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The research focused on such packing media as ceramsite, polypropylene balls, and elastic fillers, and analyzed the main characteristics of extracellular polymeric substances (EPS) in their filter biofilms. The EPS were categorized as soluble EPS (S-EPS) and bound EPS (B-EPS). The component characteristics of stratified EPS were investigated via UV-Vis spectroscopy and a three-dimensional excitation-emission matrix (3D-EEM). The results showed that the EPS content of ceramsite biofilm was 245.2 mg/g VSS, which was higher than those of elastic filler material and polypropylene ball by 1.26 and 1.51 times, respectively. The protein (PN) and polysaccharide (PS) ratio of EPS in the ceramsite filter material was highest in S-EPS and B-EPS, indicating that the EPS have a stable structure. More than 67.58% of EPS formed by ceramsite was tightly bound EPS (TB-EPS), which was beneficial to maintaining the stability of biofilms. The levels of proteins (PN) and humic substances formed by ceramsite contained in the TB-EPS were higher than those of elastic filter material and polypropylene balls. 3D-EEM fluorescence spectra revealed that TB-EPS formed by a ceramsite contained high concentrations of tryptophan, tyrosine, and humic substances. The dehydrogenase activity of biofilm formed by ceramsite filler was higher than that formed by elastic fillers and polypropylene balls.
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