Relationships between anaerobic consortia and removal efficiencies in an UASB reactor degrading 2,4 dichlorophenol (DCP)

To gain more insight into the interactions between anaerobic bacteria and reactor performances (chemical oxygen demand-COD, 2,4 dichlorophenol-2,4 DCP removals, volatile fatty acid-VFA, and methane gas productions) and how they depended on operational conditions the microbial variations in the anaerobic granular sludge from an upflow anaerobic sludge blanket (UASB) reactor treating 2,4 DCP was studied. The study was composed of two parts. In the first part, the numbers of methanogens and acedogens in the anaerobic granular sludge were counted at different COD removal efficiencies. The relationships between the numbers of methanogens, the methane gas production and VFA production were investigated. The COD removal efficiencies increased to 74% from 30% while the number of total acedogens decreased to 10 from 30 cfu ml(-1). The number of total methanogens and acedogens varied between 11 x 10(3) and 10 x 10(9)MPN g(-1) and 10 and 30 cfu ml(-1) as the 2,4 DCP removal efficiencies were obtained between 60% and 99%, respectively. It was seen that, as the number of total acedogens decreased, the COD removal efficiencies increased. However, the number of total methanogens increased as the COD removal efficiencies increased. Correlations between the bacterial number and with the removal efficiencies obtained in different operational conditions were investigated. From the results presented in this paper a high correlation between the number of bacteria, COD removals, methane gas percentage, 2,4 DCP removals and VFA was observed. In the second part, methanogen bacteria in the anaerobic granular sludge were identified. Microbial observations and biochemical tests were applied to identify the anaerobic microorganisms from the anaerobic granular sludge. In the reactor treating 2,4 DCP, Methanobacterium bryantii, Methanobacterium formicicum, Methanobrevibacter smithii, Methanococcus voltae, Methanosarcina mazei, Methanosarcina acetivorans, Methanogenium bourgense and Methanospirillum hungatei were identified.     

Investigation into the effect of high concentrations of volatile fatty acids in anaerobic digestion on methanogenic communities

A study was conducted to determine whether differences in the levels of volatile fatty acids (VFAs) in anaerobic digester plants could result in variations in the indigenous methanogenic communities. Two digesters (one operated under mesophilic conditions, the other under thermophilic conditions) were monitored, and sampled at points where VFA levels were high, as well as when VFA levels were low. Physical and chemical parameters were measured, and the methanogenic diversity was screened using the phylogenetic microarray ANAEROCHIP. In addition, real-time PCR was used to quantify the presence of the different methanogenic genera in the sludge samples. Array results indicated that the archaeal communities in the different reactors were stable, and that changes in the VFA levels of the anaerobic digesters did not greatly alter the dominating methanogenic organisms. In contrast, the two digesters were found to harbour different dominating methanogenic communities, which appeared to remain stable over time. Real-time PCR results were inline with those of microarray analysis indicating only minimal changes in methanogen numbers during periods of high VFAs, however, revealed a greater diversity in methanogens than found with the array.     

Methanogenic community structure in simultaneous methanogenesis and denitrification granular sludge

A laboratory scale up-flow anaerobic sludge bed (UASB) bioreactor fed with synthetic wastewater was operated with simultaneous methanogenesis and denitrification (SMD) granules for 235 days with a gradient decrease of C/N. Molecular cloning, qRT-PCR and T-RFLP were applied to study the methanogenic community structures in SMD granules and their changes in response to changing influent C/N. The results indicate that when C/N was 20:1, the methane production rate was fastest, and Methanosaetaceae and Methanobacteriaceae were the primary methanogens within the Archaea. The richness and evenness of methanogenic bacteria was best with the highest T-RFLP diversity index of 1.627 in the six granular sludge samples. When C/N was reduced from 20:1 to 5:1, the methanogenic activity of SMD granules decreased gradually, and the relative quantities of methanogens decreased from 36.5% to 10.9%. The abundance of Methanosaetaceae in Archaea increased from 64.5% to 84.2%, while that of Methanobacteriaceae decreased from 18.6% to 11.8%, and the richness and evenness of methanogens decreased along with the T-RFLP diversity index to 1.155, suggesting that the community structure reflected the succession to an unstable condition represented by high nitrate concentrations.

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Archaeal community composition affects the function of anaerobic co-digesters in response to organic overload

Microbial community diversity in two thermophilic laboratory-scale and three full-scale anaerobic co-digesters was analysed by genetic profiling based on PCR-amplified partial 16S rRNA genes. In parallel operated laboratory reactors a stepwise increase of the organic loading rate (OLR) resulted in a decrease of methane production and an accumulation of volatile fatty acids (VFAs). However, almost threefold different OLRs were necessary to inhibit the gas production in the reactors. During stable reactor performance, no significant differences in the bacterial community structures were detected, except for in the archaeal communities. Sequencing of archaeal PCR products revealed a dominance of the acetoclastic methanogen Methanosarcina thermophila, while hydrogenotrophic methanogens were of minor importance and differed additionally in their abundance between reactors. As a consequence of the perturbation, changes in bacterial and archaeal populations were observed. After organic overload, hydrogenotrophic methanogens (Methanospirillum hungatei and Methanoculleus receptaculi) became more dominant, especially in the reactor attributed by a higher OLR capacity. In addition, aggregates composed of mineral and organic layers formed during organic overload and indicated tight spatial relationships between minerals and microbial processes that may support de-acidification processes in over-acidified sludge.Comparative analyses of mesophilic stationary phase full-scale reactors additionally indicated a correlation between the diversity of methanogens and the VFA concentration combined with the methane yield. This study demonstrates that the coexistence of two types of methanogens, i.e. hydrogenotrophic and acetoclastic methanogens is necessary to respond successfully to perturbation and leads to stable process performance.     

Studies on the toxic effects of pentachlorophenol on the biological activity of anaerobic granular sludge

In order to explore the pathway of the anaerobic biotreatment of the wastewater containing pentachlorophenol (PCP) and ensure the normal operation of Upflow Anaerobic Sludge Blanket (UASB) reactor, the anaerobic sludge under different acclimation conditions were selected to seed and start up UASB reactors. Anaerobic toxicity assays were employed to study the biological activity, the tolerance and the capacity to degrade PCP of different anaerobic granular sludge from UASB reactors. Results showed that the anaerobic granular sludge acclimated to chlorophenols (CPs) could degrade PCP more quickly (up to 9.50mg-PCPg(-1)TVSd(-1)). And the anaerobic granular sludge without acclimation to CPs had only a little activity of degrading PCP (less than 0.07mg-PCPg(-1)TVSd(-1)). Different PCP concentrations (2, 4, 6, 8mgL(-1)) had different inhibition effects on glucose utilization, volatile fatted acidity (VFA)-degrading and methanogens activity of PCP degradation anaerobic granular sludge, and the biological activity declined with the increase in PCP concentration. The methanogens activity suffered inhibition from PCP more easily. The different acclimation patterns of seeded sludge had distinctly different effects on biological activity of the degradation of PCP of anaerobic granular sludge from UASB reactors. The biological activity of the anaerobic granular sludge acclimated to PCP only was also inhibited. This inhibition was weak compared to that of anaerobic granular sludge acclimated to CPs, further, the activity could recover more quickly in this case. In the same reactor, the anaerobic granular sludge from the mid and base layers showed higher tolerance to PCP than that from super layer or if the sludge is unacclimated to CPs, and the corresponding recovery time of the biological activity in the mid and base layers were short. Acetate-utilizing methanogens and syntrophic propinate degraders were sensitive to PCP, compared to syntrophic butyrate degraders.     

Enhancing the efficiency of nitrogen removing bacterial population to a wide range of C:N ratio (1.5:1 to 14:1) for simultaneous C & N removal

• Simultaneous C & N removal in Methammox occurs at wide C:N ratio. • Biological Nitrogen Removal at wide C:N ratio of 1.5:1 to 14:1 is not reported. • Ammonia removal shifted from mixotrophy to heterotrophy at high C:N ratio. • Acetogenic population compensated for ammonia oxidizers at high C:N ratio. • Methanogens increase the plasticity of nitrogen removers at high C:N ratio. High C:N ratio in the wastewater limits biological nitrogen removal (BNR), especially in anammox based technologies. The present study attempts to improve the COD tolerance of the BNR process by associating methanogens with nitrogen removing bacterial (NRB) populations. The new microbial system coined as ‘Methammox’, was investigated for simultaneous removal of COD (C) and ammonia (N) at C:N ratio 1.5:1 to 14:1. The ammonia removal rate (11.5 mg N/g VSS/d) and the COD removal rates (70.6 mg O/g VSS/d) of Methammox was close to that of the NRB (11.1 mg N/g VSS/d) and the methanogenic populations (77.9 mg O/g VSS/d), respectively. The activities established that these two populations existed simultaneously and independently in ‘Methammox’. Further studies in biofilm reactor fetched a balanced COD and ammonia removal (55%–60%) at a low C:N ratio (≤2:1) and high C:N ratio (≥9:1). The population abundance of methanogens was reasonably constant, but the nitrogen removal shifted from mixotrophy to heterotrophy as the C:N ratio shifted from low (C:N≤2:1) to high (C:N≥9:1). The reduced autotrophic NRB (ammonia- and nitrite-oxidizing bacteria and Anammox) population at a high C:N ratio was compensated by the fermentative group that could carry out denitrification heterotrophically. The functional plasticity of the Methammox system to adjust to a broad C:N ratio opens new frontiers in biological nitrogen removal of high COD containing wastewaters.     

激活甲烷菌的微量元素及其补充量的确定

以血清瓶为间歇反应器 ,以醋酸钙、乙醇为基质 ,研究了厌氧消化过程中甲烷菌所需要的微量金属营养元素的种类和优化组合及它们的最佳补充投加量。研究结果表明 ,厌氧消化过程中微量元素对甲烷菌有激活作用 ,最佳微量元素组合为 Fe,Co,Ni,它们的最佳补充投加分别为 1.0 mg/ L· d,0 .1mg/ L· d,0 .2 mg/ L· d