The performance of a full-scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was investigated by microbial analysis and kinetic modelling. The microbial community present in the granular sludge was detected using fluorescent in situ hybridization (FISH) and further confirmed using polymerase chain reaction. A group of 16S rRNA based fluorescent probes and primers targeting Archaea and Eubacteria were selected for microbial analysis. FISH results indicated the presence and dominance of a significant amount of Eubacteria and diverse group of methanogenic Archaea belonging to the order Methanococcales, Methanobacteriales, and Methanomicrobiales within in the UASB reactor. The influent brewery wastewater had a relatively high amount of volatile fatty acids chemical oxygen demand (COD), 2005 mg/l and the final COD concentration of the reactor was 457 mg/l. The biogas analysis showed 60-69% of methane, confirming the presence and activities of methanogens within the reactor. Biokinetics of the degradable organic substrate present in the brewery wastewater was further explored using Stover and Kincannon kinetic model, with the aim of predicting the final effluent quality. The maximum utilization rate constant U max and the saturation constant (K(B)) in the model were estimated as 18.51 and 13.64 g/l/day, respectively. The model showed an excellent fit between the predicted and the observed effluent COD concentrations. Applicability of this model to predict the effluent quality of the UASB reactor treating brewery wastewater was evident from the regression analysis (R(2) = 0.957) which could be used for optimizing the reactor performance.
Phenol, a common constituent in many industrial wastewaters is a major pollutant and has several adverse effects on the environment. The potential of various microorganisms to utilize phenol for their metabolic activity has been observed to be an effective means of remediating this toxic compound from the environment particularly wastewater. Five indigenous bacterial isolates (PD1-PD5) were obtained from phenol bearing industrial wastewater using the mineral salts medium. The isolates were further characterized based on their morphology, biochemical reactions and 16S rRNA phylogeny. The 16S rRNA sequence analysis using universal primers (27f/1492r) revealed that PD1, PD2, PD3 and PD4 were closely related to the actinomycete Rhodococcus pyrinidivorans (99%) and PD5 to Pseudomonas aeruginosa (99%). Growth kinetic patterns and phenol degradation abilities of the two representative isolates (PD1 and PD5) were also evaluated. Both the species were effective in utilizing phenol as the sole carbon source and could tolerate phenol concentrations of up to 500 to 600 mg/L. The ability of these isolates to utilize higher concentrations of phenol as their sole carbon source makes them potential candidates and better competitors in the bioremediation process.
The dominant nitrifying bacterial communities and nitrification performance of two biological nutrient removal plants were evaluated. Fluorescent in situ hybridization was used to detect and quantify the dominant nitrifying bacteria and polymerase chain reaction; cloning and sequence analysis of 16S rRNA genes was done for phylogenetic analysis. Fluorescent in situ hybridization-confocal scanning laser microscopy studies revealed the presence and dominance of Nitrosomonas-related ammonia-oxidizing bacteria (AOB) and Nitrobacter-related nitrite-oxidizing bacteria (NOB); however, a significant variation in AOB/NOB ratios was recorded. The plant with an overall higher AOB/NOB ratio (> or = 1.0) and dissolved oxygen concentration (1.8 to 2.5 mg/L) showed a higher nitrification rate. This study has also shown the co-existence and variation in phylogenetically diverse Nitrosomonas-related AOB and Nitrobacter-related NOB at these two plants. These dissimilar, distinct distribution patterns of nitrifying communities could be attributed to wastewater characteristics and the process configuration, which, in turn, would have also affected the nitrification performance of the systems.
A Kubotatrade mark submerged membrane bio-reactor was applied to treat wastewater from a sugar manufacturing industry. To achieve optimal results, fundamental and extended understanding of the microbiology is important. Fluorescence in situ hybridization was used to evaluate the microbial community present. The majority of cells visualized in the sludge flocs by staining with the DNA fluorochrome DAPI, hybridized strongly with a bacterial probe. Probes specific for the alpha-, beta-, and gamma-subclasses of proteobacteria and high G + C Gram positive bacteria were used to characterize the community structures by in situ hybridization. Sampling was carried out over 12 weeks and samples were fixed with 4% paraformaldehyde for gram positive organisms and ice cold ethanol for gram negative organisms. The activated sludge population usually constitutes about 80 to 90% of proteobacteria. However, in this study it was found that a relatively small amount of proteobacteria was present within the system. No positive hybridization signal was observed with any of the applied eubacterial family- level probes.
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