Ubwino wa njira yoperekera chakudya cha batch fermentation ndi kapangidwe kabwino ka matanki operekera chakudya

Chiyambi

Pakukula mwachangu kwa zinthu zamakono ukadaulo wa zamoyo ndi uinjiniya wa zamoyo, ukadaulo wopangira mphamvu, monga njira yaikulu yopangira mafakitale, imagwiritsidwa ntchito kwambiri m'magawo osiyanasiyana monga mankhwala, chakudya, mankhwala, ndi mphamvu. Kukonza bwino njira yophikira sikungodalira njira zapamwamba zophikira komanso sikungalekanitsidwe ndi kukonza kapangidwe kake ndi kuwongolera mwanzeru kwa thanki yophikirandi bioreactorPakati pawo, kufedwa kwa chakudya chamagulu, chifukwa cha kayendetsedwe kake ka michere, kwakhala njira imodzi yogwiritsidwa ntchito kwambiri m'mafakitale kupanga kuwiritsaNkhaniyi ifotokoza mwatsatanetsatane ubwino wa njira yothira fermentation pogwiritsa ntchito fed-batch, komanso mfundo zazikulu za thanki yophikira kapangidwe kabwino ka zinthu, kufufuza momwe mungakonzere bwino ntchito yonse komanso ubwino wa zinthu zomwe zagulitsidwa njira yopangira mphamvu kudzera mu mgwirizano wokonza njira ndi zida.

I. Makhalidwe a Njira Yopangira Kuphika kwa Batch-Feed

Kuphika chakudya chamagulu ndi njira yogwirira ntchito yomwe ili pakati pa kuphika kwamagulu ndi kuphika kosalekeza. Mfundo yake yayikulu ndikuwonjezera michere yokwanira ku malo oyamba okulira, ndipo panthawi yophika, magwero a kaboni, magwero a nayitrogeni, kapena michere ina yofunika imawonjezeredwa pang'onopang'ono kapena mosalekeza malinga ndi zofunikira za kagayidwe kachakudya ndi momwe chilengedwe chilili, motero kusunga kukula kwa tizilombo toyambitsa matenda ndi kupanga zinthu.

1. Kupewa kuletsa substrate

Mu njira yachikhalidwe yopangira fermentation, ngati kuchuluka kwa substrate kumakhala kokwera kwambiri, nthawi zambiri kumayambitsa kuwonjezeka kwa osmotic pressure kapena substrate inhibition effect, zomwe zimapangitsa kuti ntchito ya microbial metabolic ichepe. Njira yogwiritsira ntchito batch-feed imapewa vutoli mwa kuwongolera kuchuluka kwa substrate mkati mwa mlingo woyenera, kuonetsetsa kuti tizilombo toyambitsa matenda tili mu mkhalidwe wabwino kwambiri wa metabolic.

2. Kulamulira kwamphamvu kwa kagayidwe kachakudya m'thupi

Njira yodyetsera chakudya ikhoza kusinthidwa malinga ndi zofunikira zosiyanasiyana za kagayidwe kachakudya pazigawo zosiyanasiyana. Mwachitsanzo, magwero a kaboni ndi nayitrogeni amawonjezeka panthawi ya kuchulukana kwa maselo mwachangu, ndipo kupezeka kwa magwero a kaboni kumayendetsedwa panthawi ya kupanga zinthu kuti zilimbikitse kupanga kwa metabolites yachiwiri. Njira yosinthirayi yosinthira imathandizira kuwonjezera zokolola ndi kusankha kwa zinthu zomwe zikuyang'aniridwa.

3. Kukulitsa nthawi yopangira

Kuphika kwa chakudya chamagulu kumatha kukulitsa nthawi yolima ndi nthawi yosonkhanitsa zinthu, motero kumawonjezera kwambiri mphamvu yopangira ndi kuchuluka kwa zokolola za unit. thanki yophikira.

4. Kusintha kwa mitundu yosiyanasiyana zaukadaulo wa zamoyo zofunikira

Kaya ndi kupanga ma amino acid, ma organic acid, ma antibiotic, kapena zakudya zowiritsa ndi biofuels, kuyaka kwa chakudya cha batch kumatha kuyendetsedwa payekhapayekha malinga ndi makhalidwe enaake a kagayidwe kachakudya, kusonyeza kusinthasintha kwakukulu kwa njira.

II. Kusanthula ubwino wa kuwiritsa chakudya cha batch

1. Wonjezerani kuchuluka kwa zinthu ndi zokolola

Mu bioreactor, kuwongolera kuchuluka kwa carbon source supplementation kungalepheretse kuchuluka kwa zinthu zomwe zimapangidwa ndi kagayidwe kachakudya (monga ethanol, lactic acid, ndi zina zotero) ndikulimbikitsa kayendedwe ka kagayidwe kachakudya kupita ku chinthu chomwe mukufuna. Mwachitsanzo, kupanga mapuloteni ophatikizana kapena kukonzekera ma enzyme nthawi zambiri kumachitika kudzera mukuwongolera bwino chakudya, zomwe zimawonjezera kwambiri kuchuluka kwa mapuloteni ndi kuchuluka kwa kagwiritsidwe ntchito kake. bioreactor.

2. Gwiritsani ntchito zinthu zina zomwe zimayambitsa kagayidwe kachakudya mosavuta

Zina mwa zinthu zomwe zimayambitsa kagayidwe kachakudya m'thupi zimatha kuletsa kukula kwa maselo. Kudzera mu chakudya chamagulu, kuchuluka kwa zinthu zomwe zimayambitsa kagayidwe kachakudya kumatha kuyendetsedwa bwino popanda kuchepetsa kuchuluka kwa michere m'thupi, zomwe zimathandiza kuti njira yopangira mphamvu kugwira ntchito pamalo okhazikika.

3. Kuthandizira kuzindikira nzeru ndi zochita zokha

Kukula kwa zinthu zamakono sayansi ya zamoyo yalimbikitsa kukweza kwanzeru kwa njira yopangira mphamvus. Kuphika chakudya chamagulu nthawi zambiri kumaphatikizidwa ndi masensa, makina owunikira pa intaneti, ndi ma algorithms apakompyuta kuti akwaniritse kuyang'anira nthawi yeniyeni komanso kukonza bwino kuchuluka kwa chakudya kuti chikhale ndi kuchuluka kwa substrate, kuchuluka kwa mpweya wosungunuka, pH, ndi zina zotero.

4. Chepetsani zoopsa ndi ndalama zogulira

Kudyetsa chakudya chamagulu kumachepetsa mtengo woyambira wa zinthu zokulirapo komanso zakudya zomwe zimatayidwa, komanso kumachepetsa chiopsezo cha kutayika chifukwa cha kuipitsidwa kapena kulephera kwa zida. Poyerekeza ndi kuwiritsa kosalekeza, ndikoyenera kwambiri kugwiritsa ntchito mafakitale akuluakulu. Kusakaniza zinthu zina zomwe zimapangidwa ndi kagayidwe kachakudya (monga ethanol, lactic acid, ndi zina zotero), zomwe zimapangitsa kuti kagayidwe kachakudya kayende bwino kupita ku chinthu chomwe mukufuna. Mwachitsanzo, popanga mapuloteni ophatikizana kapena ma enzyme, kuwongolera bwino chakudya nthawi zambiri kumagwiritsidwa ntchito kuti kuwonjezere kuchuluka kwa mapuloteni komanso kuchuluka kwa momwe amagwiritsidwira ntchito. bioreactor.

Dongosolo la MIKEBIO Fermentation

III. Mfundo Zofunika Kwambiri Pakupanga Kukonza Bwino kwa Chotengera Chowiritsas

Kuti tigwiritse ntchito bwino ubwino wa kuwiritsa kwa batch-fed, kapangidwe kapamwamba komanso koyenera ka chotengera chofukiza ndikofunikira. Monga chinthu chachizolowezi bioreactor, chotengera chofukiza Sikuti imangokhala malo ogwiritsira ntchito kagayidwe ka tizilombo toyambitsa matenda komanso kupanga zinthu, komanso imapanga dongosolo lovuta lomwe limaphatikiza kusamutsa zinthu, kusintha mphamvu, kuyang'anira chidziwitso, komanso kuwongolera zokha. thanki yophikira Sikuti zimangofunika kuganizira kapangidwe ka makina kokha komanso kuganizira zofunikira pa ndondomeko, kuphatikiza dongosolo, ndi zomwe zikuchitika mtsogolo pakukula kwanzeru.

1. Kupanga Kusakaniza ndi Kusamutsa Zinthu Zambiri

Pa nthawi ya kuwira, kufalikira kwa mpweya ndi michere mofanana kumatsimikizira mwachindunji kuchuluka kwa kukula ndi kagayidwe kachakudya ka maselo.

• Kukonza Kapangidwe ka Stirring Blade: Impeller yachikhalidwe ya flat-blade ndi yoyenera machitidwe otsika kukhuthala, pomwe njira zophikira za high-viscosity fermentation nthawi zambiri zimagwiritsa ntchito zophikira zamtundu wa nangula kapena helical. bioreactorNthawi zambiri amagwiritsa ntchito masamba a magawo ambiri kapena masamba ophatikizana kutengera zotsatira za kuyerekezera kwa madzi kuti atsimikizire kuti kuyenda kwa vertical ndi radial kuli bwino.
• Kupititsa patsogolo Kutumiza kwa Gasi ndi Madzi: Mu kuwiritsa kwa mpweya m'madzi, kuchuluka kwa mpweya wosungunuka n'kofunika kwambiri. Mwa kuwonjezera kuchuluka kwa mpweya, kuwonjezera mitu ya mpweya yokhala ndi pobowola pang'ono, kapena kugwiritsa ntchito kusakaniza kwa jet, coefficient ya mpweya wodutsa (kLa) ikhoza kusinthidwa kwambiri. matanki opangira mphamvu Gwiritsani ntchito ngakhale mpweya wolowa m'thupi kapena kuwonjezera mpweya wokwanira kuti mukwaniritse zofunikira pakulima kwamphamvu.
• Kupewa Kupsinjika Maganizo: Kwa maselo ena osavuta kumva (monga maselo a nyama zoyamwitsa kapena bowa wina wopangidwa ndi ulusi), kusakaniza kwambiri kungayambitse kuwonongeka kwa khungu. Pakadali pano, pakufunika kulinganiza bwino pakati pa kukonza bwino kusamutsa kwa maselo ndi kuteteza ntchito ya maselo.

2. Dongosolo Lodyetsa ndi Kulamulira Kogwirizana

Dongosolo lodyetsera chakudya mu kuwiritsa kwa batch limadalira kuwongolera kolondola kwa substrate, ndipo kapangidwe ka dongosolo lodyetsera chakudya cha matanki owiritsa chakudya ndikofunikira kwambiri.

• Mapampu odyetsera zakudya olondola kwambiri: Makina amakono nthawi zambiri amakhala ndi mapampu ozungulira, mapampu a zida, kapena owongolera kuyenda kwa madzi kuti akwaniritse kudyetsa kolondola kwa milliliter.
• Kudyetsa zakudya m'njira zosiyanasiyana: Mu njira zina, sikuti magwero a kaboni okha ndi omwe amafunika kuwonjezeredwa, komanso magwero a nayitrogeni, zinthu zochepa, ndi mavitamini. thanki yophikira ayenera kukhala ndi malo olumikizirana ndi njira zambiri komanso kukhala ndi mphamvu zodzilamulira okha.
• Kugwirizana ndi kuwunika pa intaneti: Dongosolo lodyetsera nthawi zambiri limalumikizidwa ndi kuchuluka kwa okosijeni, kuyendetsa bwino mpweya, pH, ndi masensa a infrared kuti akwaniritse kulamulira kotsekedwa. Mwachitsanzo, pamene kuchuluka kwa okosijeni kutsika kwambiri, dongosololi limachepetsa mwachangu kuchuluka kwa chakudya cha kaboni kuti lipewe kusalinganika kwa kagayidwe kachakudya komwe kumachitika chifukwa cha kuchuluka kwa chakudya.

3. Dongosolo Lowongolera Kutentha ndi pH

Kagayidwe ka tizilombo toyambitsa matenda kamakhala kogwirizana kwambiri ndi kutentha ndi pH. Malo okhazikika ndiye chinthu chofunikira kwambiri kuti pakhale kupanga bwino.

• Kulamulira kutentha: Thanki yophikiraNthawi zambiri amagwiritsa ntchito majekete, ma coil, kapena zosinthira kutentha zakunja kuti azilamulira kutentha. Pakulima kwamphamvu kwambiri ndi kutulutsa kutentha kwambiri, makina osinthira kutentha akunja amatha kukwaniritsa bwino zosowa zoziziritsira mwachangu.
• Kusintha kwa pH: Pakudyetsa, kupanga asidi m'thupi kapena kupanga alkaline kumachitika kawirikawiri. thanki yophikira iyenera kukhala ndi makina owonjezera a alkaline (NaOH, madzi a ammonia) kapena acid (HCl, H2SO4), ophatikizidwa ndi ma electrode a pH omwe ali ndi mphamvu kwambiri, kuti akwaniritse malamulo enieni.
• Kapangidwe ka buffer: Kabwino kwambiri thanki yophikiras imagwiritsa ntchito zigawo za buffer mu kapangidwe ka culture medium, kuphatikiza ndi hardware control, kuti ipititse patsogolo luso la system lolimbana ndi kusinthasintha.

4. Chitsimikizo cha Ukhondo ndi Kusabereka

Njira yophikira chakudya cha gulu la chakudya imakhala ndi nthawi yayitali komanso chiopsezo chachikulu cha kuipitsidwa, kotero thanki yophikira ayenera kukhala ndi ntchito yodalirika komanso yoyera.

• Makina a CIP ndi SIP: CIP (Kuyeretsa Malo) imatha kuyeretsa mapaipi ndi thanki popanda kusokoneza, pomwe SIP (Kuyeretsa Malo) imagwiritsa ntchito nthunzi yotentha kwambiri kuti ichotse poizoni pazida. Kuphatikiza kwa zinthu ziwirizi kumathandizira kwambiri kuti ntchito igwire bwino ntchito komanso chitetezo cha kupanga.
• Kapangidwe ka malo odyetsera ndi ma valavu: Malo odyetsera ayenera kugwiritsa ntchito valavu ya diaphragm yosabala kapena chisindikizo cha nthunzi kuti apewe kuukira kwa tizilombo tating'onoting'ono takunja.
• Kusankha zinthu zosungiramo thanki: Chitsulo chosapanga dzimbiri cha 316L kapena chitsulo chapamwamba kwambiri chimasankhidwa, chomwe chili ndi kukana dzimbiri komanso kukana zinthu zotsukira, zomwe zimaonetsetsa kuti chikugwira ntchito bwino kwa nthawi yayitali.

5. Masensa ndi Kusonkhanitsa Deta

Mu anzeru njira yopangira mphamvuMasensa ndi zinthu zofunika kwambiri pakuwunika nthawi yeniyeni komanso kuwongolera molondola.

• Zinthu zofunika kuziyang'anira ndi izi: Kutentha, pH, mpweya wosungunuka, liwiro loyambitsa, kuchuluka kwa mpweya wotuluka, ndi zina zotero.
• Magawo apamwamba: Kuwunika pa intaneti kuchuluka kwa zinthu zomwe zili mu substrate (monga shuga, glutamic acid), ndi metabolites (monga lactic acid, ethanol) kwakhala chizolowezi. thanki yophikiraMa s ali ndi masensa owonera kapena masensa achilengedwe, omwe amatha kuzindikira mwachangu komanso mosawononga.
• Kuphatikiza deta ndi kuwonetsa: The thanki yophikira iyenera kukhala ndi kompyuta ndi mapulogalamu apakompyuta kuti isonkhanitse ndikusanthula deta ya sensa nthawi yeniyeni, ndikuwonetsa momwe zinthu zilili kudzera mu mawonekedwe ojambula, zomwe zimathandiza ogwiritsa ntchito kuweruza mwachangu ndikulowererapo.

6. Kukweza Luntha ndi Kudziwitsa Anthu

Kapangidwe kamtsogolo ka thanki yophikiraSikuti zidzakhala zida zakuthupi zokha, komanso zanzeru bioreactor nsanja.

• Kuwongolera Kokha: Kudzera mu PID control, fuzzy control kapena ma machine learning algorithms, kukwaniritsa kusintha kwa kudyetsa, kusakaniza, kutentha ndi pH.
• Ukadaulo wa Mapasa a Digito: Khazikitsani chitsanzo cha pa intaneti cha njira yopangira mphamvu kuyerekezera momwe zinthu zilili pasadakhale, kulosera mavuto omwe angakhalepo, ndikukwaniritsa bwino momwe zinthu zilili.
• Kuwunika Patali ndi Pulatifomu ya Mtambo: Ogwiritsa ntchito amatha kuyang'anira momwe ntchito ikuyendera thanki yophikirapadziko lonse lapansi kudzera pa intaneti, zomwe zikukweza kwambiri kusinthasintha kwa kupanga komanso kuthekera koyankha mwadzidzidzi.

7. Kukula ndi Kupanga Modular

Kupanga mafakitale nthawi zambiri kumafuna kugwira ntchito mogwirizana kwa magulu osiyanasiyana. thanki yophikiras.

• Kapangidwe ka Modular: Kamakono njira yopangira mphamvus nthawi zambiri zimakhala zokhazikika, zomwe zimathandiza kuti pakhale kuphatikiza kosinthasintha malinga ndi zosowa, zomwe zimathandiza kuti pakhale kukula kosalala kuchokera ku labotale kupita ku chiŵerengero choyendetsa kenako kupita ku kupanga kwakukulu.
• Malamulo Okulitsa: Pa nthawi yokulitsa thanki yophikiras, kusintha kwa kusamutsa mpweya ndi madzi, kuchuluka kwa mphamvu yosonkhezera ndi mphamvu yosinthira kutentha kuyenera kuganiziridwa. Kapangidwe koyenera ka chiŵerengero ndi malamulo okulitsa ndizofunikira kwambiri kuti njira yobwerezabwereza ichitike.
• Kupanga Kosinthasintha: Mitundu ina yatsopano ya thanki yophikiras gwiritsani ntchito yotayidwa bioreactors (monga machitidwe a matumba otayidwa), oyenera zosowa zazing'ono za mankhwala osiyanasiyana, zomwe zimathandizira kwambiri kusinthasintha kwa kupanga.

Kudzera mu njira zowongolera zomwe zili pamwambapa, thanki yophikiramonga bioreactorSikuti zimangokwaniritsa zofunikira pa njira yophikira yophikidwa ndi batch komanso zimathandiza kwambiri pa uinjiniya wa zamoyo ndi sayansi ya zamoyo Kaya ndikuwongolera bwino kusamutsa zinthu kapena kukwaniritsa kudyetsa mwanzeru komanso kuyang'anira patali, kapangidwe kabwino ka thanki yophikiranthawi zonse wakhala chinthu chofunikira kwambiri pakukula kwa zinthu zamakono njira yopangira mphamvus.

Dongosolo la MIKEBIO Fermentation

IV. Kugwiritsa Ntchito Pamodzi kwa Kuphika kwa Batch ndi Zipangizo Zowonjezera ndi Kukonza Bwino kwa Chotengera Chowiritsas

1. Mu gawo la mankhwala

Mu uinjiniya wa zamoyo Kupanga maantibayotiki, katemera, mapuloteni ophatikizana, ndi zina zotero, kuyaka kwa batch kudzera mu kukonza chiŵerengero cha magwero a kaboni ndi nayitrogeni kumawonjezera kuchuluka kwa mapuloteni omwe amafunidwa. Kapangidwe kabwino ka chotengera chofukizas imatsimikizira kuti njira yopangira zinthu ikuyenda bwino komanso imachepetsa chiopsezo cha kuipitsidwa.

2. Mu makampani opanga zakudya ndi zakumwa

Pakupanga mabakiteriya a lactic acid, yisiti ya mowa, ndi zina zotero, kuwonjezera kwa batch kumatha kusunga kukoma ndi khalidwe lokhazikika. chotengera chofukizas yokhala ndi kutentha koyenera komanso makina osakaniza amathandizira bwino kupangika kwa zinthu zokometsera.

3. M'magawo a mphamvu ndi chitetezo cha chilengedwe

Pakupanga mafuta a biofuel (monga bioethanol, biobutanol) ndi mapulasitiki owonongeka, kuyaka kwa batch kumathetsa mavuto a kuletsa substrate ndi kusonkhanitsa zinthu zoopsa. Kuphatikiza ndi kothandiza. chotengera chofukiza kapangidwe kake, kamapangitsa kuti pakhale chitukuko chobiriwira komanso chokhazikika.

V. Zochitika Zachitukuko Zamtsogolo

1. Luntha ndi Ukadaulo wa Mapasa a Digito: Mwa kukhazikitsa chitsanzo cha mapasa a digito cha njira yopangira mphamvu, kuyeserera kwapaintaneti komanso kukonza nthawi yeniyeni njira yophikira chakudya cha batch kumatha kuchitika.

2. Kusintha kwa Zinthu ndi Kupanga Zinthu Zambiri: Tsogolo bioreactorAdzagwiritsa ntchito kapangidwe ka modular kuti akwaniritse zofunikira zosiyanasiyana za malonda ndikukulitsa kukula mwachangu.

3. Zobiriwira, Zopanda mpweya woipa komanso Zokhalitsa: Mwa kugwiritsa ntchito bwino mphamvu zomwe zimagwiritsidwa ntchito thanki yophikirandi kuchepetsa zinyalala za zinthu zopangira, kuphatikiza uinjiniya wa zamoyo ndipo mafakitale oteteza chilengedwe adzakwezedwa.

4. Kuphatikizana kwa maphunziro osiyanasiyana: Kukula kwa maphunziro osiyanasiyana ukadaulo wa zamoyo, uinjiniya wa mankhwala, ndi ukadaulo wazidziwitso zidzapangitsa kuti kuphikidwa kwa chakudya chamagulu kulowe mu nthawi yatsopano yolondola komanso yogwira mtima.

Mapeto

Kuphika pogwiritsa ntchito zowonjezera, monga njira yophika yomwe imaphatikiza kusinthasintha ndi magwiridwe antchito, kwakhala imodzi mwa zitsanzo zazikulu zamakono sayansi ya zamoyo Kudzera mu njira zoyenera zowonjezera, phindu ndi kukhazikika kwa njira yopangira mphamvu zingawongoleredwe kwambiri. Monga njira yachizolowezi bioreactor, kapangidwe kabwino kwambiri ka thanki yophikira Sikuti imangofunika kukwaniritsa zofunikira zoyambira pakusamutsa zinthu zambiri, kusamutsa kutentha ndi kulamulira zinthu zopanda poizoni, komanso ikufunika kuphatikiza njira zanzeru ndi zochita zokha kuti ipititse patsogolo kukweza mafakitale. M'tsogolomu, ndi kupita patsogolo kosalekeza kwa sayansi ya zamoyo, kuphatikiza kwakukulu kwa kuwiritsa kwa batch yowonjezera komanso kukonza bwino thanki yophikiras ipereka malo okulirapo komanso chithandizo chaukadaulo cholimba kwambiri pakukula kwa mafakitale opanga mankhwala, chakudya ndi mphamvu.

Kumbali inayi, posankha bioreactor chofufumitsa, ndikofunikira kupeza wopanga wamba wokhala ndi mbiri yabwino komanso wamphamvu, kuti muwonetsetse kuti malonda ake ndi abwino. Jiangsu Mike Ukadaulo wa Zamoyo Co., Ltd. idakhazikitsidwa mu 2008, ikuyang'ana kwambiri pa zamoyo zida zowiritsa kafukufuku ndi chitukuko, kupanga ndi kugulitsa mabizinesi apamwamba. Kampani yathu ili ndi nsanja yoyesera yopangira fermentation, yomwe imalumikizidwa ndi luntha ndi makina odzipangira okha, ndipo yamanga makina okhwima njira yopangira mphamvu, yomwe imatha kukwaniritsa bwino njira yoyendetsera ntchito yoyendetsera ntchito yoyendetsa, kuyambitsa chitukuko cha njira zonse kuyambira kupanga njira yoyendetsera ntchito yoyendetsa mpaka kulekanitsa ndi kuyeretsa zinthu zoyendetsera ntchito yoyendetsa ntchito, ndikukhazikitsa malo ogwirira ntchito a dokotala ndi labotale yamakono.

Kampani yathu ili ndi fakitale yopangira zinthu yokhazikika, yokhala ndi zida zonse ndi zida zofunika poyikira, makamaka kuphatikiza zida zosiyanasiyana zamakina, makina ochapira a argon arc okha, makina opukutira okha, makina odulira okha ndi makina ozindikira zolakwika ndi zida zina zoposa 60 zopangira ndi kuyesa, njira yopangira yonseyi imagwira ntchito masiku ano, kuti ipatse makasitomala luso lapamwamba komanso chithandizo chaukadaulo.

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