Joonyeob Lee

Temporal variation in methanogen communities of four different full-scale anaerobic digesters treating food waste-recycling wastewater.

Methanogen communities were investigated using 454 pyrosequencing in four different full-scale anaerobic digesters treating food waste-recycling wastewater. Seasonal samples were collected for 2 years, and 24 samples were available for microbial analysis from a plug flow thermophilic (PT) digester, a continuously-stirred tank thermophilic (CT) digester, an upflow anerobic sludge blanket mesophilic (UM) digester, and a continuously-stirred tank mesophilic (CM) digester. Methanoculleus, Methanobacterium, Methanothermobacter, and Methanosaeta were revealed to be key methanogens in full-scale anaerobic digestion process treating food waste-recycling wastewater. In the PT digester, Methanoculleus was dominant (96.8%). In the CT digester, Methanoculleus was dominant (95.4%) during the first year of operation, but the dominant genus was shifted to Methanothermobacter (98.5%) due to pH increase. In the UM digester, Methanosaeta was dominant (87.2%). In the CM digester, Methanoculleus was constantly dominant (74.8%) except during CM5 when Methanosaeta was dominant (62.6%) due to the low residual acetate concentration (0.1 g/L).

Characterization of food waste-recycling wastewater as biogas feedstock.

A set of experiments was carried out to characterize food waste-recycling wastewater (FRW) and to investigate annual and seasonal variations in composition, which is related to the process operation in different seasons. Year-round samplings (n=31) showed that FRW contained high chemical oxygen demand (COD; 148.7±30.5g/L) with carbohydrate (15.6%), protein (19.9%), lipid (41.6%), ethanol (14.0%), and volatile fatty acids (VFAs; 4.2%) as major constituents. FRW was partly (62%) solubilized, possibly due to partial fermentation of organics including carbohydrate. Biodegradable portions of carbohydrate and protein were estimated from acidogenesis test by first-order kinetics: 72.9±4.6% and 37.7±0.3%, respectively. A maximum of 50% of the initial organics were converted to three major VFAs, which were acetate, propionate, and butyrate. The methane potential was estimated as 0.562L CH4/g VSfeed, accounting for 90.0% of the theoretical maximum estimated by elemental analysis.

Correlations between bacterial populations and process parameters in four full-scale anaerobic digesters treating sewage sludge.

Process parameters and bacterial populations were investigated in four full-scale anaerobic digesters treating sewage sludge. Although the four digesters were operated under similar conditions, digesters A and B had higher pH (7.2-7.4) and lipid removal efficiencies (>50%) than C and D (pH 6.1-6.4; average lipid removal <16%). Bacterial richness, diversity, and evenness were higher in digesters C and D. Among the top-populated genera, ten (group I) were more abundant in digesters A and/or B; they were putative syntrophic fatty acid or protein/amino acid-utilizers. In contrast, fifteen others (group II) were less abundant in A and/or B and included potentially dormant/dead cells originated from activated sludge. Despite the overall richness trend, the presence of the 25 genera in groups I/II was greater in digesters A and B (24) than in C and D (17); this observation suggests that group I bacteria might be essential in AD of sewage sludge.

Identifying methanogen community structures and their correlations with performance parameters in four full-scale anaerobic sludge digesters

Four full-scale mesophilic anaerobic digesters treating waste sludge were monitored to characterize methanogen communities and their relationship with process parameters. The performance of the four digesters were dissimilar with the average chemical oxygen demand removal efficiencies between 24 and 45% and differing pH. Real-time quantitative PCR showed that archaeal 16S rRNA gene concentration ([ARC]) and, more pronouncedly, its ratio to bacterial counterpart ([ARC]/[BAC]) correlated positively with the performance parameters, including the lipid removal efficiency. Pyrosequencing identified 12 methanogen genera, of which Methanolinea, Methansaeta, and Methanospirillum collectively accounted for 79.2% of total archaeal reads. However, Methanoculleus, a numerically minor (1.9±2.6%) taxa, was the most promising biomarker for positive performance, while Methanoregula was abundant in samples with poor performance. These results could be useful for the control and management of anaerobic sludge digestion.

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.

Bacteria and archaea communities in full-scale thermophilic and mesophilic anaerobic digesters treating food wastewater: Key process parameters and microbial indicators of process instability

In this study, four different mesophilic and thermophilic full-scale anaerobic digesters treating food wastewater (FWW) were monitored for 1-2years in order to investigate: 1) microbial communities underpinning anaerobic digestion of FWW, 2) significant factors shaping microbial community structures, and 3) potential microbial indicators of process instability. Twenty-seven bacterial genera were identified as abundant bacteria underpinning the anaerobic digestion of FWW. Methanosaeta harundinacea, M. concilii, Methanoculleus bourgensis, M. thermophilus, and Methanobacterium beijingense were revealed as dominant methanogens. Bacterial community structures were clearly differentiated by digesters; archaeal community structures of each digester were dominated by one or two methanogen species. Temperature, ammonia, propionate, Na+, and acetate in the digester were significant factors shaping microbial community structures. The total microbial populations, microbial diversity, and specific bacteria genera showed potential as indicators of process instability in the anaerobic digestion of FWW.

Microbial community shifts in a farm-scale anaerobic digester treating swine waste: Correlations between bacteria communities associated with hydrogenotrophic methanogens and environmental conditions

Microbial community structure in a farm-scale anaerobic digester treating swine manure was investigated during three process events: 1) prolonged starvation, and changes of 2) operating temperature (between meso- and thermophilic) and 3) hydraulic retention time (HRT). Except during the initial period, the digester was dominated by hydrogenotrophic methanogens (HMs). The bacterial community structure significantly shifted with operating temperature and HRT but not with long-term starvation. Clostridiales (26.5-54.4%) and Bacteroidales (2.5-13.7%) became dominant orders in the digester during the period of HM dominance. Abundance of diverse meso- and thermophilic bacteria increased during the same period; many of these species may be H2 producers, and/or syntrophic acetate oxidizers. Some of these species showed positive correlations with [NH4+-N] (p<0.1); this relationship suggests that ammonia was a significant parameter for bacterial selection. The bacterial niche information reported in this study can be useful to understand the ecophysiology of anaerobic digesters treating swine manure that contains high ammonia content.

Temporal variation in bacterial and methanogenic communities of three full-scale anaerobic digesters treating swine wastewater

To investigate the effects of temporal variations of process parameters on microbial community structures in the two types of full-scale anaerobic digester treating swine wastewater, three full-scale anaerobic digesters were monitored. An anaerobic filter (AF)-type digester located in Gong-Ju (GJ) showed the highest COD removal among three digesters and maintained stable efficiency. A digester in Hong-Seong (HS) was of the same type as it GJ and showed improved efficiency over the sampling period. A continuously stirred tank reactor (CSTR)-type digester in Soon-Cheon (SC) showed decreasing efficiency due to a high residual concentration of VFAs and NH4+. These process efficiencies were closely correlated to the Simpson indices of the methanogenic communities. Genera Bacillus, Methanosaeta, and Methanospirillum that have filamentous morphology were dominant in both AF-type digesters, but genera Acholeplasma, Methanosarcina, and Methanoculleus that have spherical or coccoid morphology were dominantly abundant in the CSTR-type digester. Correlation between populations suggests a possible syntrophic relationship between genera Desulfobulbus and Methanosaeta in digesters GJ and HS.

Comprehensive analysis of microbial communities in full-scale mesophilic and thermophilic anaerobic digesters treating food waste-recycling wastewater

Microbes were sampled for a year in a full-scale mesophilic anaerobic digester (MD) and a thermophilic anaerobic digester (TD) treating food waste-recycling wastewater (FRW), then microbial community structure, dynamics and diversity were quantified. In the MD, Fastidiosipila, Petrimonas, vadinBC27, Syntrophomonas, and Proteiniphilum were dominant bacterial genera; they may contribute to hydrolysis and fermentation. In the TD, Defluviitoga, Gelria and Tepidimicrobium were dominant bacteria; they may be responsible for hydrolysis and acid production. In the MD, dominant methanogens changed from Methanobacterium (17.1 ± 16.9%) to Methanoculleus (67.7 ± 17.8%) due to the increase in ammonium concentration. In the TD, dominant methanogens changed from Methanoculleus (42.8 ± 13.6%) to Methanothermobacter (49.6 ± 11.0%) due to the increase of pH. Bacteria and archaea were more diverse in the MD than in the TD. These results will guide development of microbial management methods to improve the process stability of MD and TD treating FRW.

Resource recovery using whey permeate to cultivate Phellinus linteus mycelium: Solid-state and submerged liquid fermentation

The growth characteristics of Phellinus linteus mycelium were assessed and compared under solid-state fermentation (SSF) and submerged liquid fermentation (SLF) systems on whey permeate medium. Response surface methodology was used to investigate the growth rates of mycelia under various conditions of operating temperature (TO), initial pH, and substrate concentration ([S]). The optimal growth conditions of P. linteus mycelium were determined to be 26.1°C, pH 4.6, and 60.3g of lactose/L in the SSF system, and 29.0°C, pH 5.0, and 65.3g of lactose/L in the SLF system. The maximum growth rates were predicted to be 1.92 ± 0.01 mm/d in SSF and 192.1 ± 0.0mg/L per day in SLF. Random trials were conducted to experimentally validate the evaluated optimal conditions. The differences between the modeled and observed values were only 5.3% in the SSF system and 6.1% in the SLF system. Significant engineering factors differed between the fermentation techniques; TO was significant in both cultivation systems, whereas initial pH was significant in SSF but [S] was significant in SLF. Our findings can be used to guide the operation of the bioconversion process for cultivating P. linteus mycelium using whey permeate wastewater.