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Kwangirika kwa mikorobe (MIC) nikibazo gikomeye mu nganda nyinshi, kuko gishobora guteza igihombo kinini mubukungu.Icyuma cyitwa super duplex kitagira ibyuma 2707 (2707 HDSS) gikoreshwa mubidukikije byo mu nyanja kubera imiti irwanya imiti.Ariko, kurwanya MIC ntabwo byagaragaye mubigeragezo.Ubu bushakashatsi bwasuzumye imyitwarire ya MIC 2707 HDSS yatewe na bagiteri yo mu nyanja ya aerobic Pseudomonas aeruginosa.Isesengura ry’amashanyarazi ryerekanye ko imbere ya biofilm ya Pseudomonas aeruginosa hagati ya 2216E, impinduka nziza mubushobozi bwo kwangirika no kwiyongera kwubwinshi bwa ruswa.Isesengura rya X-ray yifoto yerekana amashanyarazi (XPS) yerekanye igabanuka ryibintu bya Cr hejuru yicyitegererezo munsi ya biofilm.Isesengura ryerekanwa ryibyobo ryerekanye ko biofilm ya P. aeruginosa yabyaye ubujyakuzimu bwa 0,69 µm muminsi 14 yubushakashatsi.Nubwo ibi ari bito, byerekana ko 2707 HDSS idakingiwe rwose na MIC ya biofilm ya P. aeruginosa.
Duplex ibyuma bitagira umuyonga (DSS) bikoreshwa cyane munganda zinyuranye kubera guhuza neza imiterere yimashini nziza hamwe no kurwanya ruswa1,2.Ariko, ibibanza byaho biracyagaragara kandi bigira ingaruka kubusugire bwibyuma3,4.DSS ntishobora kurwanya mikorobe (MIC) 5,6.Nuburyo butandukanye bwa porogaramu ya DSS, haracyari ibidukikije aho kurwanya ruswa ya DSS bidahagije kugirango ukoreshwe igihe kirekire.Ibi bivuze ko ibikoresho bihenze cyane birwanya ruswa ikenewe.Jeon et al7 yasanze na super duplex ibyuma bitagira umuyonga (SDSS) bifite aho bigarukira mubijyanye no kurwanya ruswa.Kubwibyo, mubihe bimwe na bimwe, birakenewe cyane super duplex idafite ibyuma (HDSS) hamwe no kurwanya ruswa.Ibi byatumye habaho iterambere rya HDSS ivanze cyane.
Kurwanya ruswa DSS biterwa nikigereranyo cya alfa na gamma ibyiciro kandi bikagabanuka mukarere ka Cr, Mo na W 8, 9, 10 byegeranye nicyiciro cya kabiri.HDSS ikubiyemo ibintu byinshi bya Cr, Mo na N11, kubwibyo ifite imbaraga zo kurwanya ruswa kandi ifite agaciro kanini (45-50) yumubare uhwanye numubare uhwanye (PREN) ugenwa na wt.% Cr + 3.3 (wt.% Mo + 0.5 wt.% W) + 16% wt.N12.Kurwanya kwangirika kwayo guterwa nuburinganire buringaniye burimo 50% ferritic (α) na 50% austenitike (γ).HDSS ifite imiterere yubukanishi kandi irwanya ruswa ya chloride.Kunanirwa kwangirika kwagura kwagura ikoreshwa rya HDSS mubidukikije bikabije bya chloride nkibidukikije byo mu nyanja.
MIC ni ikibazo gikomeye mu nganda nyinshi nk'inganda za peteroli na gaze n'inganda14.MIC ihwanye na 20% yibyangiritse byose15.MIC ni bioelectrochemical ruswa ishobora kugaragara mubidukikije byinshi.Ibinyabuzima bigizwe hejuru yicyuma bihindura amashanyarazi, bityo bikagira ingaruka kumikorere.Abantu benshi bemeza ko ruswa ya MIC iterwa na biofilm.Ibinyabuzima bya elegitoroniki birya ibyuma kugirango babone imbaraga bakeneye kugirango babeho17.Ubushakashatsi bwa MIC buherutse kwerekana ko EET (ihererekanyabubasha rya elegitoroniki idasanzwe) ari cyo kintu kigabanya igipimo muri MIC iterwa na mikorobe ya electrogene.Zhang n'abandi.18 yerekanye ko abahuza ba electron bihutisha ihererekanyabubasha rya electron hagati ya selile sessificans ya Desulfovibrio nicyuma 304 kitagira umwanda, bikaviramo igitero gikomeye MIC.Anning n'abandi.19 na Wenzlaff n'abandi.20 berekanye ko biofilm ya bagiteri zigabanya sulfate zigabanya (SRBs) zishobora kwinjiza electron ziva mu byuma, bikaviramo gucika cyane.
DSS izwiho kwandura MIC mubitangazamakuru birimo SRBs, bagiteri zigabanya fer (IRBs), nibindi 21.Izi bagiteri zitera umwobo wa DSS munsi ya biofilms 22,23.Bitandukanye na DSS, MIC ya HDSS24 ntabwo izwi neza.
Pseudomonas aeruginosa ni bacteri ya Gram-mbi, motile, imeze nk'inkoni ikwirakwizwa cyane muri kamere25.Pseudomonas aeruginosa nayo ni itsinda rikomeye rya mikorobe mubidukikije byo mu nyanja, bigatuma MIC yibanda cyane.Pseudomonas igira uruhare runini mubikorwa byo kwangirika kandi izwi nkumukoroni wambere mugihe cya biofilm.Mahat n'abandi.28 na Yuan n'abandi.29 yerekanye ko Pseudomonas aeruginosa ikunda kongera umuvuduko wo kwangirika kwibyuma byoroheje hamwe nudusembwa mubidukikije byamazi.
Intego nyamukuru yiki gikorwa kwari ugukora ubushakashatsi ku miterere ya MIC 2707 HDSS yatewe na bacterium yo mu nyanja ya aerobic Pseudomonas aeruginosa ikoresheje uburyo bwa mashanyarazi, uburyo bwo gusesengura hejuru no gusesengura ibicuruzwa byangirika.Ubushakashatsi bwakozwe n’amashanyarazi, harimo n’ubushobozi bw’umuzunguruko (OCP), kurwanya umurongo wa polarisiyonike (LPR), amashanyarazi y’ingufu zidasanzwe (EIS), hamwe n’ibishobora kuba imbaraga za polarisiyasi, byakozwe kugira ngo bige ku myitwarire ya MIC 2707 HDSS.Isesengura ry'ingufu zikwirakwiza ingufu (EDS) ryakozwe kugirango hamenyekane ibintu bya shimi hejuru yangiritse.Byongeye kandi, X-ray yifoto yerekana amashanyarazi (XPS) yakoreshejwe kugirango hamenyekane ituze rya paside ya okiside iterwa n’ibidukikije byo mu nyanja irimo Pseudomonas aeruginosa.Ubujyakuzimu bw'ibyobo byapimwe munsi ya microscope ya lazeri (CLSM).
Imbonerahamwe 1 irerekana imiti ya 2707 HDSS.Imbonerahamwe 2 yerekana ko 2707 HDSS ifite imiterere yubukanishi ifite imbaraga zingana na 650 MPa.Ku mutini.1 yerekana microstructure ya optique yubushyuhe bwibisubizo bivurwa 2707 HDSS.Muri microstructure irimo 50% austenite na 50% ferrite yicyiciro, imirongo miremire ya austenite na ferrite idafite icyiciro cya kabiri iragaragara.
Ku mutini.2a yerekana ubushobozi bwumuzunguruko (Eocp) nigihe cyo kwerekana 2707 HDSS muri 2216E abiotic medium na P. aeruginosa umuyonga muminsi 14 kuri 37 ° C.Irerekana ko impinduka nini kandi zikomeye muri Eocp zibaho mumasaha 24 yambere.Indangagaciro za Eocp muri ibyo bihe byombi zageze kuri mV -145 (ugereranije na SCE) hafi 16 h hanyuma zigabanuka cyane, zigera kuri mV -477 (ugereranije na SCE) na -236 mV (ugereranije na SCE) kuri sample ya abiotic.na P Pseudomonas aeruginosa coupons, uko bikurikirana).Nyuma yamasaha 24, agaciro ka Eocp 2707 HDSS kuri P. aeruginosa kari gahagaze kuri mV -228 (ugereranije na SCE), mugihe agaciro kajyanye nicyitegererezo kitari ibinyabuzima cyari hafi mV -442 (ugereranije na SCE).Eocp imbere ya P. aeruginosa yari hasi cyane.
Ubushakashatsi bwamashanyarazi yubushakashatsi bwa 2707 HDSS muburyo bwa abiotic na pseudomonas aeruginosa umuyonga kuri 37 ° C:
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Imbonerahamwe 3 irerekana ibipimo bya elegitoroniki yamashanyarazi ya 2707 HDSS yerekana urugero rwa abiotic na Pseudomonas aeruginosa yanduye itangazamakuru mugihe cyiminsi 14.Ibiranga umurongo wa anode na cathode byajyanwe hanze kugirango babone amasangano atanga ubwinshi bwa ruswa (icorr), ubushobozi bwo kwangirika (Ecorr) hamwe na Tafel kumurongo (βα na βc) ukurikije uburyo busanzwe30,31.
Nkuko bigaragara ku gishushanyo.2b, guhinduka hejuru mumurongo wa P. aeruginosa byatumye Ecorr yiyongera ugereranije na abiotic curve.Agaciro ka icorr, kangana nigipimo cya ruswa, cyiyongereye kugera kuri 0.328 µA cm-2 muri sample ya Pseudomonas aeruginosa, ikaba ikubye inshuro enye ugereranije nicyitegererezo kidafite ibinyabuzima (0.087 µA cm-2).
LPR nuburyo bwa kera butangiza ibyangiza amashanyarazi kugirango isesengura ryihuse.Byakoreshejwe kandi kwiga MIC32.Ku mutini.2c yerekana polarisiyasi irwanya (Rp) nkigikorwa cyigihe cyo kwerekana.Indangagaciro ya Rp isobanura ruswa nke.Mu masaha 24 yambere, Rp 2707 HDSS yageze kuri 1955 kΩ cm2 kubigereranyo bya abiotic na 1429 kΩ cm2 kuri Pseudomonas aeruginosa.Igishushanyo 2c cyerekana kandi ko agaciro ka Rp kagabanutse vuba nyuma yumunsi umwe hanyuma ntigakomeza guhinduka muminsi 13 iri imbere.Agaciro Rp k'icyitegererezo cya Pseudomonas aeruginosa ni nka 40 kΩ cm2, kikaba kiri munsi cyane ya 450 kΩ cm2 z'icyitegererezo kidasanzwe.
Agaciro ka icorr kagereranijwe nigipimo kimwe cya ruswa.Agaciro kayo karashobora kubarwa uhereye kuri Stern-Giri ikurikira:
Kuri Zoe n'abandi.33, agaciro gasanzwe k'umusozi wa Tafel B muriyi mirimo yafashwe kuba 26 mV / dec.Igishushanyo 2d cyerekana ko icorr yicyitegererezo kitari ibinyabuzima 2707 yagumye ihagaze neza, mugihe icyitegererezo cya P. aeruginosa cyahindutse cyane nyuma yamasaha 24 yambere.Indangagaciro za icorr za P. aeruginosa ntangarugero zari gahunda yubunini burenze ubw'ubugenzuzi butari ibinyabuzima.Iyi myumvire ijyanye nibisubizo byo kurwanya polarisiyasi.
EIS nubundi buryo budasenya bukoreshwa mukuranga amashanyarazi ya chimique hejuru yubutaka.Impedance spectra hamwe no kubara ubushobozi bwa capacitance yintangarugero zagaragaye kubidukikije bya abiotic hamwe nigisubizo cya Pseudomonas aeruginosa, pasiporo ya pasiporo / biofilm irwanya Rb yakozwe hejuru yicyitegererezo, kwishyuza kwishyurwa Rct, amashanyarazi abiri yububiko Cdl (EDL) hamwe nibipimo bya QCPE Icyiciro cya buri gihe (CPE).Ibipimo byongeye gusesengurwa muguhuza amakuru ukoresheje icyerekezo kimwe (EEC).
Ku mutini.3 yerekana ibibanza bisanzwe bya Nyquist (a na b) na Bode ibibanza (a 'na b') kuburugero 2707 HDSS mubitangazamakuru bya abiotic hamwe nu muyoboro wa P. aeruginosa mugihe cyubushakashatsi butandukanye.Diameter yimpeta ya Nyquist iragabanuka imbere ya Pseudomonas aeruginosa.Umugambi wa Bode (Igishusho 3b ') werekana kwiyongera kwa impedance yose.Amakuru ajyanye nigihe cyo kwidagadura ahoraho arashobora kuboneka kuva icyiciro cya maxima.Ku mutini.4 yerekana imiterere ifatika ishingiye kuri monolayeri (a) na bilayeri (b) hamwe na EEC ihuye.CPE yinjijwe muburyo bwa EEC.Kwinjira no gutambuka kwayo kugaragazwa gutya:
Moderi ebyiri zifatika hamwe ninzira zingana zingana kugirango zihuze impedance ya sample ya 2707 HDSS:
aho Y0 nigiciro cya KPI, j numubare wibitekerezo cyangwa (-1) 1/2, ω numurongo wa buringuni, n ni indangagaciro ya KPI munsi ya imwe35.Ihererekanyabubasha ryo kurwanya ibicuruzwa (ni ukuvuga 1 / Rct) bihuye nigipimo cya ruswa.Gitoya Rct, niko igipimo cyo kwangirika27.Nyuma yiminsi 14 yubushakashatsi, Rct yintangarugero ya Pseudomonas aeruginosa yageze kuri 32 kΩ cm2, ikaba iri munsi ya 489 kΩ cm2 yintangarugero (Imbonerahamwe 4).
Amashusho ya CLSM n'amashusho ya SEM ku gishushanyo cya 5 yerekana neza ko ibinyabuzima biofilm bitwikiriye hejuru ya sample ya HDSS 2707 nyuma yiminsi 7 byuzuye.Ariko, nyuma yiminsi 14, ibinyabuzima bya biofilm byari bibi kandi selile zimwe zapfuye ziragaragara.Imbonerahamwe 5 yerekana umubyimba wa biofilm kuri 2707 HDSS nyuma yo guhura na P. aeruginosa muminsi 7 na 14.Uburebure bwa biofilm ntarengwa bwahindutse kuva 23.4 µm nyuma yiminsi 7 bugera kuri 18.9 µm nyuma yiminsi 14.Impuzandengo ya biofilm yuburebure nayo yemeje iyi nzira.Yagabanutse kuva 22.2 ± 0.7 mm nyuma yiminsi 7 igera kuri 17.8 ± 1.0 mm nyuma yiminsi 14.
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EMF yerekanye ibintu bya chimique muri biofilm nibicuruzwa byangirika kuburugero rwerekanwe na P. aeruginosa muminsi 14.Ku mutini.Igishushanyo cya 6 cyerekana ko ibikubiye muri C, N, O, na P muri biofilm nibicuruzwa byangirika biri hejuru cyane ugereranije nibyuma byera, kubera ko ibyo bintu bifitanye isano na biofilm na metabolite.Microbes ikenera gusa urugero rwa chromium na fer.Urwego rwo hejuru rwa Cr na Fe muri biofilm nibicuruzwa byangirika hejuru yicyitegererezo byerekana ko materique yicyuma yatakaje ibintu kubera kwangirika.
Nyuma yiminsi 14, ibyobo bifite na P. aeruginosa na bitagaragara byagaragaye hagati ya 2216E.Mbere yubushakashatsi, hejuru yicyitegererezo cyari cyoroshye kandi nta nenge (Ishusho 7a).Nyuma yo gukuramo no kuvanaho ibinyabuzima bya biofilm na ruswa, ibyobo byimbitse hejuru yuburorero byasuzumwe hakoreshejwe CLSM, nkuko bigaragara ku gishushanyo 7b na c.Nta mwobo ugaragara wabonetse hejuru yubugenzuzi butari ibinyabuzima (ubujyakuzimu bwa 0.02 µm).Ubujyakuzimu ntarengwa bwatewe na P. aeruginosa bwari 0.52 µm mu minsi 7 na 0,69 µm mu minsi 14, hashingiwe ku kigereranyo cy’ubujyakuzimu bwavuye mu byitegererezo 3 (hatoranijwe ubujyakuzimu 10 ntarengwa kuri buri cyitegererezo).Ibyagezweho kuri 0.42 ± 0.12 µm na 0.52 ± 0.15 µm, (Imbonerahamwe 5).Ubujyakuzimu bwimbitse ni nto ariko ni ngombwa.
(a) mbere yo kwerekanwa, (b) iminsi 14 mubidukikije, na (c) iminsi 14 mumyanda ya Pseudomonas aeruginosa.
Ku mutini.Imbonerahamwe 8 irerekana XPS yerekanwe yubuso butandukanye, kandi imiti yasesenguwe kuri buri buso yavuzwe mu mbonerahamwe ya 6. Mu mbonerahamwe ya 6, ijanisha rya atome rya Fe na Cr imbere ya P. aeruginosa (ingero A na B) zari munsi cyane ugereranije no kugenzura ibinyabuzima.(ingero C na D).Kuri sample ya P. aeruginosa, umurongo wikurikiranya kurwego rwa Cr 2p nucleus washyizwe mubice bine byimpinga zifite ingufu zihuza (BE) za 574.4, 576.6, 578.3 na 586.8 eV, zishobora kwitirirwa Cr, Cr2O3, CrO3 .na Cr (OH) 3, kimwe (Ishusho 9a na b).Kubitegererezo bitari biologiya, urwego rwibanze rwa Cr 2p rurimo impinga ebyiri nyamukuru kuri Cr (573.80 eV kuri BE) na Cr2O3 (575.90 eV kuri BE) mubishushanyo.9c na d.Itandukaniro ryibonekeje cyane hagati yintangarugero ya abiotic nicyitegererezo cya P. aeruginosa kwari ukubaho kwa Cr6 + hamwe nikigereranyo kinini cya Cr (OH) 3 (BE 586.8 eV) munsi ya biofilm.
Mugari XPS yerekana ubuso bwa sample 2707 HDSS mubitangazamakuru bibiri ni iminsi 7 na 14.
(a) Iminsi 7 ihura na P. aeruginosa, (b) iminsi 14 guhura na P. aeruginosa, (c) iminsi 7 mubidukikije, na (d) iminsi 14 mubidukikije.
HDSS yerekana urwego rwo hejuru rwo kurwanya ruswa ahantu henshi.Kim et al.2 yatangaje ko HDSS UNS S32707 yagaragaye nka DSS ivanze cyane ifite PREN irenga 45. PREN agaciro ka sample 2707 HDSS muriyi mirimo yari 49. Ibi biterwa nibirimo chromium nyinshi nibirimo byinshi molybdenum na nikel, bifite akamaro mubidukikije.n'ibidukikije birimo chloride nyinshi.Byongeye kandi, ibingana neza hamwe na microstructure idafite inenge bifite akamaro muburyo bwo gutuza no kurwanya ruswa.Nubwo, nubwo irwanya imiti myiza cyane, amakuru yubushakashatsi muri iki gikorwa yerekana ko 2707 HDSS idakingiwe rwose na biofilm ya P. aeruginosa.
Ibisubizo by'amashanyarazi byerekanaga ko igipimo cya ruswa ya 2707 HDSS mu muyoboro wa P. aeruginosa cyiyongereye cyane nyuma yiminsi 14 ugereranije n’ibidukikije.Mu gishushanyo cya 2a, igabanuka rya Eocp ryagaragaye haba mu buryo bwa abiotic ndetse no mu muyoboro wa P. aeruginosa mu masaha 24 ya mbere.Nyuma yibyo, biofilm itwikira rwose hejuru yicyitegererezo, kandi Eocp iba ihagaze neza36.Nyamara, urwego rwibinyabuzima rwa Eocp rwari hejuru cyane kurwego rwa Eocp rutari ibinyabuzima.Hariho impamvu zo kwizera ko iri tandukaniro rifitanye isano no gukora biofilm ya P. aeruginosa.Ku mutini.2d imbere ya P. aeruginosa, icorr 2707 HDSS agaciro kageze kuri 0,627 μA cm-2, ni gahunda yubunini burenze ubwo kugenzura abiotic (0.063 μA cm-2), byari bihuye nagaciro Rct yapimwe na EIS.Mu minsi mike ya mbere, indangagaciro zo kwangirika mu muhogo wa P. aeruginosa ziyongereye kubera guhuza ingirabuzimafatizo za P. aeruginosa no gukora biofilm.Ariko, iyo biofilm itwikiriye neza icyitegererezo, impedance iragabanuka.Igice cyo gukingira cyibasiwe cyane cyane no gukora biofilm na metabolite ya biofilm.Kubera iyo mpamvu, kurwanya ruswa byagabanutse igihe kandi kwizirika kwa P. aeruginosa byateje ruswa.Ibigenda mubidukikije bya abiotic byari bitandukanye.Kurwanya ruswa yo kutagenzura ibinyabuzima byari hejuru cyane ugereranije nagaciro kangana nicyitegererezo cyerekanwe na P. aeruginosa.Mubyongeyeho, kubikoresho bya abiotic, agaciro ka Rct 2707 HDSS kageze kuri 489 kΩ cm2 kumunsi wa 14, kikaba cyikubye inshuro 15 kurenza agaciro ka Rct (32 kΩ cm2) imbere ya P. aeruginosa.Niyo mpamvu, 2707 HDSS ifite imbaraga zo kurwanya ruswa mu bidukikije, ariko ntishobora kurwanya MIC ziva muri biofilm ya P. aeruginosa.
Ibisubizo birashobora kandi kugaragara uhereye kumurongo wa polarisiyasi.2b.Ishami rya Anodic ryahujwe no gukora biofilm ya Pseudomonas aeruginosa hamwe na okiside yicyuma.Kuri iki kibazo, reaction ya cathodic nigabanuka rya ogisijeni.Kubaho kwa P. aeruginosa byongereye cyane ububobere bwa ruswa, hafi ya gahunda yubunini burenze iyo kugenzura abiotic.Ibi byerekana ko biofilm ya P. aeruginosa yongerera ruswa ya 2707 HDSS.Yuan et al.29 yasanze ubucucike bwa ruswa ya Cu-Ni 70/30 yiyongereye bitewe na biofilm ya P. aeruginosa.Ibi birashobora guterwa na biocatalyse yo kugabanya ogisijeni na biofilm ya Pseudomonas aeruginosa.Iyi ndorerezi irashobora kandi gusobanura MIC 2707 HDSS muriki gikorwa.Hashobora kandi kuba ogisijeni nkeya munsi ya biofilm ya aerobic.Kubwibyo, kwanga kongera gutambutsa hejuru yicyuma hamwe na ogisijeni birashobora kuba ikintu kigira uruhare muri MIC muriki gikorwa.
Dickinson n'abandi.38 yatanze igitekerezo ko igipimo cyimiti n’amashanyarazi gishobora guterwa nigikorwa cya metabolike ya bagiteri ya sessile hejuru yicyitegererezo hamwe nimiterere yibicuruzwa byangirika.Nkuko bigaragara ku gishushanyo cya 5 nimbonerahamwe 5, umubare wutugingo nubunini bwa biofilm wagabanutse nyuma yiminsi 14.Ibi birashobora gusobanurwa neza nukuri ko nyuma yiminsi 14, inyinshi mu ngirabuzimafatizo za sessile hejuru ya 2707 HDSS zapfuye zatewe no kubura intungamubiri mu buryo bwa 2216E cyangwa kurekura ioni zifite uburozi ziva muri matrike ya 2707 HDSS.Iyi ni imbogamizi yubushakashatsi.
Muri iki gikorwa, biofilm ya P. aeruginosa yagize uruhare mu kugabanuka kwa Cr na Fe munsi ya biofilm hejuru ya 2707 HDSS (Ishusho 6).Imbonerahamwe 6 irerekana igabanuka rya Fe na Cr muri sample D ugereranije nicyitegererezo C, byerekana ko Fe na Cr byasheshwe byatewe na biofilm ya P. aeruginosa byakomeje iminsi 7 yambere.Ibidukikije 2216E bikoreshwa mu kwigana ibidukikije byo mu nyanja.Irimo 17700 ppm Cl-, igereranywa nibiri mu mazi yo mu nyanja.Kubaho 17700 ppm Cl- niyo mpamvu nyamukuru yo kugabanuka kwa Cr muri 7- na 14-iminsi ya abiotic sample yasesenguwe na XPS.Ugereranije na P. aeruginosa ntangarugero, iseswa rya Cr mu byitegererezo bya abiotic nticyari gito cyane kubera ko 2707 HDSS irwanya chlorine mu bihe bya abiotic.Ku mutini.9 yerekana ko hari Cr6 + muri firime ya passivating.Irashobora kugira uruhare mugukuraho chromium hejuru yicyuma na biofilm ya P. aeruginosa, nkuko byasabwe na Chen na Clayton.
Bitewe no gukura kwa bagiteri, indangagaciro za pH ziciriritse mbere na nyuma yo guhinga zari 7.4 na 8.2.Rero, munsi ya biofilm ya P. aeruginosa, kwangirika kwa acide kama ntigishobora kugira uruhare muriki gikorwa bitewe na pH iri hejuru cyane muburyo bwinshi.PH yuburyo butagenzura ibinyabuzima ntabwo bwahindutse cyane (kuva 7.4 yambere kugeza 7.5 yanyuma) mugihe cyiminsi 14 yikizamini.Ubwiyongere bwa pH muburyo bwo gutera inshinge nyuma yubushakashatsi bwajyanye nigikorwa cya metabolike ya P. aeruginosa kandi wasangaga bigira ingaruka zimwe kuri pH mugihe hatabayeho ibizamini.
Nkuko bigaragara ku gishushanyo cya 7, ubujyakuzimu ntarengwa bwatewe na P. aeruginosa biofilm bwari 0,69 µm, bukaba bunini cyane kuruta ubw'abiyotike (0.02 µm).Ibi bihuye namakuru yamashanyarazi yasobanuwe haruguru.Ubujyakuzimu bwa 0,69 µm burenze inshuro icumi kurenza 9.5 µm agaciro kavuzwe kuri 2205 DSS mubihe bimwe.Aya makuru yerekana ko 2707 HDSS yerekana guhangana neza na MIC kuruta 2205 DSS.Ibi ntibikwiye gutungurwa kuva 2707 HDSS ifite urwego rwisumbuye rwa Cr rutanga passivasi ndende, bigoye gutesha agaciro P. aeruginosa, kandi kubera imiterere yacyo iringaniye nta mvura yangiza yangiza itera.
Mu gusoza, ibyobo bya MIC byabonetse hejuru ya 2707 HDSS mu muhogo wa P. aeruginosa ugereranije n’ibyobo bidafite akamaro mu bidukikije.Aka kazi kerekana ko 2707 HDSS ifite imbaraga zo kurwanya MIC kuruta 2205 DSS, ariko ntabwo ikingira rwose MIC kubera biofilm ya P. aeruginosa.Ibisubizo bifasha muguhitamo ibyuma bidafite ingese hamwe nigihe cyo kubaho kubidukikije byo mu nyanja.
Coupon ya 2707 HDSS yatanzwe na kaminuza yuburaruko bushira ubuseruko (NEU) Ishuri rya Metallurgie i Shenyang, mubushinwa.Ibigize bigize 2707 HDSS bigaragara mu mbonerahamwe ya 1, yasesenguwe n’ishami rishinzwe isesengura n’ibizamini bya NEU.Ingero zose zavuwe kugirango zibonerwe igisubizo kuri 1180 ° C kumasaha 1.Mbere yo gupimwa ruswa, ibiceri bingana na 2707 HDSS ifite ubuso hejuru yubuso bwa cm 1 byahanaguwe kugeza kuri grit 2000 hamwe na silicon karbide sandpaper hanyuma bikozwemo ifu ya 0.05 µm Al2O3.Impande no hepfo birinzwe hamwe n'irangi.Nyuma yo gukama, intangarugero zogejwe namazi ya sterile deionisale hanyuma zihindurwamo 75% (v / v) Ethanol kuri 0.5 h.Baca bahumeka umwuka munsi yumucyo ultraviolet (UV) kuri 0.5 h mbere yo kuyikoresha.
Marine Pseudomonas aeruginosa strain MCCC 1A00099 yaguzwe mu kigo cyo gukusanya umuco wo mu nyanja cya Xiamen (MCCC), mu Bushinwa.Pseudomonas aeruginosa yakuze mubihe byindege kuri 37 ° C. mumashanyarazi ya ml 250 na selile 500 y ibirahuri byamashanyarazi ukoresheje marine 2216E yamazi (Qingdao Hope Biotechnology Co., Ltd., Qingdao, Mubushinwa).Hagati irimo (g / l): 19.45 NaCl, 5.98 MgCl2, 3.24 Na2SO4, 1.8 CaCl2, 0.55 KCl, 0.16 Na2CO3, 0.08 KBr, 0.034 SrCl2, 0.08 SrBr2, 0.022 H3BO3, 0.004 NaSiO3, 0016 6NH26NH3, 3.0016 NH3 umusemburo ukuramo na citrate 0.1.Autoclave kuri 121 ° C muminota 20 mbere yo gutera.Kubara selile ya sessile na planktonic hamwe na hemocytometero munsi ya microscope yoroheje kuri 400x.Ubwiyongere bwa mbere bwa planktonic Pseudomonas aeruginosa ako kanya nyuma yo gukingirwa byari selile 106 / ml.
Ibizamini by'amashanyarazi byakorewe mu ngirabuzimafatizo ya elegitoroniki itatu ya electrode ifite ubunini bwa ml 500.Urupapuro rwa platine hamwe na electrode yuzuye ya Calomel (SAE) byahujwe na reakteri binyuze muri capillaries ya Luggin yuzuyemo ibiraro byumunyu, byakoraga nka electrode.Kugira ngo hakorwe electrode ikora, insinga z'umuringa zometse kuri buri cyitegererezo kandi zipfundikirwa na epoxy resin, hasigara cm 1 z'ahantu hatarinzwe kuri electrode ikora kuruhande rumwe.Mu gupima amashanyarazi, ingero zashyizwe mu gipimo cya 2216E kandi zigashyirwa ku bushyuhe buri gihe (37 ° C) mu bwogero bw’amazi.OCP, LPR, EIS hamwe namakuru ashobora kuba afite imbaraga za polarisiyasi yapimwe hifashishijwe Autolab potentiostat (Reba 600TM, Gamry Instruments, Inc., USA).Ibizamini bya LPR byanditswe ku gipimo cya scan ya 0,125 mV s-1 mu ntera ya -5 kugeza kuri 5 mV hamwe na Eocp hamwe n’icyitegererezo cya 1 Hz.EIS yakoreshwaga hamwe na sine hejuru yumurongo wa 0.01 kugeza 10,000 Hz ukoresheje voltage ikoreshwa ya 5 mV kuri leta ihamye.Mbere yo guhanagura, electrode yari muburyo budakora kugeza igihe agaciro gahamye k'ubusa bushobora kugerwaho.Imirongo ya polarisiyasi yapimwe kuva -0.2 kugeza 1.5 V nkigikorwa cya Eocp ku gipimo cya scan ya 0.166 mV / s.Buri kizamini cyasubiwemo inshuro 3 hamwe na P. aeruginosa.
Icyitegererezo cyo gusesengura ibyuma byifashishijwe mu buryo bwa mashini hamwe na 2000 grit ya SiC itose hanyuma ikomeza gukonjeshwa hamwe na 0.05 µm Al2O3 ihagarika ifu kugirango irebe neza.Isesengura ryibyuma ryakozwe hakoreshejwe microscope optique.Ingero zashizwemo 10 wt% yumuti wa potasiyumu hydroxide 43.
Nyuma yububikwe, ibyitegererezo byogejwe inshuro 3 hamwe na saline ya fosifate (PBS) (pH 7.4 ± 0.2) hanyuma bigashyirwa hamwe na 2.5% (v / v) glutaraldehyde mumasaha 10 kugirango bikosore biofilm.Nyuma yaje kubura umwuma hamwe na Ethanol yuzuye (50%, 60%, 70%, 80%, 90%, 95% na 100% mubunini) mbere yo gukama umwuka.Hanyuma, firime ya zahabu ishyirwa hejuru yicyitegererezo kugirango itange uburyo bwo kwitegereza SEM.Amashusho ya SEM yibanze ku bibanza bifite selile nyinshi ya P. aeruginosa hejuru ya buri sample.Kora isesengura rya EDS kugirango ubone ibintu bya shimi.Ikoreshwa rya Zeiss confocal laser scanning microscope (CLSM) (LSM 710, Zeiss, Ubudage) yakoreshejwe mu gupima ubujyakuzimu.Kugira ngo harebwe ibyobo byangirika munsi ya biofilm, icyitegererezo cy’ibizamini cyabanje kozwa hakurikijwe ibipimo ngenderwaho by’Ubushinwa (CNS) GB / T4334.4-2000 kugira ngo bikureho ibicuruzwa byangirika na biofilm hejuru y’icyitegererezo.
Isesengura rya X-ray yerekana amashanyarazi (XPS, ESCALAB250 sisitemu yo gusesengura hejuru, Thermo VG, USA) isesengura ryakozwe hifashishijwe isoko ya X-ray imwe (umurongo wa Aluminium Kα ufite ingufu za 1500 eV nimbaraga za 150 W) muburyo butandukanye guhuza ingufu 0 mubihe bisanzwe bya –1350 eV.Ikirangantego kinini cyanditsweho hakoreshejwe ingufu zo kohereza 50 eV nintambwe ya 0.2 eV.
Ingero zubushakashatsi zavanyweho hanyuma zozwa buhoro hamwe na PBS (pH 7.4 ± 0.2) kuri 15 s45.Kugirango harebwe imbaraga za bagiteri ziterwa na biofilm ku ngero, biofilm yandujwe hakoreshejwe LIVE / DEAD BacLight Bacterial Viability Kit (Invitrogen, Eugene, OR, USA).Igikoresho kirimo amarangi abiri ya fluorescent: SYTO-9 irangi ryicyatsi kibisi na propidium iodide (PI) irangi ritukura rya fluorescent.Muri CLSM, fluorescent icyatsi nicyatsi gitukura byerekana selile nzima kandi zapfuye.Kubisiga irangi, ml 1 yuruvange rurimo 3 µl ya SYTO-9 na 3 µl yumuti wa PI yashizwemo iminota 20 mubushyuhe bwicyumba (23 ° C) mwijima.Nyuma yaho, ingero zanduye zasuzumwe ku burebure bubiri (488 nm kuri selile nzima na 559 nm kuri selile zapfuye) hakoreshejwe ibikoresho bya Nikon CLSM (C2 Plus, Nikon, Ubuyapani).Ubunini bwa biofilm bwapimwe muburyo bwo gusikana 3D.
Uburyo bwo gutanga iyi ngingo: Li, H. n'abandi.Microbial ruswa ya 2707 super duplex idafite ibyuma na Pseudomonas aeruginosa marine biofilm.siyanse.6, 20190. doi: 10.1038 / srep20190 (2016).
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