Mesophilic and thermophilic anaerobic co-digestion of abattoir wastewater and fruit and vegetable waste in anaerobic sequencing batch reactors

Anaerobic co-digestion of fruit and vegetable waste (FVW) and abattoir wastewater (AW) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of hydraulic retention time (HRT) and temperature variations on digesters performances were examined. At both 20 and 10 days biogas production for co-digestion was greater thanks to the improved balance of nutrients. The high specific gas productions for the different digestion processes were 0.56, 0.61 and 0.85 l g⁻¹ total volatile solids (TVS) removal for digesters treating AW, FVW and AW + FVW, respectively. At an HRT of 20 days, biogas production rates from thermophilic digesters were higher on average than from mesophilic AW, FVW and AW + FVW digestion by 28.5, 44.5 and 25%, respectively. However, at 10 days of HRT results showed a decrease of biogas production rate for AW and AW + FVW digestion processes due to the high amount of free ammonia at high organic loading rate (OLR).     

Anaerobic digestion of solid slaughterhouse waste (SHW) at laboratory scale: Influence of co-digestion with the organic fraction of municipal solid waste (OFMSW)

Mesophilic anaerobic digestion of slaughterhouse waste (SHW) and its co-digestion with the organic fraction of municipal solid waste (OFMSW) have been evaluated. These processes were carried out in a laboratory plant semi-continuously operated and two set-ups were run. The first set-up, with a hydraulic retention time (HRT) of 25 days and organic loading rate (OLR) of 1.70 kg VS m⁻³ day⁻¹ for digestion, and 3.70 kg VS m⁻³ day⁻¹ for co-digestion, was not successful. The second set-up was initiated with an HRT of 50 days and an OLR of 0.9 kg VS m⁻³ day⁻¹ for digestion and 1.85 kg VS m⁻³ day⁻¹ for co-digestion. Under these conditions, once the sludge had been acclimated to a medium with a high fat and ammonia content, it was possible to decrease the HRT while progressively increasing the OLR to the values used in the first set-up until an HRT of 25 days and OLRs of 1.70 and 3.70 kg VS m⁻³ day⁻¹, for digestion and co-digestion, respectively (the same conditions of the digesters failures previously). These digesters showed a highly stable performance, volatile fatty acids (VFAs) were not detected and long chain fatty acids (LCFAs) were undetected or only trace levels were measured in the analyzed effluent. Fat removal reached values of up to 83%. Anaerobic digestion was thus found to be a suitable technology for efficiently treating lipid and protein waste.     

Air-side ammonia stripping coupled to anaerobic digestion indirectly impacts anaerobic microbiome

Air-side stripping without a prior solid–liquid phase separation step is a feasible and promising process to control ammonia concentration in thermophilic digesters. During the process, part of the anaerobic biomass is exposed to high temperature, high pH and aerobic conditions. However, there are no studies assessing the effects of those harsh conditions on the microbial communities of thermophilic digesters. To fill this knowledge gap, the microbiomes of two thermophilic digesters (55°C), fed with a mixture of pig manure and nitrogen-rich co-substrates, were investigated under different organic loading rates (OLR: 1.1–5.2 g COD l⁻¹ day⁻¹), ammonia concentrations (0.2–1.5 g free ammonia nitrogen l⁻¹) and stripping frequencies (3–5 times per week). The bacterial communities were dominated by Firmicutes and Bacteroidetes phyla, while the predominant methanogens were Methanosarcina sp archaea. Increasing co-substrate fraction, OLR and free ammonia nitrogen (FAN) favoured the presence of genera Ruminiclostridium, Clostridium and Tepidimicrobium and of hydrogenotrophic methanogens, mainly Methanoculleus archaea. The data indicated that the use of air-side stripping did not adversely affect thermophilic microbial communities, but indirectly modulated them by controlling FAN concentrations in the digester. These results demonstrate the viability at microbial community level of air side-stream stripping process as an adequate technology for the ammonia control during anaerobic co-digestion of nitrogen-rich substrates.     

Microbial community dynamics in mesophilic anaerobic co-digestion of mixed waste

This paper identifies key components of the microbial community involved in the mesophilic anaerobic co-digestion (AD) of mixed waste at Rayong Biogas Plant, Thailand. The AD process is separated into three stages: front end treatment (FET); feed holding tank and the main anaerobic digester. The study examines how the microbial community structure was affected by the different stages and found that seeding the waste at the beginning of the process (FET) resulted in community stability. Also, co-digestion of mixed waste supported different bacterial and methanogenic pathways. Typically, acetoclastic methanogenesis was the major pathway catalysed by Methanosaeta but hydrogenotrophs were also supported. Finally, the three-stage AD process means that hydrolysis and acidogenesis is initiated prior to entering the main digester which helps improve the bioconversion efficiency. This paper demonstrates that both resource availability (different waste streams) and environmental factors are key drivers of microbial community dynamics in mesophilic, anaerobic co-digestion.     

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.     

Linkages of <Emphasis Type="Italic">Firmicutes</Emphasis> and <Emphasis Type="Italic">Bacteroidetes</Emphasis> populations to methanogenic process performance

To identify potential linkages between specific bacterial populations and process performance in anaerobic digestion, the dynamics of bacterial community structure was monitored with high-throughput sequencing in triplicate anaerobic digesters treating animal waste. Firmicutes and Bacteroidetes were found as the two most abundant populations, however, with contrasting population dynamics in response to organic overloading. Firmicutes dominated the bacterial community during stable process performance at low organic loading rate, representing over 50 % of the bacterial abundance. In contrast, the onset of organic overloading raised the relative abundance of Bacteroidetes from 20 ± 2.6 to 44 ± 3.1 %. In addition to the significant negative correlation between the relative abundance of Firmicutes and Bacteroidetes, populations of Firmicutes and Bacteroidetes were found to be linked to process parameters including organic loading rate, volatile fatty acids concentration, and methane production. Therefore, the population abundance ratio of Firmicutes to Bacteroidetes (F/B ratio) was suggested as a potential indicator for process performance. The interactions between Firmicutes and Bacteroidetes populations could be exploited to develop strategies for the prevention of performance perturbation in anaerobic digestion processes.     

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。     

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.     

Thermophilic co-digestion of organic fraction of municipal solid wastes with FOG wastes from a sewage treatment plant: reactor performance and microbial community monitoring

Working at thermophilic conditions instead of mesophilic, and also the addition of a co-substrate, are both the ways to intend to improve the anaerobic digestion of the source-collected organic fraction of municipal solid wastes (SC-OFMSW). Addition of sewage treatment plant fat, oil and grease wastes (STP-FOGW), that are nowadays sent to landfill, would represent an opportunity to recover a wasted methane potential and, moreover, improve the whole process. In this study, after a first period feeding only SC-OFMSW, a co-digestion step was performed maintaining thermophilic conditions. During the co-digestion period enhancements in biogas production (52%) and methane yield (36%) were achieved. In addition, monitoring of microbial structure by using PCR-DGGE and cloning techniques showed that bacterial community profiles clustered in two distinct groups, before and after the extended contact with STP-FOGW, being more affected by the STP-FOGW addition than the archaeal one.     

Effect of Increasing Total Solids Contents on Anaerobic Digestion of Food Waste under Mesophilic Conditions: Performance and Microbial Characteristics Analysis

The total solids content of feedstocks affects the performances of anaerobic digestion and the change of total solids content will lead the change of microbial morphology in systems. In order to increase the efficiency of anaerobic digestion, it is necessary to understand the role of the total solids content on the behavior of the microbial communities involved in anaerobic digestion of organic matter from wet to dry technology. The performances of mesophilic anaerobic digestion of food waste with different total solids contents from 5% to 20% were compared and the microbial communities in reactors were investigated using 454 pyrosequencing technology. Three stable anaerobic digestion processes were achieved for food waste biodegradation and methane generation. Better performances mainly including volatile solids reduction and methane yield were obtained in the reactors with higher total solids content. Pyrosequencing results revealed significant shifts in bacterial community with increasing total solids contents. The proportion of phylum Chloroflexi decreased obviously with increasing total solids contents while other functional bacteria showed increasing trend. Methanosarcina absolutely dominated in archaeal communities in three reactors and the relative abundance of this group showed increasing trend with increasing total solids contents. These results revealed the effects of the total solids content on the performance parameters and the behavior of the microbial communities involved in the anaerobic digestion of food waste from wet to dry technologies.     

Anaerobic digestion of municipal solid waste and agricultural waste and the effect of co-digestion with dairy cow manure

Anaerobic digestion of dairy cow manure (CM), the organic fraction of municipal solid waste (OFMSW), and cotton gin waste (CGW) was investigated with a two-phase pilot-scale anaerobic digestion (AD) system. The OFMSW and CM were digested as single wastes and as combined wastes. The single waste digestion of CM resulted in 62m(3) methane/ton of CM on dry weight basis. The single waste digestion of OFMSW produced 37m(3) methane/ton of dry waste. Co-digestion of OFMSW and CM resulted in 172m(3) methane/ton of dry waste. Co-digestion of CGW and CM produced 87m(3) methane/ton of dry waste. Comparing the single waste digestions with co-digestion of combined wastes, it was shown that co-digestion resulted in higher methane gas yields. In addition, co-digestion of OFMSW and CM promotes synergistic effects resulting in higher mass conversion and lower weight and volume of digested residual.     

Improvement of activated sludge stabilisation and filterability during anaerobic digestion by fruit and vegetable waste addition

Anaerobic co-digestion of fruit and vegetable waste (FVW) and activated sludge (AS) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of AS:FVW ratio and the organic loading rate (OLR) on digesters performances were examined. The mixtures having AS:FVW ratios of 100:00, 65:35, 35:65, by a total solid (TS) basis were operated at an hydraulic retention time (HRT) of 20d. However, 30:70, 20:80, 15:85, 10:90 and 0:100 ratios were tested at an HRT of 10d. To investigate effects of aerobic and anaerobic digestion on the sludge filterability, specific resistance to filtration (R) was also determined. Increasing FVW proportions in the feedstock significantly improved the biogas production yield. The reactor that was fed with a 30:70 ratio showed the highest VS removal and biogas production yield of 88% and 0.57 L g(-1) VS added, respectively. The filterability results showed that the anaerobic effluent was characterised by a slightly better filterability efficiency of 1.6 x 10(16) m kg(-1) than 1.74 x 10(16) m kg(-1) of aerobic effluent. However, FVW addition improved the anaerobic co-digestion effluent filterability (5.52 x 10(14) m kg(-1)).     

Effects of Temperature and Carbon-Nitrogen (C/N) Ratio on the Performance of Anaerobic Co-Digestion of Dairy Manure,Chicken Manure and Rice Straw: Focusing on Ammonia Inhibition

Anaerobic digestion is a promising alternative to disposal organic waste and co-digestion of mixed organic wastes has recently attracted more interest. This study investigated the effects of temperature and carbon-nitrogen (C/N) ratio on the performance of anaerobic co-digestion of dairy manure (DM), chicken manure (CM) and rice straw (RS). We found that increased temperature improved the methane potential, but the rate was reduced from mesophilic (30∼40°C) to thermophilic conditions (50∼60°C), due to the accumulation of ammonium nitrogen and free ammonia and the occurrence of ammonia inhibition. Significant ammonia inhibition was observed with a C/N ratio of 15 at 35°C and at a C/N ratio of 20 at 55°C. The increase of C/N ratios reduced the negative effects of ammonia and maximum methane potentials were achieved with C/N ratios of 25 and 30 at 35°C and 55°C, respectively. When temperature increased, an increase was required in the feed C/N ratio, in order to reduce the risk of ammonia inhibition. Our results revealed an interactive effect between temperature and C/N on digestion performance.     

Comparative metagenomic analysis of bacterial populations in three full-scale mesophilic anaerobic manure digesters

While the use of anaerobic digestion to generate methane as a source of bioenergy is increasing worldwide, our knowledge of the microbial communities that perform biomethanation is very limited. Using next-generation sequencing, bacterial population profiles were determined in three full-scale mesophilic anaerobic digesters operated on dairy farms in the state of Vermont (USA). To our knowledge, this is the first report of a metagenomic analysis on the bacterial population of anaerobic digesters using dairy manure as their main substrate. A total of 20,366 non-chimeric sequence reads, covering the V1-V2 hypervariable regions of the bacterial 16S rRNA gene, were assigned to 2,176 operational taxonomic units (OTUs) at a genetic distance cutoff value of 5 %. Based on their limited sequence identity to validly characterized species, the majority of OTUs identified in our study likely represented novel bacterial species. Using a naïve Bayesian classifier, 1,624 anaerobic digester OTUs could be assigned to 16 bacterial phyla, while 552 OTUs could not be classified and may belong to novel bacterial taxonomic groups that have yet to be described. Firmicutes, Bacteroidetes, and Chloroflexi were the most highly represented bacteria overall, with Bacteroidetes and Chloroflexi showing the least and the most variation in abundance between digesters, respectively. All digesters shared 132 OTUs, which as a “core” group represented 65.4 to 70.6 % of sequences in individual digesters. Our results show that bacterial populations from microbial communities of anaerobic manure digesters can display high levels of diversity despite sharing a common core substrate.     

Biodegradability and mesophilic co-digestion of municipal sludge and scum

The objective of this study is to investigate and optimize the co-digestion of scum with thickened waste activated sludge (TWAS) and primary sludge (PS) undergoing mesophilic anaerobic digestion. The effect of scum loading on the co-digestion of PS, TWAS and scum has shown to have a significant impact on the ultimate cumulative biogas production and on the specific biogas production between 20 and 40 days of digestion, while the effects of the scum holding time within the scum concentrator and temperature of the scum concentrator did not demonstrate a significant effect on the ultimate or specific biogas production. The study demonstrates that care must be taken to avoid inhibitory effects and potential souring of digesters due to scum overloading and specifically scum overloading in combination with long holding times of scum within the scum concentrator at elevated temperatures.     

Anaerobic co-digestion of food waste and piggery wastewater: focusing on the role of trace elements

The objective of this study was to evaluate the feasibility of anaerobic co-digestion of food waste and piggery wastewater, and to identify the key factors governing the co-digestion performance. The analytical results indicated that the food waste contained higher energy potential and lower concentrations of trace elements than the piggery wastewater. Anaerobic co-digestion showed a significantly improved biogas productivity and process stability. The results of co-digestion of the food waste with the different fractions of the piggery wastewater suggested that trace element might be the reason for enhancing the co-digestion performance. By supplementing the trace elements, a long-term anaerobic digestion of the food waste only resulted in a high methane yield of 0.396 m³/kg VS_added and 75.6% of VS destruction with no significant volatile fatty acid accumulation. These results suggested that the typical Korean food waste was deficient with some trace elements required for anaerobic digestion.     

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.

     

Microbial community structure and dynamics during anaerobic digestion of various agricultural waste materials

The influence of the feedstock type on the microbial communities involved in anaerobic digestion was investigated in laboratory-scale biogas reactors fed with different agricultural waste materials. Community composition and dynamics over 2 months of reactors’ operation were investigated by amplicon sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Major bacterial taxa belonged to the Clostridia and Bacteroidetes, whereas the archaeal community was dominated by methanogenic archaea of the orders Methanomicrobiales and Methanosarcinales. Correlation analysis revealed that the community composition was mainly influenced by the feedstock type with the exception of a temperature shift from 38 to 55 °C which caused the most pronounced community shifts. Bacterial communities involved in the anaerobic digestion of conventional substrates such as maize silage combined with cattle manure were relatively stable and similar to each other. In contrast, special waste materials such as chicken manure or Jatropha press cake were digested by very distinct and less diverse communities, indicating partial ammonia inhibition or the influence of other inhibiting factors. Anaerobic digestion of chicken manure relied on syntrophic acetate oxidation as the dominant acetate-consuming process due to the inhibition of aceticlastic methanogenesis. Jatropha as substrate led to the enrichment of fiber-degrading specialists belonging to the genera Actinomyces and Fibrobacter.     

微生物组学及其在厌氧消化中的研究进展

我国每年产生大量的有机废弃物,如果处置不当将会对生态、气候以及人类健康造成重大影响。厌氧消化是一种可靠的、绿色的、可持续的有机废弃物处理方式,但由于缺乏准确有效的监测手段,厌氧消化微观过程常常被视为“黑盒”。随着微生物组学的发展,学者们在菌群与运行参数关联性分析、代谢途径分析等方面有了更深入的认识。本文从“三阶段、四菌群”的厌氧消化过程出发,介绍了常用微生物组学的类型,包括:16S rRNA基因组、宏基因组、宏转录组和宏蛋白组;详细阐述了物种组成分析、α多样性分析、OTU相似性分析以及多元统计学分析等6种常用的微生物群落生物信息学分析方法;系统回顾了厌氧消化过程的微生物学研究进展,以期能为分析厌氧消化的微生物群落结构和功能、开发新的厌氧消化工艺和技术提供支持。     

Microbial community dynamics in anaerobic bioreactors and algal tanks treating piggery wastewater

Integrated biosystem is becoming a major aspect of wastewater management practice. Microbial communities in piggery wastewater sampled from anaerobic (thermophilic and mesophilic) and aerobic digesters (algal tanks) during waste remediation were analyzed by culture-independent techniques based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The use of Muyzer's 314F-GC, 518R bacterial primers, and archaeal A934F, 1309R primers followed by partial 16s rDNA sequence analysis of the main bands from DGGE revealed the presence of unknown and as yet uncultured microorganisms but also showed functional and ecologically significant denitrifying, acetogenic bacteria along with autotrophic, hydrogenotrophic, and acetoclastic methanogen archaea. Thermophilic digesters were dominated by γ-Proteobacteria, Methanothermobacter sp., while mesophilic digesters showed dominance by Firmicutes, uncultured bacteria, Methanosarcina, and Methanoculleus genera. Under aerobic conditions within algal tanks, pH rose from 7.17 to 9.32, with a significant decrease in total ammonia nitrogen, chemical oxygen demand, and soluble phosphorus levels. PCR-DGGE proved a useful tool for investigating the dynamics of microbial community in the bio-processing of piggery wastewater. Knowledge of the microbial communities involved in digestion of piggery wastewater will allow optimization of integrated biosystem by removing the main pollutants like inorganic ammonium-nitrogen, phosphorus, and pathogens from intensive farming system.