Neutral fat hydrolysis and long-chain fatty acid oxidation during anaerobic digestion of slaughterhouse wastewater

Neutral fat hydrolysis and long-chain fatty acid (LCFA) oxidation rates were determined during the digestion of slaughterhouse wastewater in anaerobic sequencing batch reactors operated at 25 degrees C. The experimental substrate consisted of filtered slaughterhouse wastewater supplemented with pork fat particles at various average initial sizes (D(in)) ranging from 60 to 450 microm. At the D(in) tested, there was no significant particle size effect on the first-order hydrolysis rate. The neutral fat hydrolysis rate averaged 0.63 +/- 0.07 d(-1). LCFA oxidation rate was modelled using a Monod-type equation. The maximum substrate utilization rate (kmax) and the half-saturation concentration (Ks) averaged 164 +/- 37 mg LCFA/L/d and 35 +/- 31 mg LCFA/L, respectively. Pork fat particle degradation was mainly controlled by LCFA oxidation rate and, to a lesser extent, by neutral fat hydrolysis rate. Hydrolysis pretreatment of fat-containing wastewaters and sludges should not substantially accelerate their anaerobic treatment. At a D(in) of 450 microm, fat particles were found to inhibit methane production during the initial 20 h of digestion. Inhibition of methane production in the early phase of digestion was the only significant effect of fat particle size on anaerobic digestion of pork slaughterhouse wastewater. Soluble COD could not be used to determine the rate of lipid hydrolysis due to LCFA adsorption on the biomass.     

Limitations of thermophilic anaerobic wastewater treatment and the consequences for process design

Thermophilic anaerobic digestion offers an attractive alternative for the treatment of medium- and high-strength wastewaters. However, literature reports reveal that thermophilic wastewater treatment systems are often more sensitive to environmental changes than the well-defined high-rate reactors at the mesophilic temperature range. Also, in many cases a poorer effluent quality is experienced while the carry over of suspended solids in the effluent is relatively high. In this paper recent achievements are discussed regarding the process stability of thermophilic anaerobic wastewater treatment systems. Laboratory experiments reveal a relatively low sensitivity to temperature changes if high-rate reactors with immobilized biomass are used. Other results show that if a staged process is applied, thermophilic reactors can be operated for prolonged periods of time under extreme loading conditions (80–100 kg chemical oxygen demand.m^-3.day^-1), while the concentrations of volatile fatty acids in the effluent remain at a low level.     

Sulfate reduction in methanogenic bioreactors

Abstract: In the anaerobic treatment of sulfate-containing wastewater, sulfate reduction interferes with methanogenesis. Both mutualistic and competitive interactions between sulfate-reducing bacteria and methanogenic bacteria have been observed. Sulfate reducers will compete with methanogens for the common substrates hydrogen, formate and acetate. In general, sulfate reducers have better growth kinetic properties than methanogens, but additional factors which may be of importance in the competition are adherence properties, mixed substrate utilization, affinity for sulfate of sulfate reducers, relative numbers of bacteria, and reactor conditions such as pH, temperature and sulfide concentration. Sulfate reducers also compete with syntrophic methanogenic consortia involved in the degradation of substrates like propionate and butyrate. In the absence of sulfate these methanogenic consortia are very important, but in the presence of sulfate they are thought to be easily outcompeted by sulfate reducers. However, at relatively low sulfate concentrations, syntrophic degradation of propionate and butyrate coupled to HZ removal via sulfate reduction rather than via methanogenesis may become important. A remarkable feature of some sulfate reducers is their ability to grow fermentatively or to grow in syntrophic association with methanogens in the absence of sulfate.     

Anaerobic digestion and wastewater treatment systems

Upflow Anaerobic Sludge Bed (UASB) wastewater (pre-)treatment systems represent a proven sustainable technology for a wide range of very different industrial effluents, including those containing toxic/inhibitory compounds. The process is also feasible for treatment of domestic wastewater with temperatures as low as 14–16° C and likely even lower. Compared to conventional aerobic treatment systems the anaerobic treatment process merely offers advantages. This especially is true for the rate of start-up. The available insight in anaerobic sludge immobilization (i.e. granulation) and growth of granular anaerobic sludge in many respects suffices for practice. In anaerobic treatment the immobilization of balanced microbial communities is essential, because the concentration of intermediates then can be kept sufficiently low.So far ignored factors like the death and decay rate of organisms are of eminent importance for the quality of immobilized anaerobic sludge. Taking these factors into account, it can be shown that there does not exist any need for phase separation when treating non- or slightly acidified wastewaters. Phase separation even is detrimental in case the acidogenic organisms are not removed from the effluent of the acidogenic reactor, because they deteriorate the settleability of granular sludge and also negatively affect the formation and growth of granular sludge. The growing insight in the role of factors like nutrients and trace elements, the effect of metabolic intermediates and end products opens excellent prospects for process control, e.g. for the anaerobic treatment of wastewaters containing mainly methanol.Anaerobic wastewater treatment can also profitably be applied in the thermophilic and psychrophilic temperature range. Moreover, thermophilic anaerobic sludge can be used under mesophilic conditions.The Expanded Granular Sludge Bed (EGSB) system particularly offers big practical potentials, e.g. for very low strength wastewaters (COD 1 g/l) and at temperatures as low as 10° C. In EGSB-systems virtually all the retained sludge is employed, while compared to UASB-systems also a substantially bigger fraction of the immobilized organisms (inside the granules) participates in the process, because an extraordinary high substrate affinity prevails in these systems. It looks necessary to reconsider theories for mass transfer in immobilized anaerobic biomass.Instead of phasing the digestion process, staging of the anaerobic reactors should be applied. In this way mixing up of the sludge can be significantly reduced and a plug flow is promoted. A staged process will provide a higher treatment efficiency and a higher process stability. This especially applies for thermophilic systems.     

Recent advances in methane fermentation technology

In the past two decades, a number of biotechnologies for anaerobic (methanogenic) wastewater treatment have been created, and practical applications of these processes are now being extended to more recalcitrant wastewaters and to wastewaters at extreme temperatures. Our knowledge of methanogenic organic degradation associated with bioreactors is also accumulating at a rapid rate. The recent advancement of such fundamental understanding is attributed to modern molecular biology techniques applied to the study of microbial communities and to continuous challenges to the cultivation of many important but recalcitrant anaerobes in bioreactors.     

Energy metabolism among eukaryotic anaerobes in light of Proterozoic ocean chemistry

Recent years have witnessed major upheavals in views about early eukaryotic evolution. One very significant finding was that mitochondria, including hydrogenosomes and the newly discovered mitosomes, are just as ubiquitous and defining among eukaryotes as the nucleus itself. A second important advance concerns the readjustment, still in progress, about phylogenetic relationships among eukaryotic groups and the roughly six new eukaryotic supergroups that are currently at the focus of much attention. From the standpoint of energy metabolism (the biochemical means through which eukaryotes gain their ATP, thereby enabling any and all evolution of other traits), understanding of mitochondria among eukaryotic anaerobes has improved. The mainstream formulations of endosymbiotic theory did not predict the ubiquity of mitochondria among anaerobic eukaryotes, while an alternative hypothesis that specifically addressed the evolutionary origin of energy metabolism among eukaryotic anaerobes did. Those developments in biology have been paralleled by a similar upheaval in the Earth sciences regarding views about the prevalence of oxygen in the oceans during the Proterozoic (the time from ca 2.5 to 0.6 Ga ago). The new model of Proterozoic ocean chemistry indicates that the oceans were anoxic and sulphidic during most of the Proterozoic. Its proponents suggest the underlying geochemical mechanism to entail the weathering of continental sulphides by atmospheric oxygen to sulphate, which was carried into the oceans as sulphate, fueling marine sulphate reducers (anaerobic, hydrogen sulphide-producing prokaryotes) on a global scale. Taken together, these two mutually compatible developments in biology and geology underscore the evolutionary significance of oxygen-independent ATP-generating pathways in mitochondria, including those of various metazoan groups, as a watermark of the environments within which eukaryotes arose and diversified into their major lineages.     

Anaerobic fixed film wastewater treatment

The anaerobic methane fermentation process has long been used in the field of wastewater engineering in sludge processing, mainly for waste stabilization and solids reduction. Recently, major advances in the fundamental understanding of the process microbiology and biochemistry, along with the development of new reactor configurations have promoted a resurgence of interest in the use of this technology for the processing of liquid industrial and municipal wastewaters. Three of these new processes, the anaerobic filter, expanded/fluidized bed, and upflow anaerobic sludge blanket, are discussed.Each of these processes is a fixed film process, which enables the attainment of high solids retention times for good system efficiency and stability, with low hydraulic retention times for system economy. Fixed film anaerobic processes are able to realize many of the benefits of anaerobic processes while overcoming many of the problems historically associated with anaerobic processes.Each of the processes is described, and examples are presented for industrial and municipal applications. Finally, the processes are qualitatively compared. At present, it is not possible to say which reactor configuration is best. In fact, the selection is often dependent on wastewater characteristics, local factors, and several other factors. More full-scale data and operating experience along with basic research needs are needed to clarify further this situation, and to design these systems optimally.     

Fate of Cryptosporidium oocysts, Giardia cysts, and microbial indicators during wastewater treatment and anaerobic sludge digestion.

The extent of reduction in selected microorganisms was tested during both aerobic wastewater treatment and anaerobic digestion of sludge at the wastewater treatment plant in Ottawa to compare the removal of two encysted pathogenic protozoa with that of microbial indicators. Samples collected included the raw wastewater, the primary effluent, the treated wastewater, the mixed sludge, the decanted liquor, and the cake. All of the raw sewage samples were positive for Cryptosporidium oocysts and Giardia cysts, as well as for the other microorganisms tested. During aerobic wastewater treatment (excluding the anaerobic sludge digestion), Cryptosporidium and Giardia were reduced by 2.96 log10 and 1.40 log10, respectively. Clostridium perfringens spores, Clostridium perfringens total counts, somatic coliphages, and heterotrophic bacteria were reduced by approximately 0.89 log10, 0.96 log10, 1.58 log10, and 2.02 log10, respectively. All of the other microorganisms were reduced by at least 3.53 log10. Sludge samples from the plant were found to contain variable densities of microorganisms. Variability in microbial concentrations was sometimes great between samples, stressing the importance of collecting a large number of samples over a long period of time. In all cases, the bacterial concentrations in the cake (dewatered biosolids) samples were high even if reductions in numbers were observed with some bacteria. During anaerobic sludge digestion, no statistically significant reduction was observed for Clostridium perfringens, Enterococcus sp., Cryptosporidium oocysts, and Giardia cysts. A 1-2 log10 reduction was observed with fecal coliforms and heterotrophic bacteria. However, the method utilized to detect the protozoan parasites does not differentiate between viable and nonviable organisms. On the other hand, total coliforms and somatic coliphages were reduced by 0.35 log10 and 0.09 log10, respectively. These results demonstrate the relative persistence of the protozoa in sewage sludge during wastewater treatment.     

Anaerobic treatment of sulphate-rich wastewaters

Until recently, biological treatment of sulphate-rich wastewater was rather unpopular because of the production of H₂S under anaerobic conditions. Gaseous and dissolved sulphides cause physical-chemical (corrosion, odour, increased effluent chemical oxygen demand) or biological (toxicity) constraints, which may lead to process failure. Anaerobic treatment of sulphate-rich wastewater can nevertheless be applied successfully provided a proper treatment strategy is selected. The strategies currently available are discussed in relation to the aim of the treatment: i) removal of organic matter, ii) removal of sulphate or iii) removal of both. Also a whole spectrum of new biotechnological applications (removal of organic chemical oxygen demand, sulphur, nitrogen and heavy metals), recently developed based on a better insight in sulphur transformations, are discussed.     

Anaerobic Treatment in Decentralised and Source-Separation-Based Sanitation Concepts

Anaerobic digestion of wastewater should be a core technology employed in decentralised sanitation systems especially when their objective is also resource conservation and reuse. The most efficient system involves separate collection and anaerobic digestion of the most concentrated domestic wastewater streams: black or brown water and solid fraction of kitchen waste. Separate collection using minimal amount of transport water besides saving this resource allows to apply a targeted treatment. A relatively small volume of digested effluent can be directly reused for fertilisation or processed when a high quality product is required. Clean nutrient production requires advanced multi-step treatment but the quality of products is risk-free. The issue of organic micro-pollutants and their accumulation in the environment is recently often addressed. Anaerobic treatment of total domestic wastewater stream can be applied as well. Treated in this way wastewater can be discharged or used for irrigation or fertilisation. The post-treatment will be usually required and its rate of complexity depends on the anaerobic effluent quality and local requirements for final effluent quality. A variety of technological solutions for treatment of domestic wastewater streams and reuse of resources is discussed in this paper.     

Sludge minimisation technologies

The treatment and disposal of excess sludge represents a bottleneck of wastewater treatment plants all over the world, due to environmental, economic, social and legal factors. There is therefore a growing interest in developing technologies to reduce the wastewater sludge generation. The goal of this paper is to present the state-of-the-art of current minimisation techniques for reducing sludge production in biological wastewater treatment processes. An overview of the main technologies is given considering three different strategies: The first option is to reduce the production of sludge by introducing in the wastewater treatment stage additional stages with a lower cellular yield coefficient compared to the one corresponding to the activated sludge process (lysis-cryptic growth, uncoupling and maintenance metabolism, predation on bacteria, anaerobic treatment). The second choice is to act on the sludge stage. As anaerobic digestion is the main process in sewage sludge treatment for reducing and stabilising the organic solids, two possibilities can be considered: introducing a pre-treatment process before the anaerobic reaction (physical, chemical or biological pre-treatments), or modifying the digestion configuration (two-stage and temperature-phased anaerobic digestion, anoxic gas flotation). And, finally, the last minimisation strategy is the removal of the sludge generated in the activated sludge plant (incineration, gasification, pyrolysis, wet air oxidation, supercritical water oxidation).     

Importance of the methanogenic archaea populations in anaerobic wastewater treatments

Methane derived from anaerobic treatment of organic wastes has a great potential to be an alternative fuel. Abundant biomass from various industries could be a source for biomethane production where combination of waste treatment and energy production would be an advantage. This article summarizes the importance of the microbial population, with a focus on the methanogenic archaea, on the anaerobic fermentative biomethane production from biomass. Types of major wastewaters that could be the source for biomethane generation such as brewery wastewater, palm oil mill effluent, dairy wastes, cheese whey and dairy wastewater, pulp and paper wastewaters and olive oil mill wastewaters in relevance to their dominant methanogenic population are fully discussed in this article.     

典型有机固废厌氧消化微生物研究现状与发展方向

经过人工富集和驯化的兼性和严格厌氧微生物是厌氧消化工艺的核心。不同厌氧消化体系中存在的问题大多可以通过改变微生物群落的代谢活性来得到有效改善。得益于微生物组学检测技术的快速发展,对厌氧消化系统中微生物多样性的认识获得了极大的拓展,同时在微生物类群间、微生物与环境的互作关系研究方面也取得了一系列新的进展。然而,有机固废厌氧消化系统中,各种微生物以及微生物和物质的相互作用构成了更为复杂的代谢网络,所以目前对这些互作关系的解析尚不完善。本文重点关注了厌氧消化过程中的典型菌群互作关系,阐述了典型有机固废厌氧消化系统中存在的问题及微生物在其中发挥的作用,最后,立足于现有组学技术推动的微生物组研究进展,对未来有机固废厌氧消化系统微生物组的研究提出展望。     

Treatment of recycled wastewaters from fishmeal factory by an anaerobic filter

Fishmeal industries processes produce effluents with high load organic matter. These effluents, after recycling and physical-chemical pretreatment, have a high organic content (5-6 g COD/l), proteins (3-5 g/l), salinity close to sea water, sodium chloride (30 g/l) and sulphate (1-3.3 g/l). An anaerobic filter was used for the treatment of this wastewater, with marine sediment as anaerobic inoculum. Anaerobic filter removed up to 70% of the influent COD concentrations at organic loading rates (OLR) of 9.5 and 14.3 (g/l d) and sulphate up to 80% at OLR of 7.1 and 14.3 (g/l d) whereas the pH ranged between 7.0 and 7.5. These results show that anaerobic filter systems are applicable to recycled wastewaters from fishmeal.     

Anaerobic fluidised bed for the purification of effluents from chemical and mechanical pulping

Anaerobic treatment has seldom been used for wastewaters from the pulp and paper industry and other branches of the chemical industry. Escape of volatile pollutants to the atmosphere, which always occurs during aerobic treatment, is avoided, and much less sludge is being produced than in an aerobic process. The greatest obstacle for using anaerobic treatment in the pulp and paper industry is the large wastewater volume, which necessitates short hydraulic detention times, because the treatment is to occur in an enclosed space. We used solid carrier particles to prevent wash-out of biomass from the reactor at high hydraulic loading, and an up-flow system in order to be able to use very small carrier particles, maximizing the surface area for biomass attachment. In this paper we describe and discuss the results obtained with this type of anaerobic reactor (fluidised bed) at bench and semitechnical scale for wastewaters from pressurized ground wood pulping and paper manufacture, sulphite pulp evaporator condensate and bleach waste. Earlier work with Kraft pulp bleaching effluent and thermomechanical pulping wastewater and evaporator condensates using anaerobic reactors is also discussed. The results obtained thus far show that there are several wastewater streams from the pulping industry, where 60 to 90% of the dissolved organic pollutants (measured as COD_Cr or TOC) was biodegraded within 4 to 24 h. The high strength waste streams (COD_Cr 2000 mg O₂ 1⁻¹) allowed organic space load of 4 to 10 kg COD_Cr m⁻³ reactor volume d⁻¹. With low strength wastes the hydraulic loading was the limiting factor.     

Low-temperature anaerobic digestion for wastewater treatment

Methanogenesis is an important biogeochemical process for the degradation of organic matter within cold environments, and is associated with the release of the potent greenhouse gas, methane. Cold methanogenesis has been harnessed, in engineered systems, as low-temperature anaerobic digestion (LTAD) for wastewater treatment and bioenergy generation. LTAD represents a nascent wastewater treatment biotechnology, which offers an attractive alternative to conventional aerobic and anaerobic processes. Successful, high-rate, LTAD of sewage and industrial wastewaters (e.g. from the brewery, food-processing and pharmaceutical sectors), with concomitant biogas generation, has been demonstrated at laboratory-scale and pilot-scale. A holistic, polyphasic approach, which integrates bioprocess, physiological and molecular biological datasets has been critical to the development of the LTAD concept.     

Ferric reduction in organic matter oxidation and its applicability for anaerobic wastewater treatment: a review and future aspects

Anaerobic treatment processes have the advantages of cost-effectiveness, energy efficiency, low sludge yield and potential of resource recovery over conventional aerobic treatment methods and have been gaining increasing attention as an approach for future wastewater management. An important feature of anaerobic processes is the use of alternative electron acceptors to oxygen, which renders treatment flexibility in using redox active elements such as iron and sulfate from other waste materials. Co-treatment of acid mine drainage and municipal wastewater, as an example, has been shown to be an effective method for removing organic materials, metals, and phosphate from the both wastes. It also suggested the applicability of ferric reduction process in wastewater treatment. Most of the previous studies on ferric reduction process and iron reducers were conducted in natural systems such as sediments, soils and groundwater. This paper reviews the significance and fundamentals of the ferric reduction process, its utility for organics oxidation, controlling factors, reaction kinetics, microbial processes of iron reduction and its ecology. The paper also evaluates the suitability and discusses future aspects of using iron reduction for wastewater treatment. Knowledge gaps are identified in this paper for developing such innovative wastewater technology and process optimization.     

Combined anaerobic ammonium and methane oxidation for nitrogen and methane removal

Anammox (anaerobic ammonium oxidation) is an environment-friendly and cost-efficient nitrogen-removal process currently applied to high-ammonium-loaded wastewaters such as anaerobic digester effluents. In these wastewaters, dissolved methane is also present and should be removed to prevent greenhouse gas emissions into the environment. Potentially, another recently discovered microbial pathway, n-damo (nitrite-dependent anaerobic methane oxidation) could be used for this purpose. In the present paper, we explore the feasibility of simultaneously removing methane and ammonium anaerobically, starting with granules from a full-scale anammox bioreactor. We describe the development of a co-culture of anammox and n-damo bacteria using a medium containing methane, ammonium and nitrite. The results are discussed in the context of other recent studies on the application of anaerobic methane- and ammonia-oxidizing bacteria for wastewater treatment.     

Use of the Static Granular Bed Reactor (SGBR) with anaerobic sludge to treat poultry slaughterhouse wastewater and kinetic modeling

Poultry slaughterhouses discharge very high amount of wastewaters and these wastewaters can be treated successfully at a very low cost using anaerobic treatment. In this study, the Static Granular Bed Reactor (SGBR), a newly developed anaerobic process which is fully anaerobic granule, and another Static Granular Bed Reactor containing both anaerobic granular biomass and non-granular biomass were employed for the treatment of poultry slaughterhouse wastewater. The objective of the use of two reactors having different types of anaerobic biomass is to evaluate whether anaerobic sludge could be used effectively instead of anaerobic granule, which is much more difficult to obtain than the other during the start up period. Average COD removal efficiencies were greater than 95% for both of the reactors. Furthermore, Grau second-order and modified Stover–Kincannon models were successfully used to develop a kinetic model of the experimental data with a high correlation coefficient (R² > 0.95).     

Inhibition of Biogas Production by Alkyl Benzene Sulfonates (LAS) in a Screening Test for Anaerobic Biodegradability

The effect of the inoculum source on the digestion of linear alkylbenzene sulfonates (LAS) under anaerobic conditions has been investigated. The potential for primary and ultimate LAS biodegradation of anaerobic sludge samples obtained from wastewater treatment plants (WWTPs) of different geographical locations was studied applying a batch test system. It was found that only 4–22% of the LAS added to the batch anaerobic digesters was primarily transformed suggesting a poor primary degradation of the LAS molecule in anaerobic discontinuous systems. Regarding ultimate biodegradation, the addition of LAS to the batch anaerobic digesters caused a reduction on the extent of biogas production. Significant differences in the inhibition extent of the biogas production were observed (4–26%) depending on the sludge used as inoculum. Effect of the surfactant on the anaerobic microorganisms was correlated with its concentration in the aqueous phase. Sorption of LAS on anaerobic sludge affects its toxicity by depletion of the available fraction of the surfactant. LAS content on sludge was related to the total amount of calcium and magnesium extractable ions. The presence of divalent cations promote the association of LAS with anaerobic sludge reducing its bioavailability and the extent of its inhibitory effect on the biogas production.