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Kwanan nan, an ƙirƙiri wani dandamalin ƙwayoyin cuta marasa sinadarai wanda ya dogara da fasahar nano ta amfani da nanostructures na ruwa na wucin gadi (EWNS). EWNS suna da babban cajin saman kuma suna cike da nau'ikan iskar oxygen mai amsawa (ROS) waɗanda zasu iya hulɗa da kuma kashe wasu ƙwayoyin cuta, gami da ƙwayoyin cuta masu ɗauke da abinci. A nan an nuna cewa halayensu yayin haɗawa za a iya gyara su kuma a inganta su don ƙara haɓaka ƙarfin maganin kashe ƙwayoyin cuta. An tsara dandamalin dakin gwaje-gwaje na EWNS don daidaita halayen EWNS ta hanyar canza sigogin haɗawa. Halayyar halayen EWNS (caji, girma da abun ciki na ROS) ta amfani da hanyoyin bincike na zamani. Bugu da ƙari, an kimanta su don yuwuwar hana ƙwayoyin cuta a kan ƙwayoyin cuta masu ɗauke da abinci kamar Escherichia coli, Salmonella enterica, Listeria innocuous, Mycobacterium paraaccidentum da Saccharomyces cerevisiae. Sakamakon da aka gabatar a nan ya nuna cewa kaddarorin EWNS za a iya daidaita su yayin haɗawa, wanda ke haifar da ƙaruwar inganci a hana aiki. Musamman, cajin saman ya ƙaru da kashi huɗu kuma nau'in iskar oxygen mai amsawa ya ƙaru. Yawan cire ƙwayoyin cuta ya dogara ne akan ƙwayoyin cuta kuma ya kama daga 1.0 zuwa 3.8 log bayan an shafe shi na minti 45 da maganin aerosol na 40,000 #/cc EWNS.
Gurɓatar ƙwayoyin cuta ita ce babbar hanyar kamuwa da cututtukan abinci da ake samu ta hanyar shan ƙwayoyin cuta ko gubarsu. A Amurka kaɗai, cututtukan da ake samu ta hanyar abinci suna haifar da cututtuka kusan miliyan 76, shigar da mutane 325,000 asibiti, da kuma mutuwar mutane 5,000 kowace shekara1. Bugu da ƙari, Ma'aikatar Aikin Gona ta Amurka (USDA) ta kiyasta cewa ƙaruwar cin amfanin gona sabo ne ke haifar da kashi 48% na duk cututtukan da aka ruwaito a cikin abinci a Amurka2. Kuɗin cututtuka da mace-mace da ƙwayoyin cuta da ake samu ta hanyar abinci a Amurka yana da yawa sosai, kamar yadda Cibiyar Kula da Cututtuka da Rigakafi (CDC) ta kiyasta a kan sama da dala biliyan 15.6 a kowace shekara3.
A halin yanzu, hanyoyin magance ƙwayoyin cuta na sinadarai4, radiation5 da thermal6 don tabbatar da amincin abinci galibi ana gudanar da su ne a wurare masu mahimmanci na kulawa (CCPs) tare da sarkar samarwa (yawanci bayan girbi da/ko lokacin marufi) maimakon ci gaba. don haka, suna iya kamuwa da gurɓatawa. 7. Ingantaccen kula da cututtukan da abinci ke haifarwa da lalacewar abinci yana buƙatar hanyoyin magance ƙwayoyin cuta waɗanda za a iya amfani da su a duk faɗin gona-da-teburi yayin da ake rage tasirin muhalli da farashi.
Kwanan nan, an ƙirƙiri wani dandamalin ƙwayoyin cuta marasa sinadarai, wanda aka gina bisa fasahar nanotechnology wanda zai iya kashe ƙwayoyin cuta na saman da na iska ta amfani da nanostructures na ruwa na wucin gadi (EWNS). An haɗa EWNS ta amfani da hanyoyi guda biyu masu layi ɗaya, electrospray da ruwa ionization (Hoto na 1a). Nazarin da aka yi a baya sun nuna cewa EWNS suna da takamaiman sifofi na zahiri da na halitta8,9,10. EWNS suna da matsakaicin electrons 10 a kowane tsari da matsakaicin girman nanoscale na 25 nm (Hoto na 1b,c)8,9,10. Bugu da ƙari, electron spin resonance (ESR) ya nuna cewa EWNS yana ɗauke da adadi mai yawa na nau'in oxygen mai amsawa (ROS), galibi hydroxyl (OH•) da superoxide (O2-) radicals (Hoto na 1c)8. EVNS yana cikin iska na dogon lokaci kuma yana iya karo da ƙananan ƙwayoyin cuta da aka dakatar a cikin iska kuma suna nan a saman, yana isar da nauyin ROS ɗinsu kuma yana haifar da rashin kunna ƙananan ƙwayoyin cuta (Hoto na 1d). Waɗannan binciken farko sun kuma nuna cewa EWNS na iya mu'amala da kuma kashe ƙwayoyin cuta daban-daban na gram-negative da gram-positive, gami da mycobacteria, a saman da kuma a cikin iska. Binciken na'urar hangen nesa ta hanyar watsawa ya nuna cewa rashin kunna ƙwayoyin cuta ya faru ne sakamakon katsewar membrane na tantanin halitta. Bugu da ƙari, nazarin shaƙatawa mai tsanani ya nuna cewa yawan allurai na EWNS ba sa haifar da lalacewar huhu ko kumburi 8.
(a) Feshin lantarki yana faruwa ne lokacin da aka yi amfani da babban ƙarfin lantarki tsakanin bututun capillary wanda ke ɗauke da ruwa da kuma na'urar lantarki mai katsewa. (b) Aiwatar da babban matsin lamba yana haifar da abubuwa biyu daban-daban: (i) feshin ruwa na lantarki da (ii) samuwar nau'ikan iskar oxygen masu amsawa (ions) da aka makale a cikin EWNS. (c) Tsarin EWNS na musamman. (d) Saboda yanayin nanoscale ɗinsu, EWNS suna da motsi sosai kuma suna iya hulɗa da ƙwayoyin cuta masu ɗauke da iska.
An kuma nuna ikon da dandamalin maganin rigakafi na EWNS ke da shi na hana ƙwayoyin cuta da ke shiga abinci a saman abinci mai sabo kwanan nan. An kuma nuna cewa ana iya amfani da ƙarfin saman EWNS tare da filin lantarki don cimma nasarar isar da sako da aka yi niyya. Bugu da ƙari, sakamakon farko na tumatir na halitta bayan an shafe minti 90 a EWNS na kimanin 50,000 #/cm3 ya kasance mai ƙarfafa gwiwa, tare da lura da ƙwayoyin cuta daban-daban na abinci kamar E. coli da Listeria 11. Bugu da ƙari, gwaje-gwajen farko na organoleptic ba su nuna wani tasirin ji ba idan aka kwatanta da tumatir mai sarrafawa. Duk da cewa waɗannan sakamakon farko na hana aiki suna ƙarfafawa ga aikace-aikacen aminci na abinci koda a ƙananan allurai na EWNS na 50,000 #/cc. duba, a bayyane yake cewa babban yuwuwar hana aiki zai fi amfani don ƙara rage haɗarin kamuwa da cuta da lalacewa.
A nan, za mu mayar da hankali kan bincikenmu kan haɓaka dandamalin samar da EWNS don ba da damar daidaita sigogin haɗa sinadarai da inganta halayen sinadaran EWNS don haɓaka ƙarfin ƙwayoyin cuta. Musamman ma, haɓakawa ya mayar da hankali kan ƙara yawan cajin saman su (don inganta isar da aka yi niyya) da abubuwan da ke cikin ROS (don inganta ingancin hana aiki). Bayyana halayen sinadarai masu inganci (girma, caji da abubuwan da ke cikin ROS) ta amfani da hanyoyin nazari na zamani kuma yi amfani da ƙananan ƙwayoyin cuta na abinci kamar E.
An haɗa EVNS ta hanyar fesawa da kuma ionization na ruwa mai tsarki (18 MΩ cm–1). Ana amfani da nebulizer na lantarki 12 don atomization na ruwa da kuma haɗa ƙwayoyin polymer da yumbu 13 da zare 14 na girman da aka sarrafa.
Kamar yadda aka yi bayani dalla-dalla a cikin wallafe-wallafen da suka gabata na 8, 9, 10, 11, a cikin wani gwaji na yau da kullun, an yi amfani da babban ƙarfin lantarki tsakanin ƙarfe mai kauri da kuma na'urar lantarki mai kauri. A lokacin wannan tsari, abubuwa biyu daban-daban sun faru: i) feshi na lantarki da ii) ionization na ruwa. Filin lantarki mai ƙarfi tsakanin lantarki biyu yana haifar da tarawar caji mara kyau a saman ruwan da aka tanada, wanda ke haifar da samuwar mazubin Taylor. Sakamakon haka, ana samun ɗigon ruwa mai caji sosai, wanda ke ci gaba da wargajewa zuwa ƙananan ƙwayoyin cuta, kamar yadda yake a cikin ka'idar Rayleigh16. A lokaci guda, filayen lantarki masu ƙarfi suna sa wasu ƙwayoyin ruwa su rarrabu kuma su cire electrons (ionize), wanda ke haifar da samuwar nau'in oxygen mai amsawa (ROS)17. An lulluɓe ROS18 da aka samar a lokaci guda a cikin EWNS (Hoto na 1c).
A hoto na 2a, an nuna tsarin samar da EWNS da aka haɓaka kuma aka yi amfani da shi a cikin haɗakar EWNS a cikin wannan binciken. An zuba ruwan da aka tsaftace a cikin kwalba mai rufe ta hanyar bututun Teflon (diamita na ciki 2 mm) zuwa cikin allurar ƙarfe mai nauyin 30G (ƙafafun ƙarfe). Ana sarrafa kwararar ruwa ta hanyar matsin iska da ke cikin kwalbar, kamar yadda aka nuna a Hoto na 2b. An ɗora allurar a kan na'urar Teflon kuma ana iya daidaita ta da hannu zuwa wani takamaiman nisa daga na'urar lantarki. Na'urar lantarki mai ƙirƙiro faifai ne na aluminum mai gogewa tare da rami a tsakiya don ɗaukar samfur. A ƙarƙashin na'urar lantarki mai ƙirƙiro akwai mazubin samfurin aluminum, wanda aka haɗa shi da sauran saitin gwaji ta hanyar tashar ɗaukar samfuri (Hoto na 2b). Don guje wa tara caji wanda zai iya kawo cikas ga aikin na'urar samfuri, duk abubuwan da ke cikin na'urar samfurin an yi su ne da wutar lantarki.
(a) Tsarin Samar da Tsarin Ruwa Mai Inganci (EWNS). (b) Sashe na gaba na samfurin da feshi na lantarki, wanda ke nuna mahimman sigogi. (c) Saitin gwaji don hana ƙwayoyin cuta aiki.
Tsarin samar da EWNS da aka bayyana a sama yana da ikon canza mahimman sigogin aiki don sauƙaƙe daidaita halayen EWNS. Daidaita ƙarfin lantarki da aka yi amfani da shi (V), nisan da ke tsakanin allura da na'urar lantarki ta counter (L), da kuma kwararar ruwa (φ) ta cikin capillary don daidaita halayen EWNS. Alamar da aka yi amfani da ita don wakiltar haɗuwa daban-daban: [V (kV), L (cm)]. Daidaita kwararar ruwa don samun mazugi Taylor mai ƙarfi na wani saiti [V, L]. Don dalilan wannan binciken, an ajiye diamita na buɗewar na'urar lantarki ta counter (D) a inci 0.5 (1.29 cm).
Saboda ƙarancin yanayin ƙasa da rashin daidaituwa, ba za a iya ƙididdige ƙarfin filin lantarki daga ƙa'idodin farko ba. Madadin haka, an yi amfani da software na QuickField™ (Svendborg, Denmark)19 don ƙididdige filin lantarki. Filin lantarki ba iri ɗaya ba ne, don haka an yi amfani da ƙimar filin lantarki a ƙarshen capillary azaman ƙimar tunani don tsari daban-daban.
A lokacin binciken, an kimanta haɗuwa da dama na ƙarfin lantarki da tazara tsakanin allura da na'urar lantarki ta counter dangane da samuwar mazubin Taylor, kwanciyar hankalin mazubin Taylor, kwanciyar hankalin samar da EWNS, da kuma sake haifuwa. An nuna haɗuwa daban-daban a cikin Tebur na Ƙarin S1.
An haɗa fitarwar tsarin samar da EWNS kai tsaye zuwa na'urar nazarin girman barbashi ta Scanning Mobility (SMPS, Model 3936, TSI, Shoreview, MN) don auna yawan barbashi, da kuma na'urar aunawa ta Aerosol Faraday (TSI, Model 3068B, Shoreview, MN).) don kwararar aerosol kamar yadda aka bayyana a cikin littafinmu na baya. An auna duka SMPS da na'urar aunawa ta aerosol a ƙimar kwararar 0.5 L/min (jimillar kwararar samfurin 1 L/min). An auna yawan adadin barbashi da kwararar aerosol na daƙiƙa 120. An maimaita ma'aunin sau 30. Dangane da ma'aunin yanzu, an ƙididdige jimlar cajin aerosol kuma an kiyasta matsakaicin cajin EWNS don jimlar adadin barbashi EWNS da aka zaɓa. Ana iya ƙididdige matsakaicin farashin EWNS ta amfani da Lissafi (1):
inda IEl shine ma'aunin wutar lantarki da aka auna, NSMPS shine ma'aunin dijital da aka auna tare da SMPS, kuma φEl shine ma'aunin kwararar kowace na'urar lantarki.
Saboda yanayin zafi (RH) yana shafar cajin saman, an kiyaye zafin jiki da (RH) a daidai lokacin gwajin a 21°C da 45%, bi da bi.
An yi amfani da na'urar duba hotuna ta ƙarfin atomic (AFM), Asylum MFP-3D (Asylum Research, Santa Barbara, CA) da kuma AC260T probe (Olympus, Tokyo, Japan) don auna girman da tsawon rayuwar EWNS. Mitar duba hotuna ta AFM ita ce 1 Hz, yankin duba hotuna ita ce 5 μm × 5 μm, da kuma layukan duba hotuna 256. An yi wa dukkan hotuna tsarin daidaitawa ta farko ta amfani da manhajar Asylum (tsawon abin rufe fuska 100 nm, iyakar 100 na dare).
An cire mazubin gwaji kuma an sanya saman mica a nisan santimita 2.0 daga na'urar lantarki ta counter na tsawon lokacin daƙiƙa 120 don guje wa taruwar ƙwayoyin cuta da samuwar ɗigon da ba su dace ba a saman mica. An fesa EWNS kai tsaye a saman mica da aka yanke (Ted Pella, Redding, CA). Hoton saman mica nan da nan bayan AFM ya yi tururi. Kusurwar hulɗar saman mica da aka yanke ba tare da an gyara ta kusa da 0° ba, don haka ana rarraba EVNS a saman mica a cikin siffar kumfa. An auna diamita (a) da tsayi (h) na ɗigon da ke yaɗuwa kai tsaye daga yanayin AFM kuma an yi amfani da shi don ƙididdige girman yaɗuwar EWNS ta amfani da hanyar da muka tabbatar a baya. Idan aka yi la'akari da cewa EWNS da ke cikin jirgin suna da girma ɗaya, ana iya ƙididdige diamita daidai ta amfani da Daidaito (2):
Dangane da hanyar da muka ƙirƙiro a baya, an yi amfani da tarkon juyawa na electron spin resonance (ESR) don gano kasancewar tsaka-tsakin radical masu ɗan gajeren lokaci a cikin EWNS. An yi amfani da iskar gas ta hanyar 650 μm Midget sparger (Ace Glass, Vineland, NJ) wanda ke ɗauke da maganin DEPMPO(5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide) mai girman 235 mM (Oxis International Inc.). Portland, Oregon). An yi duk ma'aunin ESR ta amfani da na'urar Bruker EMX spectrometer (Bruker Instruments Inc. Billerica, MA, USA) da kuma tantanin faifan lebur. An yi amfani da manhajar Acquisit (Bruker Instruments Inc. Billerica, MA, USA) don tattarawa da kuma nazarin bayanan. An gudanar da tantance halayen ROS ne kawai don saitin yanayin aiki [-6.5 kV, 4.0 cm]. An auna yawan EWNS ta amfani da SMPS bayan an yi lissafin asarar EWNS a cikin mai tasiri.
An yi amfani da na'urar lura da matakan Ozone ta amfani da na'urar lura da Ozone Monitor™ (2B Technologies, Boulder, Co) 8,9,10.
Ga duk kaddarorin EWNS, ana amfani da matsakaicin ƙimar azaman ƙimar aunawa, kuma ana amfani da daidaitaccen karkacewar azaman kuskuren aunawa. An gudanar da gwaje-gwajen T don kwatanta ƙimar halayen EWNS da aka inganta tare da ƙimar da ta dace da EWNS na tushe.
Siffa ta 2c ta nuna tsarin "jawo" na hazo na lantarki (EPES) wanda aka riga aka ƙirƙira kuma aka siffanta shi da shi wanda za a iya amfani da shi don isar da EWNS a saman. EPES yana amfani da cajin EVNS wanda za a iya "jagora" kai tsaye zuwa saman abin da aka nufa a ƙarƙashin tasirin filin lantarki mai ƙarfi. An gabatar da cikakkun bayanai game da tsarin EPES a cikin wani wallafe-wallafen da Pyrgiotakis et al. 11 suka buga kwanan nan. Don haka, EPES ya ƙunshi ɗakin PVC mai bugawa na 3D tare da ƙarshen da aka yi wa kaifi kuma ya ƙunshi faranti biyu na ƙarfe masu layi ɗaya (ƙarfe 304, mai rufi da madubi) a tsakiya 15.24 cm baya. An haɗa allunan zuwa tushen wutar lantarki mai ƙarfi na waje (Bertran 205B-10R, Spellman, Hauppauge, NY), farantin ƙasa koyaushe yana da alaƙa da ƙarfin lantarki mai kyau, kuma farantin saman koyaushe yana da alaƙa da ƙasa (ƙasa mai iyo). Bangon ɗakin an rufe shi da foil ɗin aluminum, wanda aka girka ta hanyar lantarki don hana asarar barbashi. Ɗakin yana da ƙofar ɗaukar kaya ta gaba da aka rufe wadda ke ba da damar sanya saman gwaji a kan matattarar filastik waɗanda ke ɗaga su sama da farantin ƙarfe na ƙasa don guje wa tsangwama mai ƙarfi.
An ƙididdige ingancin ajiyar EWNS a cikin EPES bisa ga wata yarjejeniya da aka ƙirƙiro a baya da aka yi cikakken bayani a cikin Ƙarin Hoto na S111.
A matsayin ɗakin sarrafawa, an haɗa ɗakin kwarara na biyu mai siffar silinda a jere zuwa tsarin EPES, inda aka yi amfani da matattarar HEPA ta tsakiya don cire EWNS. Kamar yadda aka nuna a Hoto na 2c, an tura iskar EWNS ta cikin ɗakuna biyu da aka gina a ciki. Matattarar da ke tsakanin ɗakin sarrafawa da EPES tana cire duk wani EWNS da ya rage wanda ya haifar da yanayin zafi iri ɗaya (T), ɗanɗanon dangi (RH) da matakan ozone.
An gano cewa muhimman ƙwayoyin cuta da ake samu daga abinci suna gurɓata abinci sabo kamar E. coli (ATCC #27325), alamar najasa, Salmonella enterica (ATCC #53647), ƙwayoyin cuta na abinci, Listeria harmless (ATCC #33090), maye gurbin ƙwayoyin cuta na Listeria monocytogenes, waɗanda aka samo daga ATCC (Manassas, VA) Saccharomyces cerevisiae (ATCC #4098), madadin yisti mai lalacewa, da kuma ƙwayar cuta da ba ta aiki ba, Mycobacterium paralucky (ATCC #19686).
Sayi akwatunan tumatir na innabi na halitta daga kasuwar yankinku kuma a sanyaya a cikin firiji a zafin digiri 4 na Celsius har sai an yi amfani da su (har zuwa kwana 3). Tumatir ɗin da aka gwada duk girmansu iri ɗaya ne, kimanin inci 1/2 a diamita.
An yi cikakken bayani game da ka'idojin al'ada, allurar rigakafi, fallasa, da kuma ƙididdige ƙwayoyin cuta a cikin littafinmu na baya kuma an yi cikakken bayani a cikin Ƙarin Bayanan. An kimanta ingancin EWNS ta hanyar fallasa tumatir da aka yi wa allura zuwa 40,000 #/cm3 na tsawon mintuna 45. A takaice, an yi amfani da tumatir uku don tantance ƙwayoyin cuta da suka tsira a lokacin t = minti 0. An sanya tumatir uku a cikin EPES kuma an fallasa su ga EWNS a 40,000 #/cc (tumatir da aka fallasa EWNS) kuma an sanya sauran ukun a cikin ɗakin sarrafawa (tumatir da aka sarrafa). Ba a gudanar da ƙarin sarrafa tumatir a cikin ƙungiyoyi biyu ba. An cire tumatir da tumatir da aka fallasa EWNS bayan mintuna 45 don tantance tasirin EWNS.
An gudanar da kowace gwaji sau uku. An gudanar da nazarin bayanai bisa ga ka'idar da aka bayyana a cikin Ƙarin Bayanan.
An tantance hanyoyin hana aiki ta hanyar tace samfuran EWNS da aka fallasa (minti 45 a cikin taro mai ƙarfi na EWNS 40,000 #/cm3) da samfuran ƙwayoyin cuta marasa lahani E. coli, Salmonella enterica da Lactobacillus. An gyara ƙwayoyin a cikin 2.5% glutaraldehyde, 1.25% paraformaldehyde da 0.03% picric acid a cikin 0.1 M sodium cacodylate buffer (pH 7.4) na tsawon awanni 2 a zafin ɗaki. Bayan wankewa, bayan an gama gyarawa da 1% osmium tetroxide (OsO4)/1.5% potassium ferrocyanide (KFeCN6) na tsawon awanni 2, a wanke sau 3 a cikin ruwa sannan a saka a cikin 1% uranyl acetate na tsawon awa 1, sannan a wanke sau biyu a cikin ruwa, sannan a bar shi ya bushe na tsawon mintuna 10 a cikin 50%, 70%, 90%, 100% barasa. An sanya samfuran a cikin propylene oxide na tsawon awa 1 sannan aka saka su a cikin ruwan propylene oxide na tsawon awa 1 sannan aka saka su a cikin ruwan da aka haɗa da propylene oxide da TAAP Epon (Marivac Canada Inc. St. Laurent, CA). An saka samfuran a cikin TAAB Epon kuma an yi musu polymer a zafin 60°C na tsawon awanni 48. An yanke resin granular da aka warke kuma an nuna shi ta hanyar TEM ta amfani da na'urar hangen nesa ta lantarki ta zamani JEOL 1200EX (JEOL, Tokyo, Japan) wacce aka sanye da kyamarar AMT 2k CCD (Advanced Microscopy Techniques, Corp., Woburn, Massachusetts, Amurka).
An gudanar da dukkan gwaje-gwajen a matakai uku. A kowane lokaci, an yi wa ƙwayoyin cuta wanka sau uku, wanda ya haifar da jimillar maki tara na bayanai a kowane maki, wanda aka yi amfani da matsakaicinsa azaman yawan ƙwayoyin cuta ga wannan takamaiman ƙwayoyin cuta. An yi amfani da karkacewar da aka saba amfani da ita azaman kuskuren aunawa. Duk maki suna ƙidaya.
An ƙididdige logarithm na raguwar yawan ƙwayoyin cuta idan aka kwatanta da t = 0 min ta amfani da dabarar da ke ƙasa:
inda C0 shine yawan ƙwayoyin cuta a cikin samfurin sarrafawa a lokacin 0 (watau bayan saman ya bushe amma kafin a sanya shi a cikin ɗakin) kuma Cn shine yawan ƙwayoyin cuta a saman bayan mintuna na n na fallasa.
Domin yin la'akari da lalacewar ƙwayoyin cuta ta halitta yayin fallasa na mintuna 45, an kuma ƙididdige raguwar rajista idan aka kwatanta da sarrafawa bayan mintuna 45 kamar haka:
inda Cn shine yawan ƙwayoyin cuta a cikin samfurin sarrafawa a lokacin n kuma Cn-Control shine yawan ƙwayoyin cuta masu sarrafawa a lokacin n. An gabatar da bayanai a matsayin rage log idan aka kwatanta da sarrafawa (babu fallasa EWNS).
A lokacin binciken, an kimanta haɗuwar ƙarfin lantarki da nisa tsakanin allura da na'urar lantarki ta counter dangane da samuwar mazugi na Taylor, kwanciyar hankalin mazugi na Taylor, kwanciyar hankalin samar da EWNS, da kuma sake haifuwa. An nuna haɗuwa iri-iri a cikin Teburin Ƙarin S1. An zaɓi shari'o'i biyu da ke nuna halaye masu karko da na sake haifuwa (mazugi na Taylor, samar da EWNS, da kwanciyar hankali akan lokaci) don cikakken bincike. A kan Hoto na 3 yana nuna sakamakon caji, girma, da abun ciki na ROS a duka shari'o'in. Hakanan an taƙaita sakamakon a cikin Teburi na 1. Don tunani, duka Hoto na 3 da Teburi na 1 sun haɗa da halayen EWNS8, 9, 10, 11 da aka haɗa a baya (asalin-EWNS). An sake buga ƙididdigar mahimmancin ƙididdiga ta amfani da gwajin t-jeri biyu a cikin Teburin Ƙarin S2. Bugu da ƙari, ƙarin bayanai sun haɗa da nazarin tasirin diamita ramin samfurin electrode na counter (D) da nisa tsakanin electrode na ƙasa da tip (L) (Hotunan Ƙarin S2 da S3).
(ac) Rarraba girman da aka auna ta hanyar AFM. (df) Siffar cajin saman. (g) Siffar ROS ta EPR.
Yana da mahimmanci a lura cewa ga dukkan yanayin da ke sama, ƙarfin ionization da aka auna yana tsakanin 2 zuwa 6 μA da ƙarfin lantarki tsakanin -3.8 da -6.5 kV, wanda ya haifar da amfani da wutar lantarki ƙasa da 50 mW ga wannan tsarin hulɗa na EWNS guda ɗaya. Duk da cewa an haɗa EWNS a ƙarƙashin matsin lamba mai yawa, matakan ozone sun yi ƙasa sosai, ba su taɓa wuce 60 ppb ba.
Ƙarin Hoto na S4 yana nuna filayen lantarki da aka kwaikwayi don yanayin [-6.5 kV, 4.0 cm] da [-3.8 kV, 0.5 cm] bi da bi. Ga yanayin [-6.5 kV, 4.0 cm] da [-3.8 kV, 0.5 cm], lissafin filin shine 2 × 105 V/m da 4.7 × 105 V/m, bi da bi. Ana sa ran wannan, tunda a cikin akwati na biyu rabon nisan wutar lantarki ya fi girma.
A kan hoto na 3a,b, an nuna diamita na EWNS da aka auna da AFM8. Matsakaicin diamita na EWNS da aka ƙididdige sune 27 nm da 19 nm don tsarin [-6.5 kV, 4.0 cm] da [-3.8 kV, 0.5 cm], bi da bi. Ga yanayin [-6.5 kV, 4.0 cm] da [-3.8 kV, 0.5 cm], bambancin daidaito na lissafi na rarrabawa shine 1.41 da 1.45, bi da bi, yana nuna rarraba girman kunkuntar. Matsakaicin girman da karkacewar daidaito na lissafi suna kusa da EWNS na asali, a 25 nm da 1.41, bi da bi. A kan hoto na 3c, an nuna rarraba girman EWNS na tushe da aka auna ta amfani da hanya ɗaya a ƙarƙashin yanayi ɗaya.
A hoto na 3d,e yana nuna sakamakon halayen caji. Bayanai matsakaicin ma'auni ne na ma'auni 30 na maida hankali a lokaci guda (#/cm3) da na yanzu (I). Binciken ya nuna cewa matsakaicin cajin akan EWNS shine 22 ± 6 e- da 44 ± 6 e- don [-6.5 kV, 4.0 cm] da [-3.8 kV, 0.5 cm], bi da bi. Suna da cajin saman da ya fi girma idan aka kwatanta da EWNS na asali (10 ± 2 e-), sau biyu ya fi yanayin [-6.5 kV, 4.0 cm] kuma sau huɗu ya fi [-3 .8 kV, 0.5 cm]. Hoto na 3f yana nuna cajin. bayanai don Baseline-EWNS.
Daga taswirar tattarawar adadin EWNS (Ƙarin Hotuna S5 da S6), za a iya ganin cewa yanayin [-6.5 kV, 4.0 cm] yana da ƙwayoyin cuta da yawa fiye da yanayin [-3.8 kV, 0.5 cm]. Hakanan yana da mahimmanci a lura cewa an sa ido kan yawan adadin EWNS har zuwa awanni 4 (Ƙarin Hotuna S5 da S6), inda daidaiton samar da EWNS ya nuna matakan yawan adadin ƙwayoyin cuta iri ɗaya a duka lamuran biyu.
A hoto na 3g, an nuna bakan EPR bayan an cire ingantaccen ikon sarrafa EWNS (bayan fage) a [-6.5 kV, 4.0 cm]. An kuma kwatanta bakan ROS da yanayin Baseline-EWNS a cikin wani aiki da aka buga a baya. An ƙididdige adadin EWNS da ke amsawa da tarkunan juyawa zuwa 7.5 × 104 EWNS/s, wanda yayi kama da Baseline-EWNS8 da aka buga a baya. Bakan EPR ya nuna a fili kasancewar nau'ikan ROS guda biyu, inda O2- shine nau'in da ya fi yawa kuma OH• bai cika yawa ba. Bugu da ƙari, kwatanta kai tsaye na ƙarfin kololuwar ya nuna cewa EWNS da aka inganta yana da babban abun ciki na ROS idan aka kwatanta da EWNS na asali.
A hoto na 4 yana nuna ingancin sakawa na EWNS a cikin EPES. An kuma taƙaita bayanan a cikin Tebur na I kuma an kwatanta su da ainihin bayanan EWNS. Ga duka lamuran EUNS, sakawa yana kusa da 100% ko da a ƙaramin ƙarfin lantarki na 3.0 kV. Yawanci, 3.0 kV ya isa don sakawa 100%, ba tare da la'akari da canjin cajin saman ba. A ƙarƙashin yanayi iri ɗaya, ingancin sakawa na Baseline-EWNS ya kasance 56% kawai saboda ƙarancin cajin su (matsakaicin electrons 10 a kowace EWNS).
A hoto na 5 da kuma a tebur. 2 ya taƙaita ƙimar hana aiki na ƙwayoyin cuta da aka yi wa allurar rigakafi a saman tumatir bayan an fallasa su ga kimanin 40,000 #/cm3 EWNS na tsawon mintuna 45 a yanayin da ya fi dacewa [-6.5 kV, 4.0 cm]. An yi wa allurar rigakafin E. coli da Lactobacillus marasa lahani sun nuna raguwar yawan allurar rigakafi 3.8 a lokacin fallasa na mintuna 45. A ƙarƙashin irin wannan yanayi, S. enterica ya sami raguwar 2.2-log, yayin da S. cerevisiae da M. parafortutum suka sami raguwar 1.0-log.
Na'urorin aunawa na electron (Hoto na 6) suna nuna canje-canjen jiki da EWNS ya haifar akan ƙwayoyin Escherichia coli, Streptococcus, da Lactobacillus marasa lahani wanda ya haifar da rashin aiki. Kwayoyin cuta masu sarrafawa suna da membranes na ƙwayoyin halitta marasa lahani, yayin da ƙwayoyin cuta da aka fallasa suka lalata membranes na waje.
Hoton ƙwayoyin cuta masu sarrafawa da kuma waɗanda aka fallasa ta hanyar amfani da na'urar lantarki (electron microscope) ya nuna lalacewar membrane.
Bayanan da aka tattara kan halayen sinadaran sinadarai na EWNS da aka inganta tare sun nuna cewa kaddarorin (cajin saman da abun ciki na ROS) na EWNS sun inganta sosai idan aka kwatanta da bayanan asali na EWNS da aka buga a baya8,9,10,11. A gefe guda kuma, girmansu ya kasance a cikin kewayon nanometer, yayi kama da sakamakon da aka ruwaito a baya, wanda hakan ya ba su damar kasancewa a cikin iska na dogon lokaci. Ana iya bayyana rarrabuwar da aka lura ta hanyar canje-canjen cajin saman da ke tantance girman EWNS, rashin daidaituwar tasirin Rayleigh, da yuwuwar haɗuwa. Duk da haka, kamar yadda Nielsen et al. 22 suka yi bayani dalla-dalla, babban cajin saman yana rage ƙafewa ta hanyar ƙara kuzari/tsananin digowar ruwa yadda ya kamata. A cikin wallafe-wallafenmu na baya8 an tabbatar da wannan ka'ida ta gwaji ga ƙananan droplets 22 da EWNS. Asarar caji yayin ƙarin lokaci na iya shafar girman kuma yana ba da gudummawa ga rarraba girman da aka lura.
Bugu da ƙari, cajin kowace tsari yana da kusan 22-44 e-, ya danganta da yanayin, wanda ya fi girma sosai idan aka kwatanta da EWNS na asali, wanda ke da matsakaicin cajin lantarki 10 ± 2 a kowane tsari. Duk da haka, ya kamata a lura cewa wannan shine matsakaicin cajin EWNS. Seto et al. An nuna cewa cajin ba shi da kama da juna kuma yana bin diddigin log-normal21. Idan aka kwatanta da aikinmu na baya, ninka cajin saman yana ninka ingancin ajiya a cikin tsarin EPES zuwa kusan 100%11.