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Determination of significant factors for biodegradation of microplastics by Bacillus cereus and Pseudomonas alcaligenes
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This Croatian thesis used screening experiments to identify significant factors affecting biodegradation of microplastics by Bacillus cereus and Pseudomonas alcaligenes bacteria. The study characterized optimal conditions for microbial plastic breakdown and enzyme production.
Kontinuirana proizvodnja plastičnih proizvoda povećanog opsega te neprikladno rukovanje s plastičnim otpadom rezultirali su nagomilavanjem plastičnog otpada u okolišu koji se s vremenom razlaže na manje čestice. Sve plastične čestice manje od 5 mm nazivaju se mikroplastikom. U svom sastavu mikroplastika uobičajeno sadrži aditive koji poboljšavaju svojstva plastičnih proizvoda. Međutim, kroz godine istraživanja, utvrđeni su njihovi štetni učinci na biocenozu. Stoga je od izuzetne važnosti pronaći učinkovitu metodu uklanjanja mikroplastike iz okoliša. U ovom radu ispitala se biorazgradnja polistirena (PS) i poli(vinil-klorida) (PVC) primjenom bakterijskih kultura Bacillus cereus i Pseudomonas alcaligenes u trajanju od 30 dana. Plan eksperimenata biorazgradnje dizajniran je prema Taguchi metodi prema kojem je ispitano 7 čimbenika: pH-vrijednost, temperatura, veličina MP-a, koncentracija MP-a, broj okretaja i koncentracija dodane glukoze, na dvije razine – min i max vrijednost. Tijekom procesa pratio se broj izraslih stanica bakterija (CFU), koncentracija ukupnog organskog ugljika (TOC), ukupnog ugljika (TC) i anorganskog ugljika (IC), koncentracija aniona i kationa te je provedena HPLC/MS analiza u svrhu identificiranja otpuštenih aditiva s površine MP-a ili nastalih razgradnih organskih produkata. Uzorci su okarakterizirani FTIR-ATR analizom. Na kraju svih procesa biorazgradnje ispitana je i ekotoksičnost filtrata primjenom bakterije Vibrio fischeri radi utvrđivanja postojećih potencijalnih štetnih učinaka na ispitivani organizam. Eksperimentalni rezultati obrađeni su korištenjem statističke analize varijance (ANOVA). Dobiveni rezultati ukazuju da je Bacillus cereus bolje razgradio PS, a Pseudomonas alcaligenes PVC. FTIR spektri PS-a i PVC-a snimljeni prije i nakon biorazgradnje upućuju na nastanak novih funkcionalnih skupina, gubitak postojećih te na smanjenje intenziteta pikova apsorpcijskih vrpci. Značajni čimbenici za biorazgradnju PS-a i PVC-a bakterijom Bacillus cereus su pH-vrijednost, veličina PS-a, broj okretaja, optička gustoća (OG), koncentracija dodane glukoze odnosno pH-vrijednost, temperatura, veličina PVC-a, OG i koncentracija dodane glukoze. Značajni čimbenici za biorazgradnju PS-a i PVC-a bakterijom Pseudomonas alcaligenes su veličina PS-a, broj okretaja i OG odnosno pH-vrijednost i veličina PVC-a.
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