Hyun Min Jang

Influence of thermophilic aerobic digestion as a sludge pre-treatment and solids retention time of mesophilic anaerobic digestion on the methane production, sludge digestion and microbial communities in a sequential digestion process.

In this study, the changes in sludge reduction, methane production and microbial community structures in a process involving two-stage thermophilic aerobic digestion (TAD) and mesophilic anaerobic digestion (MAD) under different solid retention times (SRTs) between 10 and 40 days were investigated. The TAD reactor (RTAD) was operated with a 1-day SRT and the MAD reactor (RMAD) was operated at three different SRTs: 39, 19 and 9 days. For a comparison, control MAD (RCONTROL) was operated at three different SRTs of 40, 20 and 10 days. Our results reveal that the sequential TAD-MAD process has about 42% higher methane production rate (MPR) and 15% higher TCOD removal than those of RCONTROL when the SRT decreased from 40 to 20 days. Denaturing gradient gel electrophoresis (DGGE) and real-time PCR results indicate that RMAD maintained a more diverse bacteria and archaea population compared to RCONTROL, due to the application of the biological TAD pre-treatment process. In RTAD, Ureibacillus thermophiles and Bacterium thermus were the major contributors to the increase in soluble organic matter. In contrast, Methanosaeta concilii, a strictly aceticlastic methanogen, showed the highest population during the operation of overall SRTs in RMAD. Interestingly, as the SRT decreased to 20 days, syntrophic VFA oxidizing bacteria, Clostridium ultunense sp., and a hydrogenotrophic methanogen, Methanobacterium beijingense were detected in RMAD and RCONTROL. Meanwhile, the proportion of archaea to total microbe in RMAD and RCONTROL shows highest values of 10.5 and 6.5% at 20-d SRT operation, respectively. Collectively, these results demonstrate that the increased COD removal and methane production at different SRTs in RMAD might be attributed to the increased synergism among microbial species by improving the hydrolysis of the rate limiting step in sludge with the help of the biological TAD pre-treatment.

Bacterial and methanogenic archaeal communities during the single-stage anaerobic digestion of high-strength food wastewater.

Single-stage anaerobic digestion (AD) was operated to treat high-strength food wastewater (FWW) derived from food waste recycling facilities at two different organic loading rates (OLRs) of 3.5 (Phase I) and 7 (Phase II) kgCOD/m(3)d. Changes in composition of microbial communities were investigated using quantitative real-time PCR (qPCR) and barcoded-pyrosequencing. At the high FWW loading rate, AD showed efficient performance (i.e., organic matter removal and methane production). Bacterial communities were represented by the phyla Bacteroidetes, Firmicutes, Synergistetes and Actinobacteria. During the entire digestion process, the relative abundance phylum Chloroflexi decreased significantly. The qPCR analysis demonstrated that the methanogenic communities shifted from aceticlastic (Methanosarcinales) to hydrogenotrophic methanogens (Methanobacteriales and Methanomicrobiales) with high increase in the proportion of syntrophic bacterial communities. Canonical correspondence analysis revealed a strong relationship between reactor performance and microbial community shifts.

Sequential dilute acid and alkali pretreatment of corn stover: sugar recovery efficiency and structural characterization.

The objectives of this study were to explore the feasibility of applying sequential dilute acid and alkali pretreatment into the hydrolysis of corn stover and to elucidate the effects of structural changes in the biomass on its enzymatic digestibility. H2SO4 used in the first step selectively hydrolyzed 74.6-77.3% of xylan and NaOH used in the second step removed 85.9-89.4% of lignin, from the raw corn stover. Compared to single dilute acid pretreatment, the proposed combined pretreatment minimized the generation of byproducts such as acetic acid, furfural and hydroxymethylfurfural in the hydrolysates, and enhanced the enzymatic hydrolysis of the solid residue. The changes in the structural features (porosity, morphology, and crystallinity) of the solid residue were strongly correlated with the enhancement of enzymatic digestibility. The overall glucose and xylose yields finally obtained after enzymatic hydrolysis reached 89.1-97.9% and 71.0-75.9%, respectively.

Microbial community structure in a thermophilic aerobic digester used as a sludge pretreatment process for the mesophilic anaerobic digestion and the enhancement of methane production

An effective two-stage sewage sludge digestion process, consisting of thermophilic aerobic digestion (TAD) followed by mesophilic anaerobic digestion (MAD), was developed for efficient sludge reduction and methane production. Using TAD as a biological pretreatment, the total volatile suspended solid reduction (VSSR) and methane production rate (MPR) in the MAD reactor were significantly improved. According to denaturing gradient gel electrophoresis (DGGE) analysis, the results indicated that the dominant bacteria species such as Ureibacillus thermophiles and Bacterium thermus in TAD were major routes for enhancing soluble organic matter. TAD pretreatment using a relatively short SRT of 1 day showed highly increased soluble organic products and positively affected an increment of bacteria populations which performed interrelated microbial metabolisms with methanogenic species in the MAD; consequently, a quantitative real-time PCR indicated greatly increased Methanosarcinales (acetate-utilizing methanogens) in the MAD, resulting in enhanced methane production.

Comprehensive microbial analysis of combined mesophilic anaerobic-thermophilic aerobic process treating high-strength food wastewater.

A combined mesophilic anaerobic-thermophilic aerobic process was used to treat high-strength food wastewater in this study. During the experimental period, most of solid residue from the mesophilic anaerobic reactor (R1) was separated by centrifugation and introduced into the thermophilic aerobic reactor (R2) for further digestion. Then, thermophilic aerobically-digested sludge was reintroduced into R1 to enhance reactor performance. The combined process was operated with two different Runs: Run I with hydraulic retention time (HRT) = 40 d (corresponding OLR = 3.5 kg COD/m(3) d) and Run II with HRT = 20 d (corresponding OLR = 7 kg COD/m(3)). For a comparison, a single-stage mesophilic anaerobic reactor (R3) was operated concurrently with same OLRs and HRTs as the combined process. During the overall digestion, all reactors showed high stability without pH control. The combined process demonstrated significantly higher organic matter removal efficiencies (over 90%) of TS, VS and COD and methane production than did R3. Quantitative real-time PCR (qPCR) results indicated that higher populations of both bacteria and archaea were maintained in R1 than in R3. Pyrosequencing analysis revealed relatively high abundance of phylum Actinobacteria in both R1 and R2, and a predominance of phyla Synergistetes and Firmicutes in R3 during Run II. Furthermore, R1 and R2 shared genera (Prevotella, Aminobacterium, Geobacillus and Unclassified Actinobacteria), which suggests synergy between mesophilic anaerobic digestion and thermophilic aerobic digestion. For archaea, in R1 methanogenic archaea shifted from genus Methanosaeta to Methanosarcina, whereas genera Methanosaeta, Methanobacterium and Methanoculleus were predominant in R3. The results demonstrated dynamics of key microbial populations that were highly consistent with an enhanced reactor performance of the combined process.

Sequential sludge digestion after diverse pre-treatment conditions: sludge removal, methane production and microbial community changes.

A lab-scale sequential sludge digestion process which consists of a mesophilic anaerobic digester (MAD) and a thermophilic aerobic digester (TAD) was developed. Thermal, thermal-alkaline and long-term alkaline pre-treatments were applied to the feed sludge to examine their effects on sludge removal and methane production. Especially after thermal-alkaline pre-treatment, high COD removal was maintained; methane production rate was also drastically increased by improving the hydrolysis step of sludge degradation. Polymerase chain reaction-denaturing gel gradient electrophoresis indicated that bacterial communities were represented by three phyla (Firmicutes, Proteobacteria, Actinobacteria) and that Clostridium straminisolvens was the major bacterial species in MAD. Quantitative real-time PCR results indicated that Methanosaeta concilli was the major archaeal species in MAD, and that Ureibacillus sp. was the most abundant bacterial species in TAD.

Quantitative and qualitative changes in antibiotic resistance genes after passing through treatment processes in municipal wastewater treatment plants

In this study, quantitative and qualitative changes in antibiotics resistance genes (ARGs) were investigated in two municipal wastewater treatment plants (WWTPs) treating pretreated livestock or industrial wastewater as well as municipal sewage. Total eight ARGs (tetX, tetM, tetA, sul1, sul2, ermB, qnrD, and blaTEM) were quantified, and their relative abundance was assessed by ARGs copies/16S rRNA gene copies. The fate of ARGs was observed to be different between two WWTPs: sul, qnrD, and blaTEM were proliferated during the treatment processes only in the WWTP1 which received pretreated livestock wastewater. Furthermore, dynamic shifts in patterns of ARGs occurrence were observed during biological, secondary sedimentation and coagulation processes. During biological treatment in both WWTPs, relative abundance of tet and ermB changed: tet increased significantly by 211.6-357.6%, while ermB decreased by 70.4-92.0%. Little variation was observed in sul, qnrD and blaTEM. Subsequently, the relative abundance of tet decreased during the secondary sedimentation and coagulation in both WWTPs: tet decreased by 56.0-86.3% during sedimentation and by 48.2-75.7% during coagulation, respectively. During the final treatment, different responses of antibiotic resistance bacteria (ARB) and ARGs to ultraviolet (UV) disinfection were found: removal efficiencies of ARB were observed in the range of 34-75%, while obvious reduction in ARGs was not observed at the UV dose of 27mJ/cm2. Although ARGs underwent various treatment processes, considerable levels of ARGs remained at discharge amounting to 4.2×1018 copies/day from WWTP1 and 5.4×1016 copies/day from WWTP2, respectively.

Effects of organic loading rates on reactor performance and microbial community changes during thermophilic aerobic digestion process of high-strength food wastewater.

To evaluate the applicability of single-stage thermophilic aerobic digestion (TAD) process treating high-strength food wastewater (FWW), TAD process was operated at four organic loading rates (OLRs) from 9.2 to 37.2 kg COD/m(3)d. The effects of OLRs on microbial community changes were also examined. The highest volumetric removal rate (13.3 kg COD/m(3)d) and the highest thermo-stable protease activity (0.95 unit/mL) were detected at OLR=18.6 kg COD/m(3)d. Denaturing gradient gel electrophoresis (DGGE) profiles and quantitative PCR (qPCR) results showed significant microbial community shifts in response to changes in OLR. In particular, DGGE and phylogenetic analysis demonstrate that the presence of Bacillus sp. (phylum of Firmicutes) was strongly correlated with efficient removal of organic particulates from high-strength food wastewater.

Characterization of antibiotic resistance genes in representative organic solid wastes: Food waste-recycling wastewater, manure, and sewage sludge

In this research, the distribution of antibiotic resistance genes (ARGs) was characterized in representative organic solid waste (OSW) in Korea: food waste-recycling wastewater (FRW), manure, and sewage sludge. The amounts of total ARG (gene copies/16S rRNA gene copies) was greatest in manure followed by sewage sludge and FRW. Interestingly, there were significantly different patterns in the diversity and mechanisms of ARGs. For example, a significant proportion of ARGs were tetracycline resistant genes in all the OSW (40.4-78.2%). β-lactam antibiotics resistant genes were higher in the FRW samples than in other types of OSW but sulfonamides resistant genes represented the greatest proportion in sludge. Regarding the characteristics of antibiotic resistance mechanisms, there was a relatively higher proportion of the ribosomal protection mechanism to tetracycline observed in the FRW and manure samples. However, tetracycline resistant genes with direct interaction were relatively higher in the sewage sludge samples. sul1 was the dominant subtype in all the OSW types and detection of ermB was observed although there was no ermC detected in sewage sludge. There were significant correlations between the occurrences of ARG subtypes: tetB and tetG in all OSW (P<0.01); tetE and tetQ only in sludge (P<0.01). The Class 1 integron-integrase gene (intI1) was significantly correlated with total ARGs only in manure and sludge (P<0.05), revealing potential horizontal gene transfer in these OSW.

Changes in bacterial and archaeal communities in anaerobic digesters treating different organic wastes

The goal of this study was to characterize microbial communities in anaerobic batch digesters treating different representative organic sources (sewage sludge, food waste, septage). Among the digesters, the anaerobic digester of food waste had the highest methanogen density, producing a peak value methane yield of 813.2mLCH4/gVS. In all the digesters, acetoclastic Methanosarcinales and hydrogenotrophic Methanomicrobiales were the most dominant methanogen groups, but their proportion among the methanogens varied depending on the organic sources. The bacteria community in the anaerobic digestion (AD) of food waste and septage was distinctly different from that found in the AD of sewage sludge (primary sludge and waste activated sludge). Shifts in both bacterial and archaeal community structures could be related to differences in chemical properties, production, and accumulation of intermediates digested from organic wastes having different characteristics. These findings could prove useful in optimizing the microbial community to enhance AD process treating organic wastes.