Aplikace nanovlákenných nosičů pro diagnostiku biomasy na kontaminované lokalitě
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Tato diplomová práce je zaměřena na aplikace nanovlákenných nosičů pro monitoring a diagnostiku biomasy v prostředí kontaminované lokality. V rámci práce byly připraveny dvě tvarové varianty nosičů s různými nánosy nanovláken (3 dtex a 10 dtex). Kromě nosičů s nanovlákny byly testovány také nosiče bez nanovláken, a to s cílem porovnat vliv nanovláken na rychlost kolonizace nosičů mikroorganismy. Všechny nosiče byly zanořeny do vrtů na reálných lokalitách s rozdílným typem kontaminace. Konkrétně se jednalo o kontaminaci chlorovanými etheny a monocyklickými aromatickými uhlovodíky BTEX. Nanovlákenné nosiče byly vzorkovány v měsíčních intervalech po dobu jednoho roku. Z odebraných vzorků byla izolována DNA sloužící jako templát pro qPCR analýzu. Vliv tvaru nosiče a nánosu nanovláken na bakteriální kolonizaci byl hodnocen s využitím qPCR analýzy cílené do oblasti 16S rDNA genu. Diagnostika specifické biomasy s biodegradabilním potenciálem probíhala pomocí kvantifikace funkčních genů kódujících klíčové enzymy nebo specifické bakteriální kmeny.Mezi jednotlivými testovanými tvarovými variantami nosičů nebyly během jejich kolonizace mikroorganismy pozorovány významné rozdíly. U nosičů s nánosem nanovláken byla zjištěna rychlejší fixace mikroorganismů na povrch a vyšší stabilita biofilmu než u nosičů vyrobených pouze z nosné nitě, tj. bez nánosu nanovláken.
This thesis focuses on the applications of nanofiber carriers for monitoring and diagnosis of biomass at the contaminated site. Within this work, carriers made of supporting thread coated by nanofiber layer were prepared. Individual carriers varied in shape (2) and nanofiber surface density (3 dtex and 10 dtex). To compare the effect of nanofibers on bacterial colonization, carriers without nanofibers, i.e. made of supporting thread only, were also tested. All carriers were submerged into wells at real sites with different contamination. Specifically, wells were contaminated by chlorinated ethenes and monocyclic aromatic hydrocarbons (BTEX).Carriers were sampled in month interval for one year. DNA was isolated from the samples and used as template for qPCR analysis. For the assessment of the influence of carrier shape and nanofiber surface density, the samples were analysed by qPCR with bacterial 16S rDNA gene-targeted primers. Diagnosis of specific biomass involved in the biodegradation was performed by quantification of functional genes encoding key enzymes or specific bacterial strains. Any significant differences between shapes of carrier in microbial growth were not observed. The results showed that nanofiber carriers supported microbial immobilization and biofilm exhibited higher stability compared to the biofilm formed on the carriers made of supporting thread only, i.e. without nanofibers.
This thesis focuses on the applications of nanofiber carriers for monitoring and diagnosis of biomass at the contaminated site. Within this work, carriers made of supporting thread coated by nanofiber layer were prepared. Individual carriers varied in shape (2) and nanofiber surface density (3 dtex and 10 dtex). To compare the effect of nanofibers on bacterial colonization, carriers without nanofibers, i.e. made of supporting thread only, were also tested. All carriers were submerged into wells at real sites with different contamination. Specifically, wells were contaminated by chlorinated ethenes and monocyclic aromatic hydrocarbons (BTEX).Carriers were sampled in month interval for one year. DNA was isolated from the samples and used as template for qPCR analysis. For the assessment of the influence of carrier shape and nanofiber surface density, the samples were analysed by qPCR with bacterial 16S rDNA gene-targeted primers. Diagnosis of specific biomass involved in the biodegradation was performed by quantification of functional genes encoding key enzymes or specific bacterial strains. Any significant differences between shapes of carrier in microbial growth were not observed. The results showed that nanofiber carriers supported microbial immobilization and biofilm exhibited higher stability compared to the biofilm formed on the carriers made of supporting thread only, i.e. without nanofibers.
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BTEX, DNA, chlorované etheny, kontaminovaná lokalita, molekulární genetika, monitoring, nanovlákna, nosiče biomasy, real-time kvantitativní PCR, vzorkování, BTEX, DNA, chlorinated ethenes, contaminated site, molecular genetics, monitoring, nanofibers, biomass carriers, real-time quantitative PCR, sampling