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Anaerobic co-digestion of various sewage sludges is a promising approach for greater recovery of energy, but the process is more complicated than mono-digestion of sewage sludge. The applicability of microbial structure analyses and gene quantification to understand microbial conditions was evaluated. The results show that information from gene analyses is useful in managing anaerobic co-digestion and damaged microbes in addition to conventional parameters like total solids, pH and biogas production. Total bacterial 16S rRNA gene copy numbers are the most useful tools for evaluating unstable anaerobic digestion of sewage sludge, rather than mcrA and total archaeal 16S rRNA gene copy numbers, and high-throughput sequencing. First order decay rates of gene copy numbers during pH failure were higher than typical decay rates of microbes in stable operation. The sequencing analyses, including multidimensional scaling, showed very different microbial structure shifts, but the results were not consistent., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, continuous operation of a pilot-scale upflow anaerobic sludge blanket (UASB) reactor for sewage treatment was conducted for 630 days to investigate the physical and microbial characteristics of the retained sludge. The UASB reactor with a working volume of 20.2 m(3) was operated at ambient temperature (16-29 °C) and seeded with digested sludge. After 180 days of operation, when the sewage temperature had dropped to 20 °C or lower, the removal efficiency of both total suspended solids (TSS) and total biochemical oxygen demand (BOD) deteriorated due to washout of retained sludge. At low temperature, the cellulose concentration of the UASB sludge increased owing to the rate limitation of the hydrolytic reaction of suspended solids in the sewage. However, after an improvement in sludge retention (settleability and concentration) in the UASB reactor, the process performance stabilized and gave sufficient results (68% of TSS removal, 75% of total BOD removal) at an hydraulic retention time (HRT) of 9.7 h. The methanogenic activity of the retained sludge significantly increased after day 246 due to the accumulation of Methanosaeta and Methanobacterium following the improvement in sludge retention in the UASB reactor. Acid-forming bacteria from phylum Bacteroidetes were detected at high frequency; thus, these bacteria may have an important role in suspended solids degradation.

The anaerobic biological treatment of sucrose-based, low-strength wastewater was investigated in expanded granular sludge bed (EGSB) reactors at low temperatures over a 300-day trial period. During the trial, the operating temperature was lowered in a stepwise manner from 20 degrees C to 5 degrees C. As a result, the reactors exhibited sufficient performances until 10 degrees C operation. The COD removal rate was 3.1-3.8 kgCOD m(-3) day(-1) at 10 degrees C. In particular, the COD removal rate increased gradually through the low-temperature operation; indeed, the later stages of the 10 degrees C operation attained a rate similar to those achieved at 20 degrees C and 15 degrees C. This finding is especially practical for applications of psychrophilic methane fermentation. Additionally, the structure of the microbial community in the granular sludge was analyzed by clone analysis based on 16S rRNA genes and fluorescence in situ hybridization (FISH). As a result, the percentage of the phylum Firmicutes, which were assumed to be Anaerobivrio sp. and Lactococcus sp., greatly increased from 0.7% to 8.0% of the total cells, especially in the surface layer of the granular sludge. These bacteria would contribute to the degradation of the sucrose substrate anaerobically at ambient temperatures. Moreover, the results suggest that a Methanospirillum species, which is a H2-utilizing methanogen, increased from 0.5% to 6.7% during the low-temperature incubation, with a significant increase of methanogenic activity from H2/CO2 at 20 degrees C. Thus, the Methanospirillum species detected in this study may have a key role as hydrogen scavenger during hydrogen-metabolism in low-temperature conditions.

The influence of the sugar content of wastewater on changes in the characteristics of the retained sludge was investigated by using two lab-scale granular sludge bed reactors at 20 degrees C. Both reactors were inoculated with granular sludge grown at 20 degrees C and were fed with synthetic wastewater containing sucrose and volatile fatty acids (VFAs). On day 70, the sucrose content of the wastewater was changed to 90% (based on wastewater COD value) for the first reactor and 0% (VFA 90%) for the second. After this change in feed composition, the COD removal efficiency became about 91% for the sucrose-fed reactor and 95% for the VFA-fed reactor. The growth yield (Yg) of the sucrose-fed sludge increased more than that of the VFA-fed sludge. Consequently, deterioration of the settleability of the sucrose-fed sludge was observed. The sucrose-degrading activity of the retained sludge obtained from the sucrose-fed reactor increased significantly from 3.7 g COD g VSS(-1) day(-1) on day 62 to 36.8 g COD g VSS(-1) day(-1) on day 230, in accordance with the predominant growth of sugar-degrading bacteria--namely, Lactococcus, Clostridium and Chloroflexi--in the retained sludge. The excessive growth of these sugar-degrading bacteria in the retained sludge caused unstable process performance in the sucrose-fed reactor at 20 degrees C.

To promptly establish anaerobic ammonium oxidation (anammox) reactors, appropriate seeding sludge with high abundance and activity of anammox bacteria was selected by quantifying 16S rRNA gene copy numbers of anammox bacteria by real-time quantitative PCR (RTQ-PCR) and batch culture experiments. The selected sludge was then inoculated into up-flow fixed-bed biofilm column reactors with nonwoven fabric sheets as biomass carrier and the reactor performances were monitored over 1 year. The anammox reaction was observed within 50 days and a total nitrogen removal rate of 26.0 kg-Nm(-3)day(-1) was obtained after 247 days. To our knowledge, such a high rate has never been reported before. Hydraulic retention time (HRT) and influent NH(4)(+) to NO(2)(-) molar ratio could be important determinant factors for efficient nitrogen removal in this study. The higher nitrogen removal rate was obtained at the shorter HRT and higher influent NH(4)(+)/NO(2)(-) molar ratio. After anammox reactors were fully developed, the community structure, spatial organization and in situ activity of the anammox biofilms were analyzed by the combined use of a full-cycle of 16S rRNA approach and microelectrodes. In situ hybridization results revealed that the probe Amx820-hybridized anaerobic anammox bacteria were distributed throughout the biofilm (accounting for more than 70% of total bacteria). They were associated with Nitrosomonas-like aerobic ammonia-oxidizing bacteria (AAOB) in the surface biofilm. The anammox bacteria present in this study were distantly related to the Candidatus Brocadia anammoxidans with the sequence similarity of 95%. Microelectrode measurements showed that a high in situ anammox activity (i.e., simultaneous consumption of NH(4)(+) and NO(2)(-)) of 4.45 g-N of (NH(4)(+)+NO(2)(-))m(-2)day(-1) was detected in the upper 800 microm of the biofilm, which was consistent with the spatial distribution of anammox bacteria.

The anaerobic ammonium oxidation (Anammox) process is a new efficient and cost effective method of ammonium removal from wastewater. Under strictly anoxic condition, ammonium is directly oxidised with nitrite as electron acceptor to dinitrogen gas. However, it is extremely difficult to cultivate Anammox bacteria due to their low growth rate. This suggests that a rapid and efficient start-up of Anammox process is the key to practical applications. To screen appropriate seeding sludge with high Anammox potential, a real-time quantitative PCR assay with newly designed primers has been developed. Thereafter, the seeding sludge with high abundance of Anammox bacteria (1.7 x 10(8) copies/mg-dry weight) was selected and inoculated into an upflow anaerobic biofilters (UABs). The UABs were operated for more than 1 year and the highest nitrogen removal rate of 24.0 kg-N m-3 day(-1) was attained. In addition, the ecophysiology of Anammox bacteria (spatial distribution and in situ activity) in biofilms was analysed by combining a full-cycle 16S rRNA approach and microelectrodes. The microelectrode measurement clearly revealed that a successive vertical zonation of the partial nitrification (NH4+ to NO2-), Anammox reaction and denitrification was developed in the biofilm in the UAB. This result agreed with the spatial distribution of corresponding bacterial populations in the biofilm. We linked the micro-scale information (i.e. single cell and/or biofilm levels) with the macro-scale information (i.e. the reactor level) to understand the details of Anammox reaction occurring in the UABs.