Pathogen survival during anaerobic digestion: Fatty acids inhibit anaerobic growth ofEscherichia coli

Summary The fate of pathogens in anaerobic digesters has been studied in a laboratory model system in which glucose-nutrient broth cultures of genetically-defined strains ofEscherichia coli received additions of fatty acids at concentrations similar to those attained during anaerobic treatment of farm wastes. Marked concentration-dependent inhibition of growth was observed for both antibiotic resistant and sensitive strains, and the effects increased with increasing chain lengths up to C₈. Survival of enteric organisms during anaerobic digestion may be limited by fatty acid toxicity.     

Ammonia inhibition in anaerobic digestion: A review

Even though ammonia is an essential nutrient for bacterial growth, it may inhibit methanogenesis during anaerobic digestion process if it is available at high concentrations. Therefore, ammonia is regarded as a potential inhibitor during anaerobic digestion, particularly when dealing with complex type of substrates such as manure or the organic fraction of municipal solid waste (OFMSW). Ammonia is produced through biological degradation of nitrogenous matter. Ammonium ion (NH₄⁺) and free ammonia (NH₃) are the two principal forms of inorganic ammonia nitrogen. Both forms can directly and indirectly cause inhibition in an anaerobic digestion system. Particularly, free ammonia (FAN) is a powerful inhibitor in an anaerobic digester above threshold concentrations. Process inhibition is related to the particular characteristics of the substrate to be anaerobically digested, pH, process temperature (mesophilic or thermophilic), type of the seed sludge (inoculum), the reactor configuration and to the concentrations of ammonium and ammonia. In this paper, ammonia inhibition in anaerobic digestion systems and the recovery efforts after inhibition are discussed. Furthermore, the impacts of ammonia inhibition on the microbial population available in anaerobic digesters, namely bacteria and Archaea, are also evaluated in detail.     

Bioaugmenting anaerobic digestion of biosolids with selected strains of Bacillus, Pseudomonas, and Actinomycetes species for increased methanogenesis and odor control

The objective of this study was to evaluate the effects of bioaugmenting anaerobic biosolids digestion with a commercial product containing selected strains of bacteria from genera Bacillus, Pseudomonas, and Actinomycetes, along with ancillary organic compounds containing various micronutrients. Specifically, the effects of the bioaugment in terms of volatile solids destruction and generation and fate of odor-causing compounds during anaerobic digestion and during storage of the digested biosolids were studied. Two bench-scale anaerobic digesters receiving primary and secondary clarifier biosolids from various full-scale biological wastewater treatment plants were operated. One of the digesters received the bioaugment developed by Organica Biotech, while the other was operated as control. The bioaugmented digester generated 29% more net CH₄ during the 8 weeks of operation. In addition, the average residual propionic acid concentration in the bioaugmented digester was 54% of that in the control. The monitoring of two organic sulfide compounds, methyl mercaptan (CH₃SH) and dimethyl sulfide (CH₃SCH₃), clearly demonstrated the beneficial effects of the bioaugmentation in terms of odor control. The biosolids digested in the bioaugmented digester generated a negligible amount of CH₃SH during 10 days of post-digestion storage, while CH₃SH concentration in the control reached nearly 300 ppm_v during the same period. Similarly, peak CH₃SCH₃ generated by stored biosolids from the bioaugmented digester was only 37% of that from the control.     

厌氧消化污泥微生物组研究方法及应用

污泥厌氧消化是在消化污泥微生物组的协调下将剩余污泥中有机物转化为甲烷的微生物过程。与传统厌氧消化过程不同,污泥厌氧消化系统的进料底物为含有大量微生物细胞及胞外多聚物等复杂大分子有机物的剩余污泥。因此,厌氧消化污泥微生物组的种群组成、功能及种群间互作关系等异常复杂,使厌氧消化污泥微生物组分析成为难点问题。但近年来高通量测序技术及生物信息学分析方法的快速发展为消化污泥微生物组研究提供了契机,并迅速推动了该研究领域的发展。本文从4个方面梳理、总结厌氧消化污泥微生物组的研究及应用现状:剩余活性污泥结构、组成及其厌氧消化;基于16SrRNA基因序列测序的微生物组研究;基于宏基因组及宏转录组分析的微生物组研究;厌氧消化污泥微生物组研究案例分析。最后我们提出了厌氧消化污泥微生物组研究亟待解决的关键科学问题。     

Robustness of archaeal populations in anaerobic co-digestion of dairy and poultry wastes

The objective of this study is to investigate the responses of methanogen populations to poultry waste addition by comparing the archaeal microbial populations in continuous anaerobic digesters with or without the addition of poultry waste as a co-substrate. Poultry waste was characterized as an organic/nitrogen-rich substrate for anaerobic digestion. Supplementing dilute dairy waste with poultry waste for anaerobic co-digestion to increase organic loading rate by 50% resulted in improved biogas production. Elevated ammonia derived from poultry waste did not lead to process inhibition at the organic loadings tested, demonstrating the feasibility of the anaerobic co-digestion of dairy and poultry wastes for improved treatment efficiency. The stability of the anaerobic co-digestion process was linked to the robust archaeal microbial community, which remained mostly unchanged in community structure following increases in organic loading and ammonia levels. Surprisingly, Crenarchaeota archaeal populations, instead of the Euryarchaeota methanogens, dominated the archaeal communities in the anaerobic digesters. The ecological functions of these abundant non-methanogen archaeal populations in anaerobic digestion remain to be identified.     

Propionibacterium species diversity in anaerobic digesters

Phenotypic characteristics of 60 strains ofPropionibacterium isolated from anaerobic hybrid digesters treating landfill leachate and a baker's yeast factory effluent were analysed using numerical taxonomy. With the use of the S_SM similarity coefficient, 92% of the anaerobic digester strains were grouped in eight major clusters. The isolates were identified by relating them to specific type strains and comparison of phenotypic characteristics. These clusters were equated with the classical speciesP. acidipropionici, P. freudenreichii, P. jensenii andP. thoenii using the current classification system. Some of the digester isolates were identified to specifies level using the current identification system, but based on overall similarity they were clustered among members of another species. Furthermore, the data indicated that there was low similarity between the digester isolates and the type strains ofP. jensenii andP. thoeni. A hypothesis is presented as to the role of these propionic acid-producing bacteria during the granulation process found in anaerobic digesters.     

Characterization of two strains of Pelobacter carbinolicus isolated from anaerobic digesters

In anaerobic industrial digesters treating wastewaters from food industry, the sludges showed high capacities to degrade ethanol. The main syntrophic ethanoldegrading organisms were characterized as Pelobacter carbinolicus. During acetoin degradation, butanediol isomers were shown to be transiently produced. In coculture with Methanobrevibacter arboriphilus, the strains oxidized not only primary monoalcohols but also 1,2- and 1,3-diols. Ecological importance as well as metabolic pathways of diol degradation are discussed.     

Anaerobic digestion foaming causes – A review

Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water companies due to significant impacts on process efficiency and operational costs. Several foaming causes have been identified over the past few years by researchers. However, the supporting experimental information is limited and in some cases absent. The present report aims to provide a detailed review of the current anaerobic digestion foaming problem and to identify gaps in knowledge regarding the theory of foam formation in anaerobic digesters.     

Effect of temperature on the performance of an anaerobic tubular reactor treating fruit and vegetable waste

This study compares the performance of anaerobic digestion of fruit and vegetable waste (FVW) in the thermophilic (55 °C) process with those under psychrophilic (20 °C) and mesophilic (35 °C) conditions in a tubular anaerobic digesters on a laboratory scale. The hydraulic retention time (HRT) ranged from 10 to 20 days, and raw fruit and vegetable waste was supplied in a semi-continuous mode at various concentrations of total solids (TS) (4, 6, 8 and 10% on dry weight). Biogas production from the experimental thermophilic digester was higher on average than from psychrophilic and mesophilic digesters by 144 and 41%, respectively. The net energy production in the thermophilic digester was 195.7 and 49.07 kJ per day higher than that for the psychrophilic and mesophilic digesters, respectively. The relation between the daily production of biogas and the temperature indicates that for the same produced quantity of biogas, the size of the thermophilic digester can be reduced with regard to that of the psychrophilic and the mesophilic digesters.     

Unexpected competitiveness of Methanosaeta populations at elevated acetate concentrations in methanogenic treatment of animal wastewater

Acetoclastic methanogenesis is a key metabolic process in anaerobic digestion, a technology with broad applications in biogas production and waste treatment. Acetoclastic methanogenesis is known to be performed by two archaeal genera, Methanosaeta and Methanosarcina. The conventional model posits that Methanosaeta populations are more competitive at low acetate levels (<1 mM) than Methanosarcina and vice versa at higher acetate concentrations. While this model is supported by an extensive body of studies, reports of inconsistency have grown that Methanosaeta were observed to outnumber Methanosarcina at elevated acetate levels. In this study, monitoring of anaerobic digesters treating animal wastewater unexpectedly identified Methanosaeta as the dominant acetoclastic methanogen population at both low and high acetate levels during organic overloading. The surprising competitiveness of Methanosaeta at elevated acetate was further supported by the enrichment of Methanosaeta with high concentrations of acetate (20 mM). The dominance of Methanosaeta in the methanogen community could be reproduced in anaerobic digesters with the direct addition of acetate to above 20 mM, again supporting the competitiveness of Methanosaeta over Methanosarcina at elevated acetate levels. This study for the first time systematically demonstrated that the dominance of Methanosaeta populations in anaerobic digestion could be linked to the competitiveness of Methanosaeta at elevated acetate concentrations. Given the importance of acetoclastic methanogenesis in biological methane production, findings from this study could have major implications for developing strategies for more effective control of methanogenic treatment processes.

     

The microbiological basis of process control in methanogenic fermentation of soluble wastes

The essential requirement for anaerobic digestion of industrial wastes is that the process should operate reliably at high performance. In the digestion of dilute, soluble wastes it is necessary to retain the active biomass within the digester at short liquid retention times for the process to be economically feasible and this is reflected in digester design. Performance of digesters can only be assessed by interpretation of measurable parameters such as pH₂, E_h, pH, volatile fatty acid concentrations, temperature, gas production, biomass content and feed rate and composition. The effects of changes in these parameters on the microbiology of methanogenic digestion and the application of this knowledge in control of the process is discussed.     

Batch and continuous biogas production from grass silage liquor

Herein batch and continuous mesophilic anaerobic digestion of grass silage liquor was studied. The continuous process was carried out in Armfield digesters with an OLR ranging from 0.851 to 1.77 kg COD m⁻³ day⁻¹. The effect of recirculation of effluent from the digester was investigated using different OLRs of grass silage liquor feed. These results showed that as the OLR increased, the methane yield decreased for the reactor with no recycle and increased for the reactor with recycle. However, the COD removal for both digesters was nearly the same at the same OLR. Overall these studies show that grass silage liquor can produce a high quality methane steam between 70% and 80% and achieve methane yields of 0.385 m³ kg⁻¹ COD.     

Different response of bacteria,archaea and fungi to process parameters in nine full-scale anaerobic digesters

Biogas production is a biotechnological process realized by complex bacterial, archaeal and likely fungal communities. Their composition was assessed in nine full-scale biogas plants with distinctly differing feedstock input and process parameters. This study investigated the actually active microbial community members by using a comprehensive sequencing approach based on ribosomal 16S and 28S rRNA fragments. The prevailing taxonomical units of each respective community were subsequently linked to process parameters. Ribosomal rRNA of bacteria, archaea and fungi, respectively, showed different compositions with respect to process parameters and supplied feedstocks: (i) bacterial communities were affected by the key factors temperature and ammonium concentration; (ii) composition of archaea was mainly related to process temperature; and (iii) relative abundance of fungi was linked to feedstocks supplied to the digesters. Anaerobic digesters with a high methane yield showed remarkably similar bacterial communities regarding identified taxonomic families. Although archaeal communities differed strongly on genus level from each other, the respective digesters still showed high methane yields. Functional redundancy of the archaeal communities may explain this effect. 28S rRNA sequences of fungi in all nine full-scale anaerobic digesters were primarily classified as facultative anaerobic Ascomycota and Basidiomycota. Since the presence of ribosomal 28S rRNA indicates that fungi may be active in the biogas digesters, further research should be carried out to examine to which extent they are important players in anaerobic digestion processes.     

Microbiological contamination of digested products from anaerobic co-digestion of bovine manure and agricultural by-products

Aims: This study was performed to investigate the microbiological contamination of digestate product (DP) obtained from the anaerobic co‐digestion of bovine manure and agricultural by‐products. Methods and results: Microbiological analyses were performed on bovine manure, fresh DP, liquid and solid fractions and stored liquid fraction of DP. A statistically significant reduction of faecal bacterial indicator was found after anaerobic digestion except for enterococci. After liquid/solid DP separation, bacteria tend to be concentrated in the solid fraction. Storage does not seem to influence the indicator parameters, except for enterococci. Escherichia coli O157:H7 and Yersinia were not found in any samples analysed. Salmonella was rarely detected in DP samples and its derivates, while Listeria monocytogenes was encountered in many samples. Conclusions: The results obtained indicate that the hygienic quality of DP is for almost all microbiological parameters better than that of the bovine manure (range of reduction 1.6–3.1 log₁₀) and suggest the need to identify specific pathogen indicators related to the hygienic characteristics of DPs. Significance and Impact of the Study: This study highlights that the anaerobic co‐digestion of bovine manure and agricultural by‐products in a field‐scale biogas plant does not increase human health risk with respect to the use of animal manure for agricultural fertilization.     

The effects of detergents on anaerobic digestion

Summary The anionic detergent sodium dodecylbenzene sulphonate (SDBS) inhibited mesophilic fermentation in anaerobic digesters. Total gas production and methanogenesis from glucose were reduced to half maximal rates at between 20 and 50 ppm SDBS during the initial phase of digestion, and over 20 days the pH declined from 7.4 to 6.0 in inhibited cultures. As well as accentuating the accumulation of propionic, iso-butyric and iso-valeric acids, a most remarkable effect of this anionic detergent observed only at high concentrations (100 ppm) was to divert the pathway of fermentation with transient accumulation of ethanol. Methanogenesis from cellulose was also inhibited. In thermophilic populations degrading glucose, SDBS was less toxic, and ethanol was not produced. Both the non-ionic detergent Tergitol (nonyl phenyl polyethylene glycol ether) and soap were virtually without effect on the mesophilic anaerobic digestion of glucose.     

Survival rates of parasite eggs in sludge during aerobic and anaerobic digestion.

The effects of mesothermic anaerobic or aerobic sludge digestion on survival of eggs from the roundworms Ascaris suum, toxocara canis, Trichuris vulpis, and Trichuris suis and from the rat tapeworm Hymenolepis diminuta were studied. Destruction of eggs throughout a 15-day treatment period, as well as their viabilities after reisolation, was analyzed. The laboratory model digesters used in this study were maintained at a 15-day retention schedule, partially simulating a continuously operating system. Ascaris eggs were destroyed in the anaerobic (23%) or aerobic (38%) digesters, and 11% Trichuris eggs were destroyed in the aerobic digesters. Trichuris eggs in anaerobic digesters and Toxocara eggs in either anaerobic or aerobic digesters were not destroyed. Destruction of eggs in digesters was correlated with the state of the eggs before subjection to the treatment processes; i.e., some Ascaris and Trichuris eggs were already embryonated in host intestinal contents or feces and hence past their most resistant stage. The viabilities of Ascaris and Toxocara eggs that survived the digestion processes were greater in anaerobically treated than in aerobically treated material. Eggs from Hymenolepis were nonviable before use in the experiments. However, they were more effectively destroyed in aerobic digesters than in anaerobic digesters.     

Characterization of microbial communities in anaerobic bioreactors using molecular probes

The microbial community structure of twenty-one single-phase and one two-phase full-scale anaerobic sewage sludge digesters was evaluated using oligonucleotide probes complementary to conserved tracts of the 16S rRNAs of phylogenetically defined groups of methanogens and sulfate-reducing bacteria. These probe results were interpreted in combination with results from traditional chemical analyses and metabolic activity assays. It was determined that methanogens in healthy mesophilic, single-phase sewage sludge digesters accounted for approximately 8–12% of the total community and thatMethanosarcinales andMethanomicrobiales constituted the majority of the total methanogen population.Methanobacteriales andMethanococcales played a relatively minor role in the digesters. Phylogenetic groups of mesophilic, Gram-negative sulfate-reducing bacteria were consistently present at significant levels:Desulfovibrio andDesulfobulbus spp. were the dominant sulfate-reducing populations,Desulfobacter andDesulfobacterium spp. were present at lower levels, andDesulfosarcina, Desulfococcus, andDesulfobotulus spp. were absent. Sulfate reduction by one or more of these populations played a significant role in all digesters evaluated in this study. In addition, sulfate-reducing bacteria played a role in favoring methanogenesis by providing their substrates. The analysis of the two-phase digester indicated that true phase separation was not accomplished: significant levels of active methanogens were present in the first phase. It was determined that the dominant populations in the second phase were different from those in the single-phase digesters.     

A meta-analysis of the microbial diversity observed in anaerobic digesters

In this study, the collective microbial diversity in anaerobic digesters was examined using a meta-analysis approach. All 16S rRNA gene sequences recovered from anaerobic digesters available in public databases were retrieved and subjected to phylogenetic and statistical analyses. As of May 2010, 16,519 bacterial and 2869 archaeal sequences were found in GenBank. The bacterial sequences were assigned to 5926 operational taxonomic units (OTUs, based on ?97% sequence identity) representing 28 known bacterial phyla, with Proteobacteria (1590 OTUs), Firmicutes (1352 OTUs), Bacteroidetes (705 OTUs), and Chloroflexi (693 OTUs) being predominant. Archaeal sequences were assigned to 296 OTUs, primarily Methanosaeta and the uncharacterized WSA2 group. Nearly 60% of all sequences could not be classified to any established genus. Rarefaction analysis indicates that approximately 60% of bacterial and 90% of archaeal diversity in anaerobic digesters has been sampled. This analysis of the global bacterial and archaeal diversity in AD systems can guide future studies to further examine the microbial diversity involved in AD and development of comprehensive analytical tools.     

Antagonistic and synergistic effects of two mineral support particles on the hydrolysis and methanation of waste water in immobilized cell anaerobic digesters

Summary The kinetic constant K ₀, determined under a range of substrate loadings, was used to compare immobilized cell anaerobic digesters treating abattoir and milk bottling waste waters. Two types of mineral support were evaluated, sepiolite and bentonite, and found to influence significantly the process kinetics. Sepiolite promoted the digestion of the milk bottling waste whereas bentonite promoted the digestion of abattoir waste. Possible reasons as to why these effects are observed are proposed based on the composition of the waste water.