la temperatura de 25°C, durata cultivării - 120 ore. The yeast culture Saccharomyces cerevisiae CNMN-Y-20, grown submerged on beer wort for 24 hours, at a temperature of 25° C, is subjected to the action of millimeter waves with a frequency of 53.3 GHz, emitted continuously, for 10 min . After irradiation, the seed material (2x106 cells ml-1), in a concentration of 5% vol., is transferred to the sterile fermentation medium YPD with the following composition, g L-1: peptone - 20.0; glucose - 20.0; yeast extract - 10 ml; drinking water - 1000 ml. Cultivation is carried out under conditions of continuous stirring (200 r.p.m.), in Erlenmayer flasks with a capacity of 1.0 L, at a temperature of 25°C, duration of cultivation - 120 hours.
Conţinutul de β-glucani constituie 19,36±0,53% în biomasa uscată, ceea ce depăşeşte cu 25,8% conţinutul fixat la cultivarea tulpinii în condiţii proxime. The content of β-glucans is 19.36±0.53% in the dry biomass, which exceeds by 25.8% the content determined when growing the stem under close conditions.
Exemplul II Example II
Cultura de levuri Saccharomyces cerevisiae CNMN-Y-20, crescută submers pe must de bere timp de 24 ore, la temperatura de 25° C este supusă acţiunii undelor milimetrice cu frecvenţa de 53,3 GHz, emise în regim continuu, timp de 20 min. După iradiere, materialul semincer (2x106 celule ml-1), în concentraţie de 5% vol., se transferă pe mediul de fermentare steril YPD cu următoarea compoziţie, g L-1: peptonă - 20,0; glucoză - 20,0; extract de drojdie - 10 ml; apă potabilă - 1000 ml. Cultivarea se efectuează în condiţii de agitare continuă (200 r.p.m.), în baloane Erlenmayer cu capacitatea de 1,0 L, la temperatura de 25°C, durata cultivării - 120 ore. The yeast culture Saccharomyces cerevisiae CNMN-Y-20, grown submerged on beer wort for 24 hours, at a temperature of 25° C, is subjected to the action of millimeter waves with a frequency of 53.3 GHz, emitted continuously, for 20 min . After irradiation, the seed material (2x106 cells ml-1), in a concentration of 5% vol., is transferred to the sterile fermentation medium YPD with the following composition, g L-1: peptone - 20.0; glucose - 20.0; yeast extract - 10 ml; drinking water - 1000 ml. Cultivation is carried out under conditions of continuous stirring (200 r.p.m.), in Erlenmayer flasks with a capacity of 1.0 L, at a temperature of 25°C, duration of cultivation - 120 hours.
Conţinutul de β-glucani constituie 19,34±0,82% în biomasa uscată, ceea ce depăşeşte cu 25,7% conţinutul fixat la cultivarea tulpinii în condiţii proxime. The content of β-glucans constitutes 19.34±0.82% in the dry biomass, which exceeds by 25.7% the content determined when growing the stem under close conditions.
Exemplul III Example III
Cultura de levuri Saccharomyces cerevisiae CNMN-Y-20, crescută submers pe must de bere timp de 24 ore, la temperatura de 25° C este supusă acţiunii undelor milimetrice cu frecvenţa de 53,3 GHz, emise în regim continuu, timp de 20 min. După iradiere, materialul semincer (2x106 celule ml-1), în concentraţie de 5% vol., se transferă în mediul nutritiv steril Rieder cu următoarea compoziţie, g L-1: glucoză - 30,0; (NH4)2SO4 - 3,0; MgSO4 ·7H2O - 0,7; KH2PO4 - 1,0; NaCl - 0,5; Ca(NO3)2 - 0,4; autolizat de drojdie - 10 ml; apă potabilă - 1000 ml. Cultivarea se efectuează în condiţii de agitare continuă (200 r.p.m.), în baloane Erlenmeyer cu capacitatea de 1,0 L, la temperatura de 25° C, durata cultivării - 120 ore. The yeast culture Saccharomyces cerevisiae CNMN-Y-20, grown submerged on beer wort for 24 hours, at a temperature of 25° C, is subjected to the action of millimeter waves with a frequency of 53.3 GHz, emitted continuously, for 20 min . After irradiation, the seminal material (2x106 cells ml-1), in a concentration of 5% vol., is transferred to the sterile Rieder nutrient medium with the following composition, g L-1: glucose - 30.0; (NH4)2SO4 - 3.0; MgSO4·7H2O - 0.7; KH2PO4 - 1.0; NaCl - 0.5; Ca(NO3)2 - 0.4; yeast autolysate - 10 ml; drinking water - 1000 ml. Cultivation is carried out under conditions of continuous agitation (200 r.p.m.), in Erlenmeyer flasks with a capacity of 1.0 L, at a temperature of 25° C, duration of cultivation - 120 hours.
Conţinutul de β-glucani constituie 26,43±0,73% în biomasa uscată, ceea ce depăşeşte cu 17,5% conţinutul fixat la cultivarea tulpinii în condiţii proxime. The content of β-glucans is 26.43±0.73% in dry biomass, which exceeds by 17.5% the content determined when growing the stem under close conditions.
Exemplul IV Example IV
Cultura de levuri Saccharomyces cerevisiae CNMN-Y-20, crescută submers pe must de bere timp de 24 ore, la temperatura de 25° C este supusă acţiunii undelor milimetrice cu frecvenţa de 53,3 GHz, emise în regim continuu, timp de 30 min. După iradiere, materialul semincer (2x106 celule ml-1), în concentraţie de 5% în bază volumetrică, se transferă în mediul nutritiv steril Rieder cu următoarea compoziţie, g L-1: glucoză - 30,0; (NH4)2SO4 - 3,0; MgSO4 ·7H2O - 0,7; KH2PO4 - 1,0; NaCl - 0,5; Ca(NO3)2 - 0,4; autolizat de drojdie - 10 ml; apă potabilă - 1000 ml. Cultivarea se efectuează în condiţii de agitare continuă (200 rot/min), în baloane Erlenmayer cu capacitatea de 1,0 L, la temperatura de 25°C, durata cultivării - 120 ore. The yeast culture Saccharomyces cerevisiae CNMN-Y-20, grown submerged on beer wort for 24 hours, at a temperature of 25° C, is subjected to the action of millimeter waves with a frequency of 53.3 GHz, emitted continuously, for 30 min . After irradiation, the seed material (2x106 cells ml-1), in a concentration of 5% on a volumetric basis, is transferred to the sterile Rieder nutrient medium with the following composition, g L-1: glucose - 30.0; (NH4)2SO4 - 3.0; MgSO4·7H2O - 0.7; KH2PO4 - 1.0; NaCl - 0.5; Ca(NO3)2 - 0.4; yeast autolysate - 10 ml; drinking water - 1000 ml. Cultivation is carried out under conditions of continuous agitation (200 rpm), in Erlenmayer flasks with a capacity of 1.0 L, at a temperature of 25°C, duration of cultivation - 120 hours.
Conţinutul de β-glucani constituie 23,99±0,91% în biomasa uscată, ceea ce depăşeşte cu 6,6% conţinutul fixat la cultivarea tulpinii în condiţii proxime. The content of β-glucans is 23.99±0.91% in dry biomass, which exceeds by 6.6% the content determined when growing the stem under close conditions.
Tabel Table
Influenţa undelor milimetrice de intensitate extraînaltă asupra biosintezei β-glucanilor la tulpina Saccharomyces cerevisiae CNMN-Y- 20 The influence of ultrahigh intensity millimeter waves on the biosynthesis of β-glucans in the strain Saccharomyces cerevisiae CNMN-Y- 20
Variante Durata iradierii,min Mediul de cultură YPD Mediul de cultură Rieder Conţinutul de β-glucani, % în B.U. X±xS % faţă de cea mai apropiată soluţie Spor, % Conţinutul de β-glucani, % în B.U. X±xS % faţă de cea mai apropiată soluţie Spor, % 53,3 GHz* 10 19,36±0,53 125,8 25,8 21,76±0,43 - - Soluţia cea mai apropiată * * 15,38±0,14 100 - 22,50±1,41 100 - 53,3 GHz* 20 19,34±0,82 125,7 25,7 26,43±0,73 117,5 17,5 Soluţia cea mai apropiată * * 15,38±0,14 100 22,50±1,41 100 - 53,3 GHz* 30 19,11±3,25 124,2 24,2 23,99±0,91 106,6 6,6 Soluţia cea mai apropiată * * 15,38±0,14 100 22,50±1,41 100 - Variants Duration of irradiation, min YPD culture medium Rieder culture medium Content of β-glucans, % in B.U. X±xS % compared to the nearest solution Spor, % The content of β-glucans, % in B.U. X±xS % compared to the closest solution Spor, % 53.3 GHz* 10 19.36±0.53 125.8 25.8 21.76±0.43 - - The closest solution * * 15.38 ±0.14 100 - 22.50±1.41 100 - 53.3 GHz* 20 19.34±0.82 125.7 25.7 26.43±0.73 117.5 17.5 The best solution close * * 15.38±0.14 100 22.50±1.41 100 - 53.3 GHz* 30 19.11±3.25 124.2 24.2 23.99±0.91 106.6 6 ,6 The closest solution * * 15.38±0.14 100 22.50±1.41 100 -
* Cultura de levuri iradiată cu unde milimetrice de intensitate extraînaltă * Yeast culture irradiated with millimeter waves of extra high intensity
** Cultura de levuri neiradiată cu unde milimetrice de intensitate extraînaltă ** Yeast culture not irradiated with extra high intensity millimeter waves
X±xS indică valoarea medie şi deviaţia standard X±xS indicates the mean value and standard deviation
1. Desai Kiran M. Bhalchandra K. Vaidya, Rekha S. Singhal, Sunil S. Bhagwat. Use of an artificial neural network in modeling yeast biomass and yield of b-glucan. Process Biochemistry, 2005, 40, p. 1617-1626 1. Desai Kiran M. Bhalchandra K. Vaidya, Rekha S. Singhal, Sunil S. Bhagwat. Use of an artificial neural network in modeling yeast biomass and yield of b-glucan. Process Biochemistry, 2005, 40, pp. 1617-1626
2. MD 4048 B1 2010.06.30 2. MD 4048 B1 2010.06.30
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| EP1877447A1 (en) * | 2005-05-05 | 2008-01-16 | Sensient Flavors Inc. | Production of beta-glucans and mannans |
| US20100190872A1 (en) * | 2005-05-05 | 2010-07-29 | Sensient Flavors Inc. | Production of beta-glucans and mannans |
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| WO2008032134A1 (en) * | 2006-09-11 | 2008-03-20 | Compana Nacional De Levaduras Levapan S.A. | Method for obtaining yeast glucan by autolysis of saccharomyces cerevisiae baker's yeast cells |
| CN101205552A (en) * | 2006-12-20 | 2008-06-25 | 天津市工业微生物研究所 | Method for preparing yeast cell wall beta-1,3-dextran |
| CN101012468A (en) * | 2007-02-07 | 2007-08-08 | 天津科技大学 | A kind of extraction process of yeast glucan |
| MD3516G2 (en) * | 2007-05-08 | 2008-09-30 | Институт Микробиологии И Биотехнологии Академии Наук Молдовы | Process for deep cultivation of Rhodotorula gracilis CNMN-Y-03 yeast strain |
| US20110250235A1 (en) * | 2008-12-18 | 2011-10-13 | Glykos Finland Oy | Production of a saccharide composition comprising glucans and mannans by alkaline and acid hydrolysis of yeast cells |
| EP2370475A1 (en) * | 2008-12-18 | 2011-10-05 | Glykos Finland Oy | Production of a saccharide composition comprising glucans and mannans by alkaline and acid hydrolysis of yeast cells |
| WO2010070207A1 (en) * | 2008-12-18 | 2010-06-24 | Glykos Finland Oy | Production of a saccharide composition comprising glucans and mannans by alkaline and acid hydrolysis of yeast cells |
| CN102257011A (en) * | 2008-12-18 | 2011-11-23 | 格莱科斯芬兰公司 | Production of a saccharide composition comprising glucans and mannans by alkaline and acid hydrolysis of yeast cells |
| MD3945C2 (en) * | 2009-04-07 | 2010-02-28 | Институт Микробиологии И Биотехнологии Академии Наук Молдовы | Process for deep cultivation of Aspergillus niger 22-19 CNMN FD 02 fungus strain |
| MD4048B1 (en) * | 2010-02-11 | 2010-06-30 | Institutul De Microbiologie Si Biotehnologie Al Academiei De Stiinte A Moldovei | Strain of yeast Saccharomyces cerevisiae - source of Beta -glucans |
| RU2011140588A (en) * | 2011-10-06 | 2013-04-27 | Общество с ограниченной ответственностью "Сервис Инжиниринг" (ООО "Сервис Инжиниринг") | METHOD FOR ACTIVATING YEAST MANNANES |
| RU2487941C2 (en) * | 2011-10-06 | 2013-07-20 | Общество с ограниченной ответственностью "Сервис Инжиниринг" (ООО "Сервис Инжиниринг") | Method to activate yeast gums |
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| MD4329B1 (en) | 2015-02-28 |
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