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He carousel e hōʻike ana i ʻekolu paheʻe i ka manawa like.E hoʻohana i nā pihi Mua a me Next no ka neʻe ʻana i ʻekolu paheʻe i ka manawa, a i ʻole e hoʻohana i nā pihi paheʻe ma ka hopena e neʻe i ʻekolu mau paheʻe i ka manawa.
I kēia mau lā, ua hoʻomohala ʻia kahi kahua antimicrobial ʻole kemika e pili ana i ka nanotechnology me ka hoʻohana ʻana i nā nanostructures wai artificial (EWNS).He kiʻekiʻe ka uku o ka EWNS a ua hoʻopiha ʻia me nā ʻano oxygen reactive (ROS) hiki ke launa pū me ka hoʻopau ʻana i kekahi mau microorganisms, me nā pathogens lawe ʻai i ka meaʻai.Eia ke hōʻike ʻia nei ko lākou mau waiwai i ka wā synthesis hiki ke hoʻoponopono maikaʻi ʻia a hoʻopaʻa ʻia e hoʻomaikaʻi hou i ko lākou hiki antibacterial.Ua hoʻolālā ʻia ke kahua hoʻokolohua EWNS e hoʻoponopono maikaʻi i nā waiwai o EWNS ma o ka hoʻololi ʻana i nā ʻāpana synthesis.ʻO ke ʻano o nā waiwai EWNS (ka uku, ka nui a me ka ʻike o ROS) me ka hoʻohana ʻana i nā ʻano loiloi hou.Eia kekahi, ua loiloi ʻia lākou no kā lākou microbial inactivation hiki ke kūʻē i ka meaʻai microorganism e like me Escherichia coli, Salmonella enterica, Listeria innocuous, Mycobacterium paraaccidentum a me Saccharomyces cerevisiae.Hōʻike nā hopena i hōʻike ʻia ma ʻaneʻi e hiki ke hoʻoponopono maikaʻi ʻia nā waiwai o EWNS i ka wā synthesis, e hopena i ka hoʻonui exponential o ka pono inactivation.ʻO ka mea nui, ua hoʻonui ʻia ka uku o ka ʻili ma kahi helu ʻehā a ua hoʻonui ʻia nā ʻano oxygen reactive.Ua hilinaʻi ʻia ka nui o ka lawe ʻana i ka microbial a mai ka 1.0 a hiki i ka 3.8 log ma hope o ka hoʻolaha ʻana o 45 mau minuke i kahi ʻano aerosol o 40,000 #/cc EWNS.
ʻO ka hoʻohaumia ʻana i ka microbial ke kumu nui o ka maʻi i lawe ʻia e ka meaʻai ma muli o ka ʻai ʻana i nā pathogens a i ʻole kā lākou toxins.Ma ʻAmelika wale nō, ʻo ka maʻi i lawe ʻia i ka meaʻai ke kumu ma kahi o 76 miliona mau maʻi, 325,000 i komo i ka haukapila, a me 5,000 mau make i kēlā me kēia makahiki1.Eia hou, ke manaʻo nei ke Keʻena ʻOihana Mahiai o ʻAmelika Hui Pū ʻIa (USDA) ʻo ka hoʻonui ʻana i ka ʻai ʻana i nā hua hou ke kuleana no 48% o nā maʻi lawe ʻai a pau i hōʻike ʻia ma United States2.He kiʻekiʻe loa ke kumukūʻai o ka maʻi a me ka make e nā pathogens lawe ʻia i ka meaʻai ma ʻAmelika Hui Pū ʻIa, i manaʻo ʻia e nā Centers for Disease Control and Prevention (CDC) ma mua o US $15.6 biliona i kēlā me kēia makahiki3.
I kēia manawa, hoʻokō ʻia nā hana antimicrobial chemical4, radiation5 a me thermal6 e hōʻoia i ka palekana o ka meaʻai ma nā wahi mana koʻikoʻi liʻiliʻi (CCP) ma ke kaulahao hana (maʻamau ma hope o ka hōʻiliʻili ʻana a/a i ka wā o ka hōʻiliʻili ʻana) ma mua o ka hoʻomau.no laila, ua maʻalahi lākou i ka hoʻohaumia ʻana.7. Pono ka ho'omalu maika'i 'ana i ka ma'i a me ka pō'ino o ka mea'ai i nā hana antimicrobial i hiki ke ho'ohana 'ia ma ka ho'omau o ka mahi'ai-a-papa me ka hō'emi 'ana i ka hopena o ke kaiapuni a me nā koina.
I kēia mau lā, ua hoʻomohala ʻia kahi kahua antimicrobial kumu ʻole kemika, nanotechnology e hiki ke hoʻopau i ka ʻili a me ka ea e hoʻohana ana i nā nanostructures wai artificial (EWNS).Ua hoʻohui ʻia ʻo EWNS me ka hoʻohana ʻana i ʻelua kaʻina hana like, electrospray a me ka ionization wai (Fig. 1a).Ua hōʻike ʻia nā haʻawina mua e loaʻa i ka EWNS kahi hoʻonohonoho kūʻokoʻa o nā waiwai kino a me ke olaola8,9,10.Loaʻa i ka EWNS ka awelika o 10 electrons no ka hale a me ka nui nanoscale awelika o 25 nm (Fig. 1b, c) 8,9,10.Eia hou, ua hōʻike ka electron spin resonance (ESR) i ka EWNS he nui ka nui o nā ʻano oxygen reactive (ROS), ka nui o nā radical hydroxyl (OH•) a me superoxide (O2-) (Fig. 1c)8.Aia ʻo EVNS i ka lewa no ka manawa lōʻihi a hiki ke hui pū me nā microorganism i hoʻokuʻu ʻia i ka lewa a hōʻike ʻia ma ka ʻili, e hāʻawi ana i kā lākou uku ROS a me ka hoʻoulu ʻana i nā microorganisms (Fig. 1d).Ua hōʻike pū ʻia kēia mau haʻawina mua e hiki i ka EWNS ke launa pū me ka hoʻopau ʻana i nā ʻano bacteria gram-negative a me ka gram-positive, me ka mycobacteria, ma ka ʻili a me ka lewa.Ua hōʻike ʻia ka microscopy electron transmission i ka hoʻopau ʻia ʻana o ka membrane cell.Eia kekahi, ua hōʻike ʻia nā haʻawina inhalation acute ʻaʻole hiki i ka nui o ka EWNS ke hōʻeha i ka ʻūmā a i ʻole ka mumū 8.
(a) Hana ʻia ka Electrospray ke hoʻohana ʻia kahi uila kiʻekiʻe ma waena o kahi paipu capillary i loko o ka wai a me kahi electrode counter.(b) ʻO ka hoʻohana ʻana i ke kaomi kiʻekiʻe ka hopena i ʻelua ʻano ʻano like ʻole: (i) electrospraying o ka wai a (ii) hoʻokumu ʻana i nā ʻano oxygen reactive (ions) i paʻa i ka EWNS.(c) ʻO ke ʻano kū hoʻokahi o EWNS.(d) Ma muli o ko lākou ʻano nanoscale, paʻa loa ʻo EWNS a hiki ke launa pū me nā pathogens airborne.
Ua hōʻike ʻia ka hiki o ka EWNS antimicrobial platform ke hoʻopau i nā microorganism i lawe ʻia i ka meaʻai ma ka ʻili o ka meaʻai hou.Ua hōʻike ʻia hoʻi e hiki ke hoʻohana ʻia ka uku o ka ʻili o EWNS i hui pū ʻia me kahi kahua uila no ka hoʻokō ʻana i ka hāʻawi ʻana.Eia kekahi, ʻo nā hualoaʻa mua no nā tōmato organik ma hope o ka hōʻike ʻana i nā minuke 90 ma kahi EWNS ma kahi o 50,000 #/cm3 he mea paipai, me nā microorganism lawe ʻai like ʻole e like me E. coli a me Listeria 11 i nānā ʻia.Eia kekahi, ʻaʻole i hōʻike ʻia nā hoʻokolohua organoleptic mua i nā hopena sensory i hoʻohālikelike ʻia me ka mālama ʻana i nā tōmato.ʻOiai ʻo kēia mau hopena inactivation mua e paipai nei no nā noi palekana meaʻai a hiki i nā haʻahaʻa haʻahaʻa EWNS o 50,000#/cc.ʻike, ua maopopo ka ʻoi aku ka maikaʻi o ka hiki ke hoʻopau ʻia e hōʻemi hou i ka pilikia o ka maʻi a me ka pala.
Ma ʻaneʻi, e kālele mākou i kā mākou noiʻi ʻana i ka hoʻomohala ʻana i kahi kahua o ka hanauna EWNS e hiki ai ke hoʻoponopono maikaʻi i nā ʻāpana synthesis a me ka hoʻomaikaʻi ʻana i nā waiwai physicochemical o EWNS e hoʻomaikaʻi i ko lākou hiki ke antibacterial.ʻO ka mea nui, ua kālele ka optimization i ka hoʻonui ʻana i ka uku o ka ʻili (e hoʻomaikaʻi i ka hāʻawi ʻia ʻana) a me ka ʻike ROS (e hoʻomaikaʻi i ka pono inactivation).E hōʻike i nā waiwai physico-chemical i hoʻopaʻa ʻia (ka nui, ka uku a me ka ʻikepili ROS) me ka hoʻohana ʻana i nā ʻano analytical hou a hoʻohana i nā microorganism meaʻai maʻamau e like me E. .
Ua hana ʻia ʻo EVNS e ka electrospraying like a me ka ionization o ka wai maʻemaʻe kiʻekiʻe (18 MΩ cm-1).Hoʻohana maʻamau ka nebulizer uila 12 no ka atomization o nā wai a me ka synthesis o ka polymer a me nā ʻāpana seramika 13 a me nā fibers 14 o ka nui kaohi.
E like me ka mea kiko'ī ma nā paʻi mua 8, 9, 10, 11, ma kahi hoʻokolohua maʻamau, ua hoʻohana ʻia kahi voli kiʻekiʻe ma waena o kahi capillary metala a me kahi electrode countered grounded.I loko o kēia kaʻina hana, ʻelua mau hanana like ʻole: i) electrospray a ii) wai ionization.ʻO ke kahua uila ikaika ma waena o nā electrodes ʻelua ke kumu o ke kūkulu ʻana i nā uku maikaʻi ʻole ma luna o ka ʻili o ka wai i hoʻopaʻa ʻia, ka hopena i ke kūkulu ʻia ʻana o nā cones Taylor.ʻO ka hopena, ua hoʻokumu ʻia nā kulu wai i hoʻopaʻa nui ʻia, e hoʻomau i ka haki ʻana i mau ʻāpana liʻiliʻi, e like me ka manaʻo Rayleigh16.I ka manawa like, hoʻomaka nā māla uila ikaika i kekahi mau molekele wai e māhele a wehe i nā electrons (ionize), kahi e alakaʻi ai i ka hoʻokumu ʻana i ka nui o nā ʻano oxygen reactive (ROS)17.Hoʻopili ʻia ʻo ROS18 i ka manawa like ma EWNS (Fig. 1c).
Ma ka fig.Hōʻike ʻo 2a i ka ʻōnaehana hanauna EWNS i kūkulu ʻia a hoʻohana ʻia i ka synthesis EWNS i kēia haʻawina.Ua hānai ʻia ka wai i hoʻomaʻemaʻe ʻia i mālama ʻia i loko o kahi hue i pani ʻia ma o kahi paipu Teflon (2 mm anawaena o loko) i loko o kahi nila kila kuhili ʻole 30G (metal capillary).Hoʻomalu ʻia ke kahe o ka wai e ke kaomi ea i loko o ka ʻōmole, e like me ka hōʻike ʻana ma ke Kiʻi 2b.Hoʻokomo ʻia ka nila ma kahi console Teflon a hiki ke hoʻoponopono lima ʻia i kahi mamao mai ka electrode counter.ʻO ka counter electrode he pahu alumini i poni ʻia me kahi puka ma waenakonu no ka laʻana.Aia ma lalo o ka electrode counter he funnel sampling aluminika, i hoʻopili ʻia me ke koena o ka hoʻonohonoho hoʻokolohua ma o kahi awa hōʻailona (Fig. 2b).No ka pale ʻana i ka hoʻonui ʻana o ka uku e hiki ke hoʻopau i ka hana sampler, ua hoʻokumu ʻia nā ʻāpana sampler a pau i ka uila.
(a) Pūnaehana Hana Nanostructure Wai ʻenehana (EWNS).(b) Keʻa-ʻāpana o ka sampler a me ka electrospray, e hōʻike ana i nā ʻāpana koʻikoʻi.(c) Hoʻonohonoho hoʻokolohua no ka hoʻopau ʻana i ka bacteria.
Hiki i ka ʻōnaehana hanauna EWNS i hōʻike ʻia ma luna nei ke hoʻololi i nā ʻāpana hana koʻikoʻi e hoʻomaʻamaʻa i ka hoʻoponopono maikaʻi ʻana i nā waiwai EWNS.E hoʻololi i ka volta i hoʻohana ʻia (V), ka mamao ma waena o ka nila a me ka electrode counter (L), a me ke kahe wai (φ) ma o ka capillary e hoʻoponopono i nā ʻano EWNS.Hoʻohana ʻia ka hōʻailona e hōʻike i nā hui like ʻole: [V (kV), L (cm)].E hoʻololi i ke kahe wai e loaʻa i kahi pahu Taylor paʻa o kekahi pūʻulu [V, L].No ke kumu o kēia noiʻi ʻana, mālama ʻia ke anawaena puka o ka electrode counter (D) ma 0.5 iniha (1.29 knm).
Ma muli o ka palena geometry a me ka asymmetry, ʻaʻole hiki ke helu ʻia ka ikaika o ke kahua uila mai nā kumu mua.Akā, ua hoʻohana ʻia ka polokalamu QuickField™ (Svendborg, Denmark)19 no ka helu ʻana i ke kahua uila.ʻAʻole like ke kahua uila, no laila ua hoʻohana ʻia ka waiwai o ke kahua uila ma ka piko o ka capillary ma ke ʻano he kumu kuhikuhi no nā hoʻonohonoho like ʻole.
I ka wā o ke aʻo ʻana, ua loiloi ʻia kekahi mau hui ʻana o ka volta a me ka mamao ma waena o ka nila a me ka electrode counter e pili ana i ka hoʻokumu ʻana o Taylor cone, Taylor cone stability, EWNS production stability, a reproducibility.Hōʻike ʻia nā hui like ʻole ma ka Papa Pākuʻi S1.
Ua hoʻopili pololei ʻia ka mea hoʻopuka o ka ʻōnaehana hanauna EWNS i kahi Scanning Mobility Particle Size Analyzer (SMPS, Model 3936, TSI, Shoreview, MN) no ke ana ʻana i ka helu particle, a me kahi Aerosol Faraday Electrometer (TSI, Model 3068B, Shoreview, MN).) no nā au aerosol ua ana ʻia e like me ka mea i wehewehe ʻia ma kā mākou paʻi mua.Hoʻohālikelike ʻia ka SMPS a me ka electrometer aerosol ma kahi kahe kahe o 0.5 L / min (ka holo ʻana o ka hāpana 1 L / min).Ua ana ʻia ka helu o nā ʻāpana a me ke kahe aerosol no 120 kekona.Hana hou ʻia ke ana ʻana he 30 mau manawa.Ma muli o nā ana o kēia manawa, ua helu ʻia ka huina aerosol a ua manaʻo ʻia ka awelika o ka uku EWNS no ka huina helu o nā ʻāpana EWNS i koho ʻia.Hiki ke helu ʻia ke kumukūʻai maʻamau o EWNS me ka hoʻohana ʻana i ka Equation (1):
ma kahi o IEl ke ana i keia manawa, NSMPS ka helu kikohoʻe i ana me ka SMPS, a φEl ke kahe kahe no ka electrometer.
Ma muli o ka pili ʻana o ka haʻahaʻa haʻahaʻa (RH) i ke kau ʻana o ka ʻili, ua mālama mau ʻia ka mahana a me (RH) i ka wā o ka hoʻokolohua ma 21°C a me 45%, kēlā me kēia.
Ua hoʻohana ʻia ka microscopy Atomic force (AFM), Asylum MFP-3D (Asylum Research, Santa Barbara, CA) a me AC260T probe (Olympus, Tokyo, Iapana) e ana i ka nui a me ke ola o ka EWNS.ʻO ke alapine scanning AFM he 1 Hz, ʻo ka wahi scanning he 5 μm × 5 μm, a me nā laina scan 256.Hoʻopili ʻia nā kiʻi āpau i ka alignment kiʻi kauoha mua me ka hoʻohana ʻana i ka polokalamu Asylum (mask range 100 nm, paepae 100 pm).
Wehe ʻia ka funnel hoʻāʻo a waiho ʻia ka ʻili mika ma kahi mamao o 2.0 knm mai ka electrode counter no ka manawa awelika o 120 s e pale aku i ka hoʻohui ʻana o nā ʻāpana a me ka hoʻokumu ʻana o nā droplets like ʻole ma ka ʻili mika.Ua pulu pololei ʻia ʻo EWNS ma luna o ka ʻili o ka mika i ʻoki hou ʻia (Ted Pella, Redding, CA).ʻO ke kiʻi o ka ʻili mika ma hope koke o ka hoʻōho ʻana o AFM.ʻO ka huina pili o ka ʻili o ka mika i hoʻololi ʻole ʻia e kokoke ana i ka 0 °, no laila ua māhele ʻia ʻo EVNS ma ka ʻili mika ma ke ʻano o ka dome.Ua ana pololei ʻia ke anawaena (a) a me ke kiʻekiʻe (h) o nā diffusing droplets mai ka topography AFM a hoʻohana ʻia no ka helu ʻana i ka volume diffusion domed EWNS me ka hoʻohana ʻana i kā mākou ala i hōʻoia mua ʻia.Ke manaʻo nei he like ka leo o ka EWNS ma luna, hiki ke helu ʻia ke anawaena like me ka hoʻohana ʻana i ka Equation (2):
Ma muli o kā mākou hana i hoʻomohala mua ʻia, ua hoʻohana ʻia kahi pahele spin electron spin resonance (ESR) e ʻike ai i ka hele ʻana o nā intermediates radical pōkole i EWNS.Ua hoʻoheheʻe ʻia nā Aerosol ma o kahi 650 μm Midget sparger (Ace Glass, Vineland, NJ) i loaʻa kahi hopena 235 mM o DEPMPO (5- (diethoxyphosphoryl) -5-methyl-1-pyrroline-N-oxide) (Oxis International Inc.).Portland, Oregon).Hoʻohana ʻia nā ana ESR a pau me ka Bruker EMX spectrometer (Bruker Instruments Inc. Billerica, MA, USA) a me kahi cell panel flat.Ua hoʻohana ʻia ka polokalamu Acquisit (Bruker Instruments Inc. Billerica, MA, USA) no ka hōʻiliʻili ʻana a me ka nānā ʻana i ka ʻikepili.ʻO ka hoʻoholo ʻana i nā hiʻohiʻona o ka ROS i hana wale ʻia no kahi hoʻonohonoho o nā kūlana hana [-6.5 kV, 4.0 cm].Ua ana ʻia nā manaʻo EWNS me ka hoʻohana ʻana i ka SMPS ma hope o ka helu ʻana i nā poho EWNS i ka hopena.
Ua nānā ʻia nā pae Ozone me ka 205 Dual Beam Ozone Monitor™ (2B Technologies, Boulder, Co)8,9,10.
No nā waiwai EWNS a pau, hoʻohana ʻia ka waiwai mean e like me ke ana ʻana, a ua hoʻohana ʻia ka ʻokoʻa maʻamau e like me ke ana ʻana.Ua hana ʻia nā hoʻāʻo T e hoʻohālikelike i nā waiwai o nā ʻano EWNS i hoʻopaʻa ʻia me nā waiwai kūpono o ka waihona EWNS.
Hōʻike ka Figure 2c i kahi ʻōnaehana "huki" electrostatic precipitation (EPES) i kūkulu mua ʻia i hiki ke hoʻohana ʻia no ka hāʻawi ʻana i ka EWNS ma ka ʻili.Hoʻohana ʻo EPES i nā koina EVNS i hiki ke "alakaʻi" pololei i ka ʻili o ka pahuhopu ma lalo o ka mana o kahi kahua uila ikaika.Hōʻike ʻia nā kikoʻī o ka ʻōnaehana EPES ma kahi paʻi hou e Pyrgiotakis et al.11 .No laila, aia ka EPES i kahi keʻena PVC i paʻi ʻia 3D me nā ʻaoʻao i hoʻopaʻa ʻia a loaʻa i ʻelua mau mea kila kuhili ʻole (304 kila kila, uhi aniani) ma ke kikowaena 15.24 knm kaawale.Ua hoʻohui ʻia nā papa i kahi puna uila kiʻekiʻe o waho (Bertran 205B-10R, Spellman, Hauppauge, NY), ua pili mau ka pā lalo i ka volta maikaʻi, a ua pili mau ka pā luna i ka honua (luna lana).Ua uhi ʻia nā paia o ke keʻena me ka alumini alumini, i hoʻopaʻa ʻia i ka uila e pale ai i ka nalo ʻana.Aia i loko o ke keʻena kahi puka hoʻouka mua i hoʻopaʻa ʻia e hiki ai ke kau ʻia nā ʻili hoʻāʻo ma luna o nā kuʻi plastik e hoʻokiʻekiʻe iā lākou ma luna o ka pā metala lalo e pale aku i ka hoʻopili uila kiʻekiʻe.
Ua helu ʻia ka maikaʻi o ka deposition o EWNS ma EPES e like me kahi protocol i kūkulu mua ʻia i kikoʻī ʻia ma ke Kiʻi S111.
Ma ke ʻano he keʻena hoʻomalu, ua hoʻopili ʻia kahi keʻena kahe cylindrical ʻelua ma ke ʻano me ka ʻōnaehana EPES, kahi i hoʻohana ʻia ai kahi kānana HEPA waena e wehe iā EWNS.E like me ka mea i hōʻike ʻia ma ka Figure 2c, ua puhi ʻia ka aerosol EWNS ma o ʻelua mau keʻena i kūkulu ʻia.Hoʻopau ka kānana ma waena o ka lumi hoʻomalu a me EPES i nā EWNS i koe e hopena i ka mahana like (T), ka haʻahaʻa pili (RH) a me nā pae ozone.
Ua ʻike ʻia nā microorganism meaʻai nui e hoʻohaumia i nā meaʻai hou e like me E. coli (ATCC #27325), fecal indicator, Salmonella enterica (ATCC #53647), meaʻai pathogen, Listeria harmless (ATCC #33090), surrogate for pathogenic Listeria monocytogenes, derived from ATCC (Manassas9 acces, Manassas, a. hū spoilage, a ʻoi aku ke kūpaʻa inactivated bacteria, Mycobacterium paralucky (ATCC #19686).
E kūʻai i nā pahu maʻamau o nā tōmato hua waina mai kou mākeke kūloko a e hoʻopaʻa i ka friji ma 4 ° C a hiki i ka hoʻohana ʻana (a hiki i 3 lā).Ua like ka nui o nā tōmato hoʻokolohua, ma kahi o 1/2 iniha ke anawaena.
ʻO ka moʻomeheu, inoculation, exposure, a me ka helu colony protocols ua kikoʻī ʻia i kā mākou paʻi mua a kikoʻī ʻia i ka ʻikepili Hoʻohui.Ua loiloi ʻia ka maikaʻi o EWNS ma ka wehe ʻana i nā tōmato inoculated i 40,000 #/cm3 no 45 mau minuke.ʻO ka pōkole, ua hoʻohana ʻia ʻekolu mau tōmato e loiloi i nā microorganism e ola nei i ka manawa t = 0 min.Hoʻokomo ʻia ʻekolu mau tōmato ma EPES a hōʻike ʻia iā EWNS ma 40,000 #/cc (EWNS exposed tomatoes) a waiho ʻia nā mea ʻekolu i koe i loko o ke keʻena hoʻomalu (control tomatoes).ʻAʻole i hoʻokō ʻia ka hana hou ʻana o nā tōmato ma nā hui ʻelua.Ua wehe ʻia nā ʻōmato i hōʻike ʻia e EWNS a me nā tōmato hoʻomalu ma hope o 45 mau minuke e loiloi i ka hopena o EWNS.
Ua hana ʻia kēlā me kēia hoʻokolohua ʻekolu.Hana ʻia ka ʻikepili ʻikepili e like me ka protocol i wehewehe ʻia ma ka ʻikepili Hoʻohui.
Ua loiloi ʻia nā ʻano hana inactivation e ka sedimentation o nā mea hōʻike EWNS i hōʻike ʻia (45 min ma 40,000 #/cm3 EWNS aerosol concentration) a me nā laʻana non-irradiated o nā bacteria ʻino ʻole E. coli, Salmonella enterica a me Lactobacillus.Ua hoʻopaʻa ʻia nā ʻāpana i 2.5% glutaraldehyde, 1.25% paraformaldehyde a me 0.03% picric acid i 0.1 M sodium cacodylate buffer (pH 7.4) no 2 mau hola ma ke ana wela.Ma hope o ka holoi ʻana, hoʻopaʻa hope me 1% osmium tetroxide (OsO4) / 1.5% potassium ferrocyanide (KFeCN6) no 2 mau hola, holoi i 3 mau manawa i ka wai a hoʻokomo i ka 1% uranyl acetate no 1 hola, a laila holoi ʻelua i ka wai, a laila dehydrate i loko no 10 mau minuke i 50%, 1,0% ka waiʻona.Hoʻokomo ʻia nā laʻana i loko o ka propylene oxide no ka hola 1 a hoʻopaʻa ʻia me kahi hui 1: 1 o propylene oxide a me TAAP Epon (Marivac Canada Inc. St. Laurent, CA).Hoʻokomo ʻia nā laʻana i loko o TAAB Epon a polymerized ma 60 ° C no 48 mau hola.Ua ʻoki ʻia ka resin granular i hoʻōla ʻia e TEM me ka hoʻohana ʻana i kahi microscope electron transmission maʻamau JEOL 1200EX (JEOL, Tokyo, Iapana) i hoʻolako ʻia me kahi kāmela AMT 2k CCD (Advanced Microscopy Techniques, Corp., Woburn, Massachusetts, USA).
Ua hana ʻia nā hoʻokolohua ʻekolu.No kēlā me kēia manawa, ua hua ʻia ka holoi maʻi bacteria ma ke ʻano papakolu, e loaʻa ana ka huina o ʻeiwa mau helu ʻikepili i kēlā me kēia kiko, ʻo ka awelika o ia mau mea i hoʻohana ʻia ma ke ʻano he kumu bacteria no kēlā microorganism.Ua hoʻohana ʻia ka ʻokoʻa maʻamau e like me ke ana hewa.helu nā helu a pau.
Ua helu ʻia ka logarithm o ka emi ʻana o ka neʻe ʻana o ka bacteria i hoʻohālikelike ʻia me ka t = 0 min me ka hoʻohana ʻana i kēia ʻano:
ʻo ka C0 ka neʻe ʻana o ka hua bacteria i ka hāpana hoʻomalu i ka manawa 0 (ʻo ia hoʻi ma hope o ka maloʻo ʻana o ka ʻili akā ma mua o ka waiho ʻia ʻana i loko o ke keʻena) a ʻo Cn ka neʻe ʻana o ka maʻi bacteria ma ka ʻili ma hope o n mau minuke o ka ʻike.
No ka hōʻemi maoli ʻana o ka maʻi bacteria i ka hoʻolaha ʻana he 45 mau minuke, ua helu pū ʻia ka hoʻohaʻahaʻa log i hoʻohālikelike ʻia me ka mana ma hope o 45 mau minuke:
kahi ʻo Cn ka ʻike o ka bacteria i ka hāpana hoʻomalu i ka manawa n a ʻo Cn-Control ka ʻike ʻana o ka bacteria mana i ka manawa n.Hōʻike ʻia ka ʻikepili ma ke ʻano he hōʻemi log i hoʻohālikelike ʻia me ka mana (ʻaʻohe hōʻike EWNS).
I ka wā o ke aʻo ʻana, ua loiloi ʻia kekahi mau hui ʻana o ka volta a me ka mamao ma waena o ka nila a me ka electrode counter e pili ana i ka hoʻokumu ʻana o Taylor cone, Taylor cone stability, EWNS production stability, a reproducibility.Hōʻike ʻia nā hui like ʻole ma ka Papa Pākuʻi S1.ʻElua mau hihia e hōʻike ana i nā waiwai paʻa a hiki ke hana hou ʻia (Taylor cone, EWNS generation, a me ke kūpaʻa i ka manawa) ua koho ʻia no ka noiʻi piha.Ma ka fig.Hōʻike ka Figure 3 i nā hopena no ka uku, ka nui, a me ka ʻike o ROS ma nā hihia ʻelua.Hōʻike pū ʻia nā hopena ma ka Papa 1. No ka ʻike ʻana, ʻo ka Figure 3 a me ka Papa 1 nā waiwai o ka EWNS8, 9, 10, 11 (baseline-EWNS) i hana mua ʻia.Hoʻopuka hou ʻia nā helu helu koʻikoʻi e hoʻohana ana i ka hoʻāʻo t-huelo ʻelua ma ka Papa Pākuʻi S2.Eia kekahi, ʻo nā ʻikepili hou e pili ana i nā haʻawina e pili ana i ka hopena o ka counter electrode sampling hole diameter (D) a me ka mamao ma waena o ka electrode ground a me ka piko (L) (Nā Kiʻi Hoʻohui S2 a me S3).
(ac) Ua ana ʻia ka nui e ka AFM.(df) ʻO ke ʻano o ka hoʻouka ʻana o ka ʻili.(g) Hōʻike ROS o ka EPR.
He mea nui nō hoʻi e hoʻomaopopo ʻia no nā kūlana i luna aʻe, ʻo ke ana ʻana o ka ionization i kēia manawa ma waena o 2 a me 6 μA a me ka uila ma waena o -3.8 a me -6.5 kV, e hopena i ka hoʻohana ʻana i ka mana ma lalo o 50 mW no kēia module hoʻopili hanauna EWNS hoʻokahi.ʻOiai ua hana ʻia ʻo EWNS ma lalo o ke kaomi kiʻekiʻe, haʻahaʻa loa nā pae ozone, ʻaʻole i ʻoi aku ma mua o 60 ppb.
Hōʻike ke Kiʻi S4 hou i nā kahua uila i hoʻohālikelike ʻia no nā hiʻohiʻona [-6.5 kV, 4.0 knm] a me [-3.8 kV, 0.5 knm].No nā hiʻohiʻona [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm], ʻo ka helu ʻana o ke kahua he 2 × 105 V/m a me 4.7 × 105 V/m.Manaʻo ʻia kēia, no ka mea, ʻoi aku ka kiʻekiʻe o ka lākiō voltage-mamao.
Ma ka fig.Hōʻike ka 3a, b i ke anawaena EWNS i ana ʻia me ka AFM8.ʻO nā anawaena EWNS i helu ʻia he 27 nm a me 19 nm no ka [-6.5 kV, 4.0 knm] a me [-3.8 kV, 0.5 knm].No nā hiʻohiʻona [-6.5 kV, 4.0 knm] a me [-3.8 kV, 0.5 knm], ʻo 1.41 a me 1.45 ka hoʻokaʻawale maʻamau geometric, e hōʻike ana i ka puʻunaue haiki.ʻO ka mean nui a me ka geometric standard deviation kokoke loa i ka baseline EWNS, ma 25 nm a me 1.41.Ma ka fig.Hōʻike ka 3c i ka mahele nui o ka base EWNS i ana ʻia me ke ʻano like ma lalo o nā kūlana like.
Ma ka fig.Hōʻike ʻo 3d,e i nā hopena o ke ʻano o ka uku.ʻO ka ʻikepili nā ana awelika o 30 mau ana like o ka ʻike (#/cm3) a me kēia manawa (I).Hōʻike ka hōʻike ʻana he 22 ± 6 e- a me 44 ± 6 e- no ka [-6.5 kV, 4.0 knm] a me [-3.8 kV, 0.5 knm] ka awelika uku ma ka EWNS.ʻOi aku ka nui o nā koina ma luna o lākou ke hoʻohālikelike ʻia me ka baseline EWNS (10 ± 2 e-), ʻelua manawa ʻoi aku ka nui ma mua o ka [-6.5 kV, 4.0 knm] hiʻohiʻona a ʻehā manawa ʻoi aku ka nui ma mua o ka [-3 .8 kV, 0.5 knm].Hōʻike ke kiʻi 3f i ka uku.ʻikepili no Baseline-EWNS.
Mai nā palapala 'āina o ka helu EWNS (Supplementary Figures S5 a me S6), hiki ke ʻike ʻia he ʻoi aku ka nui o nā ʻāpana [-6.5 kV, 4.0 cm] ma mua o ka [-3.8 kV, 0.5 knm].He mea kūpono hoʻi e hoʻomaopopo i ka nānā ʻana o ka helu EWNS a hiki i 4 mau hola (Supplementary Figures S5 a me S6), kahi i hōʻike ʻia ai ka paʻa ʻana o ka hanauna EWNS i nā pae like o ka nui o ka helu helu ma nā hihia ʻelua.
Ma ka fig.Hōʻike ʻo 3g i ka spectrum EPR ma hope o ka unuhi ʻana i ka mana EWNS i hoʻopaʻa ʻia (ka hope) ma [-6.5 kV, 4.0 knm].Ua hoʻohālikelike ʻia ka spectra ROS me ka hiʻohiʻona Baseline-EWNS ma kahi hana i paʻi mua ʻia.ʻO ka helu o EWNS e pane ana me nā pahele wili i helu ʻia he 7.5 × 104 EWNS/s, e like me ka Baseline-EWNS8 i paʻi mua ʻia.Ua hōʻike maopopo ka EPR spectra i ka loaʻa ʻana o nā ʻano ROS ʻelua, ʻo O2- ka ʻano nui a ʻo OH• ka emi ʻana o ka nui.Eia kekahi, ua hōʻike ʻia kahi hoʻohālikelike pololei o ka ikaika kiʻekiʻe i ka EWNS i hoʻopaʻa ʻia i ka ʻike ROS kiʻekiʻe loa i hoʻohālikelike ʻia i ka baseline EWNS.
Ma ka fig.Hōʻike ka 4 i ka pono o ka waiho ʻana o EWNS ma EPES.Ua hōʻuluʻulu ʻia nā ʻikepili ma ka Papa I a hoʻohālikelike ʻia me ka ʻikepili EWNS kumu.No nā hihia ʻelua o EUNS, kokoke ka deposition i 100% ʻoiai ma kahi haʻahaʻa haʻahaʻa o 3.0 kV.ʻO ka mea maʻamau, lawa ka 3.0 kV no ka waiho ʻana o 100%, me ka nānā ʻole i ka hoʻololi ʻana o ka uku.Ma lalo o nā kūlana like, ʻo 56% wale nō ka deposition efficiency o Baseline-EWNS ma muli o kā lākou uku haʻahaʻa (awelika 10 electrons no EWNS).
Ma ka fig.5 a ma ka papa.2 hōʻuluʻulu i ka waiwai inactivation o nā microorganisms inoculated ma luna o ka ili o nā tōmato ma hope o ka hoʻolaha ʻana i kahi 40,000 #/cm3 EWNS no 45 mau minuke ma ke ʻano kūpono loa [-6.5 kV, 4.0 cm].Ua hōʻike ʻo Inoculated E. coli a me Lactobacillus innocuous i ka hōʻemi nui ʻana o nā lāʻau 3.8 i ka wā o ka hōʻike ʻana i nā minuke 45.Ma lalo o nā kūlana like, ua loaʻa iā S. enterica ka emi ʻana o ka 2.2-log, ʻoiai ʻo S. cerevisiae a me M. parafortutum i hōʻemi i ka 1.0-log.
Hōʻike nā micrographs electron (Figure 6) i nā hoʻololi kino i hoʻokomo ʻia e EWNS ma luna o ka Escherichia coli, Streptococcus, a me Lactobacillus ʻinoʻino e alakaʻi ana i kā lākou hana ʻole.Loaʻa i ka bacteria control nā membrane cell paʻa, aʻo ka bacteria i hōʻike ʻia ua hōʻino nā membrane waho.
Ua hōʻike ʻia ke kiʻi microscopic electron o ka mana a me ka bacteria i hōʻike ʻia i ka pōʻino membrane.
ʻO nāʻikepili e pili ana i nā waiwai physicochemical o ka EWNS i hoʻopaʻaʻia e hōʻike pū ana i ka maikaʻi loa o nā waiwai (ka uku o ka honua a me ka ROS content) o ka EWNS i ka hoʻohālikelikeʻana i ka EWNS baseline data8,9,10,11 i paʻi muaʻia.Ma kekahiʻaoʻao, ua mau ko lākou nui i ka nanometer, e like me nā hopena i hōʻike muaʻia, e hiki ai iā lākou ke noho i ka lewa no nā manawa lōʻihi.Hiki ke wehewehe ʻia ka polydispersity i ʻike ʻia e ka hoʻololi ʻana i ka uku o ka ʻili e hoʻoholo ai i ka nui o EWNS, ka randomness o ka hopena Rayleigh, a me ka coalescence.Eia naʻe, e like me ka kikoʻī e Nielsen et al.22, hoʻemi ʻia ka hoʻokuʻu ʻana o ka ʻili kiʻekiʻe ma o ka hoʻonui ʻana i ka ikehu o ka wai.Ma kā mākou paʻi mua 8 ua hoʻopaʻa ʻia kēia kumumanaʻo no nā microdroplets 22 a me EWNS.Hiki ke hoʻololi i ka nui a me ka hāʻawi ʻana i ka hāʻawi ʻana i ka nui i ʻike ʻia.