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I kēia mau lā iho nei, ua hoʻomohala ʻia kahi kahua antimicrobial ʻaʻohe kemika e pili ana i ka nanotechnology me ka hoʻohana ʻana i nā nanostructures wai artificial (EWNS). Loaʻa i nā EWNS kahi uku kiʻekiʻe o ka ʻili a waiwai i nā ʻano oxygen reactive (ROS) hiki ke launa pū me a hoʻopau i kekahi mau microorganism, me nā pathogens i lawe ʻia e ka meaʻai. Maanei ua hōʻike ʻia e hiki ke hoʻoponopono maikaʻi ʻia ko lākou mau waiwai i ka wā o ka synthesis a hoʻonui ʻia e hoʻonui hou i ko lākou hiki antibacterial. Ua hoʻolālā ʻia ke kahua hoʻokolohua EWNS e hoʻoponopono maikaʻi i nā waiwai o EWNS ma ka hoʻololi ʻana i nā palena synthesis. Ua hana ʻia ke ʻano o nā waiwai EWNS (uku, nui, a me ka ʻike ROS) me ka hoʻohana ʻana i nā ʻano loiloi hou. Eia kekahi, ua hoʻokomo ʻia nā microorganisms meaʻai e like me Escherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium para fortitum, a me Saccharomyces cerevisiae ma luna o ka ʻili o nā tōmato hua waina organik e loiloi i ko lākou hiki ke hoʻopau microbial. Hōʻike nā hopena i hōʻike ʻia ma aneʻi hiki ke hoʻoponopono maikaʻi ʻia nā waiwai o EWNS i ka wā o ka synthesis, e hopena ana i ka hoʻonui exponential i ka pono inactivation. ʻO ke ʻano kūikawā, ua piʻi ka uku o ka ʻili ma kahi manawa o ʻehā, a ua piʻi ka nui o ROS. Ua hilinaʻi ka nui o ka wehe ʻana o ka microbial i ka microbially a mai 1.0 a 3.8 log ma hope o 45 mau minuke o ka hōʻike ʻana i kahi mahele aerosol o 40,000 #/cm3 EWNS.
ʻO ka haumia microbial ke kumu nui o nā maʻi i lawe ʻia e ka meaʻai i hoʻokumu ʻia e ka ʻai ʻana i nā pathogens a i ʻole kā lākou mau mea ʻawahia. ʻO ka maʻi i lawe ʻia e ka meaʻai ke kumu o kahi 76 miliona mau maʻi, 325,000 mau hoʻokipa maʻi, a me 5,000 mau make i kēlā me kēia makahiki ma ʻAmelika Hui Pū ʻIa wale nō1. Eia kekahi, ua manaʻo ka ʻOihana Mahiʻai o ʻAmelika Hui Pū ʻIa (USDA) ʻo ka hoʻonui ʻana i ka ʻai ʻana i nā huaʻai hou ke kuleana no 48 pakeneka o nā maʻi āpau i lawe ʻia e ka meaʻai i hōʻike ʻia ma ʻAmelika Hui Pū ʻIa2. He kiʻekiʻe loa ke kumukūʻai o ka maʻi a me ka make mai nā pathogens i lawe ʻia e 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, ua hoʻokō nui ʻia nā hana antimicrobial chemical4, radiation5 a me thermal6 e hōʻoia i ka palekana o ka meaʻai ma nā wahi kaohi koʻikoʻi palena (CCP) i loko o ke kaulahao hana (ma hope o ka ʻohi ʻana a/a i ʻole i ka wā o ka hoʻopili ʻana) ma mua o ka hoʻokō mau ʻia ʻana i ke ʻano e hiki ai i nā hua hou ke hoʻohaumia ʻia 7. Pono nā hana antimicrobial e kaohi maikaʻi i nā maʻi i lawe ʻia mai ka meaʻai a me ka palaho ʻana o ka meaʻai a loaʻa ka hiki ke hoʻopili ʻia ma waena o ka hoʻomau ʻana o ka mahiʻai-a-papa. ʻOi aku ka hopena a me ke kumukūʻai.
Ua hoʻomohala ʻia nei kahi kahua antimicrobial ʻaʻohe kemika i hoʻokumu ʻia i ka nanotechnology e hoʻopau i ka bacteria ma nā ʻili a ma ka lewa me ka hoʻohana ʻana i nā nanostructures wai artificial (EWNS). No ka synthesis o EVNS, ua hoʻohana ʻia ʻelua mau kaʻina hana like: electrospray a me ka ionization wai (Kiʻi 1a). Ua hōʻike mua ʻia ʻo EWNS he ʻano kūikawā o nā waiwai kino a me nā waiwai olaola8,9,10. Loaʻa i ka EWNS ka awelika o 10 mau electrons no kēlā me kēia ʻano a me ka nui nanometer awelika o 25 nm (Kiʻi 1b,c)8,9,10. Eia kekahi, ua hōʻike ka electron spin resonance (ESR) aia i loko o ka EWNS ka nui o nā ʻano oxygen reactive (ROS), ʻo ia hoʻi nā radical hydroxyl (OH•) a me superoxide (O2-) (Kiʻi 1c)8. Ua noho ʻo EWNS i ka lewa no ka manawa lōʻihi a hiki ke kuʻi me nā microbes i kau ʻia i ka lewa a kū ma luna o nā ʻili, e hāʻawi ana i kā lākou ukana ROS a ke kumu o ka inactivation microbial (Kiʻi 1d). Ua hōʻike pū kēia mau haʻawina mua e hiki i nā EWNS ke launa pū me nā bacteria gram-negative a me gram-positive like ʻole o ke olakino lehulehu, me ka mycobacteria, ma nā ʻili a ma ka lewa8,9. Ua hōʻike ka microscopy electron transmission ua hoʻokumu ʻia ka inactivation e ka hoʻohaunaele ʻana o ka membrane cell. Eia kekahi, ua hōʻike nā haʻawina hanu koke ʻaʻole hiki i nā dosis kiʻekiʻe o EWNS ke hōʻino i ka māmā a i ʻole ka mumū8.
(a) Hana ʻia ka electrospray ke hoʻopili ʻia kahi voltage kiʻekiʻe ma waena o kahi wai e loaʻa ana i ka capillary a me kahi counter electrode. (b) ʻO ka hoʻopili ʻana o ka voltage kiʻekiʻe e hopena i ʻelua mau hanana like ʻole: (i) electrospraying o ka wai a me (ii) hanauna o nā ʻano oxygen reactive (ions) i hoʻopaʻa ʻia i loko o ka EWNS. (c) ʻO ke ʻano kū hoʻokahi o EWNS. (d) He neʻe nui nā EWNS ma muli o ko lākou ʻano nanoscale a hiki ke launa pū me nā pathogens ea.
Ua hōʻike hou ʻia nei ka hiki o ka paepae antimicrobial EWNS ke hoʻopau i nā microorganism i lawe ʻia e ka meaʻai ma luna o ka ʻili o ka meaʻai hou. Ua hōʻike ʻia hoʻi e hiki ke hoʻohana ʻia ka uku ʻili EWNS me kahi kahua uila no ka lawe ʻana i kahi kikoʻī. ʻOi aku ka mea nui, ua ʻike ʻia kahi hopena mua e hoʻohiki ana ma kahi o 1.4 log ka hōʻemi ʻana i ka hana tōmato organik e kūʻē i nā microorganism meaʻai like ʻole e like me E. coli a me Listeria i loko o 90 mau minuke o ka hōʻike ʻana iā EWNS ma kahi nui o 50,000#/cm311. Eia kekahi, ʻaʻole i hōʻike nā hoʻokolohua loiloi organoleptic mua i ka hopena organoleptic i hoʻohālikelike ʻia me ka tōmato kaohi. ʻOiai ʻo kēia mau hopena inactivation mua e hoʻohiki i ka palekana o ka meaʻai ʻoiai ma nā dosis EWNS haʻahaʻa loa o 50,000#/cc. e nānā, ua maopopo he ʻoi aku ka maikaʻi o ka hiki ke inactivation kiʻekiʻe e hōʻemi hou i ka pilikia o ka maʻi a me ka palaho.
Maanei, e kālele mākou i kā mākou noiʻi ma ka hoʻomohala ʻana i kahi kahua hanauna EWNS e hoʻomaikaʻi i nā palena synthesis a hoʻonui i nā waiwai physicochemical o EWNS e hoʻonui i ko lākou hiki ke antibacterial. Ma ke ʻano kūikawā, ua kālele ka optimization i ka hoʻonui ʻana i kā lākou uku ʻili (e hoʻomaikaʻi i ka lawe ʻana i kuhikuhi ʻia) a me ka ʻike ROS (e hoʻomaikaʻi i ka pono inactivation). Ke ʻano o nā waiwai physico-chemical i hoʻonui ʻia (ka nui, ka uku a me ka ʻike ROS) me ka hoʻohana ʻana i nā ʻano loiloi hou a me ka hoʻohana ʻana i nā microorganism meaʻai maʻamau e like me E. coli, S. enterica, L. innocua, S. cerevisiae a me M. parafortuitum.
Ua hana ʻia ʻo EVNS ma o ka electrospraying like a me ka ionization o ka wai maʻemaʻe kiʻekiʻe (18 MΩ cm–1). Hoʻohana pinepine ʻia ka atomizer uila 12 e atomize i nā wai a me nā polymer synthetic a me nā ʻāpana keramika 13 a me nā fibers 14 o ka nui i kāohi ʻia.
E like me ka mea i hōʻike kikoʻī ʻia ma nā puke paʻi mua 8, 9, 10, 11, ma kahi hoʻokolohua maʻamau, ua hoʻopili ʻia kahi voltage kiʻekiʻe ma waena o kahi capillary metala a me kahi electrode counter grounded. I loko o kēia kaʻina hana, ʻelua mau hanana like ʻole e kū mai: 1) electrospray a me 2) ionization o ka wai. ʻO kahi kahua uila ikaika ma waena o nā electrodes ʻelua e hoʻoulu ai i nā uku maikaʻi ʻole e kūkulu ma luna o ka ʻili o ka wai i hoʻopaʻa ʻia, e hopena ana i ka hoʻokumu ʻia ʻana o nā cones Taylor. ʻO ka hopena, ua hoʻokumu ʻia nā kulu wai i hoʻopiʻi nui ʻia, e hoʻomau nei i ka haki ʻana i nā ʻāpana liʻiliʻi, e like me ke kumumanaʻo Rayleigh16. I ka manawa like, ʻo kahi kahua uila ikaika ke kumu o kekahi o nā mole wai e māhele a wehe i nā electrons (ionization), a laila e hana ana i ka nui o nā ʻano oxygen reactive (ROS)17. Ua hoʻopili ʻia nā ʻeke ROS18 i hana ʻia i ka manawa like i loko o EWNS (Kiʻi 1c).
Ma ke kiʻi 2a e hōʻike ana i ka ʻōnaehana hanauna EWNS i hoʻomohala ʻia a hoʻohana ʻia i ka synthesis EWNS ma kēia haʻawina. Ua hānai ʻia ka wai i hoʻomaʻemaʻe ʻia i mālama ʻia i loko o kahi ʻōmole pani ʻia ma o kahi paipu Teflon (2 mm ke anawaena o loko) i kahi nila kila kila 30G (capillary metala). E like me ka mea i hōʻike ʻia ma ke Kiʻi 2b, ua hoʻomalu ʻia ke kahe ʻana o ka wai e ke kaomi ea i loko o ka ʻōmole. Hoʻopili ʻia ka nila i kahi console Teflon hiki ke hoʻoponopono lima ʻia i kahi mamao mai ka electrode counter. ʻO ka electrode counter he disk alumini i wili ʻia me kahi lua ma waena no ka laʻana. Ma lalo o ka electrode counter he funnel laʻana alumini, kahi i hoʻopili ʻia i ke koena o ka hoʻonohonoho hoʻokolohua ma o kahi puka laʻana (Kiʻi 2b). Hoʻokumu ʻia nā ʻāpana sampler āpau me ka uila e pale aku i ka hōʻiliʻili ʻana o ka uku e hiki ke hoʻohaʻahaʻa i ka laʻana ʻāpana.
(a) ʻŌnaehana Hana Nanostructure Wai ʻenekinia (EWNS). (b) ʻĀpana kea o ka sampler a me ka ʻāpana electrospray e hōʻike ana i nā palena koʻikoʻi loa. (c) Hoʻonohonoho hoʻokolohua no ka hoʻopau ʻana i ka bacteria.
ʻO ka ʻōnaehana hanauna EWNS i wehewehe ʻia ma luna nei hiki ke hoʻololi i nā palena hana koʻikoʻi e hoʻomaʻamaʻa i ka hoʻonohonoho pono ʻana o nā waiwai EWNS. E hoʻoponopono i ke volta i hoʻopili ʻia (V), ka mamao ma waena o ke kui a me ka electrode counter (L), a me ke kahe wai (φ) ma o ke capillary e hoʻonohonoho pono i nā ʻano EWNS. Hoʻohana ʻia nā hōʻailona [V (kV), L (cm)] e hōʻike i nā hui like ʻole. E hoʻoponopono i ke kahe wai e loaʻa ai kahi cone Taylor paʻa o kahi set [V, L]. No nā kumu o kēia haʻawina, ua hoʻonohonoho ʻia ka puka o ka electrode counter (D) ma 0.5 ʻīniha (1.29 cm).
Ma muli o ke geometry palena a me ka asymmetry, ʻaʻole hiki ke helu ʻia ka ikaika o ke kahua uila mai nā loina mua. Akā, ua hoʻohana ʻia ka polokalamu QuickField™ (Svendborg, Denmark)19 e helu i ke kahua uila. ʻAʻole like ke kahua uila, no laila ua hoʻohana ʻia ka waiwai o ke kahua uila ma ka piko o ke capillary ma ke ʻano he waiwai kuhikuhi no nā hoʻonohonoho like ʻole.
I ka wā o ke aʻo ʻana, ua loiloi ʻia kekahi mau hui pū ʻana o ke volta a me ka mamao ma waena o ke kui a me ka counter electrode ma ke ʻano o ka hoʻokumu ʻana o Taylor cone, ke kūpaʻa o Taylor cone, ke kūpaʻa hana EWNS, a me ka hana hou ʻana. Hōʻike ʻia nā hui like ʻole ma ka Papa Hoʻohui S1.
Ua hoʻopili pololei ʻia ka hopena o ka ʻōnaehana hanauna EWNS i kahi Scanning Mobility Particle Sizer (SMPS, kumu hoʻohālike 3936, TSI, Shoreview, Minnesota) e ana i ka nui o ka ʻāpana a ua hoʻohana ʻia me kahi electrometer aerosol Faraday (TSI, kumu hoʻohālike 3068B, Shoreview, USA). MN) e ana i nā kahe aerosol, e like me ka mea i wehewehe ʻia ma kā mākou puke mua9. Ua laʻana ʻia ka SMPS a me ka electrometer aerosol ma kahi kahe o 0.5 L/min (ka nui o ke kahe laʻana 1 L/min). Ua ana ʻia nā ʻāpana a me nā kahe aerosol no 120 s. E hana hou i ke ana ʻana i 30 mau manawa. Ua helu ʻia ka nui o ka uku aerosol mai nā ana o kēia manawa, a ua manaʻo ʻia ka awelika o ka uku EWNS mai ka nui o nā ʻāpana EWNS i laʻana ʻia. Hiki ke helu ʻia ke kumukūʻai awelika o EWNS me ka hoʻohana ʻana i ka Equation (1):
kahi ʻo IEl ke au i ana ʻia, ʻo NSMPS ka helu o ka nui i ana ʻia me ka SMPS, a ʻo φEl ka nui o ke kahe i ka electrometer.
Ma muli o ka hoʻopilikia ʻana o ka haʻahaʻa pili (RH) i ka hoʻouku ʻana o ka ʻili, ua mālama ʻia ka mahana a me ka (RH) ma 21°C a me 45%, kēlā me kēia, i ka wā o ka hoʻokolohua.
Ua hoʻohana ʻia ka microscopy ikaika atomika (AFM), Asylum MFP-3D (Asylum Research, Santa Barbara, CA) a me ka probe AC260T (Olympus, Tokyo, Iapana) e ana i ka nui a me ke ola o ka EWNS. ʻO ka helu scan AFM he 1 Hz a ʻo ka wahi scan he 5 µm × 5 µm me 256 mau laina scan. Ua hoʻokomo ʻia nā kiʻi āpau i ka hoʻonohonoho kiʻi mua me ka hoʻohana ʻana i ka polokalamu Asylum (mask me kahi laulā o 100 nm a me ka paepae o 100 pm).
Wehe i ka funnel laʻana a kau i ka ʻili mica ma kahi mamao o 2.0 cm mai ka counter electrode no ka manawa awelika o 120 s e pale aku i ka hui ʻana o nā ʻāpana a me ka hoʻokumu ʻana o nā kulu like ʻole ma ka ʻili mica. Ua hoʻopili pololei ʻia ʻo EWNS i nā ʻili mica i ʻoki hou ʻia (Ted Pella, Redding, CA). Ma hope koke iho o ka sputtering, ua ʻike ʻia ka ʻili mica me ka hoʻohana ʻana iā AFM. ʻO ke kihi pili ʻili o ka mica i hoʻololi ʻole ʻia i ʻoki hou ʻia kokoke i 0°, no laila e pālahalaha ana ʻo EWNS ma luna o ka ʻili mica i loko o kahi ʻano domed20. Ua ana pololei ʻia ke anawaena (a) a me ke kiʻekiʻe (h) o nā kulu diffusing mai ka topography AFM a hoʻohana ʻia e helu i ka nui o ka diffusion domed EWNS me ka hoʻohana ʻana i kā mākou ʻano i hōʻoia mua ʻia8. Ke manaʻo nei he like ka nui o ka EVNS onboard, hiki ke helu ʻia ke anawaena like mai ka hoohalike (2):
E like me kā mākou hana i hoʻomohala mua ʻia, ua hoʻohana ʻia kahi pahele wili electron spin resonance (ESR) e ʻike i ke alo o nā mea waena radical ola pōkole ma EWNS. Ua hoʻohele ʻia nā aerosol ma o kahi hopena e loaʻa ana he 235 mM DEPMPO (5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide) (Oxis International Inc., Portland, Oregon). Ua hana ʻia nā ana EPR āpau me ka hoʻohana ʻana i kahi spectrometer Bruker EMX (Bruker Instruments Inc. Billerica, MA, USA) a me nā ʻāpana cell pālahalaha. Ua hoʻohana ʻia ka polokalamu Acquisit (Bruker Instruments Inc. Billerica, MA, USA) e hōʻiliʻili a kālailai i ka ʻikepili. Ua hana ʻia ke ʻano ROS no kahi hoʻonohonoho o nā kūlana hana [-6.5 kV, 4.0 cm]. Ua ana ʻia nā ʻano EWNS me ka hoʻohana ʻana i ka SMPS ma hope o ka noʻonoʻo ʻana i ka nalowale o EWNS i loko o ka impactor.
Ua nānā ʻia nā pae ozone me ka hoʻohana ʻana i ka 205 Dual Beam Ozone Monitor™ (2B Technologies, Boulder, Co)8,9,10.
No nā waiwai EWNS āpau, ʻo ke ana ka awelika o nā ana, a ʻo ka hewa ana ka ʻokoʻa maʻamau. Ua hana ʻia kahi hoʻāʻo-t e hoʻohālikelike i ka waiwai o ka ʻano EWNS i hoʻomaikaʻi ʻia me ka waiwai like o ke kumu EWNS.
Hōʻike ka Kiʻi 2c i kahi ʻōnaehana Electrostatic Precipitation Pass Through System (EPES) i hoʻomohala mua ʻia a wehewehe ʻia e hiki ke hoʻohana ʻia e kuhikuhi iā EWNS11 i nā ʻili. Hoʻohana ʻo EPES i kahi uku EWNS i hui pū ʻia me kahi kahua uila ikaika e "kuhikuhi" pololei i ka ʻili o ka pahuhopu. Hōʻike ʻia nā kikoʻī o ka ʻōnaehana EPES ma kahi puke hou na Pyrgiotakis et al.11. No laila, aia ʻo EPES i kahi keʻena PVC i paʻi ʻia he 3D me nā wēlau tapered e loaʻa ana ʻelua mau papa metala kila kila like (304 kila kila, poli ʻia ke aniani) ma waena he 15.24 cm ke kaʻawale. Ua hoʻopili ʻia nā papa i kahi kumu uila kiʻekiʻe o waho (Bertran 205B-10R, Spellman, Hauppauge, NY), ua maikaʻi mau ka papa lalo a ua hoʻokumu mau ʻia ka papa luna (lana). Uhi ʻia nā paia o ke keʻena me ka pepa alumini, kahi i hoʻokumu ʻia me ka uila e pale aku i ka nalowale ʻana o nā ʻāpana. Loaʻa i ke keʻena kahi puka hoʻouka mua i hoʻopaʻa ʻia e hiki ai ke kau ʻia nā ʻili hoʻāʻo ma nā ʻūlū plastik, e hāpai ana iā lākou mai ka papa metala lalo e pale aku i ka hoʻopilikia uila kiʻekiʻe.
Ua helu ʻia ka pono o ka waiho ʻana o EWNS ma EPES e like me kahi protocol i hoʻomohala mua ʻia i hōʻike kikoʻī ʻia ma ka Kiʻi Hoʻohui S111.
Ma ke ʻano he keʻena hoʻomalu, ua hoʻopili ʻia ke kahe ʻelua ma o ke keʻena cylindrical ma ke ʻano he moʻo me ka ʻōnaehana EPES me ka hoʻohana ʻana i kahi kānana HEPA waena e wehe i nā EWNS. E like me ka mea i hōʻike ʻia ma ke kiʻi 2c, ua hoʻopili ʻia ka aerosol EWNS ma o nā keʻena ʻelua i hoʻopili ʻia ma ke ʻano he moʻo. Hoʻopau ke kānana ma waena o ka lumi hoʻomalu a me EPES i nā EWNS i koe e hopena ana i ka mahana like (T), ka humidity pili (RH) a me nā pae ozone.
Ua ʻike ʻia nā microorganisms koʻikoʻi i lawe ʻia e ka meaʻai e hoʻohaumia i nā huahana hou e like me Escherichia coli (ATCC #27325), kahi hōʻailona fecal, Salmonella enterica (ATCC #53647), kahi pathogen i lawe ʻia e ka meaʻai, Listeria innocua (ATCC #33090), kahi koho ʻē aʻe i ka Listeria monocytogenes pathogenic. , Saccharomyces cerevisiae (ATCC #4098) ma ke ʻano he koho ʻē aʻe i ka hū palaho, a me Mycobacterium parafortuitous (ATCC #19686) ma ke ʻano he bacteria ola ʻoi aku ka paʻa i kūʻai ʻia mai ATCC (Manassas, Virginia).
E kūʻai ma ke ʻano maʻamau i nā pahu o nā tōmato hua waina organik mai kāu mākeke kūloko a hoʻokomo i loko o ka pahu hau ma 4°C a hiki i ka hoʻohana ʻana (a hiki i 3 lā). E koho i nā tōmato e hoʻāʻo me hoʻokahi nui, ma kahi o 1/2 ʻīniha ke anawaena.
Ua hōʻike kikoʻī ʻia nā kaʻina hana no ka hoʻoulu ʻana, ka hoʻomoe ʻana, ka hōʻike ʻana a me ka helu ʻana o nā kolone i kā mākou mau puke i hala a ua wehewehe kikoʻī ʻia ma ka ʻIkepili Hoʻohui 11. Ua loiloi ʻia ka hana EWNS ma ka hōʻike ʻana i nā tōmato i hoʻomoe ʻia i 40,000 #/cm3 no 45 mau minuke. I ka manawa pōkole, i ka manawa t = 0 min, ua hoʻohana ʻia ʻekolu tōmato e loiloi i nā microorganism ola. Ua kau ʻia ʻekolu tōmato i loko o EPES a hōʻike ʻia i EWNS ma 40,000 #/cc (nā tōmato i hōʻike ʻia e EWNS) a ua kau ʻia ʻekolu mau mea ʻē aʻe i loko o ke keʻena hoʻomalu (nā tōmato hoʻomalu). ʻAʻohe o nā hui tōmato i hoʻokau ʻia i ka hana hou. Ua wehe ʻia nā tōmato a me nā kaohi i hōʻike ʻia e EWNS ma hope o 45 mau minuke e loiloi i ka hopena o EWNS.
Ua hoʻokō ʻia kēlā me kēia hoʻokolohua ma ke ʻano ʻekolu. Ua hoʻokō ʻia ka loiloi ʻikepili e like me ke kaʻina hana i wehewehe ʻia ma ka ʻIkepili Hoʻohui.
ʻO nā laʻana bacteria E. coli, Enterobacter, a me L. innocua i hōʻike ʻia i ka EWNS (45 min, EWNS aerosol concentration 40,000 #/cm3) a ʻaʻole i hōʻike ʻia ua pelleted ʻia e loiloi i nā ʻano hana inactivation. Ua hoʻopaʻa ʻia ka precipitate no 2 mau hola ma ka mahana o ka lumi i loko o ka 0.1 M sodium cacodylate solution (pH 7.4) me kahi fixative o 2.5% glutaraldehyde, 1.25% paraformaldehyde a me 0.03% picric acid. Ma hope o ka holoi ʻana, ua hoʻopaʻa ʻia lākou me 1% osmium tetroxide (OsO4) / 1.5% potassium ferrocyanide (KFeCN6) no 2 h, holoi ʻia i 3 mau manawa me ka wai a hoʻomoʻa ʻia i loko o 1% uranyl acetate no 1 h, a laila holoi ʻia i ʻelua manawa me ka wai. Ma hope o ka dehydration 10 mau minuke o kēlā me kēia o 50%, 70%, 90%, 100% alcohol. A laila ua hoʻokomo ʻia nā laʻana i loko o ka propylene oxide no 1 hola a ua hoʻomoʻa ʻia me ka hui 1:1 o ka propylene oxide a me TAAP Epon (Marivac Canada Inc. St. Laurent, CA). Ua hoʻokomo ʻia nā laʻana i loko o TAAB Epon a ua polymerized ʻia ma 60°C no 48 mau hola. Ua ʻoki ʻia ka resin granular i hoʻōla ʻia a ua nānā ʻia e TEM me ka hoʻohana ʻana i ka JEOL 1200EX (JEOL, Tokyo, Iapana), kahi microscope electron hoʻoili maʻamau i lako me kahi kāmela AMT 2k CCD (Advanced Microscopy Techniques, Corp., Woburn, MA, USA).
Ua hoʻokō ʻia nā hoʻokolohua āpau ma ke ʻano he ʻekolu. No kēlā me kēia manawa, ua hoʻopili ʻia nā holoi bacteria ma ke ʻano he ʻekolu, a ua loaʻa iā ia he ʻeiwa mau helu ʻikepili no kēlā me kēia kiko, a ua hoʻohana ʻia ka awelika o ia mau mea ma ke ʻano he nui bacteria no kēlā meaola. Ua hoʻohana ʻia ka ʻokoʻa maʻamau ma ke ʻano he hewa ana. Helu ʻia nā helu āpau.
Ua helu ʻia ka logarithm o ka emi ʻana o ka nui o nā bacteria i hoʻohālikelike ʻia me t = 0 min me ka hoʻohana ʻana i kēia ʻano hana:
kahi ʻo C0 ka nui o ka bacteria i loko o ka hāpana kaohi i ka manawa 0 (ʻo ia hoʻi ma hope o ka maloʻo ʻana o ka ʻili akā ma mua o ke kau ʻia ʻana i loko o ke keʻena) a ʻo Cn ka nui o ka bacteria ma luna o ka ʻili ma hope o n mau minuke o ka hōʻike ʻana.
No ka hoʻoponopono ʻana i ka hōʻino kūlohelohe o nā bacteria i ka wā hōʻike ʻia he 45 mau minuke, ua helu ʻia hoʻi ka Log-Reduction i hoʻohālikelike ʻia me ka kaohi ma 45 mau minuke penei:
Ma kahi o Cn ka nui o nā bacteria i loko o ka hāpana kaohi i ka manawa n a ʻo Cn-Control ka nui o nā bacteria kaohi i ka manawa n. Hōʻike ʻia nā ʻikepili ma ke ʻano he hōʻemi log i hoʻohālikelike ʻia me ka kaohi (ʻaʻohe hōʻike EWNS).
I ka wā o ke aʻo ʻana, ua loiloi ʻia kekahi mau hui pū ʻana o ke voltage a me ka mamao ma waena o ke kui a me ka counter electrode ma ke ʻano o ka hoʻokumu ʻana o Taylor cone, ka paʻa o Taylor cone, ka paʻa o ka hana ʻana o EWNS, a me ka hana hou ʻana. Hōʻike ʻia nā hui like ʻole ma ka Papa Hoʻohui S1. Ua koho ʻia ʻelua mau hihia no kahi haʻawina piha e hōʻike ana i nā waiwai paʻa a hiki ke hana hou ʻia (Taylor cone, hana ʻana o EWNS, a me ke kūpaʻa i ka hala ʻana o ka manawa). Ma ke kiʻi. 3 hōʻike i nā hopena ma ka uku, ka nui a me ka ʻike o ROS no ʻelua mau hihia. Ua hōʻuluʻulu pū ʻia nā hopena ma ka Papa 1. No ka ʻike, ʻo ka Kiʻi 3 a me ka Papa 1 e hoʻokomo i nā waiwai o ka EWNS8, 9, 10, 11 i hoʻohui mua ʻia (baseline-EWNS). Ua paʻi hou ʻia nā helu koʻikoʻi helu e hoʻohana ana i kahi t-test ʻelua-tailed ma ka Papa Hoʻohui S2. Eia kekahi, ʻo ka ʻikepili hou aku e komo pū me nā haʻawina e pili ana i ka hopena o ke anawaena lua laʻana o ka counter electrode (D) a me ka mamao ma waena o ka electrode honua a me ka piko o ka nila (L) (Nā Kiʻi Hoʻohui S2 a me S3).
(a–c) Ka hoʻolaha nui ʻana o ka AFM. (d – f) Ke ʻano o ka hoʻouku ʻili. (g) Ke ʻano o ROS a me ESR.
He mea nui hoʻi e hoʻomaopopo no nā kūlana āpau i luna, aia nā au ionization i ana ʻia ma ka pae o 2-6 µA, a aia nā voltages ma ka pae o -3.8 a -6.5 kV, e hopena ana i ka hoʻohana ʻana i ka mana no kēia EWNS hoʻokahi-terminal ma lalo o 50 mW. . module hanauna. ʻOiai ua synthesized ʻia ʻo EWNS ma lalo o ke kaomi kiʻekiʻe, ua haʻahaʻa loa nā pae ozone, ʻaʻole i ʻoi aku ma mua o 60 ppb.
Hōʻike ka Kiʻi Hoʻohui S4 i nā kahua uila i hoʻohālikelike ʻia no nā hiʻohiʻona [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm], kēlā me kēia. Ua helu ʻia nā kahua e like me nā hiʻohiʻona [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm] ma ke ʻano he 2 × 105 V/m a me 4.7 × 105 V/m, kēlā me kēia. Pono kēia e manaʻo ʻia, ʻoiai ʻoi aku ka kiʻekiʻe o ka lakio o ke uila i ka mamao ma ka lua o ka hihia.
Ma ke kiʻi 3a,b e hōʻike ana i ke anawaena EWNS i ana ʻia me ka AFM8. Ua helu ʻia nā anawaena EWNS awelika no nā hiʻohiʻona [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm] he 27 nm a me 19 nm, kēlā me kēia. ʻO nā ʻokoʻa maʻamau geometric o nā hoʻolaha no nā hihia [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm] he 1.41 a me 1.45, kēlā me kēia, e hōʻike ana i kahi hoʻolaha nui haiki. ʻO ka nui awelika a me ka ʻokoʻa maʻamau geometric ua kokoke loa i ka baseline-EWNS, ʻo ia hoʻi he 25 nm a me 1.41, kēlā me kēia. Ma ke kiʻi 3c e hōʻike ana i ka hoʻolaha nui o ka baseline EWNS i ana ʻia me ka hoʻohana ʻana i ke ʻano like ma lalo o nā kūlana like.
Ma ke kiʻi 3d, hōʻike ʻo e i nā hopena o ke ʻano o ka hoʻouku ʻana. ʻO nā ʻikepili he mau ana awelika o 30 mau ana like o ka noʻonoʻo ʻana (#/cm3) a me ke au (I). Hōʻike ka loiloi ʻo ka hoʻouku awelika ma ka EWNS he 22 ± 6 e- a me 44 ± 6 e- no [-6.5 kV, 4.0 cm] a me [-3.8 kV, 0.5 cm], kēlā me kēia. Ke hoʻohālikelike ʻia me Baseline-EWNS (10 ± 2 e-), ʻoi aku ke kiʻekiʻe o kā lākou hoʻouku ʻili, ʻelua manawa o ka hiʻohiʻona [-6.5 kV, 4.0 cm] a ʻehā manawa o ka [-3 .8 kV, 0.5 cm]. Hōʻike ʻo 3f i ka ʻikepili uku EWNS kumu.
Mai nā palapala ʻāina o ka nui o ka helu EWNS (Nā Kiʻi Hoʻohui S5 a me S6), hiki ke ʻike ʻia he nui aku ka nui o nā ʻāpana ma ke kahua [-6.5 kV, 4.0 cm] ma mua o ke kahua [-3.8 kV, 0.5 cm]. Pono hoʻi e hoʻomaopopo ʻia ua nānā ʻia nā nui o ka helu EWNS a hiki i 4 mau hola (Nā Kiʻi Hoʻohui S5 a me S6), kahi i hōʻike ai ke kūpaʻa o ka hanauna EWNS i nā pae like o nā nui o ka helu ʻāpana ma nā hihia ʻelua.
Hōʻike ka Kiʻi 3g i ka spectrum EPR ma hope o ka hoʻemi ʻana o ka mana (hope) no ka EWNS i hoʻonui ʻia ma [-6.5 kV, 4.0 cm]. Hoʻohālikelike ʻia ka spectrum ROS me ka baseline EWNS i loko o kahi pepa i paʻi mua ʻia. ʻO ka helu i helu ʻia o EWNS e pane ana me ka pahele wili he 7.5 × 104 EWNS/s, kahi like me ka Baseline-EWNS8 i paʻi mua ʻia. Ua hōʻike maopopo ka spectra EPR i ke alo o ʻelua ʻano ROS, kahi i lanakila ai ʻo O2-, ʻoiai ʻo OH• i loaʻa i kahi liʻiliʻi. Eia kekahi, ua hōʻike ka hoʻohālikelike pololei ʻana o nā ikaika kiʻekiʻe he kiʻekiʻe loa ka ʻike ROS o ka EWNS i hoʻonui ʻia i hoʻohālikelike ʻia me ka baseline EWNS.
Ma ke kiʻi 4 e hōʻike ana i ka pono o ka waiho ʻana o EWNS ma EPES. Ua hōʻuluʻulu ʻia nā ʻikepili ma ka Papa I a ua hoʻohālikelike ʻia me ka ʻikepili EWNS mua. No nā hihia EUNS ʻelua, ua kokoke ka waiho ʻana i ka 100% ʻoiai ma kahi volta haʻahaʻa o 3.0 kV. ʻO ka maʻamau, ua lawa ka 3.0 kV e hoʻokō i ka waiho ʻana o 100% me ka nānā ʻole i ka loli o ka uku ʻili. Ma lalo o nā kūlana like, ʻo ka pono o ka waiho ʻana o ka Baseline-EWNS he 56% wale nō ma muli o ka uku haʻahaʻa (ʻawelike 10 mau electrons no kēlā me kēia EWNS).
Hōʻuluʻulu manaʻo ʻo Kiʻi 5 a me ka Papa 2 i ke kekelē o ka hoʻopau ʻana o nā microorganism i hoʻokomo ʻia ma luna o ka ʻili o nā tōmato ma hope o ka hoʻopili ʻia ʻana i kahi 40,000 #/cm3 EWNS no 45 mau minuke ma lalo o ke kūlana kūpono [-6.5 kV, 4.0 cm]. Ua hōʻike ʻo E. coli a me L. innocua i hoʻokomo ʻia i kahi emi nui o 3.8 log ma hope o 45 mau minuke o ka hoʻopili ʻia ʻana. Ma lalo o nā kūlana like, ua hōʻike ʻo S. enterica i kahi emi haʻahaʻa o 2.2 logs, ʻoiai ʻo S. cerevisiae lāua ʻo M. parafortuitum i hōʻike i kahi emi 1.0 log.
Nā kiʻi uila uila (Kiʻi 6) e hōʻike ana i nā loli kino i hoʻokumu ʻia e EWNS i loko o nā pūnaewele E. coli, Salmonella enterica, a me L. innocua e alakaʻi ana i ka hoʻopau ʻana. Ua hōʻike nā bacteria kaohi i nā membrane cell i hoʻopau ʻia, ʻoiai ua hōʻino nā bacteria i hōʻike ʻia i nā membrane waho.
Ua hōʻike ke kiʻi microscope electron o ka kaohi a me nā bacteria i hōʻike ʻia i ka hōʻino ʻana o ka membrane.
Hōʻike pū nā ʻikepili ma nā waiwai physicochemical o ka EWNS i hoʻonui ʻia ua hoʻomaikaʻi nui ʻia nā waiwai EWNS (ka uku ʻili a me ka ʻike ROS) i hoʻohālikelike ʻia me ka ʻikepili baseline EWNS i paʻi mua ʻia8,9,10,11. Ma ka ʻaoʻao ʻē aʻe, ua noho ko lākou nui ma ka pae nanometer, kahi like loa me nā hopena i paʻi mua ʻia, e ʻae ana iā lākou e noho i ka lewa no ka manawa lōʻihi. Hiki ke wehewehe ʻia ka polydispersity i ʻike ʻia e nā loli i ka uku ʻili, ka mea e hoʻoholo ai i ka nui o ka hopena Rayleigh, randomness, a me ka hiki ke hui pū ʻana o EWNS. Eia naʻe, e like me ka mea i hōʻike kikoʻī ʻia e Nielsen et al.22, ʻo ka uku ʻili kiʻekiʻe e hōʻemi ana i ka mahu ma ka hoʻonui pono ʻana i ka ikehu ʻili/ka tension o ka hāʻule wai. Ua hōʻoia ʻia kēia kumumanaʻo no nā microdroplets22 a me EWNS i kā mākou puke mua8. Hiki i ka nalowale o ka overtime ke hoʻopilikia i ka nui a hāʻawi i ka hoʻolaha nui i ʻike ʻia.
Eia kekahi, ʻo ka uku no kēlā me kēia hale ma kahi o 22–44 e-, ma muli o nā kūlana, ʻoi aku ka kiʻekiʻe ma mua o ka EWNS kumu, nona ka uku awelika o 10 ± 2 mau electrons no kēlā me kēia hale. Eia nō naʻe, pono e hoʻomaopopo ʻia ʻo ia ka uku awelika o EWNS. ʻO Seto et al. Ua hōʻike ʻia ʻaʻole like ka uku a hahai i kahi hoʻolaha log-normal21. Ke hoʻohālikelike ʻia me kā mākou hana ma mua, ʻo ka pālua ʻana o ka uku ʻili e pāpālua i ka pono o ka waiho ʻana ma ka ʻōnaehana EPES i aneane 100%11.