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Indawo yokhathalelo lwempilo engcolileyo idlala indima ebalulekileyo ekusasazekeni kwezinto eziphilayo ezinganyangekiyo ngamayeza amaninzi (MDR) kunye ne-C. difficile. Injongo yolu phononongo yayikukuvavanya isiphumo se-ozone eveliswa yi-dielectric barrier discharge (DBD) plasma reactor kwisenzo se-vancomycin-resistant Enterococcus faecalis (VRE), i-carbapenem-resistant Klebsiella pneumoniae (CRE), i-carbapenem-resistant Imiphumo ye-antibacterial yezinto ezahlukeneyo ezingcoliswe yi-Pseudomonas spp. Pseudomonas aeruginosa (CRPA), i-carbapenem-resistant Acinetobacter baumannii (CRAB) kunye ne-Clostridium difficile spores. Izinto ezahlukeneyo ezingcoliswe yi-VRE, CRE, CRPA, CRAB kunye ne-C. difficile spores zanyangwa nge-ozone kumanqanaba ahlukeneyo kunye namaxesha okuvezwa. I-Atomic force microscopy (AFM) ibonise utshintsho lomphezulu webhaktheriya emva konyango lwe-ozone. Xa idosi ye-500 ppm ozone isetyenziswa kwi-VRE nakwi-CRAB kangangemizuzu eli-15, kwabonwa ukwehla kwe-log10 malunga ne-2 okanye ngaphezulu kwintsimbi engagqwaliyo, ilaphu kunye nomthi, kwaye kwabonwa ukwehla kwe-log10 kwiglasi nakwiplastiki. Ii-spores ze-C. difficile zifunyenwe zikwazi ukumelana ne-ozone ngaphezu kwazo zonke ezinye izinto eziphilayo ezivavanyiweyo. Kwi-AFM, emva konyango nge-ozone, iiseli zebhaktiriya zadumba zaza zatshintsha. I-ozone eveliswa yi-DBD Plasma Reactor sisixhobo esilula nesixabisekileyo sokucoca i-MDRO kunye ne-C. difficile spores, ezaziwa ngokuba ziintsholongwane eziqhelekileyo zosulelo oluhambelana nokhathalelo lwempilo.
Ukuvela kwezinto eziphilayo ezinganyangekiyo ngamayeza amaninzi (MDR) kubangelwa kukusetyenziswa gwenxa kwee-antibiotics ebantwini nakwizilwanyana kwaye kuchongwe yi-World Health Organisation (WHO) njengosongelo olukhulu kwimpilo yoluntu1. Ngokukodwa, amaziko ezempilo ajongene nokuvela nokusasazeka kwee-MRO. Ii-MRO eziphambili yi-methicillin-resistant Staphylococcus aureus kunye ne-vancomycin-resistant enterococcus (VRE), i-extended-spectrum beta-lactamase-producing enterobacteria (ESBL), i-Pseudomonas aeruginosa enganyangekiyo ngamayeza amaninzi, i-Acinetobacter baumannii enganyangekiyo ngamayeza amaninzi, kunye ne-carbapenem-resistant Enterobacter (CRE). Ukongeza, usulelo lweClostridium difficile luyimbangela ephambili yorhudo oluhambelana nezempilo, olubeka umthwalo omkhulu kwinkqubo yezempilo. I-MDRO kunye ne-C. difficile zidluliselwa ngezandla zabasebenzi bezempilo, iindawo ezingcolisiweyo, okanye ngqo ukusuka komnye umntu ukuya komnye. Izifundo zakutshanje zibonise ukuba iindawo ezingcolisiweyo kwiindawo zokhathalelo lwempilo zidlala indima ebalulekileyo ekudluliseleni i-MDRO kunye ne-C. difficile xa abasebenzi bezempilo (ii-HCW) bedibana neendawo ezingcolisiweyo okanye xa izigulana zidibana ngqo neendawo ezingcolisiweyo 3,4. iindawo ezingcolisiweyo kwiindawo zokhathalelo lwempilo zinciphisa ukwanda kosulelo lwe-MLRO kunye ne-C. difficile okanye ukungena kwikoloni5,6,7. Ngenxa yenkxalabo yehlabathi malunga nokwanda kokuxhathisa iintsholongwane, kuyacaca ukuba uphando oluthe kratya luyadingeka kwiindlela kunye neenkqubo zokucoca kwiindawo zokhathalelo lwempilo. Kutshanje, iindlela zokucoca i-terminal ezingadibaniyo, ngakumbi izixhobo ze-ultraviolet (UV) okanye iinkqubo ze-hydrogen peroxide, ziye zaqatshelwa njengeendlela ezithembisayo zokucoca. Nangona kunjalo, ezi zixhobo ze-UV okanye ze-hydrogen peroxide ezifumaneka kurhwebo azibizi nje kuphela, ukucoca i-UV kusebenza kuphela kwiindawo eziveziweyo, ngelixa ukucoca i-hydrogen peroxide kwi-plasma kufuna ixesha elide lokucoca ngaphambi komjikelo olandelayo wokucoca i-5.
I-Ozone ineempawu ezaziwayo zokulwa nokugqwala kwaye inokuveliswa ngexabiso eliphantsi8. Ikwaziwa nokuba inobungozi kwimpilo yabantu, kodwa inokubola ngokukhawuleza ibe yioksijini8. Ii-reactors ze-plasma ze-Dielectric barrier discharge (DBD) zezona zivelisi ze-ozone zixhaphakileyo9. Izixhobo ze-DBD zikuvumela ukuba wenze i-plasma enobushushu obuphantsi emoyeni kwaye uvelise i-ozone. Ukuza kuthi ga ngoku, ukusetyenziswa kwe-ozone ngokuyintloko kunqunyelwe ekubulaleni iintsholongwane emanzini edamini lokudada, amanzi okusela kunye nelindle10. Izifundo ezininzi zibike ukusetyenziswa kwayo kwiindawo zonyango8,11.
Kolu phononongo, sisebenzise i-compact DBD plasma ozone generator ukubonisa ukusebenza kwayo ekucoceni i-MDRO kunye ne-C. difficile, kwanaleyo ifakwe kwiimathiriyeli ezahlukeneyo ezisetyenziswa kakhulu kwiindawo zonyango. Ukongeza, inkqubo yokubulala iintsholongwane kwi-ozone icacisiwe kusetyenziswa imifanekiso ye-atomic force microscopy (AFM) yeeseli eziphathwe nge-ozone.
Iintlobo zesifo zifunyenwe kwi-isolates zeklinikhi ze: VRE (SCH 479 kunye ne-SCH 637), Klebsiella pneumoniae engamelaniyo ne-carbapenem (CRE; SCH CRE-14 kunye ne-DKA-1), Pseudomonas aeruginosa engamelana ne-carbapenem (CRPA; 54 kunye ne-83) kunye ne-carbapenem engamelana ne-bacteria. iibacteria Pseudomonas aeruginosa (CRPA; 54 kunye ne-83). I-Acinetobacter baumannii engamelana nayo (CRAB; F2487 kunye ne-SCH-511). I-C. difficile ifunyenwe kwi-National Pathogen Culture Collection (NCCP 11840) ye-Korea Agency for Disease Control and Prevention. Yahlulwe kwisigulana eMzantsi Korea ngo-2019 kwaye yafunyaniswa ukuba ikwi-ST15 kusetyenziswa i-multilocus sequence typing. Umhluzi weBrain Heart Infusion (BHI) (BD, Sparks, MD, USA) ofakwe i-VRE, CRE, CRPA kunye ne-CRAB uxutywe kakuhle waza wafakwa kwi-incubator kuma-37° C. iiyure ezingama-24.
I-C. difficile yafakwa imicu kwi-agar yegazi kangangeeyure ezingama-48. Emva koko kongezwa iikholoni ezininzi kwi-5 ml yomhluzi wentliziyo yobuchopho kwaye yafakwa phantsi kweemeko ze-anaerobic iiyure ezingama-48. Emva koko, inkcubeko yashukunyiswa, yongezwa i-5 ml ye-95% ye-ethanol, yashukunyiswa kwakhona yaze yashiywa kubushushu begumbi imizuzu engama-30. Emva kokufakwa kwi-centrifugation kwi-3000 g imizuzu engama-20, lahla i-supernatant kwaye umise i-pellet equlethe ii-spores kwaye ubulale iintsholongwane kwi-0.3 ml yamanzi. Iiseli eziphilayo zabalwa ngokuhlwayela imbewu ye-spiral ye-bacterial cell suspension kwiiplate ze-agar yegazi emva kokuxutywa okufanelekileyo. Ukudibanisa iigram kuqinisekisile ukuba i-85% ukuya kwi-90% yezakhiwo zebhaktiriya yayizii-spores.
Olu phononongo lulandelayo lwenziwe ukuze kuphandwe imiphumo ye-ozone njengesibulali-ntsholongwane kwiindawo ezahlukeneyo ezingcoliswe yi-MDRO kunye ne-C. difficile spores, ezaziwa ngokuba zibangela usulelo olunxulumene nokhathalelo lwempilo. Lungisa iisampulu zentsimbi engagqwaliyo, ilaphu (umqhaphu), iglasi, iplastiki (i-acrylic), kunye nomthi (ipayini) olinganisa isentimitha enye ngesentimitha enye. Hlambulula iintsholongwane kwiikhuphoni ngaphambi kokusetyenziswa. Zonke iisampulu zihlanjululwe nge-autoclaving ngaphambi kokuba zosulelwe yibhaktheriya.
Kolu phononongo, iiseli zebhaktiriya zasasazwa kwiindawo ezahlukeneyo ze-substrate kunye nakwiiplate ze-agar. Iiphaneli emva koko ziyacocwa ngokuzifaka kwi-ozone ixesha elithile nakwizinga elithile kwigumbi elivaliweyo. Kumfanekiso 1 kukho umfanekiso wezixhobo zokucoca i-ozone. Ii-reactor ze-DBD plasma zenziwe ngokuncamathisela ii-electrode zentsimbi engenasici ezivulekileyo neziveziweyo ngaphambili nangasemva kweepleyiti ze-alumina (dielectric) ezinobukhulu obuyi-1 mm. Kwii-electrode ezivulekileyo, indawo yokuvuleka kunye nomngxuma yayiyi-3 mm kunye ne-0.33 mm, ngokulandelanayo. I-electrode nganye inomfanekiso ongqukuva onobubanzi obuyi-43 mm. Umbane ophezulu we-voltage ephezulu (GBS Elektronik GmbH Minipuls 2.2) wasetyenziselwa ukusebenzisa i-voltage ye-sinusoidal emalunga ne-8 kV peak ukuya kwi-frequency ye-12.5 kHz kwii-electrode ezivulekileyo ukuvelisa i-plasma emaphethelweni ee-electrode. Ekubeni le ndlela yokubulala iintsholongwane ngegesi, ukubulala iintsholongwane kwenziwa kwigumbi elahlulwe ngobuninzi libe ziindawo eziphezulu nezisezantsi, eziqulathe iisampulu ezingcoliswe ziintsholongwane kunye neejenereyitha zeplasma, ngokwahlukeneyo. Igumbi eliphezulu lineendawo ezimbini zokukhupha nokukhupha i-ozone eseleyo. Ngaphambi kokusebenzisa kuvavanyo, utshintsho kwixesha loxinzelelo lwe-ozone kwigumbi emva kokuvula ukufakwa kweplasma lulinganiswe ngokwe-absorption spectrum yomgca we-spectral we-253.65 nm wesibane se-mercury.
(a) Isicwangciso sokuseta uvavanyo lokubulala iintsholongwane kwiibhaktheriya kwizinto ezahlukeneyo kusetyenziswa i-ozone eveliswa kwi-DBD plasma reactor, kunye (b) ukuxinana kwe-ozone kunye nexesha lokuveliswa kweplasma kwigumbi lokubulala iintsholongwane. Umfanekiso wenziwe kusetyenziswa i-OriginPro version 9.0 (OriginPro software, Northampton, MA, USA; https://www.originlab.com).
Okokuqala, ngokubulala iintsholongwane kwiiseli zebhaktiriya ezibekwe kwiipleyiti ze-agar nge-ozone, ngelixa kutshintshwa uxinano lwe-ozone kunye nexesha lokunyanga, kwafunyaniswa uxinano olufanelekileyo lwe-ozone kunye nexesha lokunyanga lokususa ungcoliseko lwe-MDRO kunye ne-C. difficile. Ngexesha lenkqubo yokubulala iintsholongwane, igumbi liqala lihlanjululwe ngomoya ojikelezayo kwaye emva koko lizaliswe yi-ozone ngokuvula iyunithi yeplasma. Emva kokuba iisampulu ziphathwe nge-ozone ixesha elimiselweyo, kusetyenziswa ipompo ye-diaphragm ukususa i-ozone eseleyo. Ukulinganisa kusetyenziswe isampulu yenkcubeko epheleleyo yeeyure ezingama-24 (~ 108 CFU/ml). Iisampulu zokumiswa kweeseli zebhaktiriya (20 μl) zaqala zaxutywa ngokulandelelana izihlandlo ezilishumi nge-sterile saline, kwaye emva koko ezi sampuli zasasazwa kwiipleyiti ze-agar ezicociweyo nge-ozone kwigumbi. Emva koko, iisampulu eziphindaphindiweyo, ezibandakanya iisampulu ezivezwe kwaye zingavezwanga kwi-ozone, zafakwa kwi-incubator kwi-37°C iiyure ezingama-24 kwaye zabalwa iikholoni ukuvavanya ukusebenza kokubulala iintsholongwane.
Ngaphezu koko, ngokweemeko zokubulala iintsholongwane ezichazwe kuphononongo olungentla, isiphumo sokucoca obu buchwephesha kwi-MDRO kunye ne-C. difficile sivavanywe kusetyenziswa iikhuphoni zezinto ezahlukeneyo (intsimbi engagqwaliyo, ilaphu, iglasi, iplastiki kunye neekhuphoni zomthi) ezisetyenziswa kakhulu kumaziko ezonyango. Kwasetyenziswa iinkcubeko ezipheleleyo zeeyure ezingama-24 (~108 cfu/ml). Iisampuli zokumiswa kweeseli zebhaktheriya (20 μl) zaxutywa ngokulandelelana izihlandlo ezilishumi nge-sterile saline, emva koko iikhuphoni zaxutywa kwezi broths zixutywe ukuze kuvavanywe ukungcola. Iisampuli ezisusiweyo emva kokuntywiliselwa kumhluzi wokuxuba zafakwa kwizitya zePetri ezingeyongozi kwaye zomiswa kubushushu begumbi iiyure ezingama-24. Beka isiciko sesitya sepetri kwisampuli kwaye usibeke ngononophelo kwigumbi lovavanyo. Susa isiciko kwisitya sePetri kwaye utyhile isampuli kwi-500 ppm ozone imizuzu eli-15. Iisampuli zokulawula zafakwa kwikhabhinethi yokhuseleko lwebhayoloji kwaye azizange zityhileke kwi-ozone. Emva nje kokuvezwa kwi-ozone, iisampuli kunye neesampuli ezingafakwanga mitha (oko kukuthi ulawulo) zaxutywa ne-sterile saline kusetyenziswa umxube we-vortex ukwahlula iintsholongwane kumphezulu. I-eluted suspension yaxutywa ngokulandelelana izihlandlo ezili-10 nge-sterile saline, emva koko inani leebhaktheriya ezixutyiweyo lamiselwa kwiiplate ze-agar yegazi (zebhaktheriya ye-aerobic) okanye iiplate ze-agar yegazi ze-anaerobic zeBrucella (zeClostridium difficile) zaza zafakwa kwi-incubator kwi-37°C kangangeeyure ezingama-24. okanye phantsi kweemeko ze-anaerobic iiyure ezingama-48 kwi-37°C ngokuphindwe kabini ukumisela uxinzelelo lokuqala lwe-inoculum. Umahluko kumanani eebhaktheriya phakathi kolawulo olungatyhilekanga kunye neesampuli eziveziweyo wabalwa ukunika ukwehla kwerekhodi kumanani eebhaktheriya (oko kukuthi, ukusebenza kakuhle kwe-sterilizing) phantsi kweemeko zovavanyo.
Iiseli zebhayoloji kufuneka zingasebenzi kwipleyiti yokufota ye-AFM; ngoko ke, kusetyenziswa idiski ye-mica ethe tyaba nefanayo enesikali esirhabaxa esincinci kunobukhulu beseli njengesiseko. Ububanzi kunye nobukhulu beediski babuyi-20 mm kunye ne-0.21 mm, ngokulandelelana. Ukuze zibambe iiseli ngokuqinileyo kumphezulu, umphezulu we-mica ugqunywe nge-poly-L-lysine (200 µl), nto leyo eyenza ukuba itshajwe kakuhle kwaye i-membrane yeseli itshajwe kakubi. Emva kokugqunywa nge-poly-L-lysine, iidiski ze-mica zahlanjwa kathathu ngamanzi e-1 ml deionized (DI) kwaye zomiswa ngomoya ubusuku bonke. Emva koko, iiseli zebhaktiriya zafakwa kumphezulu we-mica ogqunywe nge-poly-L-lysine ngokuthatha isisombululo sebhaktiriya esinyibilikisiweyo, sashiywa imizuzu engama-30, kwaye umphezulu we-mica wahlanjwa nge-1 ml yamanzi anyibilikisiweyo.
Isiqingatha seesampuli siphathwe nge-ozone kwaye imo yomphezulu weepleyiti ze-mica ezigcwele i-VRE, i-CRAB kunye ne-C. ii-spores ezinzima zabonwa kusetyenziswa i-AFM (XE-7, iinkqubo zepaki). Indlela yokusebenza ye-AFM imiselwe kwimo yokuthepha, eyindlela eqhelekileyo yokufota iiseli zebhayoloji. Kwizilingo, kwasetyenziswa i-microcantilever eyenzelwe imo yokungaqhagamshelani (OMCL-AC160TS, OLYMPUS Microscopy). Imifanekiso ye-AFM yarekhodwa ngokusekelwe kwisantya sokuskena kweprobe se-0.5 Hz nto leyo eyabangela isisombululo somfanekiso see-pixels ezingama-2048 × 2048.
Ukuze sifumanise iimeko apho ii-reactor ze-DBD plasma zisebenza khona ekubulaleni iintsholongwane, senze uthotho lweemvavanyo sisebenzisa zombini i-MDRO (VRE, CRE, CRPA, kunye ne-CRAB) kunye ne-C. difficile ukutshintsha uxinzelelo lwe-ozone kunye nexesha lokuvezwa. Kumfanekiso 1b ubonisa ijika lexesha loxinzelelo lwe-ozone kwimeko nganye yovavanyo emva kokuvula isixhobo se-plasma. Uxinzelelo lwanda ngokwe-logarithmic, lwafikelela kwi-300 kunye ne-500 ppm emva kwemizuzu eyi-1.5 kunye ne-2.5, ngokulandelelana. Uvavanyo lokuqala nge-VRE lubonise ukuba ubuncinci obufunekayo ukuze kususwe iintsholongwane ngokufanelekileyo yi-300 ppm ozone imizuzu eli-10. Ke ngoko, kuvavanyo olulandelayo, i-MDRO kunye ne-C. difficile zavezwa kwi-ozone kumanqanaba amabini ahlukeneyo (300 kunye ne-500 ppm) kunye namaxesha amabini ahlukeneyo okuvezwa (imizuzu eli-10 kunye ne-15). Ukusebenza kakuhle kokubulala iintsholongwane kwidosi nganye ye-ozone kunye nexesha lokuvezwa kubalwe kwaye kwaboniswa kwiTheyibhile 1. Ukuvezwa kwi-ozone engama-300 okanye ama-500 ppm imizuzu eli-10–15 kubangele ukwehla ngokubanzi kwe-VRE ye-log10 okanye ngaphezulu. Eli nqanaba liphezulu lokubulawa kweebhaktheriya nge-CRE lifezekiswe ngemizuzu eli-15 yokuvezwa kwi-ozone engama-300 okanye ama-500 ppm. Ukwehla okuphezulu kwe-CRPA (> 7 log10) kufezekisiwe ngokuchanabeka kwi-500 ppm ye-ozone kangangemizuzu eli-15. Ukwehla okuphezulu kwe-CRPA (> 7 log10) kufezekisiwe ngokuchanabeka kwi-500 ppm ye-ozone kangangemizuzu eli-15. Высокое снижение CRPA (> 7 log10) было достигнуто при воздействии 500 частей на миллион озона в течение 15 минут. Ukwehla okuphezulu kwe-CRPA (> 7 log10) kufezekisiwe ngokuchanabeka kwi-ozone eyi-500 ppm imizuzu eli-15.暴露于500 ppm 的臭氧15 分钟后，可大幅降低CRPA (> 7 log10).暴露于500 ppm 的臭氧15 分钟后，可大幅降低CRPA (> 7 log10). Существенное снижение CRPA (> 7 log10) после 15-минутного воздействия озона с концентрацией 500 ppm. Ukuncipha okukhulu kwe-CRPA (> 7 log10) emva kwemizuzu eli-15 yokuchatshazelwa yi-500 ppm ozone.Ukubulawa okungathandekiyo kweebhaktheriya zeCRAB kwi-300 ppm ozone; nangona kunjalo, kwi-500 ppm ozone, bekukho ukuncipha kwe-> 1.5 log10. nangona kunjalo, kwi-500 ppm ozone, bekukho ukuncipha kwe-> 1.5 log10. однако при концентрации озона 500 частей на миллион наблюдалось снижение > 1,5 log10. nangona kunjalo, kwi-ozone concentration ye-500 ppm, ukwehla kwe->1.5 log10 kwabonwa.然而，在500 ppm 臭氧下，减少了> 1.5 log10.然而，在500 ppm 臭氧下，减少了> 1.5 log10. Однако при концентрации озона 500 частей на миллион наблюдалось снижение >1,5 log10. Nangona kunjalo, kwi-ozone concentration ye-500 ppm, ukwehla kwe->1.5 log10 kwabonwa. Ukuveza ii-spores ze-C. difficile kwi-300 okanye i-500 ppm ozone kuphumele ekunciphiseni kwe-> 2.5 log10. Ukuveza ii-spores ze-C. difficile kwi-300 okanye i-500 ppm ozone kuphumele ekunciphiseni kwe-> 2.5 log10. Воздействие на споры C. difficile озона с концентрацией 300 okanye 500 частей на миллион приводило к снижению > 2,5 log10. Ukuvezwa kwe-C. difficile spores kwi-300 okanye i-500 ppm ozone kubangele ukuncipha kwe->2.5 log10.将艰难梭菌孢子暴露于300 或500 ppm的臭氧中导致> 2.5 log10 减少。 300 或500 ppm 的臭氧中导致> 2.5 log10 减少。 Воздействие на споры C. difficile озона с концентрацией 300 okanye 500 частей на миллион приводило к снижению >2,5 log10. Ukuvezwa kwe-C. difficile spores kwi-300 okanye i-500 ppm ozone kubangele ukuncipha kwe->2.5 log10.
Ngokusekelwe kuvavanyo olungasentla, kufunyenwe imfuneko eyaneleyo yokuvala iintsholongwane ngedosi ye-500 ppm ozone kangangemizuzu eli-15. Iispores ze-VRE, CRAB kunye ne-C. difficile ziye zavavanywa ukuze kubonwe isiphumo sokubulala iintsholongwane se-ozone kwizinto ezahlukeneyo kuquka intsimbi engatyiwayo, ilaphu, iglasi, iplastiki kunye nomthi osetyenziswa rhoqo ezibhedlele. Ukusebenza kwazo kokubulala iintsholongwane kuboniswe kwiTheyibhile 2. Izinto eziphilayo zovavanyo zivavanywe kabini. Kwi-VRE nakwi-CRAB, i-ozone ayisebenzanga kakuhle kwiindawo zeglasi kunye neplastiki, nangona ukuncipha kwe-log10 malunga ne-factor ye-2 okanye ngaphezulu kwabonwa kwiindawo zentsimbi engatyiwayo, ilaphu kunye nomthi. Iispores ze-C. difficile zifunyenwe zichasana kakhulu nonyango lwe-ozone kunezinye izinto eziphilayo ezivavanyiweyo. Ukuze kufundwe ngokwezibalo isiphumo se-ozone kwisiphumo sokubulala izinto ezahlukeneyo ngokuchasene ne-VRE, CRAB, kunye ne-C. difficile, iimvavanyo ze-t zisetyenziselwe ukuthelekisa umahluko phakathi kwenani le-CFU nge-milliliter kumaqela olawulo kunye novavanyo kwizinto ezahlukeneyo (Umzobo 2). Iintlobo zibonise umahluko obalulekileyo ngokwezibalo, kodwa umahluko obalulekileyo ubonwe kwi-VRE kunye ne-CRAB spores kune-C. difficile spores.
Isicwangciso esichazayo semiphumo ye-ozone ekubulaweni kweebhaktheriya kwizinto ezahlukeneyo (a) i-VRE, (b) i-CRAB, kunye (c) ne-C. difficile.
Imifanekiso ye-AFM yenziwe kwiispores ze-VRE, CRAB, kunye ne-C. difficile eziphathwe nge-ozone nezinganyangwanga ukuze kufundwe ngokweenkcukacha inkqubo yokubulala iintsholongwane kwigesi ye-ozone. Kumfanekiso 3a, c kunye no-e kubonisa imifanekiso ye-AFM yeespores ze-VRE, CRAB kunye ne-C. difficile ezinganyangwanga, ngokwahlukeneyo. Njengoko kubonwe kwimifanekiso ye-3D, iiseli zigudile kwaye azinasiphako. Imifanekiso 3b, d kunye no-f ibonisa iispores ze-VRE, CRAB kunye ne-C. difficile emva konyango lwe-ozone. Azizange zinciphise kuphela ubungakanani bazo zonke iiseli ezivavanyiweyo, kodwa umphezulu wazo waba rhabaxa ngokubonakalayo emva kokuvezwa kwi-ozone.
Imifanekiso ye-AFM yee-VRE, MRAB kunye ne-C. difficile spores ezinganyangwanga (a, c, e) kunye (b, d, f) ezinyangwe nge-500 ppm ozone imizuzu eli-15. Imifanekiso ithathwe kusetyenziswa iPark Systems XEI version 5.1.6 (XEI Software, Suwon, Korea; https://www.parksystems.com/102-products/park-xe-bio).
Uphando lwethu lubonisa ukuba i-ozone eveliswa zizixhobo ze-plasma ze-DBD ibonisa amandla okususa ii-spores ze-MDRO kunye ne-C. difficile ngempumelelo, ezaziwa ngokuba zizizathu eziphambili zosulelo oluhambelana nezempilo. Ukongeza, kuphononongo lwethu, ekubeni ungcoliseko lokusingqongileyo olune-MDRO kunye ne-C. difficile spores lunokuba ngumthombo wosulelo oluhambelana nezempilo, isiphumo sokubulala iintsholongwane se-ozone sifunyenwe siphumelele kwizinto ezisetyenziswa kakhulu kwiindawo zesibhedlele. Uvavanyo lokususa iintsholongwane lwenziwe kusetyenziswa izixhobo ze-plasma ze-DBD emva kokungcola okwenziweyo kwezinto ezifana nentsimbi engagqwaliyo, ilaphu, iglasi, iplastiki kunye nomthi one-MDRO kunye ne-C. difficile spores. Ngenxa yoko, nangona isiphumo sokususa iintsholongwane sahluka ngokuxhomekeke kwizinto, amandla okususa iintsholongwane e-ozone ayamangalisa.
Izinto ezichukunyiswa rhoqo kumagumbi esibhedlele zifuna ukucoca rhoqo, ngendlela ephantsi. Indlela eqhelekileyo yokucoca ezo zinto kukucoca ngesandla ngesibulali-ntsholongwane esimanzi esifana ne-quaternary ammonium compound 13. Nokuba kulandelwa ngokungqongqo iingcebiso zokusetyenziswa kwezibulali-ntsholongwane, i-MPO kunzima ukuyisusa ngokucoca kwendalo (ngokuqhelekileyo ukucoca ngesandla)14. Ke ngoko, kufuneka ubuchwepheshe obutsha, njengeendlela ezingachukumisiyo. Ngenxa yoko, kubekho umdla kwizibulali-ntsholongwane zegesi, kubandakanya i-hydrogen peroxide kunye ne-ozone10. Inzuzo yezibulali-ntsholongwane zegesi kukuba zinokufikelela kwiindawo kunye nezinto ezingenakufikelelwa ziindlela zemveli zesandla. I-hydrogen peroxide isandula ukusetyenziswa kwiindawo zonyango, nangona kunjalo i-hydrogen peroxide ngokwayo inetyhefu kwaye kufuneka iphathwe ngokweenkqubo zokuphatha ezingqongqo. Ukucoca nge-plasma nge-hydrogen peroxide kufuna ixesha elide lokucoca ngaphambi komjikelo olandelayo wokubulala iintsholongwane. Ngokwahlukileyo koko, i-ozone isebenza njengearhente ebanzi yokulwa neentsholongwane, esebenza ngokuchasene neebhaktheriya kunye neentsholongwane ezichasene nezinye izibulali-ntsholongwane8,11,15. Ukongeza, i-ozone inokuveliswa ngexabiso eliphantsi emoyeni ongaphakathi kwaye ayifuni iikhemikhali ezinobungozi ezongezelelweyo ezinokushiya unyawo oluyingozi kwindalo esingqongileyo, kuba ekugqibeleni iyaqhekeka ibe yioksijini. Nangona kunjalo, isizathu sokuba i-ozone ingasetyenziswa kakhulu njengesibulali-ntsholongwane sesi silandelayo. I-ozone iyingozi kwimpilo yabantu, ngoko ke uxinzelelo lwayo aludluli kwi-0.07 ppm ngokomyinge ngaphezu kweeyure ezi-816, ngoko ke izibulali-ntsholongwane ze-ozone ziye zaphuhliswa kwaye zafakwa kwimarike, ikakhulu ekucoceni iigesi zokukhupha umoya. Kunokwenzeka nokuphefumla igesi kwaye kuvelise ivumba elibi emva kokususa ungcoliseko5,8. I-ozone yayingasetyenziswa ngokukhutheleyo kwiindawo zonyango. Nangona kunjalo, i-ozone ingasetyenziswa ngokukhuselekileyo kwiindawo zokucoca ungcoliseko kunye neenkqubo ezifanelekileyo zokungenisa umoya, kwaye ukususwa kwayo kunokukhawuleziswa kakhulu ngokusebenzisa i-catalytic converter. Kolu phononongo, sibonisa ukuba izibulali-ntsholongwane ze-ozone zeplasma zinokusetyenziselwa ukubulala iintsholongwane kwiindawo zempilo. Siphuhlise isixhobo esinamandla aphezulu okubulala iintsholongwane, ukusebenza okulula kunye nenkonzo ekhawulezayo kwizigulane ezibhedlele. Ukongeza, siphuhlise iyunithi elula yokubulala iintsholongwane esebenzisa umoya ongqongileyo ngaphandle kweendleko ezongezelelweyo. Ukuza kuthi ga ngoku, akukho lwazi lwaneleyo malunga neemfuno eziphantsi ze-ozone zokungasebenzi kwe-MDRO. Izixhobo ezisetyenzisiweyo kolu phando lwethu kulula ukuzibeka kwaye zinexesha elifutshane lokusebenza kwaye kulindeleke ukuba zibe luncedo ekubulaleni rhoqo izixhobo.
Indlela esebenza ngayo i-ozone ekubulaleni iintsholongwane ayicacanga ngokupheleleyo. Izifundo ezininzi zibonise ukuba i-ozone yonakalisa ii-membrane zeseli zebhaktheriya, nto leyo ekhokelela ekuvuthweni ngaphakathi kweseli kunye nokuphela kwe-cell lysis17,18. I-ozone inokuphazamisana nomsebenzi we-enzymatic weseli ngokuphendula ngamaqela e-thiol kwaye inokutshintsha iziseko ze-purine kunye ne-pyrimidine kwi-nucleic acids. Olu phononongo lubonise imo ye-VRE, CRAB, kunye ne-C. difficile spores ngaphambi nasemva konyango lwe-ozone kwaye lufumanise ukuba azizange zehle ngobukhulu kuphela, kodwa zaba nzima kakhulu kumphezulu, nto leyo ebonisa umonakalo okanye ukugqwala kwe-membrane engaphandle. kwaye izinto zangaphakathi ngenxa yegesi ye-ozone zinamandla amakhulu okuxilisa. Lo monakalo unokukhokelela ekungasebenzini kweseli, kuxhomekeke kubukhali botshintsho lweseli.
Ii-spores ze-C. difficile kunzima ukuzisusa kumagumbi esibhedlele. Ii-spores zihlala kwiindawo apho zichitha khona i-10,20. Ukongeza, kolu phononongo, nangona ukwehla okuphezulu kwe-logarithmic ngokuphindwe kalishumi kwinani leebhaktheriya kwiiplate ze-agar kwi-500 ppm ozone imizuzu eli-15 yayiyi-2.73, isiphumo sokubulala iibhaktheriya se-ozone kwizinto ezahlukeneyo eziqulethe ii-spores ze-C. difficile sinciphisiwe. Ke ngoko, amaqhinga ahlukeneyo anokuqwalaselwa ukunciphisa usulelo lwe-C. difficile kwiindawo zokhathalelo lwempilo. Ukusetyenziswa kumagumbi e-C. difficile ahlukeneyo kuphela, kunokuba luncedo ukulungisa ixesha lokuvezwa kunye nobukhali bonyango lwe-ozone. Ukongeza, kufuneka sikhumbule ukuba indlela yokususa ukungcola kwi-ozone ayinakuyithatha indawo yokucoca okuqhelekileyo ngesandla ngamayeza okubulala iintsholongwane kunye namaqhinga okulwa neentsholongwane, kwaye inokusebenza kakhulu ekulawuleni i-C. difficile 5. Kolu phononongo, ukusebenza kwe-ozone njengesibulali-ntsholongwane kwahluka kwiintlobo ezahlukeneyo ze-MPO. Ukusebenza kakuhle kunokuxhomekeka kwizinto ezininzi ezifana nenqanaba lokukhula, udonga lweseli, kunye nokusebenza kakuhle kweendlela zokulungisa21,22. Isizathu sempembelelo eyahlukileyo yokubulala iintsholongwane ye-ozone kumphezulu wento nganye isenokuba kukwenziwa kwe-biofilm. Izifundo zangaphambili zibonise ukuba i-E. faecium kunye ne-E. faecium zonyusa ukumelana nokusingqongileyo xa zikhona njenge-biofilms23, 24, 25. Nangona kunjalo, olu phononongo lubonisa ukuba i-ozone inempembelelo ebalulekileyo yokubulala iintsholongwane kwi-MDRO kunye ne-C. difficile spores.
Isithintelo sophando lwethu kukuba sivavanye impembelelo yokugcinwa kwe-ozone emva kokulungiswa. Oku kunokukhokelela ekuqikeleleni ngokugqithisileyo inani leeseli zebhaktheriya ezisebenzayo.
Nangona olu phononongo lwenziwe ukuze kuvavanywe ukusebenza kakuhle kwe-ozone njengesibulali-ntsholongwane esibhedlele, kunzima ukuqukanisa iziphumo zethu kuzo zonke iindawo zesibhedlele. Ke ngoko, kufuneka uphando olongezelelekileyo ukuze kuphandwe ukuba le ntsholongwane ye-DBD ozone iyasebenza na kwaye ihambelana njani nendawo yesibhedlele yokwenyani.
I-ozone eveliswa zii-DBD plasma reactors inokuba yinto elula nexabisekileyo yokususa ungcoliseko kwi-MDRO kunye ne-C. difficile. Ngoko ke, unyango lwe-ozone lunokuthathwa njengendlela esebenzayo yokubulala iintsholongwane kwindawo yesibhedlele.
Iiseti zedatha ezisetyenzisiweyo kunye/okanye ezihlalutyiweyo kolu phando lwangoku ziyafumaneka kubabhali abafanelekileyo xa beceliwe ngokufanelekileyo.
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