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Posachedwapa, nsanja ya antimicrobial yopanda mankhwala yochokera ku nanotechnology pogwiritsa ntchito madzi opangira madzi (EWNS) yapangidwa.EWNS ili ndi chiwongolero chapamwamba kwambiri ndipo imakhala ndi mitundu yowonjezereka ya okosijeni (ROS) yomwe ingagwirizane ndi kusokoneza tizilombo toyambitsa matenda, kuphatikizapo tizilombo toyambitsa matenda.Apa zikuwonetsedwa kuti katundu wawo panthawi ya kaphatikizidwe amatha kusinthidwa bwino ndikuwongoleredwa kuti apititse patsogolo kuthekera kwawo kwa antibacterial.Pulatifomu ya labotale ya EWNS idapangidwa kuti ikonze bwino zinthu za EWNS posintha magawo a kaphatikizidwe.Makhalidwe a katundu wa EWNS (malipiro, kukula ndi zomwe zili mu ROS) pogwiritsa ntchito njira zamakono zowunikira.Kuphatikiza apo, adawunikidwa chifukwa cha kuthekera kwawo kwa tizilombo toyambitsa matenda motsutsana ndi tizilombo toyambitsa matenda monga Escherichia coli, Salmonella enterica, Listeria innocuous, Mycobacterium paraaccidentum ndi Saccharomyces cerevisiae.Zotsatira zomwe zaperekedwa apa zikuwonetsa kuti katundu wa EWNS akhoza kusinthidwa bwino panthawi ya kaphatikizidwe, zomwe zimapangitsa kuti pakhale kuwonjezeka kwakukulu kwa kusagwira ntchito bwino.Makamaka, kuchuluka kwa mpweya kumawonjezeka ndi gawo la zinayi ndipo mitundu yowonjezereka ya okosijeni inakula.Mlingo wochotsa tizilombo tating'onoting'ono udali wodalira pang'ono ndipo umachokera ku 1.0 mpaka 3.8 log pambuyo pa kuwonekera kwa mphindi 45 ku mlingo wa aerosol wa 40,000 #/cc EWNS.
Kuipitsidwa ndi tizilombo tating'onoting'ono ndizomwe zimayambitsa matenda obwera chifukwa cha zakudya zomwe zimayambitsidwa ndi kuyamwa kwa tizilombo toyambitsa matenda kapena poizoni wawo.Ku United States kokha, matenda obwera chifukwa cha zakudya amadwalitsa anthu pafupifupi 76 miliyoni, ogonekedwa m’chipatala 325,000, ndipo anthu 5,000 amafa chaka chilichonse.Kuphatikiza apo, dipatimenti ya zaulimi ku United States (USDA) ikuyerekeza kuti 48% ya matenda onse obwera chifukwa cha chakudya ku United States2.Mtengo wa matenda ndi imfa zomwe zimayambitsidwa ndi tizilombo toyambitsa matenda ku United States ndizokwera kwambiri, zomwe zikuyerekezeredwa ndi Centers for Disease Control and Prevention (CDC) zoposa US $ 15.6 biliyoni pachaka3.
Pakali pano, njira zophatikizirapo mankhwala opha tizilombo 4, radiation5 ndi thermal6 pofuna kuwonetsetsa kuti chitetezo cha chakudya chikuchitika pamlingo wocheperako (CCPs) motsatira njira yopangira (nthawi zambiri pambuyo pokolola komanso/kapena pakupakira) osati mosalekeza.motero, amatha kuipitsidwa.7. Kuwongolera bwino kwa matenda obwera chifukwa cha chakudya komanso kuwonongeka kwa chakudya kumafuna njira zothana ndi tizilombo toyambitsa matenda zomwe zitha kugwiritsidwa ntchito popitilira famu ndikuchepetsa kuwononga chilengedwe komanso ndalama.
Posachedwapa, pulogalamu ya antimicrobial yopanda mankhwala, ya nanotechnology yapangidwa yomwe imatha kusokoneza mabakiteriya apansi ndi mpweya pogwiritsa ntchito madzi opangira madzi (EWNS).EWNS idapangidwa pogwiritsa ntchito njira ziwiri zofanana, electrospray ndi ionization yamadzi (mkuyu 1a).Kafukufuku wam'mbuyomu awonetsa kuti EWNS ili ndi zida zapadera zakuthupi ndi zachilengedwe8,9,10.EWNS ndi avareji 10 ma elekitironi pa dongosolo ndi pafupifupi nanoscale kukula 25 nm (Mkuyu. 1b, c)8,9,10.Kuonjezera apo, electron spin resonance (ESR) inasonyeza kuti EWNS ili ndi mitundu yambiri ya okosijeni (ROS), makamaka hydroxyl (OH•) ndi superoxide (O2-) radicals (Mkuyu 1c)8.EVNS ili mumlengalenga kwa nthawi yayitali ndipo imatha kugundana ndi tizilombo toyambitsa matenda tomwe timayimitsidwa mlengalenga ndikukhalapo pamtunda, kupereka malipiro awo a ROS ndikupangitsa kuti tizilombo toyambitsa matenda tizilombo toyambitsa matenda (mkuyu 1d).Maphunziro oyambirirawa adawonetsanso kuti EWNS imatha kuyanjana ndi kuyambitsa mabakiteriya osiyanasiyana a gram-negative ndi gram-positive, kuphatikizapo mycobacteria, pamtunda ndi mlengalenga.Ma electron microscopy adawonetsa kuti kusakhazikikako kudachitika chifukwa cha kusokonezeka kwa nembanemba ya cell.Kuonjezera apo, kafukufuku wovuta kwambiri wa inhalation wasonyeza kuti mlingo waukulu wa EWNS suyambitsa kuwonongeka kwa mapapo kapena kutupa 8 .
(a) Electrospray imachitika pamene mphamvu yamagetsi ikugwiritsidwa ntchito pakati pa chubu la capillary lomwe lili ndi madzi ndi electrode.(b) Kugwiritsa ntchito kuthamanga kwakukulu kumabweretsa zochitika ziwiri zosiyana: (i) electrospraying ya madzi ndi (ii) kupanga mitundu yowonjezereka ya okosijeni (iyoni) yomwe ili mu EWNS.(c) Mapangidwe apadera a EWNS.(d) Chifukwa cha mawonekedwe awo a nanoscale, EWNS ndi yothamanga kwambiri ndipo imatha kuyanjana ndi tizilombo toyambitsa matenda.
Kuthekera kwa nsanja ya EWNS antimicrobial kuletsa tizilombo toyambitsa matenda pazakudya zatsopano kwawonetsedwanso posachedwa.Zawonetsedwanso kuti mtengo wapamtunda wa EWNS kuphatikiza ndi gawo lamagetsi ukhoza kugwiritsidwa ntchito kuti akwaniritse zomwe akufuna.Komanso, zotsatira zoyambirira za tomato wachilengedwe pambuyo powonekera kwa mphindi 90 pa EWNS pafupifupi 50,000 #/cm3 zinali zolimbikitsa, ndi tizilombo toyambitsa matenda osiyanasiyana monga E. coli ndi Listeria 11.Kuphatikiza apo, mayeso oyambilira a organoleptic sanawonetse zotsatira zomveka poyerekeza ndi kuwongolera tomato.Ngakhale zotsatira zoyambilira zoyambitsa izi ndi zolimbikitsa pakugwiritsa ntchito chitetezo chazakudya ngakhale pamiyeso yotsika kwambiri ya EWNS ya 50,000 #/cc.onani, zikuwonekeratu kuti kuthekera kwakukulu koyambitsa kutha kukhala kopindulitsa kwambiri kuti muchepetse chiopsezo cha matenda ndi kuwonongeka.
Pano, tiyang'ana kafukufuku wathu pakupanga nsanja ya EWNS kuti athe kukonza bwino magawo ophatikizika ndi kukhathamiritsa kwa physicochemical properties a EWNS kuti apititse patsogolo mphamvu zawo za antibacterial.Makamaka, kukhathamiritsa kwayang'ana pa kukulitsa mtengo wawo wapamtunda (kuti apititse patsogolo kasamalidwe kamene akuwongolera) ndi zomwe zili mu ROS (kupititsa patsogolo kusagwira ntchito bwino).Khazikitsani ma physico-chemical properties (kukula, mtengo ndi ROS) pogwiritsa ntchito njira zamakono zowunikira ndikugwiritsa ntchito tizilombo toyambitsa matenda monga E. .
EVNS inapangidwa ndi electrospraying panthawi imodzi ndi ionization ya madzi oyera kwambiri (18 MΩ cm-1).Magetsi nebulizer 12 amagwiritsidwa ntchito kwa atomization zakumwa ndi kaphatikizidwe wa polima ndi ceramic particles 13 ndi ulusi 14 wa kukula ankalamulira.
Monga momwe zafotokozedwera m'mabuku am'mbuyomu 8, 9, 10, 11, pakuyesa wamba, mphamvu yamagetsi yayikulu idayikidwa pakati pa capillary yachitsulo ndi electrode yotsatsira.Panthawiyi, zochitika ziwiri zosiyana zimachitika: i) electrospray ndi ii) ionization yamadzi.Munda wamagetsi wamphamvu pakati pa maelekitirodi awiriwa umapangitsa kuti mitengo yoyipa ipangike pamwamba pamadzi opindika, zomwe zimapangitsa kuti ma cone a Taylor apangidwe.Zotsatira zake, madontho amadzi odzaza kwambiri amapangidwa, omwe amapitirizabe kusweka kukhala tinthu tating'onoting'ono, monga momwe Rayleigh theory16.Nthawi yomweyo, minda yamagetsi yamphamvu imapangitsa kuti mamolekyu ena amadzi agawike ndikuchotsa ma electron (ionize), zomwe zimapangitsa kuti pakhale mitundu yambiri ya okosijeni (ROS)17.Imodzi kwaiye ROS18 anali encapsulated mu EWNS (mkuyu 1c).
Pa mkuyu.2a ikuwonetsa dongosolo la m'badwo wa EWNS lopangidwa ndikugwiritsidwa ntchito mu kaphatikizidwe ka EWNS mu kafukufukuyu.Madzi oyeretsedwa osungidwa mu botolo lotsekedwa amadyetsedwa kudzera mu chubu cha Teflon (2 mm mkati mwake) mu singano ya 30G yachitsulo chosapanga dzimbiri (chitsulo capillary).Kuthamanga kwa madzi kumayendetsedwa ndi mphamvu ya mpweya mkati mwa botolo, monga momwe tawonetsera pa Chithunzi 2b.Singano imayikidwa pa cholembera cha Teflon ndipo imatha kusinthidwa pamanja mtunda wina kuchokera pamagetsi amagetsi.The counter electrode ndi opukutidwa aluminiyamu litayamba ndi dzenje pakati pa zitsanzo.Pansi pa kauntala elekitirodi ndi aluminiyamu sampling funnel, amene olumikizidwa kwa ena khwekhwe kuyesera kudzera doko chitsanzo (mkuyu. 2b).Pofuna kupewa kuchuluka kwa ndalama zomwe zingasokoneze ntchito ya sampler, zigawo zonse za sampler zimakhazikika pamagetsi.
(a) Engineered Water Nanostructure Generation System (EWNS).(b) Gawo lodutsa la sampler ndi electrospray, kusonyeza magawo ofunika kwambiri.(c) Kukonzekera koyeserera koyambitsa mabakiteriya.
Dongosolo la EWNS lomwe lafotokozedwa pamwambapa limatha kusintha magawo ogwiritsira ntchito kuti athandizire kukonza bwino kwa katundu wa EWNS.Sinthani mphamvu yogwiritsira ntchito (V), mtunda wa pakati pa singano ndi electrode ya counter (L), ndi kutuluka kwa madzi (φ) kupyolera mu capillary kuti mukonze bwino makhalidwe a EWNS.Chizindikiro chomwe chimagwiritsidwa ntchito kuyimira mitundu yosiyanasiyana: [V (kV), L (cm)].Sinthani kayendedwe ka madzi kuti mupeze khola lokhazikika la Taylor la seti inayake [V, L].Pazifukwa za kafukufukuyu, kabowo kakang'ono ka kauntala elekitirodi (D) kanasungidwa mainchesi 0.5 (1.29 cm).
Chifukwa cha zochepa za geometry ndi asymmetry, mphamvu yamagetsi yamagetsi sangathe kuwerengedwa kuchokera ku mfundo zoyambirira.M'malo mwake, pulogalamu ya QuickField™ (Svendborg, Denmark)19 idagwiritsidwa ntchito kuwerengera gawo lamagetsi.Munda wamagetsi siwofanana, kotero kuti mtengo wamagetsi pamphepete mwa capillary unagwiritsidwa ntchito ngati chidziwitso cha masanjidwe osiyanasiyana.
Pa kafukufukuyu, mitundu ingapo ya ma voltage ndi mtunda pakati pa singano ndi ma electrode owerengera adawunikidwa malinga ndi mapangidwe a Taylor cone, Taylor cone kukhazikika, kukhazikika kwa kupanga kwa EWNS, komanso kuberekana.Kuphatikizika kosiyanasiyana kumawonetsedwa mu Supplementary Table S1.
Kutulutsa kwamtundu wa EWNS kunalumikizidwa mwachindunji ndi Scanning Mobility Particle Size Analyzer (SMPS, Model 3936, TSI, Shoreview, MN) poyezera kuchuluka kwa chiwerengero cha tinthu, komanso Aerosol Faraday Electrometer (TSI, Model 3068B, Shoreview, MN).) kwa mafunde a aerosol adayesedwa monga momwe tafotokozera m'buku lathu lapitalo.Zonse za SMPS ndi electrometer ya aerosol zomwe zimatengedwa pamtunda wa 0.5 L/mphindi (chitsanzo chonse chotuluka 1 L/mphindi).Kuchuluka kwa tinthu tating'onoting'ono komanso kutuluka kwa aerosol kunayesedwa kwa masekondi 120.Muyeso umabwerezedwa 30 nthawi.Kutengera muyeso wapano, mtengo wa aerosol wonse umawerengedwa ndipo pafupifupi mtengo wa EWNS umayerekezedwa pa chiwerengero choperekedwa cha tinthu tating'ono ta EWNS.Mtengo wapakati wa EWNS ukhoza kuwerengedwa pogwiritsa ntchito Equation (1):
kumene IEl ndi yomwe imayezedwa panopa, NSMPS ndi chiwerengero cha digito choyesedwa ndi SMPS, ndipo φEl ndi mlingo wothamanga pa electrometer.
Chifukwa chinyezi chachibale (RH) chimakhudza mtengo wa pamwamba, kutentha ndi (RH) zinkasungidwa nthawi zonse panthawi yoyesera pa 21 ° C ndi 45%, motsatira.
Atomic force microscopy (AFM), Asylum MFP-3D (Asylum Research, Santa Barbara, CA) ndi AC260T probe (Olympus, Tokyo, Japan) anagwiritsidwa ntchito poyeza kukula ndi moyo wa EWNS.Kusanthula kwafupipafupi kwa AFM kunali 1 Hz, malo ojambulira anali 5 μm × 5 μm, ndi mizere ya scan 256.Zithunzi zonse zidayang'aniridwa ndi 1st makonzedwe azithunzi pogwiritsa ntchito pulogalamu ya Asylum (mask range 100 nm, threshold 100pm).
Kuyesa faniyo anachotsedwa ndipo mica pamwamba anaikidwa pa mtunda wa 2.0 masentimita kuchokera kauntala elekitirodi kwa avereji nthawi 120 s kupewa tinthu agglomeration ndi mapangidwe kusakhazikika m'malovu pa mica pamwamba.EWNS idapopera mwachindunji pamwamba pa mica yodulidwa kumene (Ted Pella, Redding, CA).Chithunzi cha mica pamwamba patangopita AFM sputtering.Kokona yolumikizana ya pamwamba pa mica yosasinthidwa kumene ili pafupi ndi 0 °, kotero EVNS imagawidwa pamtunda wa mica ngati dome.M'mimba mwake (a) ndi kutalika (h) kwa madontho osakanikirana adayezedwa mwachindunji kuchokera ku AFM topography ndipo amagwiritsidwa ntchito kuwerengera kuchuluka kwa EWNS domed diffusion pogwiritsa ntchito njira yathu yomwe idatsimikiziridwa kale.Pongoganiza kuti EWNS yomwe ili m'bwalo ili ndi voliyumu yofanana, kukula kofananako kumatha kuwerengedwa pogwiritsa ntchito Equation (2):
Kutengera njira yathu yomwe idapangidwa kale, electron spin resonance (ESR) spin msampha idagwiritsidwa ntchito kuti azindikire kukhalapo kwapakati kwakanthawi kochepa mu EWNS.Ma aerosols adaphulika kudzera mu 650 μm Midget sparger (Ace Glass, Vineland, NJ) yomwe ili ndi yankho la 235 mM la DEPMPO(5-(diethoxyphosphoryl) -5-methyl-1-pyrroline-N-oxide) (Oxis International Inc.).Portland, Oregon).Miyezo yonse ya ESR inkagwiritsidwa ntchito pogwiritsa ntchito Bruker EMX spectrometer (Bruker Instruments Inc. Billerica, MA, USA) ndi cell panel flat.Pulogalamu ya Acquisit (Bruker Instruments Inc. Billerica, MA, USA) inagwiritsidwa ntchito kusonkhanitsa ndi kusanthula deta.Kutsimikiza kwa mawonekedwe a ROS kunkachitika kokha pazigawo zogwirira ntchito [-6.5 kV, 4.0 cm].Kukhazikika kwa EWNS kunayesedwa pogwiritsa ntchito SMPS pambuyo powerengera zotayika za EWNS mu impactor.
Miyezo ya ozoni idawunikidwa pogwiritsa ntchito 205 Dual Beam Ozone Monitor™ (2B Technologies, Boulder, Co)8,9,10.
Pazinthu zonse za EWNS, mtengo wamtengo wapatali umagwiritsidwa ntchito ngati muyeso woyezera, ndipo kupatuka kokhazikika kumagwiritsidwa ntchito ngati cholakwika cha kuyeza.Mayeso a T adachitidwa kuti afanizire mayendedwe a EWNS okongoletsedwa ndi ma EWNS oyambira.
Chithunzi 2c chikuwonetsa makina opangidwa kale komanso odziwika bwino a electrostatic precipitation (EPES) "chikoka" chomwe chingagwiritsidwe ntchito popereka EWNS pamtunda.EPES imagwiritsa ntchito ndalama za EVNS zomwe zingathe "kutsogoleredwa" molunjika pamwamba pa chandamale mothandizidwa ndi mphamvu yamagetsi yamphamvu.Tsatanetsatane wa dongosolo la EPES laperekedwa m'mabuku aposachedwa a Pyrgiotakis et al.11 .Chifukwa chake, EPES imakhala ndi chipinda cha PVC chosindikizidwa cha 3D chokhala ndi malekezero opindika ndipo chimakhala ndi zitsulo ziwiri zofanana (304 zitsulo zosapanga dzimbiri, zopaka galasi) mbale zachitsulo pakati pa 15.24 cm motalikirana.matabwa anali olumikizidwa kwa kunja mkulu voteji gwero (Bertran 205B-10R, Spellman, Hauppauge, NY), pansi mbale nthawi zonse olumikizidwa kwa voteji zabwino, ndi mbale pamwamba nthawi zonse kugwirizana pansi (yoyandama pansi).Makoma a chipindacho amaphimbidwa ndi zojambulazo za aluminiyamu, zomwe zimakhala ndi magetsi kuti zisawonongeke.Chipindacho chili ndi chitseko chotsekera chakutsogolo chomwe chimalola kuti malo oyeserera ayikidwe pamapulasitiki omwe amawakweza pamwamba pa chitsulo chapansi kuti apewe kusokoneza kwamagetsi.
Kugwiritsa ntchito bwino kwa EWNS mu EPES kunawerengedwa molingana ndi protocol yomwe idapangidwa kale yofotokozedwa mu Supplementary Figure S111.
Monga chipinda chowongolera, chipinda chachiwiri cha cylindrical flow flow chinalumikizidwa motsatizana ndi dongosolo la EPES, momwe fyuluta yapakatikati ya HEPA idagwiritsidwa ntchito kuchotsa EWNS.Monga momwe chithunzi 2c chikusonyezera, aerosol ya EWNS idapopedwa kudzera m'zipinda ziwiri zomangidwamo.Fyuluta pakati pa chipinda chowongolera ndi EPES imachotsa EWNS iliyonse yotsalira chifukwa cha kutentha komweko (T), chinyezi chapafupi (RH) ndi ma ozone.
Tizilombo toyambitsa matenda tomwe timapezeka m'zakudya tapezeka kuti tikuipitsa zakudya zatsopano monga E. coli (ATCC #27325), fecal indicator, Salmonella enterica (ATCC #53647), tizilombo toyambitsa matenda, Listeria wopanda vuto (ATCC #33090), surrogate for pathogenic Listeria monocytogenes,Mavisiamytogenes,Mavisiamytogenes,Mavisiamy, ATCC (ATCC #4098), choloweza mmalo mwa yisiti yowonongeka, ndi bakiteriya wosamva wosamva, Mycobacterium paralucky (ATCC #19686).
Gulani mabokosi achisawawa a tomato wamphesa kuchokera kumsika kwanu ndikusunga mufiriji pa 4°C mpaka mugwiritse ntchito (mpaka masiku atatu).Tomato woyesera anali onse ofanana kukula, pafupifupi 1/2 inchi m'mimba mwake.
Chikhalidwe, katemera, kuwonekera, ndi kuwerengera kwa koloni zafotokozedwa mwatsatanetsatane m'buku lathu lapitalo ndikufotokozedwa mwatsatanetsatane mu Supplementary Data.Kuchita bwino kwa EWNS kudawunikidwa povumbulutsa tomato wothiridwa ku 40,000 #/cm3 kwa mphindi 45.Mwachidule, tomato atatu adagwiritsidwa ntchito kuyesa tizilombo tomwe tikukhalamo panthawi t = 0 min.Tomato atatu adayikidwa mu EPES ndipo adawonekera kwa EWNS pa 40,000 # / cc (EWNS poyera tomato) ndipo atatu otsalawo adayikidwa mu chipinda chowongolera (tomato control).Zina processing wa tomato m'magulu onsewa sizinachitike.EWNS-poyera tomato ndi kulamulira tomato anachotsedwa patatha mphindi 45 kuti aone zotsatira za EWNS.
Kuyesera kulikonse kunkachitika katatu.Kusanthula kwa data kunachitika molingana ndi protocol yofotokozedwa mu Supplementary Data.
Njira zosagwiritsa ntchito zidayesedwa ndi kusungunuka kwa zitsanzo za EWNS (45 min pa 40,000 #/cm3 EWNS aerosol concentration) ndi zitsanzo zopanda ma radiation za mabakiteriya opanda vuto E. coli, Salmonella enterica ndi Lactobacillus.Tinthu tating'onoting'ono ta 2.5% glutaraldehyde, 1.25% paraformaldehyde ndi 0.03% picric acid mu 0.1 M sodium cacodylate buffer (pH 7.4) kwa maola awiri kutentha.Pambuyo kutsuka, positi kukonza ndi 1% osmium tetroxide (OsO4)/1.5% potaziyamu ferrocyanide (KFeCN6) kwa 2 hours, kusamba katatu m'madzi ndi incubate mu 1% uranyl acetate kwa ola 1, ndiye kusamba kawiri m'madzi, ndiye kuchepetsa madzi kwa mphindi 10, 0% 0% 50% mowa.Zitsanzozo zinayikidwa mu propylene oxide kwa ola la 1 ndikuyika ndi 1: 1 osakaniza a propylene oxide ndi TAAP Epon (Marivac Canada Inc. St. Laurent, CA).Zitsanzozo zidaphatikizidwa mu TAAB Epon ndikupangidwa ndi polymer pa 60 ° C kwa maola 48.Utoto wa granular wochiritsidwa unadulidwa ndikuwonetseredwa ndi TEM pogwiritsa ntchito maikulosikopu yamagetsi yamagetsi ya JEOL 1200EX (JEOL, Tokyo, Japan) yokhala ndi kamera ya AMT 2k CCD (Advanced Microscopy Techniques, Corp., Woburn, Massachusetts, USA).
Zoyesera zonse zidachitika mwapatatu.Pa nthawi iliyonse, mabakiteriya amatsuka katatu, zomwe zimapangitsa kuti pakhale mfundo zisanu ndi zinayi pa mfundo iliyonse, pafupifupi yomwe imagwiritsidwa ntchito ngati mabakiteriya a tizilombo toyambitsa matenda.Kupatuka kokhazikika kunagwiritsidwa ntchito ngati cholakwika choyezera.Mfundo zonse zimawerengera.
Logarithm ya kuchepa kwa kuchuluka kwa mabakiteriya poyerekeza ndi t = 0 min idawerengedwa pogwiritsa ntchito njira iyi:
kumene C0 ndi kuchuluka kwa mabakiteriya mu chitsanzo chowongolera pa nthawi 0 (ie pambuyo pouma pamwamba koma asanaikidwe m'chipinda) ndipo Cn ndi kuchuluka kwa mabakiteriya pamtunda pambuyo pa n maminiti akuwonekera.
Kuwerengera kuwonongeka kwachilengedwe kwa mabakiteriya pakuwonekera kwa mphindi 45, kutsika kwa chipika poyerekeza ndi kuwongolera pambuyo pa mphindi 45 kudawerengedwanso motere:
kumene Cn ndi kuchuluka kwa mabakiteriya mu chitsanzo chowongolera pa nthawi n ndi Cn-Control ndi kuchuluka kwa mabakiteriya olamulira panthawi n.Deta imaperekedwa ngati kuchepetsedwa kwa chipika poyerekeza ndi kuwongolera (palibe kuwonekera kwa EWNS).
Pa kafukufukuyu, mitundu ingapo ya ma voltage ndi mtunda pakati pa singano ndi ma electrode owerengera adawunikidwa malinga ndi mapangidwe a Taylor cone, Taylor cone kukhazikika, kukhazikika kwa kupanga kwa EWNS, komanso kuberekana.Kuphatikizika kosiyanasiyana kumawonetsedwa mu Supplementary Table S1.Milandu iwiri yowonetsa zinthu zokhazikika komanso zopangika (Taylor cone, EWNS generation, ndi kukhazikika pakapita nthawi) adasankhidwa kuti aphunzire mokwanira.Pa mkuyu.Chithunzi 3 chikuwonetsa zotsatira za mtengo, kukula, ndi zomwe zili mu ROS muzochitika zonsezi.Zotsatira zikuwonetsedwanso mu Table 1. Kufotokozera, zonse Chithunzi 3 ndi Table 1 zikuphatikizapo katundu wa EWNS8, 9, 10, 11 (zoyambira-EWNS) zomwe zinapangidwa kale.Kuwerengera kufunikira kowerengera pogwiritsa ntchito kuyesa kwa michira iwiri kumasindikizidwanso mu Supplementary Table S2.Kuonjezera apo, deta yowonjezera imaphatikizapo maphunziro a zotsatira za kauntala electrode sampling hole diameter (D) ndi mtunda pakati pa electrode pansi ndi nsonga (L) (Supplementary Figures S2 ndi S3).
(ac) Kugawa kwakukula kuyeza ndi AFM.(df) Mawonekedwe a pamwamba.(g) Makhalidwe a ROS a EPR.
Ndikofunikiranso kuzindikira kuti pazikhalidwe zonse zomwe zili pamwambazi, ionization panopa yoyezedwa inali pakati pa 2 ndi 6 μA ndi voteji pakati pa -3.8 ndi -6.5 kV, zomwe zinapangitsa kuti mphamvu yogwiritsira ntchito mphamvu yosachepera 50 mW pa gawo limodzi la EWNS generation contact module.Ngakhale kuti EWNS idapangidwa mopanikizika kwambiri, milingo ya ozone inali yotsika kwambiri, yosapitilira 60 ppb.
Supplementary Figure S4 ikuwonetsa magawo amagetsi ofananirako a [-6.5 kV, 4.0 cm] ndi [-3.8 kV, 0.5 cm] motsatana.Pazithunzi za [-6.5 kV, 4.0 cm] ndi [-3.8 kV, 0.5 cm], kuwerengera kwamunda ndi 2 × 105 V/m ndi 4.7 × 105 V/m, motero.Izi zikuyembekezeka, chifukwa chachiwiri chiŵerengero cha ma voltage-distance ndi apamwamba kwambiri.
Pa mkuyu.3a,b ikuwonetsa kukula kwa EWNS kuyesedwa ndi AFM8.Ma diameter a EWNS omwe anawerengedwa anali 27 nm ndi 19 nm pa [-6.5 kV, 4.0 cm] ndi [-3.8 kV, 0.5 cm], motero.Pazithunzi za [-6.5 kV, 4.0 cm] ndi [-3.8 kV, 0.5 cm], zopatuka za geometric za magawowa ndi 1.41 ndi 1.45, motsatana, zomwe zikuwonetsa kugawa kocheperako.Kukula kwapakati komanso kusiyanasiyana kwa geometric ndizoyandikira kwambiri EWNS yoyambira, pa 25 nm ndi 1.41, motsatana.Pa mkuyu.3c ikuwonetsa kukula kwa magawo a EWNS omwe amayezedwa pogwiritsa ntchito njira yomweyo pansi pamikhalidwe yomweyi.
Pa mkuyu.3d, e ikuwonetsa zotsatira za mawonekedwe amalipiro.Deta ndi miyeso yapakati ya 30 miyeso yokhazikika nthawi imodzi (#/cm3) ndi yapano (I).Kusanthula kukuwonetsa kuti mtengo wapakati pa EWNS ndi 22 ± 6 e- ndi 44 ± 6 e- kwa [-6.5 kV, 4.0 cm] ndi [-3.8 kV, 0.5 cm], motero.Amakhala ndi zolipiritsa zapamwamba kwambiri poyerekeza ndi EWNS yoyambira (10 ± 2 e-), kuwirikiza kawiri kuposa [-6.5 kV, 4.0 cm] ndi kuwirikiza kanayi kuposa [-3 .8 kV, 0.5 cm].Chithunzi 3f chikuwonetsa mtengo.data ya Baseline-EWNS.
Kuchokera pamapu owerengera a nambala ya EWNS (Supplementary Figures S5 ndi S6), zitha kuwoneka kuti [-6.5 kV, 4.0 cm] ili ndi tinthu tambiri kuposa [-3.8 kV, 0.5 cm].Ndizofunikanso kudziwa kuti chiwerengero cha chiwerengero cha EWNS chinayang'aniridwa mpaka maola a 4 (Supplementary Figures S5 ndi S6), kumene kukhazikika kwa m'badwo wa EWNS kunasonyeza milingo yofanana ya chiwerengero cha tinthu pazochitika zonsezi.
Pa mkuyu.3g imawonetsa mawonekedwe a EPR pambuyo pochotsa zowongolera za EWNS (kumbuyo) pa [-6.5 kV, 4.0 cm].Mawonekedwe a ROS adafanizidwanso ndi zochitika za Baseline-EWNS mu ntchito yomwe idasindikizidwa kale.Chiwerengero cha EWNS chochita ndi misampha yozungulira chinawerengedwa kukhala 7.5 × 104 EWNS / s, yomwe ili yofanana ndi yomwe idasindikizidwa kale Baseline-EWNS8.Mawonekedwe a EPR adawonetsa bwino kukhalapo kwa mitundu iwiri ya ROS, pomwe O2- ndiyomwe ndiyomwe ili yamitundu yambiri ndi OH• kukhala yocheperako.Kuphatikiza apo, kuyerekeza kwachindunji kwamphamvu kwambiri kunawonetsa kuti EWNS yokongoletsedwa inali ndi ROS yapamwamba kwambiri poyerekeza ndi EWNS yoyambira.
Pa mkuyu.4 ikuwonetsa magwiridwe antchito a EWNS mu EPES.Zomwe zimafotokozedwanso mwachidule mu Table I ndikufaniziridwa ndi deta yoyambirira ya EWNS.Pazochitika zonse ziwiri za EUNS, kuyikako kuli pafupi ndi 100% ngakhale pamagetsi otsika a 3.0 kV.Nthawi zambiri, 3.0 kV ndi yokwanira 100% kuyika, mosasamala kanthu za kusintha kwa mtengo wapamtunda.Pansi pamikhalidwe yomweyi, magwiridwe antchito a Baseline-EWNS anali 56% okha chifukwa cha mtengo wawo wotsika (pafupifupi ma electron 10 pa EWNS).
Pa mkuyu.5 ndi mu table.2 mwachidule inactivation mtengo wa tizilombo inoculated padziko tomato pambuyo kukhudzana ndi za 40,000 #/cm3 EWNS kwa mphindi 45 pa akadakwanitsira akafuna [-6,5 kV, 4.0 cm].Innoculated E. coli ndi Lactobacillus innocuous anasonyeza kuchepa kwakukulu kwa zipika za 3.8 panthawi ya 45 mphindi.Pazifukwa zomwezo, S. enterica inali ndi kuchepa kwa 2.2-log, pamene S. cerevisiae ndi M. parafortutum anali ndi kuchepa kwa 1.0-log.
Ma electron micrographs (Chithunzi 6) akuwonetsa kusintha kwa thupi komwe kunayambitsidwa ndi EWNS pa Escherichia coli, Streptococcus, ndi maselo a Lactobacillus omwe amachititsa kuti asagwiritsidwe ntchito.Mabakiteriya owongolera anali ndi nembanemba zama cell, pomwe mabakiteriya owonekera adawononga nembanemba zakunja.
Kujambula kwa electron microscopic kulamulira ndi mabakiteriya owonekera kunavumbula kuwonongeka kwa nembanemba.
Deta ya physicochemical properties of optimized EWNS pamodzi imasonyeza kuti katundu (pamwamba pa pamwamba ndi ROS content) za EWNS zinali zabwino kwambiri poyerekeza ndi deta yoyambirira ya EWNS yofalitsidwa kale8,9,10,11.Kumbali ina, kukula kwawo kunakhalabe mumtundu wa nanometer, mofanana kwambiri ndi zotsatira zomwe zanenedwa kale, zomwe zimawalola kukhala mlengalenga kwa nthawi yaitali.Kuwoneka kwa polydispersity kumatha kufotokozedwa ndi kusintha kwa mtengo wapamtunda komwe kumatsimikizira kukula kwa EWNS, kusasinthika kwa Rayleigh effect, ndi kuthekera kolumikizana.Komabe, monga momwe Nielsen et al.22, mtengo wapamwamba wapamtunda umachepetsa kutuluka kwa nthunzi powonjezera mphamvu yamadzi / kupsinjika kwamadzi.M'buku lathu lapitalo8 chiphunzitsochi chinatsimikiziridwa moyesera kwa ma microdroplets 22 ndi EWNS.Kutayika kwa ndalama panthawi yowonjezereka kungakhudzenso kukula ndikuthandizira kugawidwa kwa kukula komwe kumawonedwa.