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Muva nje, inkundla yemithi elwa namagciwane engenawo amakhemikhali esekelwe ku-nanotechnology esebenzisa ama-nanostructures amanzi okwenziwa (EWNS) iye yasungulwa.I-EWNS inokushajwa okuphezulu kwendawo futhi igcwele uhlobo lwe-oksijini esebenzayo (ROS) engasebenzisana futhi yenze inani lezilwanyana ezincanyana lisebenze, okuhlanganisa amagciwane atholakala ekudleni.Lapha kuboniswa ukuthi izakhiwo zabo ngesikhathi sokuhlanganiswa zingacutshungulwa kahle futhi zenziwe ngcono ukuze kuthuthukiswe amandla abo e-antibacterial.Inkundla yelabhorethri ye-EWNS yakhelwe ukulungisa kahle izakhiwo ze-EWNS ngokushintsha amapharamitha wokuhlanganisa.Ukuhlukaniswa kwezakhiwo ze-EWNS (inkokhelo, usayizi nokuqukethwe kwe-ROS) kusetshenziswa izindlela zesimanje zokuhlaziya.Ngaphezu kwalokho, zahlolwa ukuthi azikwazi yini ukungasebenzi kwe-microbial ngokumelene namagciwane atholakala ekudleni njenge-Escherichia coli, i-Salmonella enterica, i-Listeria innocuous, i-Mycobacterium paraaccidentum ne-Saccharomyces cerevisiae.Imiphumela evezwe lapha ibonisa ukuthi izici ze-EWNS zingacushwa kahle ngesikhathi sokuhlanganiswa, okuholela ekwenyukeni okukhulu kokungasebenzi kahle kokusebenza.Ikakhulukazi, ukushaja okungaphezulu kwanda ngesilinganiso sezine futhi izinhlobo ze-oxygen esebenzayo zanda.Izinga lokususwa kwe-microbial lalincike ku-microbially futhi lalisuka ku-1.0 ukuya kulogi engu-3.8 ngemva kokuchayeka kwamaminithi angu-45 kudosi ye-aerosol engu-40,000 #/cc EWNS.
Ukungcoliswa kwamagciwane kuyimbangela eyinhloko yokugula okubangelwa ukudla okubangelwa ukungenwa amagciwane noma ubuthi bawo.E-United States kuphela, ukugula okubangelwa ukudla kubangela izifo ezingaba yizigidi ezingu-76, abantu abalaliswa esibhedlela abangu-325 000, futhi bafe abangu-5 000 unyaka ngamunye.Ngaphezu kwalokho, uMnyango Wezolimo wase-United States (i-USDA) ulinganisela ukuthi ukusetshenziswa okwandisiwe kwemikhiqizo emisha kubangela u-48% wazo zonke izifo ezitholakala ngokudla ezibikiwe e-United States2.Izindleko zezifo nokufa okubangelwa amagciwane atholakala ekudleni e-United States ziphezulu kakhulu, zilinganiselwa i-Centers for Disease Control and Prevention (CDC) ngaphezu kwe-US $ 15.6 billion ngonyaka3.
Njengamanje, ukungenelela kwamakhemikhali4, ama-radiation5 kanye ne-thermal6 kwe-antimicrobial ukuze kuqinisekiswe ukuphepha kokudla kwenziwa kakhulu ezindaweni zokulawula ezibucayi (CCPs) eduze kochungechunge lokukhiqiza (imvamisa ngemva kokuvuna kanye/noma phakathi nokupakishwa) kunokuba kwenziwe ngokuqhubekayo.ngakho-ke, bathambekele ekungcolisweni.7. Ukulawulwa okungcono kokugula okubangelwa ukudla kanye nokonakala kokudla kudinga ukungenelela kwemithi elwa namagciwane okungenzeka kusetshenziswe kuyo yonke ipulazi kuya etafuleni ngenkathi kunciphisa umthelela nezindleko zemvelo.
Muva nje, kusungulwe inkundla ye-antimicrobial engenamakhemikhali, esekwe ku-nanotechnology engakwazi ukuvala amagciwane angaphezulu nasomoya kusetshenziswa ama-nanostructures amanzi okwenziwa (EWNS).I-EWNS yahlanganiswa kusetshenziswa izinqubo ezimbili ezihambisanayo, i-electrospray kanye ne-ionization yamanzi (Fig. 1a).Ucwaningo lwangaphambilini lubonise ukuthi i-EWNS inesethi ehlukile yezakhiwo zomzimba nezebhayoloji8,9,10.I-EWNS inesilinganiso sama-electron angu-10 isakhiwo ngasinye kanye nosayizi we-nanoscale ophakathi kuka-25 nm (Fig. 1b,c)8,9,10.Ukwengeza, i-electron spin resonance (ESR) ibonise ukuthi i-EWNS iqukethe inani elikhulu lezinhlobo ze-oxygen esebenzayo (ROS), ikakhulukazi i-hydroxyl (OH•) ne-superoxide (O2-) radicals (Fig. 1c)8.I-EVNS isemoyeni isikhathi eside futhi ingangqubuzana nama-microorganisms amiswe emoyeni futhi akhona phezulu, alethe inkokhelo yawo ye-ROS futhi abangele ukungasebenzi kwama-microorganisms (Fig. 1d).Lezi zifundo zakuqala ziphinde zabonisa ukuthi i-EWNS ingasebenzisana futhi yenze amagciwane ahlukahlukene egram-negative kanye negram-positive, kuhlanganise ne-mycobacteria, ezindaweni kanye nasemoyeni.I-microscopy ye-electron yokudlulisela ibonise ukuthi ukungasebenzi kwakubangelwa ukuphazamiseka kwe-membrane yeseli.Ukwengeza, ucwaningo lwe-acute inhalation luye lwabonisa ukuthi imithamo ephezulu ye-EWNS ayibangeli umonakalo wamaphaphu noma ukuvuvukala 8.
(a) I-Electrospray yenzeka lapho kusetshenziswa amandla kagesi aphezulu phakathi kweshubhu le-capillary eliqukethe uketshezi kanye ne-counter electrode.(b) Ukusetshenziswa komfutho ophezulu kubangela izenzakalo ezimbili ezihlukene: (i) ukufafazwa kwamanzi nge-electro kanye (ii) nokwakheka kwezinhlobo zomoya-mpilo osebenzayo (ama-ion) avaleleke ku-EWNS.(c) Isakhiwo esiyingqayizivele se-EWNS.(d) Ngenxa yobunjalo bayo be-nanoscale, i-EWNS ihamba kakhulu futhi ingasebenzisana namagciwane ahamba emoyeni.
Ikhono lenkundla ye-EWNS ye-antimicrobial yokuvala ama-microorganisms atholakala ekudleni endaweni yokudla okusha nawo abonisiwe muva nje.Kuphinde kwaboniswa ukuthi ukushaja okungaphezulu kwe-EWNS kuhlanganiswe nenkundla kagesi kungasetshenziswa ukufeza ukulethwa okuhlosiwe.Ngaphezu kwalokho, imiphumela yokuqala yotamatisi we-organic ngemva kokuchayeka imizuzu engu-90 ku-EWNS engaba ngu-50,000 #/cm3 yayikhuthaza, kuphawulwe amagciwane ahlukahlukene atholakala ekudleni njenge-E. coli kanye ne-Listeria 11.Ngaphezu kwalokho, ukuhlolwa kokuqala kwe-organoleptic akuzange kubonise imiphumela yezinzwa uma kuqhathaniswa nokulawula utamatisi.Nakuba le miphumela yokuqala yokungasebenzi ikhuthaza izicelo zokuphepha kokudla ngisho nakumadosi e-EWNS aphansi kakhulu angu-50,000#/cc.bheka, kusobala ukuthi amandla aphezulu okungasebenzi angaba yinzuzo eyengeziwe ekunciphiseni ingozi yokutheleleka kanye nokonakala.
Lapha, sizogxila ocwaningweni lwethu ekuthuthukisweni kwenkundla yesizukulwane se-EWNS ukuze sikwazi ukulungisa kahle amapharamitha wokuhlanganisa kanye nokwenza kahle kwezakhiwo ze-physicochemical ze-EWNS ukuze kuthuthukiswe amandla azo okulwa namagciwane.Ikakhulukazi, ukwenza kahle kugxile ekwandiseni ukushaja kwazo okungaphezulu (ukuthuthukisa ukulethwa okuhlosiwe) nokuqukethwe kwe-ROS (ukuthuthukisa ukusebenza kahle kokungasebenzisi).Faka izici ezithuthukisiwe ze-physico-chemical (usayizi, ukushaja nokuqukethwe kwe-ROS) usebenzisa izindlela zokuhlaziya zesimanje futhi usebenzise ama-microorganisms okudla avamile afana no-E. .
I-EVNS yahlanganiswa nge-electrospraying kanyekanye kanye ne-ionization yamanzi ahlanzekile aphezulu (18 MΩ cm–1).I-nebulizer kagesi engu-12 ijwayele ukusetshenziselwa ukwenza i-athomu yoketshezi kanye nokuhlanganiswa kwezinhlayiya ze-polymer ne-ceramic 13 kanye namafayibha angu-14 osayizi olawulwayo.
Njengoba kuchaziwe ekushicilelweni kwangaphambilini 8, 9, 10, 11, ekuhlolweni okuvamile, kwasetshenziswa amandla kagesi aphezulu phakathi kwe-capillary yensimbi kanye ne-grounded counter electrode.Phakathi nale nqubo, kwenzeka izinto ezimbili ezihlukene: i) i-electrospray kanye nee) ionization yamanzi.Inkambu eqinile kagesi phakathi kwama-electrode amabili ibangela ukuthi amashaji angemahle akheke phezu kwamanzi ajiyile, okuholela ekwakhekeni kwama-Taylor cones.Ngenxa yalokho, kwakheka amaconsi amanzi ashajwe kakhulu, aqhubeka ehlukana abe yizinhlayiya ezincane, njengakumbono kaRayleigh16.Ngesikhathi esifanayo, amasimu kagesi aqinile abangela ukuthi amanye ama-molecule amanzi ahlukane futhi akhiphe ama-electron (ionize), okuholela ekwakhekeni kwenani elikhulu lezinhlobo ze-oxygen esebenzayo (ROS)17.I-ROS18 ekhiqizwe ngesikhathi esisodwa yafakwa ku-EWNS (Fig. 1c).
Emkhiwaneni.I-2a ikhombisa uhlelo lokuzala lwe-EWNS olwakhiwe futhi lwasetshenziswa ekuhlanganiseni kwe-EWNS kulolu cwaningo.Amanzi ahlanzekile agcinwe ebhodleleni elivaliwe ayephakelwa ngeshubhu le-Teflon (ububanzi obuyi-2 mm bangaphakathi) abe inaliti yensimbi engenasici engu-30G (i-metal capillary).Ukugeleza kwamanzi kulawulwa umfutho womoya ngaphakathi ebhodleleni, njengoba kuboniswe kuMfanekiso 2b.Inaliti ifakwe kukhonsoli ye-Teflon futhi ingalungiswa ngesandla ibe ibanga elithile ukusuka ku-electrode yokubala.I-electrode yekhawunta iyidiski ye-aluminium epholishiwe enembobo phakathi nendawo ukuze kuthathwe amasampula.Ngezansi kwekhawunta ye-electrode kunefaneli yesampula ye-aluminium, exhunywe kukho konke ukusetha kokuhlola ngembobo yesampula (Fig. 2b).Ukuze ugweme ukunqwabelana kwezindleko okungase kuphazamise ukusebenza kwamasampula, zonke izingxenye zesampula zisekelwe ngogesi.
(a) Uhlelo Lokukhiqiza Nanostructure Yamanzi Enjiniyela (EWNS).(b) Ingxenye ephambanayo yesampula kanye ne-electrospray, ebonisa amapharamitha abaluleke kakhulu.(c) Ukusethwa kokuhlola kokungasebenzi kwamagciwane.
Isistimu yokukhiqiza ye-EWNS echazwe ngenhla iyakwazi ukushintsha amapharamitha angukhiye wokusebenza ukuze kube lula ukuhlela kahle izakhiwo ze-EWNS.Lungisa i-voltage esetshenzisiwe (V), ibanga phakathi kwenaliti kanye ne-electrode yokubala (L), nokugeleza kwamanzi (φ) nge-capillary ukuze ulungise kahle izici ze-EWNS.Uphawu olusetshenziselwa ukumela inhlanganisela ehlukene: [V (kV), L (cm)].Lungisa ukugeleza kwamanzi ukuze uthole ikhoni ka-Taylor ezinzile yesethi ethile [V, L].Ngezinjongo zalolu cwaningo, ububanzi bokuvuleka kwe-electrode yekhawunta (D) bugcinwe kumayintshi angu-0.5 (1.29 cm).
Ngenxa yejometri elinganiselwe kanye ne-asymmetry, amandla enkambu kagesi awakwazi ukubalwa kusukela kuzimiso zokuqala.Esikhundleni salokho, isofthiwe ye-QuickField™ (Svendborg, Denmark)19 yasetshenziswa ukubala indawo kagesi.Inkambu kagesi ayifani, ngakho-ke inani lenkambu kagesi ekugcineni kwe-capillary lisetshenziswe njengenani elibhekisela ekucushweni okuhlukahlukene.
Phakathi nocwaningo, izinhlanganisela eziningana zamandla kagesi kanye nebanga phakathi kwenaliti kanye ne-electrode yokubala zihlolwe ngokokwakheka kwe-Taylor cone, ukuqina kwe-Taylor cone, ukuzinza kokukhiqiza kwe-EWNS, kanye nokuzala kabusha.Inhlanganisela ehlukahlukene ikhonjiswe kokuthi Ithebula Le-Supplementary S1.
Ukukhishwa kwesistimu yokukhiqiza ye-EWNS kuxhunywe ngokuqondile ku-Scanning Mobility Particle Size Analyzer (SMPS, Model 3936, TSI, Shoreview, MN) ukuze kukale inombolo yezinhlayiyana, kanye ne-Aerosol Faraday Electrometer (TSI, Model 3068B, Shoreview, MN).) emisinga ye-aerosol ikalwe njengoba kuchazwe ekushicilelweni kwethu kwangaphambilini.Kokubili i-SMPS ne-electrometer ye-aerosol okuthathwe ngesilinganiso sokugeleza esingu-0.5 L/min (isamba sokugeleza kwesampula 1 L/min).Ukugxiliswa kwenombolo yezinhlayiya nokugeleza kwe-aerosol kukalwe imizuzwana engu-120.Isilinganiso siphindwe izikhathi ezingu-30.Ngokusekelwe ezilinganisweni zamanje, inani lokushaja kwe-aerosol liyabalwa futhi inkokhiso emaphakathi ye-EWNS ilinganiselwa enanini elinikeziwe lezinhlayiya ze-EWNS ezikhethiwe.Isilinganiso sezindleko ze-EWNS singabalwa kusetshenziswa i-Equation (1):
lapho i-IEl iyinani lamanje elilinganisiwe, i-NSMPS iyingqikithi yedijithali elinganiswa nge-SMPS, futhi u-φEl izinga lokugeleza nge-electrometer ngayinye.
Ngenxa yokuthi umswakama ohlobene (RH) uthinta ukushajwa kwendawo, izinga lokushisa kanye (RH) zigcinwe zingashintshi ngesikhathi sokuhlolwa ku-21°C no-45%, ngokulandelana.
I-Atomic force microscopy (AFM), i-Asylum MFP-3D (Asylum Research, Santa Barbara, CA) kanye ne-AC260T probe (Olympus, Tokyo, Japan) zisetshenziswe ukukala usayizi nempilo yonke ye-EWNS.Imvamisa yokuskena ye-AFM yayingu-1 Hz, indawo yokuskena yayingu-5 μm × 5 μm, kanye nemigqa yokuskena engu-256.Zonke izithombe zibe ngaphansi kokuqondisa kwesithombe se-oda lokuqala kusetshenziswa isofthiwe ye-Asylum (ibanga lemaski 100 nm, i-threshold 100 pm).
Ifaneli yokuhlola ikhishiwe futhi i-mica surface yabekwa ebangeni elingu-2.0 cm ukusuka ku-electrode yekhawunta isikhathi esimaphakathi samasekhondi angu-120 ukuze kugwenywe ukuhlangana kwezinhlayiyana kanye nokwakheka kwamaconsi angajwayelekile endaweni ye-mica.I-EWNS yafuthwa ngokuqondile phezu kwe-mica esanda kusikwa (Ted Pella, Redding, CA).Isithombe sendawo ye-mica ngokushesha ngemva kokugwaza kwe-AFM.I-engeli yokuxhumana yendawo ye-mica engashintshiwe esanda kusikwa iseduze no-0°, ngakho-ke i-EVNS isakazwa endaweni ye-mica ngendlela yedome.Ububanzi (a) nobude (h) bamaconsi asakazekayo kukalwa ngokuqondile kusimo sendawo se-AFM futhi kwasetshenziswa ukubala ivolumu yokusabalalisa eholwa yi-EWNS kusetshenziswa indlela yethu egunyazwe ngaphambilini.Uma kucatshangwa ukuthi i-EWNS ebhodini inevolumu efanayo, ububanzi obulinganayo bungabalwa kusetshenziswa i-Equation (2):
Ngokusekelwe endleleni yethu eyakhiwe ngaphambilini, i-electron spin resonance (ESR) spin trap yasetshenziswa ukuze kutholwe ukuba khona kwama-radical intermediate wesikhashana ku-EWNS.Ama-aerosol afakwe ibhamuza nge-Midget sparger engu-650 μm (Ace Glass, Vineland, NJ) equkethe isixazululo esingu-235 mM se-DEPMPO(5-(diethoxyphosphoryl) -5-methyl-1-pyrroline-N-oxide) (Oxis International Inc.).Portland, Oregon).Zonke izilinganiso ze-ESR zenziwa kusetshenziswa i-spectrometer ye-Bruker EMX (Bruker Instruments Inc. Billerica, MA, USA) kanye neseli lephaneli eliyisicaba.Isofthiwe ye-Acquisit (Bruker Instruments Inc. Billerica, MA, USA) isetshenziselwe ukuqoqa nokuhlaziya idatha.Ukunqunywa kwezici ze-ROS kwenziwa kuphela ngesethi yezimo zokusebenza [-6.5 kV, 4.0 cm].Ukugxiliswa kwe-EWNS kukalwe kusetshenziswa i-SMPS ngemva kokubala ukulahlekelwa kwe-EWNS ku- impactor.
Amazinga e-ozone aqashwe kusetshenziswa i-205 Dual Beam Ozone Monitor™ (2B Technologies, Boulder, Co)8,9,10.
Kuzo zonke izici ze-EWNS, inani elimaphakathi lisetshenziswa njengevelu yokulinganisa, futhi ukuchezuka okujwayelekile kusetshenziswa njengephutha lokulinganisa.Ukuhlolwa kwe-T kwenziwa ukuze kuqhathaniswe amanani ezibaluli ezithuthukisiwe ze-EWNS namanani ahambisanayo esisekelo se-EWNS.
Umfanekiso 2c ukhombisa uhlelo lwe-electrostatic precipitation (EPES) oluthuthukiswe ngaphambilini futhi olunezimpawu “lokudonsa” olungasetshenziselwa ukulethwa okuhlosiwe kwe-EWNS phezulu.I-EPES isebenzisa izindleko ze-EVNS ezingase "ziqondiswe" ngokuqondile ebusweni bethagethi ngaphansi kwethonya lenkambu kagesi enamandla.Imininingwane yohlelo lwe-EPES yethulwa ekushicilelweni kwakamuva nguPyrgiotakis et al.11 .Ngakho-ke, i-EPES iqukethe ikamelo le-PVC eliphrintiwe le-3D elineziphetho ezicijile futhi liqukethe insimbi engagqwali ehambisanayo engu-304 (insimbi engagqwali engu-304, ehlanganiswe nesibuko) amapuleti ensimbi phakathi nendawo eyi-15.24 cm ukuhlukana.Amabhodi ayexhunywe kumthombo wamandla kagesi aphezulu wangaphandle (Bertran 205B-10R, Spellman, Hauppauge, NY), ipuleti eliphansi lalihlale lixhunywe ku-voltage eqondile, futhi ipuleti eliphezulu lalihlale lixhunywe emhlabathini (umhlabathi ontantayo).Izindonga zekamelo zimbozwe nge-aluminium foil, esekelwe ngogesi ukuvimbela ukulahleka kwezinhlayiyana.Igumbi linomnyango olayishiwe ovaliwe ongaphambili ovumela izindawo zokuhlola ukuthi zibekwe ezitendi zepulasitiki eziziphakamisa ngaphezu kwepuleti lensimbi elingaphansi ukuze kugwenywe ukuphazamiseka kwamandla kagesi aphezulu.
Ukusebenza kahle kokufakwa kwe-EWNS ku-EPES kubalwe ngokomthetho olandelwayo owenziwe ngaphambilini ochazwe ku-Supplementary Figure S111.
Njengegumbi lokulawula, igumbi lesibili lokugeleza kwe-cylindrical lixhunywe ochungechungeni ohlelweni lwe-EPES, lapho isihlungi se-HEPA esiphakathi sasetshenziselwa ukususa i-EWNS.Njengoba kukhonjisiwe kuMfanekiso 2c, i-aerosol ye-EWNS yampontshwa emagumbini amabili akhelwe ngaphakathi.Isihlungi esiphakathi kwegumbi lokulawula ne-EPES sisusa noma iyiphi i-EWNS esele eholela ezingeni lokushisa elifanayo (T), umswakama ohlobene (RH) namazinga we-ozone.
Amagciwane abalulekile atholakala ekudleni atholwe ukuthi angcolise ukudla okusha okufana ne-E. coli (ATCC #27325), inkomba ye-fecal, i-Salmonella enterica (ATCC #53647), i-pathogen etholakala ekudleni, i-Listeria engenabungozi (ATCC #33090), i-surrogate ye-pathogenic Listeria monocytogenes, i-Mavisiamytogenes, i-Mavisiamy, i-ATCC, i-Mavisiamy (ATCC #4098), esikhundleni seyeast spoilage, kanye ne-bacterium engazweli kakhudlwana, i-Mycobacterium paralucky (ATCC #19686).
Thenga amabhokisi angahleliwe otamatisi wamagilebhisi wemvelo emakethe yangakini bese uwafaka efrijini ku-4°C uze usetshenziswe (kufika ezinsukwini ezi-3).Utamatisi wokuhlola wonke wawunosayizi ofanayo, cishe u-1/2 intshi ububanzi.
Isiko, ukujova, ukuchayeka, kanye nezinqubo zokubala zekholoni kuchazwe kabanzi ekushicileleni kwethu kwangaphambilini futhi kunemininingwane ku-Supplementary Data.Ukusebenza kwe-EWNS kwahlaziywa ngokudalula utamatisi ogonyelwe ku-40,000 #/cm3 imizuzu engu-45.Kafushane, kwasetshenziswa utamatisi abathathu ukuhlola ama-microorganisms asaphila ngesikhathi t = 0 min.Amatamatisi amathathu afakwe ku-EPES futhi avezwa ku-EWNS ku-40,000 #/cc (EWNS evezwe utamatisi) kanti amathathu asele afakwe ekamelweni lokulawula (utamatisi wokulawula).Ukucubungula okwengeziwe kwamatamatisi kuwo womabili amaqembu akuzange kwenziwe.Utamatisi ovezwe yi-EWNS kanye notamatisi olawulayo akhishwe ngemva kwemizuzu engu-45 ukuhlola umphumela we-EWNS.
Ukuhlolwa ngakunye kwenziwa ngokuphindwe kathathu.Ukuhlaziywa kwedatha kwenziwa ngokuvumelana nephrothokholi echazwe ku-Supplementary Data.
Izindlela zokungasebenzi zihlolwe ngokuchithwa kwe-sedimentation yamasampuli e-EWNS aveziwe (45 min at 40,000 #/cm3 EWNS aerosol concentration) kanye namasampuli angenawo imisebe yebhaktheriya engenangozi i-E. coli, i-Salmonella enterica ne-Lactobacillus.Izinhlayiya zilungiswe ku-2.5% glutaraldehyde, 1.25% paraformaldehyde kanye ne-0.03% ye-picric acid ku-0.1 M i-sodium cacodylate buffer (pH 7.4) amahora angu-2 ekamelweni lokushisa.Ngemuva kokugeza, lungisa nge-1% osmium tetroxide (OsO4)/1.5% potassium ferrocyanide (KFeCN6) amahora angu-2, geza izikhathi ezi-3 emanzini futhi ufake i-uranyl acetate engu-1% ihora elingu-1, bese ugeza kabili emanzini, bese ukhipha amanzi emanzini imizuzu engu-10, 0 0 0% 0% 500, utshwala.Amasampula abe esefakwa ku-propylene oxide ihora elingu-1 futhi afakwe ingxube engu-1:1 ye-propylene oxide ne-TAAP Epon (Marivac Canada Inc. St. Laurent, CA).Amasampuli ashumekwe ku-TAAB Epon futhi enziwe i-polymerized ku-60°C amahora angu-48.I-resin ye-granular ephulukisiwe yasikwa futhi yabonwa yi-TEM kusetshenziswa isibonakhulu se-electron yokudlulisa i-JEOL 1200EX (JEOL, Tokyo, Japan) efakwe ikhamera ye-AMT 2k CCD (Advanced Microscopy Techniques, Corp., Woburn, Massachusetts, USA).
Zonke izivivinyo zenziwa ngokuphindwe kathathu.Ephuzwini ngalinye lesikhathi, ukugezwa kwamagciwane kwafakwa imbewu ngokuphindwe kathathu, okuholele engqikithini yamaphoyinti edatha ayisishiyagalolunye ngephuzu ngalinye, isilinganiso sawo esisetshenziswe njengokugxiliswa kwebhaktheriya kuleyo microorganism ethile.Ukuchezuka okujwayelekile kusetshenziswe njengephutha lokulinganisa.Wonke amaphuzu abalwa.
I-logarithm yokuncipha kokuhlangana kwamagciwane uma kuqhathaniswa no-t = 0 min ibalwe kusetshenziswa ifomula elandelayo:
lapho i-C0 iwukuhlangana kwamagciwane kusampula yokulawula ngesikhathi 0 (okungukuthi ngemva kokuba ubuso bomile kodwa ngaphambi kokubekwa egunjini) kanye ne-Cn ukugcwala kwamagciwane endaweni ngemva kwemizuzu engu-n yokuchayeka.
Ukulandisa ngokucekelwa phansi kwemvelo kwamabhaktheriya ngesikhathi sokuchayeka kwemizuzu engu-45, ukuncishiswa kwelogi uma kuqhathaniswa nokulawula ngemva kwemizuzu engu-45 nakho kubalwe ngale ndlela elandelayo:
lapho i-Cn iwukugcwala kwamabhaktheriya kusampula yokulawula ngesikhathi n kanye ne-Cn-Control ukugxila kokulawula amagciwane ngesikhathi n.Idatha yethulwa njengokwehliswa kwelogi uma kuqhathaniswa nokulawula (akukho ukuchayeka kwe-EWNS).
Phakathi nocwaningo, izinhlanganisela eziningana zamandla kagesi kanye nebanga phakathi kwenaliti kanye ne-electrode yokubala zihlolwe ngokokwakheka kwe-Taylor cone, ukuqina kwe-Taylor cone, ukuzinza kokukhiqiza kwe-EWNS, kanye nokuzala kabusha.Inhlanganisela ehlukahlukene ikhonjiswe kokuthi Ithebula Le-Supplementary S1.Izimo ezimbili ezibonisa izakhiwo ezizinzile nezikwazi ukukhiqiza kabusha (i-Taylor cone, isizukulwane se-EWNS, nokuzinza ngokuhamba kwesikhathi) zikhethelwe ucwaningo olunzulu.Emkhiwaneni.Umfanekiso wesi-3 ubonisa imiphumela yenkokhiso, usayizi, nokuqukethwe kwe-ROS kuzo zombili izimo.Imiphumela iphinde ikhonjiswe kuThebula 1. Njengereferensi, kokubili uMdwebo 3 kanye neThebula 1 kuhlanganisa izici ze-EWNS8, 9, 10, 11 (eyisisekelo-EWNS) eyahlanganiswa ngaphambilini engathuthukisiwe.Izibalo zokubaluleka kwezibalo zisebenzisa i-t-test enemisila emibili zishicilelwa kabusha ku-Supplementary Table S2.Ukwengeza, idatha eyengeziwe ihlanganisa izifundo zomphumela we-counter electrode isampula yembobo ububanzi (D) kanye nebanga phakathi kwe-electrode yomhlabathi kanye nethiphu (L) (Izibalo Ezingeziwe S2 kanye ne-S3).
(ac) Ukusatshalaliswa kosayizi kukalwa nge-AFM.(df) Isici sokushaja okungaphezulu.(g) Ukucaciswa kwe-ROS kwe-EPR.
Kubalulekile futhi ukuqaphela ukuthi kuzo zonke lezi zimo ezingenhla, i-ionization yamanje elinganisiwe yayiphakathi kuka-2 no-6 μA kanye ne-voltage phakathi kuka- -3.8 no -6.5 kV, okuholela ekusetshenzisweni kwamandla okungaphansi kuka-50 mW kule module yokuxhumana yesizukulwane esisodwa se-EWNS.Nakuba i-EWNS yahlanganiswa ngaphansi kwengcindezi ephezulu, amazinga e-ozone ayephansi kakhulu, awazange adlule ku-60 ppb.
Umfanekiso Owengeziwe we-S4 ubonisa izinkambu zikagesi ezilingisiwe zezimo [-6.5 kV, 4.0 cm] kanye no-[-3.8 kV, 0.5 cm], ngokulandelana.Ngezimo ze-[-6.5 kV, 4.0 cm] kanye ne-[-3.8 kV, 0.5 cm], izibalo zenkambu ziyi-2 × 105 V/m kanye no-4.7 × 105 V/m, ngokulandelana.Lokhu kulindeleke, njengoba esimweni sesibili isilinganiso sebanga le-voltage siphezulu kakhulu.
Emkhiwaneni.3a,b ikhombisa ububanzi be-EWNS obukalwa nge-AFM8.Isilinganiso esibaliwe samadiayimitha e-EWNS sasingu-27 nm no-19 nm kuma-scheme angu- [-6.5 kV, 4.0] kanye no-[-3.8 kV, 0.5 cm], ngokulandelana.Kuzimo ze-[-6.5 kV, 4.0 cm] kanye nezingu-[-3.8 kV, 0.5 cm], ukuchezuka okujwayelekile kwejiyomethri kokusatshalaliswa kungu-1.41 no-1.45, ngokulandelana, okubonisa ukusabalalisa kosayizi omncane.Kokubili usayizi wencazelo kanye nokuchezuka okujwayelekile kwejiyomethri kusondele kakhulu ku-EWNS yesisekelo, ku-25 nm no-1.41, ngokulandelanayo.Emkhiwaneni.I-3c ibonisa ukusatshalaliswa kosayizi wesisekelo se-EWNS esilinganiswa kusetshenziswa indlela efanayo ngaphansi kwezimo ezifanayo.
Emkhiwaneni.3d, e ikhombisa imiphumela yokushaja abalingisi.Idatha izilinganiso ezimaphakathi zezilinganiso ezingama-30 ngesikhathi esisodwa zokugxilisa ingqondo (#/cm3) kanye nezamanje (I).Ukuhlaziywa kukhombisa ukuthi inkokhiso emaphakathi ku-EWNS ingu-22 ± 6 e- kanye no-44 ± 6 e- ku-[-6.5 kV, 4.0 cm] kanye no-[-3.8 kV, 0.5 cm], ngokulandelana.Anamashaji angaphezulu kakhulu uma kuqhathaniswa nesisekelo se-EWNS (10 ± 2 e-), amakhulu ngokuphindwe kabili kunesimo se-[-6.5 kV, 4.0 cm] futhi amakhulu ngokuphindwe kane kune-[-3 .8 kV, 0.5 cm].Umfanekiso 3f ukhombisa inkokhiso.idatha ye-Baseline-EWNS.
Kusukela kumabalazwe wokugxilisa ingqondo wenombolo ye-EWNS (Izibalo Ezingeziwe S5 kanye ne-S6), kungabonakala ukuthi isimo se-[-6.5 kV, 4.0 cm] sinezinhlayiya eziningi kakhulu kunesimo [-3.8 kV, 0.5 cm].Kubalulekile futhi ukuqaphela ukuthi ukugxiliswa kwenombolo ye-EWNS kwaqashwa kwaze kwafika emahoreni angu-4 (Izibalo Ezingeziwe S5 kanye ne-S6), lapho ukuzinza kwesizukulwane se-EWNS kubonise amazinga afanayo okugxilwa kwezinhlayiyana zezinhlayiyana kuzo zombili izimo.
Emkhiwaneni.I-3g ibonisa i-spectrum ye-EPR ngemva kokukhipha isilawuli se-EWNS esithuthukisiwe (ingemuva) kokuthi [-6.5 kV, 4.0 cm].I-spectra ye-ROS iphinde yaqhathaniswa ne-Baseline-EWNS scenario emsebenzini oshicilelwe ngaphambilini.Inombolo ye-EWNS esabelayo nge-spin traps ibalwe yaba ngu-7.5 × 104 EWNS/s, efana ne-Baseline-EWNS8 eshicilelwe ngaphambilini.I-spectra ye-EPR ibonise ngokucacile ukuba khona kwezinhlobo ezimbili ze-ROS, i-O2- iwuhlobo oluhamba phambili futhi i-OH• ingeningi kakhulu.Ukwengeza, ukuqhathanisa okuqondile kokuqina okuphezulu kubonise ukuthi i-EWNS eyenziwe kahle inokuqukethwe kwe-ROS okuphezulu kakhulu uma kuqhathaniswa ne-EWNS eyisisekelo.
Emkhiwaneni.4 ikhombisa ukusebenza kahle kwe-EWNS ku-EPES.Idatha iphinde ifinyezwe kuThebula I futhi iqhathaniswe nedatha ye-EWNS yasekuqaleni.Kuzo zombili izimo ze-EUNS, ukubekelwa phansi kusondele ku-100% ngisho nakugesi ophansi ongu-3.0 kV.Ngokujwayelekile, i-3.0 kV yanele u-100% wokufakwa, kungakhathaliseki ukuthi ishaja ishintshile kanjani.Ngaphansi kwezimo ezifanayo, ukusebenza kahle kokufakwa kwe-Baseline-EWNS kwakungama-56% kuphela ngenxa yokushajwa kwawo okuphansi (isilinganiso sama-electron ayi-10 nge-EWNS ngayinye).
Emkhiwaneni.5 futhi etafuleni.2 ifingqa inani lokungasebenzi kwama-microorganisms afakwe ebusweni bukatamatisi ngemva kokuchayeka ku-40,000 #/cm3 EWNS imizuzu engu-45 kumodi enkulu [-6.5 kV, 4.0 cm].I-E. coli e-inoculated kanye ne-Lactobacillus engenacala ibonise ukuncipha okukhulu kwamalogi angu-3.8 ngesikhathi sokuchayeka kwamaminithi angu-45.Ngaphansi kwezimo ezifanayo, i-S. enterica ibe nokwehla okungu-2.2-log, kuyilapho i-S. cerevisiae ne-M. parafortutum ibe nokwehla okungu-1.0-log.
Ama-electron micrographs (Umfanekiso 6) abonisa izinguquko ezingokomzimba ezidalwe yi-EWNS kumaseli angenabungozi we-Escherichia coli, Streptococcus, kanye ne-Lactobacillus okuholela ekungasebenzini kwawo.Ibhaktheriya elilawulayo lalinolwelwesi lwamaseli olungaguquki, kuyilapho amabhaktheriya ayeveziwe ayewonakalise ulwelwesi lwangaphandle.
I-electronic imaging microscopic yokulawula kanye namagciwane aveziwe aveze ukulimala kolwelwesi.
Idatha yezakhiwo ze-physicochemical ye-EWNS ethuthukisiwe ngokuhlangene ibonisa ukuthi izakhiwo (izindleko ezingaphezulu nokuqukethwe kwe-ROS) ze-EWNS zithuthukiswe kakhulu uma kuqhathaniswa nedatha yesisekelo ye-EWNS eshicilelwe ngaphambili8,9,10,11.Ngakolunye uhlangothi, usayizi wabo wahlala ebangeni le-nanometer, efana kakhulu nemiphumela ebikwe ngaphambilini, ebavumela ukuthi bahlale emoyeni isikhathi eside.I-polydispersity ephawuliwe ingachazwa ngoshintsho lwenkokhiso yendawo enquma usayizi we-EWNS, ukungahleliwe komphumela we-Rayleigh, kanye nokuhlangana okungenzeka.Nokho, njengoba kuchazwe nguNielsen et al.22, ukushaja okuphezulu kwendawo kunciphisa ukuhwamuka ngokwandisa ngempumelelo amandla angaphezulu/ukushuba kokwehla kwamanzi.Ekushicileleni kwethu okwedlule8 le theory yaqinisekiswa ngokuhlolwa kuma-microdroplets 22 kanye ne-EWNS.Ukulahlekelwa kwenkokhiso ngesikhathi esengeziwe kungase futhi kuthinte usayizi futhi kube nomthelela ekusabalaliseni kosayizi obukiwe.