In the present study, the effect of three intermittent aeration (IA) cycles on treatment performance and microbial diversity was investigated in an integrated fixed film activated sludge (IFAS) reactor treating municipal wastewater. The results showed that IA strategies were able to achieve efficient removal of organics and nitrogen ranging between 90 and 95% and 70 and 80%, respectively, however the phosphorus removal was found to be inversely proportional to the duration of aeration off time in each IA cycle. The microscopic analysis revealed that the suspended and attached biomass had compact morphology and open floc structure, respectively. For each gram of volatile suspended solids, 165 and 148 mg of extracellular polymeric substances (EPS) were extracted from attached and suspended biomass, respectively, constituting carbohydrates (∼24%), proteins (∼31%), humic acids (∼28%), DNA (∼2%) and unknown substances (∼12%). The microbial diversities of suspended biomass in IFAS reactor were investigated using culture-dependent approach, which confirmed the presence of Clostridium spp., Pseudomonas spp., Bacillus spp., Escherichia coli spp., Nitrosococcus spp., Streptococcus spp., Acinetobacter spp., Betaproteobacteria outliers, Klebsiella pneumoniae, Klebsiella aerogenes, Serratia marcescens, Micrococcus, Proteus vulgaris spp., Actinomycetes spp., and Actinobacteria including Micromonospora spp. and Streptomyces spp. Molecular tools for diversity analyses were used for ammonia and nitrite oxidizer identification, such as Nitrospira and Nitrosococcus species. Denitrifiers include the species of Pseudomonas, Betaproteobacteria and Flavobacterium. Acinetobacter, Betaproteobacteria and Gammaproteobacteria were responsible for the phosphorus removal in the present system. Overall, the system performed efficiently showing Proteobacteria (59%), Acinetobacter (12%) and Bacteroidetes (11%) as the dominant bacterial groups. However, the dominance of the bacterial diversity varied with the IA cycle time numerating the maximum percentage of bacterial species during IA1 phase i.e. 2.5 h aeration/0.5 h non-aeration.
Keywords: 16S rRNA; EPS; IFAS; SEM; microbial community; nutrient removal.