{"id":8449,"date":"2020-08-24T14:13:41","date_gmt":"2020-08-24T13:13:41","guid":{"rendered":"https:\/\/www.vtei.cz\/?p=8449"},"modified":"2024-07-17T10:51:48","modified_gmt":"2024-07-17T09:51:48","slug":"preparation-and-evaluation-of-nanofibrous-materials-and-their-use-in-waste-water-treatment","status":"publish","type":"post","link":"https:\/\/www.vtei.cz\/en\/2020\/08\/preparation-and-evaluation-of-nanofibrous-materials-and-their-use-in-waste-water-treatment\/","title":{"rendered":"Preparation and evaluation of nanofibrous materials and their use in waste water treatment"},"content":{"rendered":"<h4><i class=\"fa fa-exclamation-circle fa-3x pull-left\"><\/i> This article is available in Czech only. For translation or more information on this topic, please contact author.<\/h4>\n<p>&nbsp;<\/p>\n<h2>Souhrn<\/h2>\n<p>Nanomateri\u00e1ly z\u00edskaly velkou pozornost v\u00a0r\u016fzn\u00fdch oblastech d\u00edky sv\u00fdm unik\u00e1tn\u00edm vlastnostem. Jednou z\u00a0mo\u017en\u00fdch aplikac\u00ed nanovl\u00e1ken, d\u00edky jejich specifick\u00e9mu povrchu, je nosi\u010d biomasy, tedy imobilizace mikroorganism\u016f n\u00e1sledovan\u00e1 tvorbou biofilmu. Proto\u017ee vlastnosti nanovl\u00e1kenn\u00e9ho nosi\u010de maj\u00ed v\u00fdznamn\u00fd dopad na rychlost mikrobi\u00e1ln\u00ed kolonizace a\u00a0funk\u010dnost biofilmu, variabilita parametr\u016f p\u0159i p\u0159\u00edprav\u011b nanovl\u00e1ken (materi\u00e1l, modifikace, struktura povrchu apod.) umo\u017e\u0148uje vytvo\u0159it specifick\u00e9 prost\u0159ed\u00ed pro dan\u00e9 bakterie.<\/p>\n<p>Hlavn\u00edm c\u00edlem t\u00e9to pr\u00e1ce je zkoum\u00e1n\u00ed vlastnost\u00ed nanovl\u00e1ken p\u0159ipraven\u00fdch r\u016fzn\u00fdmi zvl\u00e1k\u0148ovac\u00edmi technologiemi a\u00a0hodnocen\u00ed jejich aplikace p\u0159i biologick\u00e9m \u010di\u0161t\u011bn\u00ed odpadn\u00edch vod. Jako materi\u00e1l byly pro v\u00fdrobu nanovl\u00e1ken vyu\u017eity dva typy polymer\u016f, polyuretan a\u00a0polyvinylbutyral, kter\u00e9 byly zvl\u00e1kn\u011bny za obdobn\u00fdch podm\u00ednek. Pro p\u0159\u00edpravu nanovl\u00e1kenn\u00fdch materi\u00e1l\u016f bylo pou\u017eito p\u011bt zvl\u00e1k\u0148ovac\u00edch technologi\u00ed: zvl\u00e1k\u0148ov\u00e1n\u00ed st\u0159\u00eddav\u00fdm proudem, zvl\u00e1k\u0148ov\u00e1n\u00ed stejnosm\u011brn\u00fdm proudem z\u00a0ty\u010dky,\u00a0z\u00a0jehly a\u00a0ze struny (technologie Nanospider<sup>TM<\/sup>) a\u00a0elektro-odst\u0159ediv\u00e9 zvl\u00e1k\u0148ov\u00e1n\u00ed. Anal\u00fdza struktur a\u00a0morfologie nanovl\u00e1ken v\u010detn\u011b drsnosti povrchu byla provedena pomoc\u00ed skenovac\u00ed elektronov\u00e9 mikroskopie (SEM) a\u00a0pomoc\u00ed konfok\u00e1ln\u00ed mikroskopie, zat\u00edmco biologick\u00e1 rozlo\u017eitelnost nanovl\u00e1ken byla stanovena normovan\u00fdm respirometrick\u00fdm testem (stanoven\u00ed BSK). Na z\u00e1klad\u011b v\u00fdsledk\u016f z\u00a0t\u011bchto anal\u00fdz byl zvolen nejvhodn\u011bj\u0161\u00ed nanovl\u00e1kenn\u00fd materi\u00e1l pro biologick\u00e9 aplikace\u00a0\u2013 polyvinylbutyral p\u0159ipraven\u00fd pomoc\u00ed technologie Nanospider<sup>TM<\/sup>, kter\u00fd byl n\u00e1sledn\u011b testov\u00e1n v\u00a0biologick\u00e9m laboratorn\u00edm reaktoru jako nosi\u010d biomasy. B\u011bhem provozu bioreaktoru byl sledov\u00e1n pr\u016fb\u011bh nitrifikace, resp. r\u016fst nitrifika\u010dn\u00edch bakteri\u00ed pomoc\u00ed respirometrie a\u00a0molekul\u00e1rn\u011b-genetick\u00fdch metod (FISH a\u00a0real-time qPCR). V\u00fdsledky anal\u00fdz ukazuj\u00ed, \u017ee zvolen\u00fd PVB nosi\u010d v\u00fdrazn\u011b podporoval r\u016fst biomasy, kde bylo pomoc\u00ed molekul\u00e1rn\u011b genetick\u00fdch metod nalezeno zna\u010dn\u00e9 zastoupen\u00ed nitrifika\u010dn\u00edch bakteri\u00ed. Nav\u00edc, chemismus reaktoru v\u00a0pr\u016fb\u011bhu experimentu potvrzoval vysokou \u00fa\u010dinnost nitrifika\u010dn\u00edho procesu.<\/p>\n<h2>\u00davod<\/h2>\n<p>Materi\u00e1ly z\u00a0nanovl\u00e1ken (netkan\u00e9 textilie, scaffoldy a\u00a0jin\u00e9 3D struktury) byly ji\u017e pou\u017eity v\u00a0mnoha komer\u010dn\u00edch a\u00a0v\u00fdzkumn\u00fdch oblastech\u00a0\u2013 filtrace, biotechnologie, r\u016fzn\u00e9 environment\u00e1ln\u00ed aplikace, farmaceutick\u00fd pr\u016fmysl, kosmetika, tk\u00e1\u0148ov\u00e9 in\u017een\u00fdrstv\u00ed a\u00a0medic\u00edna [1\u20133]. K\u00a0p\u0159\u00edprav\u011b nanovl\u00e1ken lze pou\u017e\u00edt \u0159adu dostupn\u00fdch technik, jako je separace f\u00e1z\u00ed, drawing, samoskladba, chemick\u00e9 depozice z\u00a0plynn\u00e9 f\u00e1ze, meltblown, elektrospinning [4] atd. Elektrostatick\u00e9 zvl\u00e1k\u0148ov\u00e1n\u00ed je v\u00a0sou\u010dasnosti zna\u010dn\u011b roz\u0161\u00ed\u0159en\u00e9 a\u00a0d\u00e1v\u00e1 mo\u017enost vyr\u00e1b\u011bt nanovl\u00e1kna v\u00a0pr\u016fmyslov\u00e9m m\u011b\u0159\u00edtku [5]. S\u00a0p\u0159\u00edpravou nanovl\u00e1kenn\u00fdch materi\u00e1l\u016f a\u00a0jejich budouc\u00ed aplikac\u00ed jsou spojena t\u0159i z\u00e1kladn\u00ed t\u00e9mata: (1) mo\u017enost ekonomicky v\u00fdhodn\u00e9 velkoobjemov\u00e9 v\u00fdroby, (2) vhodn\u00e9 nastaven\u00ed a\u00a0reprodukovatelnost ve v\u0161ech f\u00e1z\u00edch v\u00fdroby a\u00a0(3) bezpe\u010dnostn\u00ed a\u00a0environment\u00e1ln\u00ed atributy.<\/p>\n<p>Nanovl\u00e1kna p\u0159ipraven\u00e1 pomoc\u00ed elektrostatick\u00e9ho zvl\u00e1k\u0148ov\u00e1n\u00ed maj\u00ed vysok\u00fd pom\u011br plochy povrchu k\u00a0objemu, zna\u010dnou p\u00f3rovitost, dobr\u00e9 mechanick\u00e9 vlastnosti a\u00a0velmi specifick\u00e9 povrchov\u00e9 vlastnosti [1]. Mo\u017enost zpracov\u00e1n\u00ed \u0161irok\u00e9 \u0161k\u00e1ly polymer\u016f poskytuje prostor pro manipulaci a\u00a0kontrolu povrchov\u00e9 struktury nanomateri\u00e1l\u016f, pr\u016fm\u011br\u016f vl\u00e1ken a\u00a0p\u00f3rovitosti [6]. Pr\u016fm\u011br vl\u00e1ken a\u00a0dal\u0161\u00ed vlastnosti lze ovlivnit koncentrac\u00ed polymeru, molekulovou hmotnost\u00ed nebo koncentrac\u00ed \u010di viskozitou roztoku, nastaven\u00edm elektrick\u00e9ho nap\u011bt\u00ed, zvolen\u00fdmi technick\u00fdmi parametry za\u0159\u00edzen\u00ed (zvl\u00e1k\u0148ovac\u00ed tryska, pr\u016fm\u011br elektrody, vzd\u00e1lenost elektrod apod.) a\u00a0parametry okoln\u00edho prost\u0159ed\u00ed [7, 8].<\/p>\n<p>Polyvinylbutyral (PVB) je netoxick\u00fd polymer bez z\u00e1pachu a\u00a0\u0161etrn\u00fd k\u00a0\u017eivotn\u00edmu prost\u0159ed\u00ed a\u00a0je vhodnou organickou slo\u017ekou pro v\u00fdrobu organick\u00fdch\/anorganick\u00fdch hybridn\u00edch kompozit\u016f. PVB je \u0161iroce pou\u017e\u00edv\u00e1n v\u00a0mnoha aplikac\u00edch kv\u016fli n\u00edzk\u00fdm n\u00e1klad\u016fm na v\u00fdrobu, flexibilit\u011b, dobr\u00e9 p\u0159ilnavosti a\u00a0kv\u016fli jeho dlouhodob\u00e9 stabilit\u011b. Nav\u00edc vykazuje dobr\u00e9 mechanick\u00e9 a\u00a0vynikaj\u00edc\u00ed dielektrick\u00e9 vlastnosti a\u00a0je odoln\u00fd v\u016f\u010di vlhkosti (stabilita ve vodn\u00e9m prost\u0159ed\u00ed) [9]. Polyuretan (PUR) m\u00e1 \u0161irokou \u0161k\u00e1lu r\u016fzn\u00fdch chemick\u00fdch modifikac\u00ed, na kter\u00fdch z\u00e1vis\u00ed jeho v\u00fdsledn\u00e9 mechanick\u00e9 a\u00a0fyzik\u00e1ln\u011b-chemick\u00e9 vlastnosti. Lze p\u0159ipravit PUR biologicky stabiln\u00ed a\u00a0z\u00e1rove\u0148 biokompatibiln\u00ed. Tyto polymery byly vybr\u00e1ny pro jejich rozd\u00edln\u00e9 struktur\u00e1ln\u00ed vlastnosti, pr\u016fmyslovou produkci (\u0161irok\u00e9 vyu\u017eit\u00ed) a\u00a0dobr\u00e9 mechanick\u00e9 vlastnosti.<\/p>\n<p>Nej\u010dast\u011bji se charakterizace nanovl\u00e1kenn\u00fdch materi\u00e1l\u016f zaji\u0161\u0165uje pomoc\u00ed skenovac\u00ed elektronov\u00e9 mikroskopie (SEM). Jen m\u00e1lo prac\u00ed v\u0161ak zkoum\u00e1 vliv procesn\u00edch parametr\u016f na morfologii v\u00fdsledn\u00e9 nanovl\u00e1kenn\u00e9 s\u00edt\u011b [7], s\u00a0v\u00fdjimkou hodnocen\u00ed pr\u016fm\u011bru vl\u00e1ken [5, 8, 10\u201312]. Za \u00fa\u010delem zji\u0161t\u011bn\u00ed specifick\u00fdch vlastnost\u00ed nanomateri\u00e1l\u016f je nezbytn\u00e9 pochopit ovlivn\u011bn\u00ed distribuce a\u00a0struktury nanovl\u00e1ken samotn\u00fdm zvl\u00e1k\u0148ovac\u00edm procesem. Nap\u0159\u00edklad integrita povrchu m\u00e1 zna\u010dn\u00fd vliv na adhezi bun\u011bk, jejich r\u016fst a\u00a0proliferaci, zat\u00edmco vnit\u0159n\u00ed struktura nanovl\u00e1ken ovliv\u0148uje uvol\u0148ov\u00e1n\u00ed inkorporovan\u00fdch biologicky aktivn\u00edch l\u00e1tek a\u00a0proliferaci bun\u011bk do vnit\u0159n\u00ed struktury materi\u00e1lu [6]. Charakterizace nanovl\u00e1ken je v\u00a0sou\u010dasnosti pova\u017eov\u00e1na za jednu z\u00a0nejzaj\u00edmav\u011bj\u0161\u00edch oblast\u00ed v\u00fdzkumu nanovl\u00e1ken. Z\u00a0tohoto d\u016fvodu jsme provedli podrobnou anal\u00fdzu p\u0159ipraven\u00fdch nanovl\u00e1ken pomoc\u00ed obrazov\u00e9 anal\u00fdzy sn\u00edmk\u016f SEM s\u00a0c\u00edlem posoudit z\u00e1kladn\u00ed struktur\u00e1ln\u00ed vlastnosti nanovl\u00e1ken d\u016fle\u017eit\u00e9 pro biologick\u00e9 interakce.<\/p>\n<p>Konkr\u00e9tn\u011b pro biologick\u00e9 aplikace (nap\u0159. nosi\u010de biomasy) nanovl\u00e1kenn\u00fdch struktur je nutn\u00e9 zn\u00e1t jejich detailn\u00ed charakteristiku, aby bylo mo\u017en\u00e9 odhadnout chov\u00e1n\u00ed bakteri\u00ed (biofilm) na jejich povrchu, respektive v povrchov\u00e9 struktu\u0159e. Z\u00e1kladn\u00edmi parametry jsou povrchov\u00fd n\u00e1boj, p\u00f3rovitost, morfologie povrchu, kter\u00e1 siln\u011b souvis\u00ed s bun\u011b\u010dnou adhez\u00ed a dal\u0161\u00ed specifick\u00e9 fyzik\u00e1ln\u011b-mechanick\u00e9 vlastnosti [13, 14]. Nap\u0159\u00edklad u procesu nitrifikace a denitrifikace v odpadn\u00edch vod\u00e1ch doch\u00e1z\u00ed k zna\u010dn\u00e9mu ovliv\u0148ov\u00e1n\u00ed aktivit mikroorganism\u016f typem biomasy, ve kter\u00e9 se bakterie nach\u00e1z\u00ed (aktivovan\u00fd kal\/biofilm na nosi\u010d\u00edch), a tedy k\u00a0ovliv\u0148ov\u00e1n\u00ed \u00fa\u010dinnosti zm\u00edn\u011bn\u00fdch proces\u016f. V\u00fdhody pou\u017eit\u00ed t\u011bchto typ\u016f nosi\u010d\u016f v\u00a0oblasti \u010di\u0161t\u011bn\u00ed odpadn\u00edch vod jsou pops\u00e1ny ve studii Sharma a\u00a0kol. [15]. V\u00a0sou\u010dasn\u00e9 dob\u011b je pou\u017eit\u00ed nosi\u010d\u016f biomasy jedna z\u00a0mo\u017enost\u00ed, jak zvy\u0161ovat \u00fa\u010dinnosti st\u00e1vaj\u00edc\u00edch \u010cOV bez nutnosti finan\u010dn\u011b n\u00e1ro\u010dn\u00fdch stavebn\u00edch \u00faprav objekt\u016f.<\/p>\n<h2>Materi\u00e1ly a\u00a0metody<\/h2>\n<h3>Pou\u017eit\u00e9 polymery<\/h3>\n<p>V\u00a0t\u00e9to studii byly pou\u017eity dva typy polymer\u016f\u00a0\u2013 PVB a\u00a0PUR. Desetiprocentn\u00ed hm. roztok PVB (Mowitals B 60 H, Kuraray America Inc., USA; pr\u016fm\u011brn\u00e1 molekulov\u00e1 hmotnost 60 000 amu) byl p\u0159ipraven v\u00a0etanolu (9: 1 Et-OH: PVB). Nejvhodn\u011bj\u0161\u00ed polymern\u00ed roztok PUR (tj. nejvhodn\u011bj\u0161\u00ed parametry pro elektrostatick\u00e9 zvl\u00e1k\u0148ov\u00e1n\u00ed, zejm\u00e9na pokud jde o\u00a0viskozitu) byl vybr\u00e1n z\u00a0rozmez\u00ed koncentrac\u00ed PUR v\u00a0dimethylformamidu (18\u201335 % hm.). Nem\u016f\u017eeme poskytnout podrobn\u011bj\u0161\u00ed informace o\u00a0koncentrac\u00edch pou\u017eit\u00fdch v\u00a0tomto bod\u011b, proto\u017ee jsou p\u0159edm\u011btem pr\u016fmyslov\u00e9 ochrany.<\/p>\n<h3>Pou\u017eit\u00e9 zvl\u00e1k\u0148ovac\u00ed metody<\/h3>\n<p>Pro p\u0159\u00edpravu nanovl\u00e1ken bylo pou\u017eito p\u011bt typ\u016f zvl\u00e1k\u0148ovac\u00edch metod (elektrospinning) se stejnosm\u011brn\u00fdm a\u00a0st\u0159\u00eddav\u00fdm zdrojem nap\u011bt\u00ed (viz <em>tabulky 1<\/em> a\u00a0<em>2<\/em>). Podstatou elektrospinningu je tvorba nanovl\u00e1ken z\u00a0polymern\u00edho roztoku za p\u0159\u00edtomnosti elektrick\u00fdch sil. Metody se li\u0161\u00ed zdrojem nap\u011bt\u00ed, typem elektrod a\u00a0kolektor\u016f, uspo\u0159\u00e1d\u00e1n\u00edm za\u0159\u00edzen\u00ed, produktivitou, strukturou p\u0159ipraven\u00fdch nanovl\u00e1ken a\u00a0dal\u0161\u00edmi specifick\u00fdmi parametry.<\/p>\n<h5>Tabulka 1. Procesn\u00ed parametry pro p\u0159\u00edpravu PVB nanovl\u00e1ken<br \/>\nTable 1. Process parameters for the preparation of PVB nanofibers<\/h5>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-1.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"195\" class=\"alignnone size-full wp-image-8435 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-1.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-1.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-1-300x73.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-1-768x187.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/195;\" \/><\/a>\n<h5>Tabulka 2. Procesn\u00ed parametry pro p\u0159\u00edpravu PUR nanovl\u00e1ken<br \/>\nTable 2. Process parameters for the preparation of PUR nanofibers<\/h5>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-2.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"195\" class=\"alignnone size-full wp-image-8437 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-2.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-2.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-2-300x73.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-2-768x187.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/195;\" \/><\/a>\n<h3>Mikroskopick\u00e1 charakterizace nanovl\u00e1ken<\/h3>\n<p>Sn\u00edmky SEM byly vyfoceny pomoc\u00ed mikroskopu UHR FE-SEM Carl Zeiss ULTRA Plus (Carl Zeiss, N\u011bmecko) p\u0159i zv\u011bt\u0161en\u00ed 12\u20131 000 000\u00d7 v\u00a0re\u017eimu SE a\u00a0p\u0159i akcelera\u010dn\u00edm nap\u011bt\u00ed 0,02\u201330 kV. Vzorky nanovl\u00e1ken (bez podkladov\u00e9 vrstvy) byly pe\u010dliv\u011b na\u0159ez\u00e1ny na \u010dtverce o\u00a0velikosti 5 \u00d7 5 mm a\u00a0p\u0159ipevn\u011bny k\u00a0ter\u010d\u016fm pomoc\u00ed lepic\u00ed p\u00e1sky. Na vzorky byla nanesena zlat\u00e1 vrstva (p\u0159\u00edstroj Coorum Q150R ES, Quorum Technologies, UK), aby byla zaji\u0161t\u011bna dostate\u010dn\u00e1 vodivost pro v\u0161echny vzorky.<\/p>\n<p>Anal\u00fdza obrazu byla provedena v\u00a0programu Matlab [16] a\u00a0zahrnovala \u0161est z\u00e1kladn\u00edch krok\u016f, kter\u00e9 byly naprogramov\u00e1ny v\u00a0r\u00e1mci t\u00e9to pr\u00e1ce. Konfok\u00e1ln\u00ed mikroskopie byla provedena na vysoce v\u00fdkonn\u00e9m 3D optick\u00e9m mikroskopu S\u00a0neox (Sensofar metrology, \u0160pan\u011blsko) s\u00a0objektivem EPI 20X v35, kter\u00fd umo\u017e\u0148uje bezkontaktn\u00ed optick\u00e9 3D profilov\u00e1n\u00ed. Neox pou\u017e\u00edv\u00e1 senzor s\u00a0vysok\u00fdm rozli\u0161en\u00edm 1 360 \u00d7 1 024 pixel\u016f (pozorovan\u00e1 plocha 850,08 \u00d7 709,32 \u00b5m). Vzorky nanovl\u00e1ken (bez podkladu) byly na\u0159ez\u00e1ny na \u010dtverce 10 \u00d7 10 mm a\u00a0p\u0159ilepeny k\u00a0laboratorn\u00edmu skl\u00ed\u010dku pomoc\u00ed lepic\u00ed p\u00e1sky, \u010d\u00edm\u017e byla zaji\u0161t\u011bna stabilita vzork\u016f pro vyhodnocen\u00ed povrchu. Na vzorky byla nanesena zlat\u00e1 vrstva pro sn\u00ed\u017een\u00ed transparentnosti nanovl\u00e1ken.<\/p>\n<h3>Biodegradace nanovl\u00e1ken<\/h3>\n<p>Biologick\u00e1 rozlo\u017eitelnost nanovl\u00e1ken byla stanovena pomoc\u00ed respirometrie na z\u00e1klad\u011b m\u011b\u0159en\u00ed biologick\u00e9 spot\u0159eby kysl\u00edku (BSK) v\u00a0p\u0159\u00edtomnosti aerobn\u00edch mikroorganism\u016f ve vodn\u00e9m prost\u0159ed\u00ed podle EN ISO 1485. Hodnota BSK byla m\u011b\u0159ena pomoc\u00ed respirometru Micro-Oxymax (Columbus Instruments Int., USA). K\u00a0testu byla pou\u017eita destilovan\u00e1 voda (vodivost 3,0 S\u00b7cm<sup>-1<\/sup>) a standardn\u00ed testovac\u00ed medium (podle normy). Bakteri\u00e1ln\u00ed inokulum bylo p\u0159ipraveno odsazen\u00edm aktivovan\u00e9ho kalu odebran\u00e9ho z \u010dist\u00edrny odpadn\u00edch vod (Liberec, \u010cesk\u00e1 republika; vzorkovan\u00e9 na konci nitrifika\u010dn\u00ed n\u00e1dr\u017ee). Hmotnost su\u0161iny kalu byla 4,3 g\u00b7l<sup>-1<\/sup> a\u00a0hodnota KTJ (kolonie tvo\u0159\u00edc\u00ed jednotky) byla stanovena na 100\u00a0000 KTJ\u00b7ml<sup>-1<\/sup>. Vzorky byly m\u011b\u0159eny v duplik\u00e1tu po dobu 28 dn\u016f. V\u00fdsledn\u00e9 hodnoty biodegradace <em>Dt<\/em> pro ka\u017ed\u00fd vzorek byly vypo\u010dteny jako (BSK * 100)\/TSK, kde TSK je teoretick\u00e1 spot\u0159eba kysl\u00edku v\u00a0mg\u00b7g<sup>-1<\/sup>.<\/p>\n<h3>Laboratorn\u00ed biologick\u00fd reaktor<\/h3>\n<p>Model biologick\u00e9ho reaktoru (<em>obr. 1<\/em>) tvo\u0159\u00ed otev\u0159en\u00e1 silnost\u011bnn\u00e1 sklen\u011bn\u00e1 n\u00e1doba (\u00d8 24 cm) o\u00a0objemu 18 litr\u016f. Na dn\u011b n\u00e1doby jsou po stran\u00e1ch instalov\u00e1ny provzdu\u0161\u0148ovac\u00ed elementy napojen\u00e9 na dmychadlo (AirMac, DB40, 48 l\u00b7min<sup>-1<\/sup>), kter\u00e9 zaji\u0161\u0165uje homogenn\u00ed provzdu\u0161\u0148ovan\u00ed syst\u00e9mu. P\u0159ed zah\u00e1jen\u00edm provozu byl bioreaktor inokulov\u00e1n aktivovan\u00fdm kalem z\u00a0nitrifika\u010dn\u00ed n\u00e1dr\u017ee komun\u00e1ln\u00ed \u010cOV Liberec. Do reaktoru byla jako n\u00e1tok p\u0159iv\u00e1d\u011bna modelov\u00e1 odpadn\u00ed voda obsahuj\u00edc\u00ed NH<sub>4<\/sub>Cl jako zdroj dus\u00edku a\u00a0fosf\u00e1tov\u00fd pufr pro stabilizaci pH a\u00a0jako zdroj fosforu (PO<sub>4<\/sub><sup>3-<\/sup>). Celkov\u00fd objem vody v reaktoru byl 8 l (p\u0159i maxim\u00e1ln\u00edm napln\u011bn\u00ed). Kontinu\u00e1ln\u00ed provoz byl zaji\u0161\u0165ov\u00e1n peristaltick\u00fdmi \u010derpadly (Watson Marlow Sci-Q323). Na dn\u011b n\u00e1doby byly v nerezov\u00fdch r\u00e1me\u010dc\u00edch um\u00edst\u011bny testovan\u00e9 nosi\u010de biomasy o velikosti 55 \u00d7 55 mm. Cel\u00fd syst\u00e9m nosi\u010d\u016f byl nep\u0159etr\u017eit\u011b pono\u0159en.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-1.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"445\" class=\"alignnone size-full wp-image-8419 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-1.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-1.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-1-300x167.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-1-768x427.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/445;\" \/><\/a>\n<h6>Obr. 1. Sch\u00e9ma prototypu biologick\u00e9ho reaktoru pro testov\u00e1n\u00ed nosi\u010d\u016f biomasy<br \/>\nFig. 1. Schema of a\u00a0prototype biological reactor for testing biomass carriers<\/h6>\n<p>Biologick\u00fd modul pracuje jako SBR (Sequencing Batch Reactor) ve dvou opera\u010dn\u00edch cyklech b\u011bhem dne. Prvn\u00ed t\u0159i dny prob\u00edhalo v\u00a0bioreaktoru cyklick\u00e9 opakov\u00e1n\u00ed n\u00e1sleduj\u00edc\u00edch proces\u016f: n\u00e1tok + aerace (10,5 h), n\u00e1tok + sedimentace (0,5\u00a0h) a\u00a0odtah odpadn\u00ed vody na polovinu objemu reaktoru (0,5 h). Po odkalen\u00ed bioreaktoru (po t\u0159ech dnech od spu\u0161t\u011bn\u00ed) byla aerace spu\u0161t\u011bna nep\u0159etr\u017eit\u011b a\u00a0k\u00a0sedimentaci ji\u017e nedoch\u00e1zelo. D\u00e1le byla udr\u017eov\u00e1na pouze biomasa p\u0159isedl\u00e1\/narostl\u00e1 na nosi\u010d\u00edch, tj. bez suspendovan\u00e9ho kalu. Teplota vody byla stabiln\u011b v\u00a0rozmez\u00ed 19 a\u017e 22 \u00b0C a\u00a0pH v\u00a0neutr\u00e1ln\u00ed oblasti, nanejv\u00fd\u0161 v\u00a0m\u00edrn\u011b kyselen\u00e9. Koncentrace jednotliv\u00fdch slou\u010denin dus\u00edku a\u00a0fosforu a\u00a0CHSK byly m\u011b\u0159eny pomoc\u00ed kyvetov\u00fdch test\u016f (od spole\u010dnosti Lange) na spektrofotometru HACH DR 6000 UV-VIS od spole\u010dnosti Hach-Lange.<\/p>\n<p>V\u00a0pr\u016fb\u011bhu experimentu byl v\u00fdvoj nitrifika\u010dn\u00edch bakteri\u00ed, tj. amoniak oxiduj\u00edc\u00edch (AOB) a\u00a0dusitan oxiduj\u00edc\u00edch (NOB) bakteri\u00ed, v\u00a0biofilmu na nanovl\u00e1knech pozorov\u00e1n pomoc\u00ed real\u2011time kvantitativn\u00ed polymer\u00e1zov\u00e9 \u0159et\u011bzov\u00e9 reakce (qPCR). Na konci experimentu byla nav\u00edc p\u0159\u00edtomnost AOB a\u00a0NOB na nanovl\u00e1knech analyzov\u00e1na pomoc\u00ed metody fluorescen\u010dn\u00ed in situ hybridizace (FISH) a\u00a0aktivita nitrifika\u010dn\u00ed mikrofl\u00f3ry byla sledov\u00e1na pomoc\u00ed respirometrick\u00e9ho m\u011b\u0159en\u00ed.<\/p>\n<h3>Metody molekul\u00e1rn\u011b-genetick\u00e9<\/h3>\n<p>Pro detekci p\u0159\u00edtomnosti AOB a\u00a0NOB v\u00a0biofilmu na nanovl\u00e1knech z\u00a0bioreaktoru pomoc\u00ed molekul\u00e1rn\u011b-genetick\u00fdch metod, real-time qPCR a\u00a0FISH, byla vyt\u0159ep\u00e1na biomasa ze \u010dtverce nanovl\u00e1ken o\u00a0velikosti\u00a06\u00a0\u00d7 6\u00a0mm pou\u017eit\u00edm ultrazvuku (15\u00a0min, 35 kHz, 20 \u00b0C). Z\u00a0vyt\u0159epan\u00e9 biomasy byla pro \u00fa\u010dely real-time PCR provedena extrakce bakteri\u00e1ln\u00ed DNA, zat\u00edmco pro FISH anal\u00fdzu prob\u011bhla fixace a\u00a0permeabilizace bun\u011bk.<\/p>\n<p>Extrakce DNA z\u00a0biomasy byla provedena pomoc\u00ed kitu specializovan\u00e9ho na vysoce inhibovan\u00e9 vzorky, NucleoSpin Soil (Macherey-Nagel, N\u011bmecko). Koncentrace DNA byla m\u011b\u0159ena pomoc\u00ed fluorometru Qubit\u00ae 2.0. Z\u00a0izolovan\u00e9 DNA byla provedena real-time qPCR anal\u00fdza na p\u0159\u00edstroji LightCycler\u00ae 480\u00a0(Roche). Jako fluorescen\u010dn\u00ed zdroj bylo pou\u017eito fluorogenn\u00ed interkala\u010dn\u00ed barvivo typu SYBR Green. C\u00edlem qPCR anal\u00fdzy bylo na jednotliv\u00fdch nosi\u010d\u00edch detekovat jak celkov\u00e9 bakteri\u00e1ln\u00ed o\u017eiven\u00ed, tak p\u0159\u00edtomnost funk\u010dn\u00edch gen\u016f a\u00a0kl\u00ed\u010dov\u00e9ho mikrobi\u00e1ln\u00edho konsorcia pod\u00edlej\u00edc\u00edch se na obou kroc\u00edch nitrifikace, oxidace amoniaku (AOB) a\u00a0dusitanu (NOB). Informace k\u00a0testovan\u00fdm marker\u016fm pro real-time qPCR jsou uvedeny v\u00a0<em>tabulce 3<\/em>.<\/p>\n<h5>Tabulka 3. Seznam testovan\u00fdch marker\u016f<br \/>\nTable 3. List of tested markers<\/h5>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-3.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"325\" class=\"alignnone size-full wp-image-8439 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-3.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-3.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-3-300x122.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-3-768x312.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/325;\" \/><\/a>\n<p>Protokol metody FISH sest\u00e1val ze \u010dty\u0159 z\u00e1kladn\u00edch krok\u016f: (1) fixace a\u00a0permeabilizace bun\u011bk, (2) hybridizace fluorescen\u010dn\u00edch sond specifick\u00fdch pro EUB, AOB a\u00a0NOB, (3) vymyt\u00ed nenav\u00e1zan\u00fdch sond a\u00a0(4) mikroskopick\u00e1 anal\u00fdza. Detekce ozna\u010den\u00fdch bun\u011bk byla provedena pomoc\u00ed fluorescen\u010dn\u00edho mikroskopu ZEISS Axio Imager.M2 s\u00a0kamerou AxioCamICc1. Kvantifikace ozna\u010den\u00fdch bun\u011bk na sn\u00edmc\u00edch z\u00a0fluorescen\u010dn\u00edho mikroskopu byla provedena pomoc\u00ed programu Matlab (MathWorks). Vyhodnocen\u00ed p\u0159\u00edtomnosti a\u00a0mno\u017estv\u00ed specifick\u00fdch bakteri\u00ed (AOB, NOB a\u00a0EUB) bylo prov\u00e1d\u011bno na z\u00e1klad\u011b pom\u011bru ploch bun\u011bk s\u00a0\u010dervenou fluorescenc\u00ed (AOB nebo NOB) k\u00a0oblastem bun\u011bk se zelenou fluorescenc\u00ed (EUB). Z\u00a0ka\u017ed\u00e9ho vzorku bylo po\u0159\u00edzeno cca 20 sn\u00edmk\u016f. Pou\u017eit\u00e9 FISH sondy jsou shrnuty v\u00a0<em>tabulce 4.<\/em><\/p>\n<h5>Tabulka 4. Seznam a\u00a0popis pou\u017eit\u00fdch FISH sond<br \/>\nTable 4. List and description of used FISH probes<\/h5>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-4.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"504\" class=\"alignnone size-full wp-image-8441 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-4.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-4.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-4-300x189.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-4-768x484.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/504;\" \/><\/a>\n<h3>Metoda respirometrick\u00e1<\/h3>\n<p>Syst\u00e9m respirometru Micro-Oxymax (Columbus Instruments) se v\u00a0z\u00e1kladu skl\u00e1d\u00e1 z\u00a0\u010derpadla vzork\u016f, roz\u0161i\u0159ovac\u00edho rozhran\u00ed a\u00a0individu\u00e1ln\u00edch plynov\u00fdch \u010didel pro ka\u017ed\u00fd m\u011b\u0159en\u00fd plyn. Pr\u016ftoky plyn\u016f mohou b\u00fdt zvoleny od 100 do 500\u00a0ml\u00b7min<sup>-1<\/sup> a\u00a0rozsahy senzor\u016f jsou pro CO<sub>2<\/sub> 0\u20130,9 % a\u00a0pro O<sub>2<\/sub> 15\u201321,5 %.<\/p>\n<p>Pro respirometrick\u00e9 m\u011b\u0159en\u00ed byl vzorek nanovl\u00e1kenn\u00e9ho nosi\u010de z\u00a0bioreaktoru (\u010dtverec 55\u00a0\u00d7\u00a055\u00a0mm) um\u00edst\u011bn do l\u00e1hve obsahuj\u00edc\u00ed 200 ml BSM m\u00e9dia, 20\u00a0mg\u00b7l<sup>-1<\/sup> N-NH<sub>4<\/sub><sup>+<\/sup> a\u00a010 ml fosf\u00e1tov\u00e9ho pufru. P\u0159ipraven\u00e9 l\u00e1hve byly n\u00e1sledn\u011b um\u00edst\u011bny na respirometr, kde byl po zad\u00e1n\u00ed parametr\u016f m\u011b\u0159en\u00ed cel\u00fd syst\u00e9m ut\u011bsn\u011bn, a\u00a0lahve se vzorky byly um\u00edst\u011bny na t\u0159epa\u010dku, kter\u00e1 zaji\u0161\u0165ovala prom\u00edch\u00e1v\u00e1n\u00ed obsahu b\u011bhem m\u011b\u0159en\u00ed. Vyhodnocen\u00ed respirac\u00ed prob\u00edhalo v\u00a0programu Matlab (The\u00a0Mathworks, Inc.).<\/p>\n<h2>V\u00fdsledky a\u00a0diskuse<\/h2>\n<h3>SEM a\u00a0anal\u00fdza obrazu<\/h3>\n<p>Vizualizace struktury nosi\u010d\u016f pomoc\u00ed SEM (<em>obr. 2<\/em>) a\u00a0n\u00e1sledn\u00e1 anal\u00fdza obrazu se v\u00a0t\u00e9to studii uk\u00e1zaly jako kl\u00ed\u010dov\u00e9 pro pochopen\u00ed interakc\u00ed mezi bu\u0148kami a\u00a0nosi\u010dem. Anal\u00fdza SEM sn\u00edmk\u016f (<em>obr. 3<\/em>) p\u0159isp\u011bla k\u00a0rozli\u0161en\u00ed z\u00e1kladn\u00edch rozd\u00edl\u016f mezi testovan\u00fdmi nanovl\u00e1kenn\u00fdmi materi\u00e1ly a\u00a0umo\u017enila stanovit nejvhodn\u011bj\u0161\u00ed polymer a\u00a0metodu pro p\u0159\u00edpravu vhodn\u00e9ho nosi\u010de biomasy.<\/p>\n<p><strong> <a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"1204\" class=\"alignnone size-full wp-image-8421 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2-199x300.jpg 199w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2-680x1024.jpg 680w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-2-768x1156.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/1204;\" \/><\/a><\/strong><\/p>\n<h6>Obr. 2. SEM sn\u00edmky nanovl\u00e1kenn\u00fdch nosi\u010d\u016f PUR (vlevo) a PVB (vpravo) z metod (a, A) AC elektrospinning, (b, B) ty\u010dka (DC), (c, C) Nanospider<sup>TM<\/sup> (DC), (d, D) jehla (DC) a\u00a0(e, E) elektro-odst\u0159ediv\u00e9 zvl\u00e1k\u0148ov\u00e1n\u00ed (DC); m\u011b\u0159\u00edtko odpov\u00edd\u00e1 5 \u00b5m<br \/>\nFig. 2. SEM images of nanofiber carriers PUR (left) and PVB (right) from methods (a, A) AC electrospinning, (b, B) rod (DC), (c, C) Nanospider<sup>TM<\/sup> (DC), (d, D) needle (DC) and (e, E) electro-forcespinning (DC); the scale corresponds to 5 \u00b5m<\/h6>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-3.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"306\" class=\"alignnone size-full wp-image-8423 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-3.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-3.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-3-300x115.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-3-768x294.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/306;\" \/><\/a>\n<h6>Obr. 3. V\u00fdsledky anal\u00fdzy obrazu SEM: (a) pr\u016fm\u011br vl\u00e1ken (v\u00a0nanometrech), (b) p\u00f3rovitost vl\u00e1ken (v\u00a0procentech); v\u0161echna data jsou zobrazena jako pr\u016fm\u011brn\u00e9 hodnoty se standardn\u00ed odchylkou<br \/>\nFig. 3. Results of SEM image analysis: (a) fiber diameter (nanometers), (b) fiber porosity (percent); all data are displayed as mean values with standard deviation<\/h6>\n<p>Nejmen\u0161\u00ed pr\u016fm\u011br m\u011bla u\u00a0obou polymer\u016f nanovl\u00e1kna vyroben\u00e1 metodou DC elektrospinning z\u00a0jehly, co\u017e bylo pravd\u011bpodobn\u011b zp\u016fsobeno horizont\u00e1ln\u00ed konfigurac\u00ed jehlov\u00e9 elektrody, kter\u00e1 m\u00e1 nav\u00edc mal\u00fd pr\u016fm\u011br. Polymer proch\u00e1z\u00ed \u00fazk\u00fdm prostorem na v\u00fdstupu z\u00a0elektrody (jehly), a\u00a0proto je ji\u017e v\u00a0tomto bod\u011b tvarov\u00e1n. Jak\u00e9koli sn\u00ed\u017een\u00ed vnit\u0159n\u00edho pr\u016fm\u011bru jehly zvy\u0161uje povrchov\u00e9 nap\u011bt\u00ed roztoku a\u00a0m\u00e1 za n\u00e1sledek men\u0161\u00ed kapi\u010dky, co\u017e zp\u016fsobuje sn\u00ed\u017een\u00ed rychlosti let\u00edc\u00edho nanovl\u00e1kna a\u00a0poskytuje del\u0161\u00ed \u010dasov\u00fd \u00fasek pro ta\u017een\u00ed, resp. tv\u00e1\u0159en\u00ed vl\u00e1kna. Naopak nejsiln\u011bj\u0161\u00ed nanovl\u00e1kna byla z\u00edsk\u00e1na z\u00a0odst\u0159ediv\u00e9ho elektrostatick\u00e9ho zvl\u00e1k\u0148ov\u00e1n\u00ed s\u00a0PVB polymerem, kde nedoch\u00e1zelo k\u00a0tak v\u00fdrazn\u00e9mu ta\u017een\u00ed vl\u00e1ken jako v\u00a0p\u0159\u00edpad\u011b metod ostatn\u00edch (krat\u0161\u00ed dr\u00e1ha letu vl\u00e1kna). Jin\u00e9 zp\u016fsoby zvl\u00e1k\u0148ov\u00e1n\u00ed nevykazovaly znateln\u00e9 zm\u011bny pr\u016fm\u011br\u016f nanovl\u00e1ken. PUR nanovl\u00e1kna m\u011bla tak\u00e9 tendenci b\u00fdt kompaktn\u011bj\u0161\u00ed (tj. hust\u011bj\u0161\u00ed uspo\u0159\u00e1d\u00e1n\u00ed) a\u00a0tvo\u0159ila koherentn\u011bj\u0161\u00ed vrstvu obsahuj\u00edc\u00ed m\u00e9n\u011b viditeln\u00e9 defekty, tj. mezery mezi vl\u00e1kny byly rovnom\u011brn\u011bj\u0161\u00ed.<\/p>\n<p>Por\u00e9znost PUR polymeru dos\u00e1hla hodnot v rozmez\u00ed 38 a\u017e 45 %. Velikost p\u00f3r\u016f byla m\u00edrn\u011b v\u011bt\u0161\u00ed p\u0159i pou\u017eit\u00ed AC a odst\u0159ediv\u00e9ho elektrostatick\u00e9ho zvl\u00e1k\u0148ov\u00e1n\u00ed. PVB nanovl\u00e1kenn\u00e9 struktury vykazovaly drsn\u011bj\u0161\u00ed uspo\u0159\u00e1d\u00e1n\u00ed povrchu a chaoti\u010dt\u011bj\u0161\u00ed \u0159azen\u00ed jednotliv\u00fdch vl\u00e1ken, co\u017e pravd\u011bpodobn\u011b souviselo s\u00a0p\u00f3rovitost\u00ed struktur. Nejni\u017e\u0161\u00ed porozity a\u00a0velikosti p\u00f3r\u016f pro PVB bylo dosa\u017eeno na Nanospideru, av\u0161ak v\u00fdsledky porozit se glob\u00e1ln\u011b p\u0159\u00edli\u0161 neli\u0161\u00ed.<\/p>\n<p>Vyhodnocen\u00ed d\u00e1le uk\u00e1zalo, \u017ee typ zvolen\u00e9ho polymeru i\u00a0pou\u017eit\u00e1 zvl\u00e1k\u0148ovac\u00ed technologie m\u011bly v\u00fdrazn\u00fd vliv na uspo\u0159\u00e1danost nanovl\u00e1kenn\u00e9 struktury. Nejvy\u0161\u0161\u00ed linearita vl\u00e1ken byla dosa\u017eena odst\u0159ediv\u00fdm elektrostatick\u00fdm zvl\u00e1k\u0148ov\u00e1n\u00edm s\u00a0PVB polymerem, zat\u00edmco nejvy\u0161\u0161\u00ed stupe\u0148 neuspo\u0159\u00e1danosti vykazovaly PUR nanovl\u00e1kna p\u0159ipraven\u00e1 AC zvl\u00e1k\u0148ov\u00e1n\u00edm.<\/p>\n<h3>Konfok\u00e1ln\u00ed mikroskopie<\/h3>\n<p>Chov\u00e1n\u00ed bun\u011bk na polymern\u00edch materi\u00e1lech (nap\u0159. adsorpce a\u00a0proliferace) z\u00e1vis\u00ed do zna\u010dn\u00e9 m\u00edry na povrchov\u00fdch vlastnostech materi\u00e1lu, v\u010detn\u011b sm\u00e1\u010divosti, hydrofility\/hydrofobity, chemick\u00e9m slo\u017een\u00ed, distribuci povrchov\u00e9ho n\u00e1boje, tuhosti a\u00a0zejm\u00e9na drsnosti povrchu. P\u0159i studiu povrchu nanovl\u00e1kenn\u00fdch struktur pomoc\u00ed konfok\u00e1ln\u00ed mikroskopie lze hodnotit jejich makro-geometrii (tvar, zavln\u011bn\u00ed povrchu), mikro-geometrii (drsnost povrchu) a\u00a0nano-geometrii (drsnost jednotliv\u00fdch vl\u00e1ken). Tato metoda je pova\u017eov\u00e1na za u\u017eite\u010dnou a\u00a0nedestruktivn\u00ed mo\u017enost charakterizace povrchu \u0161irok\u00e9 \u0161k\u00e1ly materi\u00e1l\u016f. \u010clenitost povrchu PVB nanovl\u00e1ken je zobrazena na <em>obr. 4<\/em>.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-4.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"697\" class=\"alignnone size-full wp-image-8425 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-4.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-4.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-4-300x261.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-4-768x669.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/697;\" \/><\/a>\n<h6>Obr. 4. Sn\u00edmky z\u00a0konfok\u00e1ln\u00ed mikroskopie povrchu nanovl\u00e1kenn\u00fdch nosi\u010d\u016f PUR (a\u2013e) a\u00a0PVB (f\u2013j) z\u00a0metod (a, f) AC elektrospinning, (b, g) ty\u010dka (DC), (c, h) Nanospider<sup>TM<\/sup> (DC), (d, i) jehla (DC) a\u00a0(e, j) elektro-odst\u0159ediv\u00e9 zvl\u00e1k\u0148ov\u00e1n\u00ed (DC); horn\u00ed vrstvy nanovl\u00e1ken jsou zobrazeny \u010derven\u011b, zat\u00edmco spodn\u00ed vrstvy nanovl\u00e1ken jsou modr\u00e9<br \/>\nFig. 4. Confocal microscopy images of the surface of nanofiber carriers PUR (a\u2013e) and PVB (f\u2013j) from methods (a, f) AC electrospinning, (b, g) rod (DC), (c, h) Nanospider<sup>TM<\/sup> (DC), (d, i) needle (DC) and (e, j) electro-forcespinning (DC); the top layers of nanofibers are shown in red, while the bottom layers of nanofibers are blue<\/h6>\n<p>Hlavn\u00ed rozd\u00edl mezi nanovl\u00e1kenn\u00fdmi strukturami PUR a\u00a0PVB z\u00a0r\u016fzn\u00fdch zvl\u00e1k\u0148ovac\u00edch metod byl v\u00a0drsnosti povrchu nanovl\u00e1ken (<em>obr. 5<\/em>), jak ukazuje konfok\u00e1ln\u00ed mikroskopie. Ve v\u011bt\u0161in\u011b p\u0159\u00edpad\u016f m\u011bl PUR hlad\u0161\u00ed povrch, co\u017e m\u016f\u017ee b\u00fdt d\u00e1no jeho specifick\u00fdmi vlastnostmi (viskoelastick\u00fd a\u00a0vykazuje slab\u0161\u00ed intramolekul\u00e1rn\u00ed s\u00edly). Naproti tomu PVB je amorfn\u00ed termoplastick\u00fd polymer, jeho\u017e nanovl\u00e1kenn\u00e1 vrstva vykazovala drsn\u011bj\u0161\u00ed povrch, tj. nanovl\u00e1kna vy\u010dn\u00edvala v\u00edce na povrch a\u00a0netvo\u0159ila hladkou rovnom\u011brnou vrstvu. To bylo pravd\u011bpodobn\u011b zp\u016fsobeno vy\u0161\u0161\u00ed p\u0159\u00edtomnost\u00ed deformovan\u00fdch nanovl\u00e1ken ve vrstv\u011b PVB v\u00a0kombinaci s\u00a0jejich chaotick\u00fdm uspo\u0159\u00e1d\u00e1n\u00edm. Nejedn\u00e1 se tedy pouze o\u00a0plo\u0161nou nerovnom\u011brnost povrchu, ale o\u00a0bodov\u00e9 v\u00fdstupy (\u201enano\u201c a\u00a0\u201emikro\u201c drsnost povrchu), kter\u00e9 jsou d\u016fle\u017eit\u00e9 pr\u00e1v\u011b pro adhezi mikroorganism\u016f k\u00a0substr\u00e1tu [17]. Konfok\u00e1ln\u00ed mikroskopie i\u00a0anal\u00fdza SEM uk\u00e1zaly, \u017ee z\u00a0DC metod maj\u00ed vrstvy kompaktn\u011bj\u0161\u00ed a\u00a0jemn\u011bj\u0161\u00ed povrchy ne\u017e z\u00a0metody AC a\u00a0odst\u0159ediv\u00e9ho elektrostatick\u00e9ho zvl\u00e1k\u0148ov\u00e1n\u00ed.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-5.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"561\" class=\"alignnone size-full wp-image-8427 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-5.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-5.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-5-300x210.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-5-768x539.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/561;\" \/><\/a>\n<h6>Obr. 5. Drsnost povrchu nanovl\u00e1ken (Sa) stanoven\u00e1 z\u00a0anal\u00fdzy sn\u00edmk\u016f konfok\u00e1ln\u00ed mikroskopie je zobrazena jako pr\u016fm\u011brn\u00e1 hodnota se standardn\u00ed odchylkou<br \/>\nFig. 5. Nanofiber surface roughness (Sa) determined from confocal microscopy image analysis is shown as the mean value with standard deviation<\/h6>\n<h3>Biodegradace nanovl\u00e1kenn\u00fdch nosi\u010d\u016f<\/h3>\n<p>Rychlost degradace polymeru z\u00e1vis\u00ed p\u0159ev\u00e1\u017en\u011b na jeho chemick\u00e9 struktu\u0159e, p\u0159\u00edtomnosti hydrolytick\u00fdch nestabiln\u00edch vazeb, \u00farovni hydrofility\/hydrofobity, krystalick\u00e9 nebo amorfn\u00ed morfologii, pom\u011bru jednotliv\u00fdch pom\u011br\u016f slo\u017eek kopolymeru nebo jeho molekulov\u00e9 hmotnosti [10]. O\u010dek\u00e1v\u00e1 se, \u017ee bun\u011b\u010dn\u00e1 proliferace i\u00a0biologick\u00e1 rozlo\u017eitelnost budou tak\u00e9 z\u00e1viset na uspo\u0159\u00e1d\u00e1n\u00ed a\u00a0geometrii nanovl\u00e1kenn\u00fdch materi\u00e1l\u016f [18].<\/p>\n<p>Biodegradace v\u00a0t\u00e9to studii byla obecn\u011b velmi n\u00edzk\u00e1 pro oba polymery (ze v\u0161ech metod). Byla prok\u00e1z\u00e1na stabilita obou polymer\u016f v\u00a0inokulovan\u00e9m vodn\u00e9m prost\u0159ed\u00ed a\u00a0na z\u00e1klad\u011b v\u00fdsledk\u016f lze jako jedin\u00fd, av\u0161ak nepatrn\u00fd, rozd\u00edl zm\u00ednit zanedbateln\u011b vy\u0161\u0161\u00ed stabilitu PUR oproti PVB (v\u00a0jednotk\u00e1ch tis\u00edcin procent). Biodegradace v\u0161ech p\u0159ipraven\u00fdch nosi\u010d\u016f tedy byla z\u00a0makro-hlediska zanedbateln\u00e1, a\u00a0tedy tyto nosi\u010de se jev\u00ed jako vhodn\u00e9 pro pou\u017e\u00edv\u00e1n\u00ed ve vodn\u00e9m prost\u0159ed\u00ed.<\/p>\n<h3>Biologick\u00e9 testov\u00e1n\u00ed nosi\u010d\u016f<\/h3>\n<p>Na z\u00e1klad\u011b v\u00fd\u0161e zm\u00edn\u011bn\u00e9 charakterizace nanovl\u00e1ken byla pro testov\u00e1n\u00ed v\u00a0bioreaktoru vybr\u00e1na PVB nanovl\u00e1kna p\u0159ipraven\u00e1\u00a0Nanospiderem jako nejvhodn\u011bj\u0161\u00ed materi\u00e1l z\u00a0hlediska predikovan\u00e9 proliferace bun\u011bk, n\u00e1ro\u010dnosti p\u0159\u00edpravy a\u00a0ekonomick\u00fdch parametr\u016f. P\u0159i v\u00fdb\u011bru nejvhodn\u011bj\u0161\u00ed zvl\u00e1k\u0148ovac\u00ed metody bylo nutn\u00e9 zohlednit mo\u017enost pr\u016fmyslov\u00e9 produkce nanovl\u00e1ken Nanospiderem, co\u017e v\u00fdrazn\u011b sni\u017euje celkov\u00e9 n\u00e1klady na v\u00fdrobu. PVB je dnes velmi roz\u0161\u00ed\u0159en\u00fd polymer, jeho\u017e cena umo\u017e\u0148uje p\u0159\u00edpravu levn\u00fdch, ale z\u00e1rove\u0148 velmi kvalitn\u00edch materi\u00e1l\u016f. V\u00fdrobn\u00ed cena vrstvy PVB nanovl\u00e1ken vyroben\u00e9 pomoc\u00ed Nanospideru se pohybuje v\u00a0\u0159\u00e1dech korun za 1 m<sup>2<\/sup>. Nav\u00edc, nanovl\u00e1kna PVB p\u0159ipraven\u00e1 t\u00edmto typem zvl\u00e1k\u0148ov\u00e1n\u00ed maj\u00ed vhodn\u00e9 struktur\u00e1ln\u00ed vlastnosti a\u00a0p\u0159\u00edprava zvl\u00e1k\u0148ovac\u00edho roztoku je velmi snadn\u00e1 (prost\u00e9 rozpu\u0161t\u011bn\u00ed polymeru PVB v\u00a0etanolu v\u00a0dan\u00e9m pom\u011bru). Naopak v\u00a0p\u0159\u00edpad\u011b PUR je p\u0159\u00edprava zvl\u00e1k\u0148ovac\u00edho roztoku podstatn\u011b slo\u017eit\u011bj\u0161\u00ed (nastaven\u00ed zvl\u00e1k\u0148ovac\u00edch parametr\u016f, precizn\u00ed nam\u00edch\u00e1n\u00ed polymern\u00edho roztoku a\u00a0n\u00edzk\u00e1 zvl\u00e1knitelnost metodou Nanospider) a\u00a0cena jednotliv\u00fdch slo\u017eek je vy\u0161\u0161\u00ed. Ekonomicky p\u0159ijateln\u00e1 varianta v\u00fdroby nanovl\u00e1kenn\u00fdch nosi\u010d\u016f je tak\u00e9 pomoc\u00ed metody AC elektrospinning. Vrstvy z\u00a0t\u00e9to metody jsou v\u0161ak v\u00edce neuspo\u0159\u00e1dan\u00e9 a\u00a0jejich mechanick\u00e1 stabilita je podstatn\u011b ni\u017e\u0161\u00ed. U\u00a0ostatn\u00edch metod se cena p\u0159\u00edli\u0161 navy\u0161uje d\u00edky n\u00edzk\u00e9 produktivit\u011b a\u00a0n\u00e1ro\u010dn\u011bj\u0161\u00ed obsluze za\u0159\u00edzen\u00ed.<\/p>\n<p>Porozita na \u00farovni 45 % u\u00a0PVB je p\u0159ijateln\u00e1 pro os\u00eddlov\u00e1n\u00ed bakteriemi a\u00a0z\u00e1rove\u0148 nen\u00ed p\u0159\u00edli\u0161 velk\u00e1, aby se mikroorganismy dost\u00e1valy hluboko do vnit\u0159n\u00ed struktury, kde se zna\u010dn\u011b sni\u017euje kontakt s\u00a0polutanty a\u00a0doch\u00e1z\u00ed k\u00a0nahromad\u011bn\u00ed neaktivn\u00edho biofilmu. Z\u00a0hlediska povrchu je PVB oproti PUR jednozna\u010dn\u011b v\u00fdhodn\u011bj\u0161\u00ed materi\u00e1l pro v\u00fdrobu nosi\u010de biomasy, jeliko\u017e PUR je sp\u00ed\u0161e hlad\u0161\u00ed bez v\u00fdrazn\u011bj\u0161\u00edch bodov\u00fdch v\u00fdstup\u016f. Bioreaktor s\u00a0vybran\u00fdmi PVB nosi\u010di byl v\u00a0provozu dva m\u011bs\u00edce a\u00a0byl sledov\u00e1n n\u00e1r\u016fst biofilmu na nosi\u010d\u00edch, resp. aktivita mikroorganism\u016f.<\/p>\n<h4>Chemismus reaktoru<\/h4>\n<p>Pr\u016fb\u011b\u017en\u00e9 fyzik\u00e1ln\u011b-chemick\u00e9 testov\u00e1n\u00ed reaktoru (<em>tabulka 5<\/em>) slou\u017eilo ke sledov\u00e1n\u00ed \u00fa\u010dinnosti nitrifikace.<\/p>\n<p>C\u00edlem chemick\u00e9 anal\u00fdzy bylo sledov\u00e1n\u00ed rozd\u00edln\u00fdch koncentrac\u00ed amoniak\u00e1ln\u00edho dus\u00edku ve vstupn\u00ed (modelov\u00e9) vod\u011b a\u00a0v\u00a0odtoku. Z\u00a0poklesu koncentrace N-NH<sub>4<\/sub><sup>+<\/sup> iont\u016f a\u00a0z\u00a0n\u00e1r\u016fstu N-NO<sub>3<\/sub><sup>&#8211;<\/sup> iont\u016f bylo mo\u017en\u00e9 sledovat m\u00edru oxidace amoniak\u00e1ln\u00edho dus\u00edku na dusi\u010dnany. Sledov\u00e1n\u00edm koncentrace dusitan\u016f byla ov\u011b\u0159ov\u00e1na kompletnost nitrifikace.<\/p>\n<p>Koncentrace N-NH<sub>4<\/sub><sup>+<\/sup> v\u00a0modelov\u00e9 vod\u011b (n\u00e1tok) kol\u00edsala mezi 9,5 a\u00a015,0 mg\u00b7l<sup>-1<\/sup>. Koncentrace N-NH<sub>4<\/sub><sup>+<\/sup> v\u00a0odtoku z\u00a0reaktoru byla typicky men\u0161\u00ed ne\u017e 0,5 mg\u00b7l<sup>-1<\/sup>, co\u017e ukazuje na vy\u0161\u0161\u00ed ne\u017e 95% \u00fa\u010dinnost oxidace amoniaku. Kompletn\u00ed nitrifikace byla prok\u00e1z\u00e1na zanedbatelnou koncentrac\u00ed N-NO<sub>2<\/sub><sup>&#8211;<\/sup> iont\u016f v\u00a0reaktoru (&lt; 0,1 mg\u00b7l<sup>-1<\/sup>).<\/p>\n<h5>Tabulka 5. V\u00fdsledky anal\u00fdz chemismu\u00a0bioreaktoru v\u00a0t\u00fddenn\u00edch intervalech<br \/>\nTable 5. Results of bioreactor chemistry analyzes at weekly intervals<\/h5>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-5.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"561\" class=\"alignnone size-full wp-image-8443 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-5.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-5.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-5-300x210.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-tabulka-5-768x539.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/561;\" \/><\/a>\n<h4>V\u00fdsledky molekul\u00e1rn\u00ed genetiky<\/h4>\n<p>Ve v\u0161ech vzorc\u00edch PVB nanovl\u00e1ken odebran\u00fdch z\u00a0bioreaktoru bylo celkov\u00e9 bakteri\u00e1ln\u00ed o\u017eiven\u00ed na vysok\u00e9 \u00farovni. V\u00a0pr\u016fb\u011bhu experimentu byl na v\u0161ech nanovl\u00e1knech detekov\u00e1n v\u00a0n\u00edzk\u00e9m mno\u017estv\u00ed funk\u010dn\u00ed gen k\u00f3duj\u00edc\u00ed kl\u00ed\u010dov\u00fd enzym oxidace amoniaku (<em>amoA<\/em>) specifick\u00fd pro AOB <em>Nitrosomonas<\/em>. N\u00e1podobn\u011b bylo mno\u017estv\u00ed funk\u010dn\u00edch gen\u016f specifick\u00fdch pro NOB <em>Nitrobacter<\/em> (<em>NxrB1<\/em>, <em>nxrA<\/em>) na v\u0161ech nanovl\u00e1knech v\u00a0pr\u016fb\u011bhu experimentu velmi n\u00edzk\u00e9 a\u017e na mezi detekce. Markery specifick\u00e9 pro NOB <em>Nitrospira<\/em> (<em>nxrB<\/em>, NSR) byly, na rozd\u00edl od NOB <em>Nitrobacter<\/em>, detekov\u00e1ny ji\u017e ve velk\u00e9m mno\u017estv\u00ed na v\u0161ech nanovl\u00e1knech od\u00a0prvn\u00edho odb\u011bru.<\/p>\n<p>N\u00e1r\u016fst celkov\u00e9 a\u00a0testovan\u00e9 specifick\u00e9 bakteri\u00e1ln\u00ed biomasy na jednotliv\u00fdch nanovl\u00e1knech v\u00a0pr\u016fb\u011bhu experimentu byl vyj\u00e1d\u0159en pomoc\u00ed relativn\u00ed kvantifikace (<em>obr. 6<\/em>), jen\u017e popisuje relativn\u00ed zm\u011bnu mno\u017estv\u00ed dan\u00e9ho markeru v\u016f\u010di referen\u010dn\u00edmu vzorku, kter\u00fdm je v\u00a0tomto p\u0159\u00edpad\u011b vzorek z\u00a01. odb\u011bru. Hodnota relativn\u00ed kvantifikace referen\u010dn\u00edho vzorku je 1. Pokud je hodnota relativn\u00ed kvantifikace vzorku 2, znamen\u00e1 to, \u017ee v\u00a0tomto vzorku je dvakr\u00e1t v\u00edce c\u00edlov\u00e9 DNA ne\u017e v\u00a0referen\u010dn\u00edm vzorku. V\u00a0p\u0159\u00edpad\u011b specifick\u00e9ho funk\u010dn\u00edho genu <em>amoA<\/em> (AOB <em>Nitrosomonas<\/em>) byl pozorov\u00e1n postupn\u00fd n\u00e1r\u016fst v\u00a0pr\u016fb\u011bhu experimentu u\u00a0v\u0161ech nanovl\u00e1ken, stejn\u011b tak v\u00a0p\u0159\u00edpad\u011b NOB <em>Nitrospira<\/em> (NSR, <em>nxrB<\/em>). V\u00a0pr\u016fb\u011bhu experimentu se testovan\u00e1 bakteri\u00e1ln\u00ed biomasa v\u00fdrazn\u011b nem\u011bnila, co\u017e indikuje, \u017ee v\u00a01.\u00a0odb\u011bru, tedy po 14\u00a0dnech od zano\u0159en\u00ed nanovl\u00e1ken, byl na povrchu nanovl\u00e1ken ji\u017e stabiln\u00ed biofilm. Mno\u017estv\u00ed testovan\u00fdch marker\u016f se v\u00fdrazn\u011b neli\u0161ilo mezi jednotliv\u00fdmi kvadruplik\u00e1ty.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-6.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"633\" class=\"alignnone size-full wp-image-8429 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-6.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-6.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-6-300x237.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-6-768x608.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/633;\" \/><\/a>\n<h6>Obr. 6. Relativn\u00ed kvantifikace celkov\u00e9 a\u00a0specifick\u00e9 bakteri\u00e1ln\u00ed biomasy na jednotliv\u00fdch kvadruplik\u00e1tech v\u00a0pr\u016fb\u011bhu experimentu v\u016f\u010di prvn\u00edmu odb\u011bru<br \/>\nFig. 6. Relative quantification of total and specific bacterial biomass for individual quadruplicates during the experiment compared to the first sampling<\/h6>\n<p>V\u00fdsledky FISH anal\u00fdzy posledn\u00edho odb\u011bru potvrzuj\u00ed z\u00e1v\u011bry z\u00a0qPCR anal\u00fdzy. Ze sn\u00edmk\u016f (<em>obr. 7<\/em>) je patrn\u00e9 n\u00edzk\u00e9 celkov\u00e9 mno\u017estv\u00ed AOB ve vzorc\u00edch. Naopak zna\u010dn\u00e9 zastoupen\u00ed \u010derven\u00e9 barvy (NOB) k\u00a0zelen\u00e9 (EUB) ve\u00a0sn\u00edmc\u00edch je pro NOB, kdy se jednotliv\u00e9 kvadruplik\u00e1ty p\u0159\u00edli\u0161 neli\u0161\u00ed.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"1196\" class=\"alignnone size-full wp-image-8431 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7-201x300.jpg 201w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7-685x1024.jpg 685w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-7-768x1148.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/1196;\" \/><\/a>\n<h6>Obr. 7. Uk\u00e1zka sn\u00edmk\u016f z\u00a0FISH anal\u00fdzy posledn\u00edho odb\u011bru, z\u00a0ka\u017ed\u00e9 sady sn\u00edmk\u016f vybr\u00e1n jeden od ka\u017ed\u00e9ho z\u00a0kvadruplik\u00e1tu pro AOB (vlevo) a\u00a0NOB (vpravo)<br \/>\nFig. 7. Images from the FISH analysis of the last sampling, one from each set of images selected from each of the quadruplicates for AOB (left) and NOB (right)<\/h6>\n<p>Celkov\u011b ni\u017e\u0161\u00ed mno\u017estv\u00ed AOB oproti NOB je b\u011b\u017en\u00fd jev, kter\u00fd se vyskytuje na \u010cOV. Pom\u011br AOB\/NOB hraje z\u00e1sadn\u00ed roli v\u00a0optimalizaci nitrifika\u010dn\u00edho procesu. V\u00a0p\u0159\u00edpad\u011b rychlej\u0161\u00edho r\u016fstu AOB oproti NOB bude doch\u00e1zet k\u00a0akumulaci dusitan\u016f v\u00a0syst\u00e9mu, co\u017e povede k\u00a0vytv\u00e1\u0159en\u00ed toxick\u00e9ho prost\u0159ed\u00ed. Na\u0161t\u011bst\u00ed bylo prok\u00e1z\u00e1no, \u017ee NOB maj\u00ed dominantn\u00ed zastoupen\u00ed v\u00a0nitrifika\u010dn\u00edm procesu a\u00a0re\u00e1ln\u00e1 situace neodpov\u00edd\u00e1 teoretick\u00fdm p\u0159edpoklad\u016fm, podle kter\u00fdch by m\u011bly v\u00a0syst\u00e9mu p\u0159eva\u017eovat AOB [19, 20].<\/p>\n<h4>Respirometrick\u00e9 testy<\/h4>\n<p>Respirometrie s\u00a0nosi\u010di PVB (po dvoum\u011bs\u00ed\u010dn\u00edm testov\u00e1n\u00ed v\u00a0biologick\u00e9m reaktoru) byla provedena v\u00a0kvadruplik\u00e1tu s\u00a0kontroln\u00edm vzorkem. V\u00fdsledky rychlosti spot\u0159eby O<sub>2<\/sub> (<em>obr. 8<\/em>) ukazovaly dobrou aktivitu nitrifika\u010dn\u00edch bakteri\u00ed oproti blanku (pouze medium bez inokula). S\u00a0ohledem na slabou vrstvu biofilmu na nosi\u010d\u00edch byla \u00fa\u010dinnost tohoto syst\u00e9mu zna\u010dn\u00e1, co\u017e bylo potvrzeno stanovenou koncentrac\u00ed N\u2011NH<sub>4<\/sub><sup>+<\/sup> ve vzorc\u00edch po ukon\u010den\u00ed respirometrie, kter\u00e1 byla ni\u017e\u0161\u00ed ne\u017e\u00a01. Nanovl\u00e1kenn\u00e9 nosi\u010de nevykazovaly po ukon\u010dov\u00e1n\u00ed experimentu viditeln\u00e9 po\u0161kozen\u00ed, tj. struktura se z\u00a0makro-pohledu nezm\u011bnila oproti p\u016fvodn\u00edmu stavu.<\/p>\n<a href=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-8.jpg\" rel=\"shadowbox[sbpost-8449];player=img;\"><img decoding=\"async\" width=\"800\" height=\"408\" class=\"alignnone size-full wp-image-8433 lazyload\" data-src=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-8.jpg\" alt=\"\" data-srcset=\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-8.jpg 800w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-8-300x153.jpg 300w, https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/08\/Havlicek-8-768x392.jpg 768w\" data-sizes=\"(max-width: 800px) 100vw, 800px\" src=\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\" style=\"--smush-placeholder-width: 800px; --smush-placeholder-aspect-ratio: 800\/408;\" \/><\/a>\n<h6>Obr. 8. Rychlost spot\u0159eby kysl\u00edku p\u0159i z\u00e1v\u011bre\u010dn\u00e9m testov\u00e1n\u00ed PVB nosi\u010d\u016f na respirometru<br \/>\nFig. 8. Oxygen consumption rate during the final testing of PVB carriers on the respirometer<\/h6>\n<h2>Z\u00e1v\u011br<\/h2>\n<p>V\u00a0r\u00e1mci v\u00fdvoje nanovl\u00e1kenn\u00fdch struktur je velmi d\u016fle\u017eit\u00e9 detailn\u011b analyzovat a\u00a0charakterizovat dan\u00e9 materi\u00e1ly. V\u00a0p\u0159\u00edpad\u011b testov\u00e1n\u00ed t\u011bchto struktur v\u00a0syst\u00e9mech \u010di\u0161t\u011bn\u00ed odpadn\u00edch vod (nosi\u010de biomasy) je d\u016fle\u017eit\u00e1 i\u00a0spr\u00e1vn\u00e1 interpretace a\u00a0hodnocen\u00ed r\u016fstu biofilmu na testovan\u00fdch nosi\u010d\u00edch. Biologick\u00e9 testy jsou zalo\u017eeny na odli\u0161n\u00fdch principech a\u00a0v\u00fdsledky anal\u00fdz r\u016fzn\u00fdch metod jsou velmi \u010dasto slo\u017eit\u011b porovnateln\u00e9. Metoda respirometrick\u00e1 hodnot\u00ed aktivitu specifick\u00e9 skupiny mikroorganism\u016f v\u00a0pr\u016fb\u011bhu m\u011b\u0159en\u00ed a\u00a0d\u00e1v\u00e1 celkov\u00fd n\u00e1hled o\u00a0stavu sledovan\u00fdch bakteri\u00ed. Oproti tomu metody molekul\u00e1rn\u011b genetick\u00e9 poskytuj\u00ed detailn\u00ed n\u00e1hled do biofilmu na \u00farovni DNA a\u00a0sleduj\u00ed tedy konkr\u00e9tn\u00ed mikroorganismy (kvalitu i\u00a0kvantitu), kter\u00e9 se zde nach\u00e1zej\u00ed. Anal\u00fdza SEM sn\u00edmk\u016f p\u0159ipraven\u00fdch materi\u00e1l\u016f se uk\u00e1zala jako nezbytn\u00e1 pro charakterizaci nanovl\u00e1kenn\u00e9 vrstvy. Pomoc\u00ed t\u00e9to anal\u00fdzy byly stanoveny parametry, tj. pr\u016fm\u011br vl\u00e1ken a\u00a0porozita, pro ur\u010den\u00ed nejvhodn\u011bj\u0161\u00edho materi\u00e1lu pro sledovan\u00e9 biologick\u00e9 aplikace. Pomoc\u00ed konfok\u00e1ln\u00ed mikroskopie bylo mo\u017en\u00e9 detailn\u011b zobrazit drsnost povrchu, kter\u00e1 je z\u00e1sadn\u00ed pro adhezi \u010dist\u00edrensk\u00fdch bakteri\u00ed, a\u00a0vybrat tak vhodn\u00fd materi\u00e1l pro testov\u00e1n\u00ed.<\/p>\n<p>Z\u00a0v\u00fd\u0161e uveden\u00e9ho vypl\u00fdv\u00e1, \u017ee kl\u00ed\u010dov\u00fdmi prvky pro v\u00fdb\u011br vhodn\u00e9ho materi\u00e1lu k\u00a0p\u0159\u00edprav\u011b nosi\u010d\u016f biomasy jsou v\u00fdsledn\u00e9 struktury nanovl\u00e1ken, jejich povrch a\u00a0stabilita ve vodn\u00e9m prost\u0159ed\u00ed. Pr\u016fm\u011br vl\u00e1ken rozhoduje o\u00a0povrchov\u00e9 plo\u0161e a\u00a0mechanick\u00fdch vlastnostech, co\u017e jsou d\u016fle\u017eit\u00e9 parametry jak pro adhezi bakteri\u00ed, tak pro \u017eivotnost samotn\u00e9ho nosi\u010de. Charakteristiky povrchu jsou z\u00e1sadn\u00ed pro bakteri\u00e1ln\u00ed adhezi a\u00a0prostupnost bakteri\u00ed a\u00a0jejich extracelul\u00e1rn\u00ed matrix t\u011bsn\u011b pod povrch (zabudov\u00e1n\u00ed biofilmu do povrchu). S\u00a0t\u00edmto souvis\u00ed i\u00a0porozita nanovl\u00e1kenn\u00e9ho nosi\u010de, kter\u00e1 mus\u00ed umo\u017enit bakteri\u00edm vstup do vnit\u0159n\u00ed struktury nosi\u010de, ale pouze do ur\u010dit\u00e9 hloubky. V\u00a0p\u0159\u00edpad\u011b p\u0159\u00edli\u0161 velk\u00fdch p\u00f3r\u016f doch\u00e1z\u00ed sm\u011brem dovnit\u0159 k\u00a0zahu\u0161\u0165ov\u00e1n\u00ed biofilmu a\u00a0ve vzd\u00e1len\u011bj\u0161\u00edch vrstv\u00e1ch ji\u017e nedoch\u00e1z\u00ed ke kontaktu s\u00a0polutanty ve vod\u011b, co\u017e vede k\u00a0\u00fabytku bakteri\u00e1ln\u00ed populace. V\u00a0t\u00e9to f\u00e1zi m\u016f\u017ee doch\u00e1zet ke zvy\u0161ov\u00e1n\u00ed hmotnosti nosi\u010de, co\u017e je ne\u017e\u00e1douc\u00ed. Na z\u00e1klad\u011b vyjmenovan\u00fdch parametr\u016f a\u00a0s\u00a0ohledem na ekonomickou n\u00e1ro\u010dnost byl jako ide\u00e1ln\u00ed nosi\u010d vybr\u00e1n materi\u00e1l z\u00a0PVB nanovl\u00e1ken p\u0159ipraven\u00fd metodou Nanospider. Pr\u016fm\u011br nanovl\u00e1ken (cca 400 nm) v\u00a0tomto p\u0159\u00edpad\u011b zaji\u0161\u0165uje dobrou mechanickou stabilitu vrstvy a\u00a0z\u00e1rove\u0148 vl\u00e1kna tvo\u0159\u00ed dostate\u010dn\u011b jemnou vnit\u0159n\u00ed strukturu, co\u017e je pro bakterie v\u00fdhodn\u00e9. Porozita t\u00e9to vrstvy je v\u00a0rozmez\u00ed, kdy bakterie a\u00a0jejich produkty nepronikaj\u00ed p\u0159\u00edli\u0161 hluboko do struktury nosi\u010de, ale z\u00e1rove\u0148 v\u00a0bl\u00edzkosti povrchu jim vytv\u00e1\u0159\u00ed vhodn\u00e9 prost\u0159ed\u00ed. Povrch je dostate\u010dn\u011b \u010dlenit\u00fd a\u00a0umo\u017e\u0148uje v\u00fdbornou bakteri\u00e1ln\u00ed adhezi.<\/p>\n<p>Biologick\u00e9 testov\u00e1n\u00ed v\u00a0bioreaktoru a\u00a0n\u00e1sledn\u00e9 hodnocen\u00ed biofilmu uk\u00e1zalo, \u017ee velmi specifick\u00fd povrch PVB nanovl\u00e1ken p\u0159ipraven\u00fdch technologi\u00ed Nanospider umo\u017enuje velmi kvalitn\u00ed adhezi \u010dist\u00edrensk\u00fdch bakteri\u00ed, kter\u00e9 jsou v\u00a0takov\u00e9m biofilmu aktivn\u00ed a\u00a0jsou \u010d\u00e1ste\u010dn\u011b chr\u00e1n\u011bny p\u0159ed nep\u0159\u00edzniv\u00fdmi vlivy. V\u00fdsledky byly potvrzeny v\u0161emi hodnot\u00edc\u00edmi metodami. Dal\u0161\u00ed v\u00fdvoj je sm\u011b\u0159ov\u00e1n do aplikovatelnosti t\u011bchto nosi\u010d\u016f biomasy na poloprovozn\u00ed testov\u00e1n\u00ed, kde mus\u00ed b\u00fdt br\u00e1n ohled p\u0159edev\u0161\u00edm na ekonomickou p\u0159\u00ednosnost.<\/p>\n<h3>Pod\u011bkov\u00e1n\u00ed<\/h3>\n<p><em>Tato pr\u00e1ce byla podpo\u0159ena z\u00a0projektu Studentsk\u00e9 grantov\u00e9 sout\u011b\u017ee (SGS-2019-3023) na Technick\u00e9 univerzit\u011b v\u00a0Liberci v\u00a0roce 2020.<\/em><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Nanomaterials have attracted great attention in diverse areas because of their unique properties. One of the possible nanofiber applications, due to their high surface area, is a\u00a0biomass carrier, thus the immobilization of the microbial cells followed by the biofilm formation.<\/p>\n","protected":false},"author":8,"featured_media":8446,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[89],"tags":[1963,1957,1964,1965,1960,1966,1962],"coauthors":[1948,1949,1950],"class_list":["post-8449","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-water-technology-water-supply-waste-water-treatment","tag-biomass-carriers","tag-fish","tag-nanofiber-characterization","tag-nitrification","tag-real-time-qpcr","tag-respirometry","tag-sem"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/8449","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=8449"}],"version-history":[{"count":2,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/8449\/revisions"}],"predecessor-version":[{"id":30596,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/8449\/revisions\/30596"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/8446"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=8449"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=8449"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=8449"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=8449"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}