Microbial activity and community structure in a lake sediment used for psychrophilic anaerobic wastewater treatment

Aims: The aim of the study was to investigate the feasibility of a continuous reactor for psychrophilic anaerobic wastewater treatment by using the sludge from cold natural environment. Methods and Results: Six sludge samples (S1–S6) were collected from different cold natural locations to select sludge with high anaerobic microbial activity under low temperatures. After a 225‐day incubation, the maximum specific methane production rate of a waterfowl lake sediment (S1) at 15°C (70·5 mLCH₄ gVSS^?1 day^?1) was much higher than all other samples. S1 was thus chosen as the seed sludge for the reactor treating synthetic brewery wastewater at 15°C, by immobilizing the micro‐organisms on polyurethane foam carriers. The chemical oxygen demand (COD) removal efficiency reached over 80% after 240‐day operation at an organic loading rate of 5·3 kg m^?3 day^?1, and significant enrichment of biomass was observed. Clone libraries of the microbial communities in the inoculum had high diversities for both archaea and bacteria. Along with a decrease in microbial community diversities, the dominant bacteria (79·5%) at the end of the operation represented the phylum Firmicutes, while the dominant archaeon (41·5%) showed a similarity of 98% with the psychrotolerant methanogen Methanosarcina lacustris. Conclusions: The possibility of using anaerobic micro‐organisms from cold environments in anaerobic wastewater treatment under psychrophilic conditions is supported by these findings. Significance and Impact of the Study: This study enriches the theory on microbial community and the application on anaerobic treatment of sludge from cold natural environments.     

Anaerobic treatment of the municipal wastewater under psychrophilic conditions

The article discusses possibilities of municipal wastewater treatment in various types of anaerobic reactors (UASB, UAF, AnSBR). The temperature during corresponding laboratory experiments varied between 9–23?°C. Real wastewater from WWTP Bratislava-Petr?alka and synthetic substrate were used in the experiments. The HRT values for reactors were found to be 10–46?h. Treatment of municipal wastewater with the AnSBR and UAF system has shown more positive results in laboratory scale studies then the UASB system. The mean removal efficiency of COD depended on the type of anaerobic reactor, temperature, used HRT and was found between 37–48% (UASB), 56–88% (AnSBR) and 46–90% (UAF).     

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.     

Biofilm reactors in anaerobic wastewater treatment

Attached biofilm reactors provide the means for implementing energy-efficient anaerobic wastewater treatment at full scale. Progress has been made in the development of fixed, expanded and fluidized bed anaerobic processes by addressing fundamental reactor design issues. Several new biofilm reactor concepts have evolved from recent studies.     

Long-term, high-rate anaerobic biological treatment of whey wastewaters at psychrophilic temperatures

Two laboratory-scale anaerobic hybrid reactors, R1 and R2, treated low- (1 kg COD m-3) and high-strength (10 kg COD m-3) whey-based wastewaters, respectively, in a 500-day trial. The chemical oxygen demand (COD) removal efficiencies of R1 averaged 70-80%, at organic loading rates of 0.5-1.3 kg COD m-3 day-1, between 20 and 12 degrees C. The COD removal efficiencies of R2 exceeded 90%, at organic loading rates up to 13.3 kg COD m-3 day-1, between 20 and 14 degrees C. Lowering the operating temperature of R2 to 12 degrees C resulted in a decrease in COD removal efficiency, to between 50% and 60%, and a disintegration of granular sludge. The decline in performance, and granule disintegration, was reversed by decreasing the organic loading rate of R2 to 6.6 kg m-3 day-1. Specific methanogenic activity profiles revealed mesophilic (37 degrees C) temperature optima for biomass in both reactors, even after 500 days of psychrophilic operation, although the development of psychrotolerance in the biomass was noted.     

High-rate anaerobic treatment of wastewater at low temperatures

Anaerobic treatment of a volatile fatty acid (VFA) mixture was investigated under psychrophilic (3 to 8 degrees C) conditions in two laboratory-scale expanded granular sludge bed reactor stages in series. The reactor system was seeded with mesophilic methanogenic granular sludge and fed with a mixture of VFAs. Good removal of fatty acids was achieved in the two-stage system. Relative high levels of propionate were present in the effluent of the first stage, but propionate was efficiently removed in the second stage, where a low hydrogen partial pressure and a low acetate concentration were advantageous for propionate oxidation. The specific VFA-degrading activities of the sludge in each of the modules doubled during system operation for 150 days, indicating a good enrichment of methanogens and proton-reducing acetogenic bacteria at such low temperatures. The specific degradation rates of butyrate, propionate, and the VFA mixture amounted to 0.139, 0.110, and 0.214 g of chemical oxygen demand g of volatile suspended solids-1 day-1, respectively. The biomass which was obtained after 1.5 years still had a temperature optimum of between 30 and 40 degrees C.     

Thermophilic anaerobic treatment of sulphur rich forest industry wastewater

Thermophilic anaerobic treatment of sulphur-rich paper mill wastewater (0.8-3.1 gCOD/l, 340–850 mgSO₄/l; COD:SO₄ 3.4-5.3) was studied in three laboratory-scale, upflow anaerobic sludge blanket (UASB) reactors and in bioassays. The reactors were inoculated with non-adapted thermophilic granular sludge. In the bioassays, no inhibition of the inoculum was detected and about 62% COD removal (sulphide stripped) was obtained. About 70 to 80% of the removed COD was methanised. In the reactors, up to 60–74% COD removal (effluent sulphide stripped) was obtained at loading rates up to 10–30 kgCOD/m³d and hydraulic retention times down to 6 to 2 hours. The effluent total sulphide was up to 150–250 mg/l. Sulphide inhibition could not be confirmed from the reactor performances. The results from bioassays suggested that both the inoculum and sludge from the UASB reactor used acetate mainly for methane production, while sulphide was produced from hydrogen or its precursors.     

Anaerobic treatment of low-strength municipal wastewater by a two-stage pilot plant under psychrophilic conditions

A two-stage anaerobic treatment pilot plant was tested for the treatment of raw domestic wastewater under temperatures ranging from 21 to 14 degrees C. The plant consisted of a hydrolytic upflow sludge bed (HUSB) digester (25.5m3) followed by an upflow anaerobic sludge blanket (UASB) digester (20.36m3). The hydraulic retention time (HRT) varied from 5.7 to 2.8h for the first stage (HUSB digester) and from 13.9 to 6.5h for the second stage (UASB digester). Total suspended solids (TSS), total chemical oxygen demand (TCOD), and biochemical oxygen demand (BOD) removals ranged from 76% to 89%, from 49% to 65%, and from 50% to 77%, respectively, for the overall system. The percentage of influent COD converted to methane was 36.1%, the hydrolysis of influent volatile suspended solids (VSS) reached 59.7% and excess biomass was 21.6% of the incoming VSS. Plant performance was influenced by the wastewater concentration and temperature, yet better results were obtained for influent COD higher than 250mg/l.     

Stability and reproducibility of low-temperature anaerobic biological wastewater treatment

The reproducibility of low-temperature anaerobic biological wastewater treatment trials was evaluated. Two identical anaerobic expanded granular sludge bed bioreactors were used to treat synthetic volatile fatty acid-based industrial wastewater under ambient conditions (18-20 degrees C) and to investigate the effect of various environmental perturbations on reactor performance and microbial community dynamics, which were assessed by chemical oxygen demand removal or effluent volatile fatty acid determination and terminal restriction fragment length polymorphism analysis, respectively. Methanogenic activity was monitored using specific methanogenic activity assays. Reactor performance and microbial community dynamics were each well replicated between Reactor 1 and Reactor 2. Archaeal dynamics, in particular, were associated with reactor operating parameters. Terminal restriction fragment length polymorphism data suggested dynamic acetoclastic and hydrogenophilic methanogenic populations and were in agreement with temporal specific methanogenic activity data. Putative psychrophilic populations were observed in anaerobic bioreactor sludge for the first time.     

Determining specific biomass activity in anaerobic wastewater treatment processes

An experimental method for the measurement of specific gas production rate was developed and tested with biomass samples taken from anaerobic fluidized bed reactors, operating with a variety of carriers with molasses, condensate from cellulose production and brewery wastewater as feeds. The method is based on reactor sampling and offline gas volume measurement during a known time interval. Important factors are biomass and liquid sampling under oxygen-free conditions, using the liquid from the reactor as substrate, providing sufficient mixing and maintaining the physical integrity of the biomass. The method was developed in such a way that small samples (20 ml) were taken under anaerobic conditions (poising agent) for short-term (2–3 min.) gas rate measurements in a small fluidized bed (25 ml) batch reactor with U-tube. Biomass content was measured by an instrumental nitrogen method (Dumas), followed by weight determination of the carrier. The gas rates measured with the test system, and their dependence on substrate concentration, were in good agreement with those directly measured from the continuous fluidized bed reactor. Additions of molasses and acetate to the sample proved that the influence of concentration on the biomass activity can be obtained only by operating the continuous reactor at the concentration levels of interest. Comparison between the reactors showed large differences in the specific activity and the total reactor activity. It was found when comparing two reactors, that the values of the specific and the total activities permitted the calculation of the relative biomass quantities. In this way the influence of the carrier-type could be evaluated.     

嗜冷产甲烷菌及其在废水厌氧处理中的应用

嗜冷产甲烷菌对于自然界的碳素循环具有非常重要的意义,近年来引起了国内外学者的广泛关注.利用嗜冷产甲烷菌实现低温厌氧生物处理过程,可从本质上突破低温厌氧工艺的技术瓶颈,进而大大拓展厌氧生物处理技术的应用范围并降低废水处理的成本.本文针对研究者广泛关注的热点问题,从分离培养及生理生化特性、适冷机制和分子生物学研究几个方面,对嗜冷产甲烷菌的研究进展进行了全面的综述,并对其在低温厌氧生物处理技术中的应用前景进行了分析和展望.     

Brewery wastewater treatment using anaerobic inverse fluidized bed reactors

Two anaerobic inverse fluidized bed reactors were utilized to evaluate organic matter removal from brewery wastewater, applying different OLR and testing two support materials. Hydrodynamic tests varying liquid flow and solid concentration were developed on the supports in order to establish operational conditions. A batch colonization stage was applied using 25% active volume of extendosphere and triturated polyethylene as support materials. The reactors were subsequently operated continuously with stepwise increments in organic loading rate until limiting conditions was reached. For the supports studied, IFBR technology was suitable for organic matter removal present in brewery wastewater with COD removal efficiencies greater than 90%. The reactor with triturated polyethylene support showed an excellent COD removal with OLR values up to 10 g COD/Ld, whereas the reactor with extendosphere support had an excellent hydrodynamic and biologic behavior working with OLR values up to 70 g COD/Ld.     

Feasibility assay in phase-separated anaerobic treatment of cheese industry wastewater

This research was conducted as a part of continuous development of innovative bioprocess technology for the treatment of high strength wastewater. Mixed cheese processing wastewater was tested for the feasibility of phase separated anaerobic digestion in batch mode. Three concentrations of soluble chemical oxygen demand (SCOD) made by dilution of raw wastewater were tested for acidification of organics in the wastewater at two pHs, 6.0 and neutral. More than 95% of fat, 97% of soluble protein and 100% of lactose in the mixed waste were acidified in the acidogenic phase. Three different concentrations of artificial substrate consisting of a mixture of short chain organic acids, acetic, propionic, butyric, and valeric acids, along with alcohol were investigated for waste stabilization in methanogenic phase experiments. More than 95% of SCOD reduction was achieved in the methanogenic phase. This translated that 73.5–83.8% of organics in the wastewater could be recovered as energy (methane gas) instead of massive production of sludge.     

Engineering application of anaerobic ammonium oxidation process in wastewater treatment

Anaerobic ammonium oxidation (Anammox), a promising biological nitrogen removal process, has been verified as an efficient, sustainable and cost-effective alternative to conventional nitrification and denitrification processes. To date, more than 110 full-scale anammox plants have been installed and are in operation, treating industrial NH₄ ⁺-rich wastewater worldwide, and anammox-based technologies are flourishing. This review the current state of the art for engineering applications of the anammox process, including various anammox-based technologies, reactor selection and attempts to apply it at different wastewater plants. Process control and implementation for stable performance are discussed as well as some remaining issues concerning engineering application are exposed, including the start-up period, process disturbances, greenhouse gas emissions and especially mainstream anammox applications. Finally, further development of the anammox engineering application is proposed in this review.     

Protein degradation during anaerobic wastewater treatment: derivation of stoichiometry

The stoichiometry of reactions that describe protein degradation in anaerobic treatment systems were investigated. A methodology was developed to describe protein degradation to organic acids using a single reaction step. The reactions for individual amino acid fermentation and their mediating organisms were reviewed. The dominant fermentation pathways were selected based on a number of assumptions. Using the amino acid content of a model protein, it was then possible to determine stoichiometric coefficients for each major organic acid product in the overall degradation of the protein. The theoretical coefficients were then compared to those determined from two experimental runs on a continuously-fed, well-mixed, laboratory-scale anaerobic wastewater treatment system. In general, the coefficients compared well thus validating the use of a single reaction step for the overall catabolic reaction of protein degradation to organic acids. Furthermore, even when the protein concentration in feed or the feed flow rate was doubled, the amino acid fermentation pathways were found to occur predominantly by only one pathway. Although the choice of Stickland reactions over uncoupled degradation provided good comparisons, an electron balance showed that only about 40% of the amino acids could have proceeded coupled to other amino acid reactions. Uncoupled degradation of the remaining amino acids must have relied on the uptake of hydrogen produced from these reactions by hydrogen-consuming methane bacteria.     

Development of an improved anaerobic filter for municipal wastewater treatment

Development of an improved reactor configuration of anaerobic filter was carried out for the elimination of clogging of filter media. The experiments over different hydraulic retention times (HRTs) indicated that the HRT of 12 h was the most appropriate one for the system studied while treating the municipal wastewater, which resulted 90% and 95% BOD and COD reduction, respectively. Reduction up to 95% in suspended solids concentration could be achieved without any pretreatment. The specific biogas yield obtained was 0.35 m(3) CH(4)/kgCODr with 70% of CH(4) content in the biogas generated from the system at the HRT of 12 h. Operational problems such as clogging of filter media were not observed throughout the period of study over 600 d.