Modeling interactions of Clostridium cadaveris and Clostridium sporogenes in anaerobic acidogenesis of glucose and peptone.

This study focuses on developing a mathematical model to assess interaction among acidogenic bacteria during the anaerobic degradation of two substrates. Clostridium cadaveris and Clostridium sporogenes were cultured in various combinations with glucose and peptone. Parameter estimates are given for both conventional Monod parameters from single substrate-single species cultures and sum kinetics with interaction parameters obtained from dual substrate-single species cultures. The presence of multiple substrates led to both inhibitory and enhancing effects on biodegradation rates for dual substrates compared to single substrate cultures. A new model of interspecies interaction was developed within the framework of Lotka-Volterra incorporating substrate interaction parameters, with a focus on accuracy, realism, simplicity, and biological significance. The model demonstrated competitive interaction for resource sharing and the additional non-linearity parameter eliminated the constraint of the linear relationship between growth rate and population density.

Highly efficient and stable deep-blue organic light-emitting diode using phosphor-sensitized thermally activated delayed fluorescence.

Phosphorescent and thermally activated delayed fluorescence (TADF) blue organic light-emitting diodes (OLEDs) have been developed to overcome the low efficiency of fluorescent OLEDs. However, device instability, originating from triplet excitons and polarons, limits blue OLED applications. Here, we develop a phosphor-sensitized TADF emission system with TADF emitters to achieve high efficiency and long operational lifetime. Peripheral carbazole moieties are introduced in conventional multi-resonance-type emitters containing one boron atom. The triplet exciton density of the TADF emitter is reduced by facilitating reverse intersystem crossing, and the Förster resonant energy transfer rate from phosphor sensitizer is enhanced by high absorption coefficient of the emitters. The emitter exhibited an operational lifetime of 72.9 hours with Commission Internationale de L'Eclairage chromaticity coordinate y = 0.165, which was 6.6 times longer than those of devices using conventional TADF emitters.

Effect of fish waste augmentation on anaerobic co-digestion of sludge with food waste.

The effect of sudden augmentation with fish waste (FW) on an operating anaerobic digester was investigated. Fifteen repeated FW spikes (FWS) composed of 1% or 5% FW per working volume of digester were suddenly fed into semi-continuous operation of a mixture of sludge and food waste. Overall process efficiency was not inhibited by FW augmentation. The bacterial community were clustered differently in the 5% FWS treatment than in the control and 1% FWS. Protein-degrading bacteria (Porphyromonadacea, Family XI, and Family XII) were commonly found in the 5% FWS treatment. Their proportions positively correlated with numbers of other bacteria and dominant methanogens (Methanosaeta and Methanospirillum), showing their important role in FWS digestion. FWS caused a shift of bacteria community, but an increase in archaeal concentration. Therefore, sudden addition of an appropriate amount of FW to existing digesters did not provoke process failure. This result contributes an ecologically-benign method to process FW.

Startup of Demo-Scale Anaerobic Digestion Plant Treating Food Waste Leachate: Process Instability and Recovery.

A demo-scale (600 m3 working volume) anaerobic digester treating food waste leachate was monitored during its startup period. The operation strategy was adjusted twice (i.e., three distinct phases) during the operation to recover the process from instability. During the first phase, the organic loading rate (OLR) > 2.7 kg chemical oxygen demand (COD)/m3∙day corresponded to volatile fatty acid (VFA) accumulation along with a decreasing pH, resulting in the drop in biogas yield to 0.43 ± 0.9 m3/kg CODin. During phase 2, fast recovery of this process was aimed at using a sequencing batch operation. One batch cycle (5 to 2 days) consisted of the combined drawing and feeding step (5 h), the reacting step (91 to 17 h), and the settling step (24 h). The duration of the reacting step was determined for each cycle such that (1) the biogas production ceased before the cycle end and (2) the residual VFA concentration was < 1 g/L. In total, 11 cycles were operated with a gradual increase in biogas yield to 0.55 m3/kg CODin with the absence of any sign of system disturbance. After phase 2, the digester was fed at the designed OLR of 4.1 ± 0.3 kg COD/m3∙day. The biogas yield was elevated to 0.58 ± 0.2 m3/kg CODin during phase 3 with the residual VFA concentration maintained at 2.2 ± 0.6 g/L. Methanogen populations, as determined by real-time PCR, did not change significantly throughout the period. These results imply that the adaptation of this process to the OLR of ca. 4 kg COD/m3∙day was not due to the increase in methanogen population but due to the elevation of its activity. Overall, this study suggests that the sequencing batch operation with adjustable cycle duration can be one successful recovery strategy for biogas plants under system instability.

Effect of different microbial seeds on batch anaerobic digestion of fish waste.

Initial microbial compositions would be the precursor for the efficient anaerobic digestion (AD) of fish waste (FW). A mesophilic batch test was conducted using four seeds collected from different digesters treating various combinations of substrates to investigate their effects on FW degradation. Key microbial groups were identified by 16s rRNA gene-based metagenomics analysis. Among four, the seed from the digester co-digesting livestock manure, food waste, and food wastewater showed the best performance and obtained the highest methane yield (350.5 ± 5.2 mL/gVSadded) and lowest lag phase (0.6 ± 0.1 d). Proteiniphilum, Aminobacterium, dgA-11 gut group, and Syntrophomonas were dominant bacterial genera identified in FW degradation. Methanosaeta was the dominant methanogen in the best performing seed and microbial network analysis revealed its contribution to achieving the highest CH4 yield. Obtained results could be useful in selecting microbial seed sources to avoid system imbalance in full-scale digesters that treat FW.

Tracking microbial community shifts during recovery process in overloaded anaerobic digesters under biological and non-biological supplementation strategies.

Anaerobic digestion encounters operational instability due to fluctuations in organic loading. Propionic acid (HPr) is frequently accumulated due to its unfavorable reaction thermodynamics. Here, 'specific' bioaugmentation using HPr enrichment cultures (three different injection regimes of quantity and frequency) was compared with 'non-specific' bioaugmentation using anaerobic sludge, and with non-biological supplementation of magnetite or coenzyme M. The specific bioaugmentation treatments showed superior recovery responses during continuous feeding after a peak overload. A 'one-shot' bioaugmentation with enrichment showed the best remediation, with ~25% recovery time and >10% CH4 conversion efficiency compared to the control. Consecutive bioaugmentation showed evidence of increased stability of the introduced community. Families Synergistaceae, Syntrophobacteraceae, and Kosmotogaceae were likely responsible for HPr-oxidation, in potential syntrophy with Methanoculleus and Methanobacterium. The different supplementation strategies can be considered to reduce the effect of start-up or overload in anaerobic digesters based on the availability of supplementation resources.

Effects of inhibitions by sodium ion and ammonia and different inocula on acetate-utilizing methanogenesis: Methanogenic activity and succession of methanogens.

Acetate-fed anaerobic sequential batch experiments with four different inhibitory conditions (non-inhibitory (Lo), sodium-ion inhibitory (Na), ammonia inhibitory (Am), combined inhibitory (Hi)) were conducted using thirteen different inocula to investigate the inhibition effects by sodium-ion and ammonia and different inocula on acetate-utilizing methanogenesis and succession of methanogens. Sodium-ion and ammonia significantly extended lag-time λ and reduced specific-methanogenic-activity RCH4, and caused synergistic inhibition. The inhibition differed according to the initial methanogen community structures: the inhibition effects on λ and RCH4 were strongest ininocula with Methanosaeta concilii dominant and weakest in inocula with Methanoculleus bourgensis dominant. These inhibitory conditions determined the succession of methanogens: the most competitive methanogens were Methanosaeta concilii in Lo, Methanosarcina sp. in Na, Methanosarcina sp. and Methanoculleus bourgensis in Am, Methanoculleus bourgensis in Hi. This study provides valuable information for microbial management and optimization for AD processes treating wastewater that is rich in protein and/or salt.

Shift in methanogenic community in protein degradation using different inocula.

Anaerobic digestion (AD) of protein-rich wastes is problematic due to production of ammonia and hydrogen sulfide. In this work, eight inocula were used in batch AD of solutions of gelatin and gluten at 3 g COD substrate/1g VSS inoculum. AD plants from which inocula originated were treating food waste or food wastewater, wastewater sludge, or a combination of them. Inocula were evaluated by fitting methane production data using the modified Gompertz model. Sequencing of 16 s rRNA of microorganisms showed that Methanoculleus was dominant in inocula from plants that were treating food waste, and Methanosaeta was dominant in the others. The maximum methane production rate varied by a factor of three for each substrate: 2.734-7.438 mLCH4 gCOD-1 d-1 for gelatin, and 1.950 to 5.532 mLCH4 gCOD-1 d-1 for gluten. This study demonstrates that inoculum must be chosen appropriately when treating proteinaceous waste by AD.

Long-term enrichment of anaerobic propionate-oxidizing consortia: Syntrophic culture development and growth optimization.

Propionate is a quantitatively important methanogenic intermediate in anaerobic digesters and only limited number of microbes can utilize it under syntrophic association with methanogens. The syntrophic propionate oxidizing bacterias are known to be slow growers due to the low energy yield. Thus, propionate get accumulated frequently in anaerobic digesters and it negatively affect digester performance. In this study, propionate degrading consortia from four different seeding sources were enriched in sequential bath mode in two phases; first adaption phase with 1 g/L of propionate concentration and later, high-strength phase with 3 g/L. From 16s rRNA gene based metagenomics analysis of the former phase, four syntrophic microbial groups, Syntrophaceae, Syntrophomonadaceae, Methanobacterium and Methanosaeta were found to be dominant with complete degradation of propionate. The substrate accelerated microbial shifts were observed at high-strength phase with significant decrease of Syntrophaceae up to 26.9 %. Using Response Surface Methodology, pH 6.8-6.9 and temperature 34.5-34.9 °C were found to be optimum growth conditions for the propionate degradation culture. Observed results could be useful to improve degradation efficiencies and obtained enriched culture can be used to recover propionate-accumulated digesters by bio-augmentation.

Shift in bacterial diversity in acidogenesis of gelatin and gluten seeded with various anaerobic digester inocula.

The aim of this study was to investigate divergence of bacteria degrading model proteins of food-processing wastewater. Gelatin and gluten were used as substrate to represent animal and plant proteins from food wastes, respectively. The inocula were obtained from eight full-scale anaerobic digestion reactors. Food-to-microorganism ratio was 3 g chemical oxygen demand equivalent of substrate per 1 g volatile suspended solids of inoculum. A first-order reaction model revealed reaction constants ranged 1.34 ≤ k ≤ 2.30 d-1 for gelatin and 0.63 ≤ k ≤ 1.69 d-1 for gluten. Metagenomic analysis of 16s rRNA sequences showed that dominant bacteria after gelatin degradation batch were different for each inocula. Klebsiella aerogenes, Hathewaya, Peptoclostridium, or Clostridium collagenovorans were most abundant. Klebsiella aerogenes was the most abundant species after gluten degradation for all inocula.

Monitoring microbial community structure and variations in a full-scale petroleum refinery wastewater treatment plant.

This research investigated the process efficiency and microbial communities and their diversity in a full-scale wastewater treatment plant (WWTP) fed with petroleum refining wastewater (PRW) that contained toxic hydrocarbon contaminants and carcinogens. Process parameters and bacterial community structures were monitored for six months to create a link between microbial dynamics and influent characteristics of petrochemical wastewater. The WWTP showed a stable process with efficiencies >70% for both soluble chemical oxygen demand (SCOD) and benzene removal. More than 30 genera were identified by metagenomic analysis, and the bacterial populations changed significantly during the operation period. Among them, genera Sulfuritalea (11.9 ± 3.5%), Ottowia (4.3 ± 2.2%), Thauera (3.1 ± 7.2%) and Hyphomicrobium (1.3 ± 0.7%) were dominant and important bacterial genera that may have been responsible for the degradation of aromatic compounds such as benzene and phenol.

Biokinetics of protein degrading Clostridium cadaveris and Clostridium sporogenes in batch and continuous mode of operations.

A quantitative real-time polymerase chain reaction (QPCR) was applied to estimate biokinetic coefficients of Clostridium cadaveris and Clostridium sporogenes, which utilize protein as carbon source. Experimental data of changes in peptone concentration and 16S rRNA gene copy numbers of C. cadaveris and C. sporogenes were fitted to model. The fourth-order Runge-Kutta approximation with non-linear least squared analysis was employed to solve the ordinary differential equations to estimate biokinetic coefficients. The maximum specific growth rate (µmax), half saturation concentration (Ks), growth yield (Y), and decay coefficient (Kd) of C. cadaveris and C.sporogenes were 0.73 ± 0.05 and 1.35 ± 0.32 h-1, 6.07 ± 1.52 and 5.67 ± 1.53 g/L, 2.25 ± 0.75 × 1010 and 7.92 ± 3.71 × 109 copies/g, 0.002 ± 0.003 and 0.002 ± 0.001 h-1, respectively. The theoretical specific growth rate of C. sporogenes always exceeded than that of C. cadaveris at peptone concentration higher than 3.62 g/L. When the influent peptone concentration was 5.0 g/L, the concentration of C.cadaveris gradually decreased to the steady value of 2.9 × 1010 copies/mL at 4 hours HRT, which indicates 67.1% of the initial population reduction, but the wash out occurred at 1.9 and 3.2 hours HRTs. The 16S rRNA gene copy numbers of C. sporogenes gradually decreased to steady values ranging from 1.1 × 1010 to 2.9 × 1010 copies/mL. C. sporogenes species was predicted to wash out at an HRT of 1.6 h.

A snapshot of microbial community structures in 20 different field-scale anaerobic bioreactors treating food waste.

This study aimed to identify significant factors shaping the microbial populations in biogas plants treating food waste (FW). Twenty full-scale anaerobic acidogenic/methanogenic bioreactors, located at 11 FW treatment facilities, were compared to find patterns in their microbial community structures and potential interactions with the process parameters. Temperature, hydraulic retention time, and organic loading rate were design parameters that systematically influenced the microbial communities. The latter two clearly separated the acidogenic and methanogenic bioreactors. Lactobacillus was the dominant (69.7 ±â€¯19.8%) bacteria in the acidogenic reactors, while hydrogen-utilizing methanogens, such as Methanoculleus (65.1 ±â€¯33.5%), were the dominant archaea in most methanogenic digesters. Defluiviitoga was the dominant (82.7 ±â€¯1.4%) bacteria in the thermophilic digesters, but was also the most abundant (33.1-33.6%) bacteria in dry mesophilic digesters. The two bioreactor categories had lower bacterial diversities, and also higher propionate concentrations (>5 g/L in 4 out of 5 cases), which may impose potential risks for the management of such digesters. The current 'snapshot' of the microbial communities suggests several bacterial and archaeal taxa as potential indicators of bioreactor categories and/or process variables.

Magnetite as an enhancer in methanogenic degradation of volatile fatty acids under ammonia-stressed condition.

Anaerobic batch tests with a 22 full-factorial design of ammonia (1.5, 6.5 g N/L) and magnetite concentrations (0, 20 mmol/L) were conducted separately for methanogenic degradation of acetate, propionate, and butyrate (volatile fatty acids (VFAs)) to 1) quantify the effect of magnetite as an enhancer in methanogenic degradation of each of the VFAs in conditions without ammonia stress (1.5 g N/L) and with ammonia stress (6.5 g N/L), and 2) identify methanogenic consortia that are related to such enhancement. Among the three VFAs, methanogenic degradation of propionate was the least feasible (57% lower specific methanogenic activity RCH4 and three times longer lag time λ than acetate degradation). At low ammonia concentration, only propionate showed improvement in RCH4 (46%) with supplementation of magnetite. In the ammonia-stressed condition without magnetite, RCH4 decreased by 38-58% and λ increased 2.2-8.8 times for all VFAs; magnetite supplementation significantly alleviated these effects. These results demonstrate that magnetite supplementation effectively increases methanogenic degradation of the VFAs even under ammonia-stressed conditions. 16S metagenomic sequencing revealed that distinctive methanogenic consortia were active in the different combinations of substrate, ammonia and magnetite. Alkaliphilus, Hyphomonadaceae SWB02 and Clostridia DTU014, Clostridia D8A-2, Christensenellaceae R-7 group and Rikenellaceae DMER64 were identified as potential syntrophic bacteria that can establish magnetite-mediated direct electron transfer with methanogens (Methanosaeta concilii, Methanosaeta harundinacea, Methanolinea tarda, Methanoculleus bourgensis and Methanosarcina spp.) during methanogenic degradation of VFAs. The results may be useful as a reference to develop effective strategies using magnetite supplementation to remediate anaerobic digestion processes that have been afflicted by VFA accumulation and ammonia inhibition.

Microbial community structure in full scale anaerobic mono-and co-digesters treating food waste and animal waste.

Five mesophilic full-scale anaerobic digesters treating food waste (FW-digester), animal waste (AW-digester), and co-substrate of food waste and animal waste (CO-digesters) were monitored identify bacterial and archaeal communities and to quantify the effect of substrate characteristics on them, and to identify 'core' microorganism. The substrate characteristics and microbial communities of the FW-digester, AW-digester, and CO-digesters were statistically different. Organic concentration and [Na+] were identified as major variations that effect microbial community. Methanogen community was more diverse in AW-digester than in FW-digester. Methanogen community in CO-digester was as diverse as in AW-digester, and the most dominant species was Methanoculleus bourgensis same as in FW-digester. Twenty-one bacterial genera and four methanogen species were found in all digesters as a consequence of their metabolic versatility to degrade organic and inhibitor compounds. The results implied that these core microorganisms may contribute to maintaining a stable microbial ecosystem.

Single and combined inhibition of Methanosaeta concilii by ammonia, sodium ion and hydrogen sulfide.

Single and combined inhibition of lag time λ and specific methanogenic activity RCH4 of Methanosaeta concilii by NH3, Na+ and H2S were investigated using inhibition tests with a single inhibitor and a 33 full-factorial experiment of NH3, Na+ and H2S concentrations (1.5 ≤ total ammonia nitrogen (TAN)/L ≤ 4.5 g, 1 ≤ Na+/L ≤ 4.3 g, 14.2 ≤ total hydrogen sulfide sulfur (THSS)/L ≤ 836 mg). All three inhibitors significantly increased λ and reduced RCH4 of M. concilii. The half-maximal inhibitory concentrations of NH3, Na+ and H2S for M. concilii were 6.4 g TAN/L, 5.2 g Na+/L and 1.6 g THSS/L. Partial cubic models adequately approximated the corresponding response surfaces of λ and RCH4 from the 33 full-factorial experiment. The inhibitors inhibited RCH4 synergistically, but inhibited λ in a complex manner. The combination of NH3 and Na+ showed the strongest synergistic inhibition of both λ and RCH4.

Development of an interspecies interaction model: An experiment on Clostridium cadaveris and Clostridium sporogenes under anaerobic condition.

The specific primer and probe sets for quantifying Clostridium cadaveris and Clostridium sporogenes using a quantitative real-time PCR were designed. Each primer and probe set detected only the target species very specifically. The two species were cultivated in pure and mixed culture in batch mode with glucose as the only carbon source. The designed QPCR sets were used successfully to estimate the biokinetic parameters of each species in pure culture: i.e., maximum specific growth rate µmax, half saturation concentration Ks, growth yield Y, and decay coefficient Kd. of C. cadaveris and C. sporogenes were 0.311 ±â€¯0.020 and 0.360 ±â€¯0.019 h-1, 4.241 ±â€¯1.653 and 5.171 ±â€¯1.097 g/L, 0.301 ±â€¯0.065 and 0.199 ±â€¯0.037 1011 copies/g, 0.005 ±â€¯0.043 and 0.009 ±â€¯0.025 h-1, respectively. The effect of interspecific interaction of on substrate consumption rate and microbial growth was evaluated using mixed culture; curve fitting and comparison of coefficients detected increase in substrate consumption rate but decrease in microbial growth rate; these results imply interspecific interaction effect. A new model was of the interspecific interaction was developed, with focus on accuracy, realism, simplicity and biological significance. This interspecific interaction model may be extended to more-complex bioprocesses such as biological wastewater treatment systems and anaerobic digestion.

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 ±â€¯16.9%) to Methanoculleus (67.7 ±â€¯17.8%) due to the increase in ammonium concentration. In the TD, dominant methanogens changed from Methanoculleus (42.8 ±â€¯13.6%) to Methanothermobacter (49.6 ±â€¯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.

Microbial communities underpinning mesophilic anaerobic digesters treating food wastewater or sewage sludge: A full-scale study.

Ten mesophilic full-scale anaerobic digesters treating food wastewater (FW-digesters) or sewage sludge (SL-digesters) were monitored for 1 year to investigate: (1) microbial communities underpinning FW-digesters and SL-digesters, (2) the effects of total ammonia-nitrogen concentration [TAN] and Na+ concentration [Na+] on variations of these communities. [TAN] and [Na+] in the digester varied among digesters: 1.7-6.5 g TAN/L and 1.0-3.6 g Na+/L for the FW-digesters, and 0.1-2.2 g TAN/L and 0.1-1.2 g Na+/L for the SL-digesters; [TAN] negatively correlated with the process efficiency of the FW-digesters. Microbial communities were less diverse in the FW-digesters than in the SL-digesters. The FW- and SL-digesters formed very distinct microbial community structures; [TAN] and [Na+] in the digester were the critical factors shaping these structures. Immigrant bacteria from influent sludge significantly influence the bacterial communities of the SL-digesters. Methanoculleus might be tolerant to high ammonia in AD of such organic wastewater.