Effects of dilution on dissolved oxygen depletion and microbial populations in the biochemical oxygen demand determination

The biochemical oxygen demand (BOD) value is still a key parameter that can determine the level of organics, particularly the content of biodegradable organics in water. In this work, the effects of sample dilution, which should be done inevitably to get appropriate dissolved oxygen (DO) depletion, on the measurement of 5-day BOD (BOD₅), was investigated with and without seeding using natural and synthetic water. The dilution effects were also evaluated for water samples taken in different seasons such as summer and winter because water temperature can cause a change in the types of microbial species, thus leading to different oxygen depletion profiles during BOD testing. The predation phenomenon between microbial cells was found to be dependent on the inorganic nutrients and carbon sources, showing a change in cell populations according to cell size after 5-day incubation. The dilution of water samples for BOD determination was linked to changes in the environment for microbial growth such as nutrition. The predation phenomenon between microbial cells was more important with less dilution. BOD₅ increased with the specific amount of inorganic nutrient per microbial mass when the natural water was diluted. When seeding was done for synthetic water samples, the seed volume also affected BOD due to the rate of organic uptake by microbes. BOD₅ increased with the specific bacterial population per organic source supplied at the beginning of BOD measurement. For more accurate BOD measurements, specific guidelines on dilution should be established.     

Immobilized multi-species based biosensor for rapid biochemical oxygen demand measurement

To improve the practicability of rapid biochemical oxygen demand (BOD) method, we proposed a stable BOD sensor based on immobilizing multi-species BODseed for wastewater monitoring in the flow system. The activation time of the biofilm was greatly shortened for the biofilm prepared by BODseed in the organic-inorganic hybrid material. Some influence factors such as temperature, pH, and concentration of phosphate buffer solution (PBS) were investigated in detail in which high tolerance to environment was validated for the BOD sensor permitted a wide pH and PBS concentration ranges. The minimum detectable BOD was around 0.5 mg/l BOD under the optimized 1.0 mg/ml BODseed immobilized concentration. The as-prepared BOD sensor exhibited excellent stability and reproducibility for different samples. Furthermore, the as-prepared BOD biosensor displayed a notable advantage in indiscriminate biodegradation to different organic compounds and their mixture, similar to the character of conventional BOD(5) results. The results of the BOD sensor method are well agreed with those obtained from conventional BOD(5) method for wastewater samples. The proposed rapid BOD sensor method should be promising in practical application of wastewater monitoring.     

Diversity and assembly patterns of activated sludge microbial communities: A review

Understanding diversity and assembly patterns of microbial communities in activated sludge (AS) is pivotal for addressing fundamental ecological questions and wastewater treatment engineering. Recent applications of molecular methods especially high-throughput sequencing (HTS) have led to the explosion of information about AS community diversity, including the identification of uncultured taxa, and characterization of low-abundance but environmentally important populations such as antibiotic resistant bacteria and pathogens. Those progresses have facilitated the leverage of ecological theories in describing AS community assembly. The lognormal species abundance curve has been applied to estimate AS microbial richness. Taxa-area and taxa-time relationships (TAR and TTR) have been observed for AS microbial communities. Core AS microbial communities have been identified. Meanwhile, the roles of both deterministic and stochastic processes in shaping AS community structures have been examined. Nonetheless, it remains challenging to define tempo-spatial scales for reliable identification of community turnover, and find tight links between AS microbial structure and wastewater treatment plant (WWTP) functions. To solve those issues, we expect that future research will focus on identifying active functional populations in AS using omics- methods integrated with stable-isotope probing (SIP) with the development of bioinformatics tools. Developing mathematic models to understand AS community structures and utilize information on AS community to predict the performance of WWTPs will also be vital for advancing knowledge of AS microbial ecology and environmental engineering.     

The operating line for activated sludge

The mass balance for O2 on an activated sludge process has been re-arranged to identify the O2 limited boundary for any given treatment installation and for any combination of feed flow rate and input bCOD. The boundary is described graphically as the operating line.     

Microbial fuel cell-based biosensor for fast analysis of biodegradable organic matter

The current study was made to develop a biosensor based on a single-chamber microbial fuel cell in which anaerobes were retained in the anode compartment separated from the cathode compartment by a proton exchange membrane. In the sensor a replaceable anaerobic consortium was used for analyzing biodegradable organic matter. The anaerobes acted as biocatalysts in oxidizing organic matter and transferring electrons to the anode. The biocatalysts were renewed for each sample analysis by replacing the old anaerobic consortium with an equal amount of fresh one. A glucose standard solution was used as the target substrate. To obtain the maximum sensor output, the MFC-based sensor system was optimized using an 800 Ω resistor as the load to the external electric circuit and 25 mM phosphate buffer with 50 mM NaCl as catholyte in the aerobic compartment. The temperature of anaerobic compartment was maintained at optimal 37 °C. The cell potential across the electrodes increased with increasing loading of glucose. The sensor response was linear against concentration of glucose up to 25 g l⁻¹. The detection limit was found as 0.025 g l⁻¹. The microbial fuel cell with replaceable anaerobic consortium could be used as a biosensor for on-line monitoring of organic matter.     

A flow injection analysis system with encapsulated high-density <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> cells for rapid determination of biochemical oxygen demand

The biochemical oxygen demand (BOD) determination was studied using a novel flow injection analysis (FIA) system with encapsulated Saccharomyces cerevisiae cells and an oxygen electrode and was compared with conventional 5-day BOD tests. S. cerevisiae cells were packed in a calcium alginate capsule at a dry cell weight of 250 g/l of capsule core. The level of dissolved oxygen (DO) was reduced due to the enhanced respiratory activity of the microbial cells when the injected nutrient passed through the bioreactor. The decrease in DO (ΔDO) was intensified with the amount of microbial cells packed in the bioreactor. However, the specific ΔDO decreased as the amount of cells loaded in the bioreactor increased. The ΔDO value was dependent on the pH and temperature of the mobile phase and reached its maximum value at 35°C and pH 7–8. Also, ΔDO became larger at longer response times as the flow rate of the mobile phase decreased. The measurement of ΔDO was repeated more than six times consecutively using a 20-ppm standard glucose and glutamic acid solution, which confirmed the reproducibility with a standard deviation of 0.95%. A strong linear correlation between ΔDO and BOD was also observed. The 5-day BOD values of actual water and wastewater samples were in accordance with the BOD values obtained by this FIA method using encapsulated S. cerevisiae cells. Unlike the cell-immobilized bead system, there was no contamination of the bioreactor resulting from any leak of yeast cells from the sensor capsules during BOD measurements.     

Electricity generation using chocolate industry wastewater and its treatment in activated sludge based microbial fuel cell and analysis of developed microbial community in the anode chamber

Feasibility of using chocolate industry wastewater as a substrate for electricity generation using activated sludge as a source of microorganisms was investigated in two-chambered microbial fuel cell. The maximum current generated with membrane and salt bridge MFCs was 3.02 and 2.3 A/m², respectively, at 100 Ω external resistance, whereas the maximum current generated in glucose powered MFC was 3.1 A/m². The use of chocolate industry wastewater in cathode chamber was promising with 4.1 mA current output. Significant reduction in COD, BOD, total solids and total dissolved solids of wastewater by 75%, 65%, 68%, 50%, respectively, indicated effective wastewater treatment in batch experiments. The 16S rDNA analysis of anode biofilm and suspended cells revealed predominance of β-Proteobacteria clones with 50.6% followed by unclassified bacteria (9.9%), α-Proteobacteria (9.1%), other Proteobacteria (9%), Planctomycetes (5.8%), Firmicutes (4.9%), Nitrospora (3.3%), Spirochaetes (3.3%), Bacteroides (2.4%) and γ-Proteobacteria (0.8%). Diverse bacterial groups represented as members of the anode chamber community.     

A yeast co-culture-based biosensor for determination of waste water contamination levels

Artificial microbial co-cultures were formed to develop the receptor element of a biosensor for assessment of biological oxygen demand (BOD). The co-cultures possessed broad substrate specificities and enabled assays of water and fermentation products within a broad BOD range (2.4–80 mg/dm³) with a high correlation to the standard method (R = 0.9988). The use of the co-cultures of the yeasts Pichia angusta, Arxula adeninivorans and Debaryomyces hansenii immobilized in N-vinylpyrrolidone-modified poly(vinyl alcohol) enabled developing a BOD biosensor possessing the characteristics not inferior to those in the known biosensors. The results are indicative of a potential of using these co-cultures as the receptor element base in prototype models of instruments for broad application.     

A spectrophotometric biochemical oxygen demand determination method using 2,6-dichlorophenolindophenol as the redox color indicator and the eukaryote Saccharomyces cerevisiae

A method to determine the spectrophotometric biochemical oxygen demand (BOD(sp)) was studied with high sensitivity and reproducibility by employing 2,6-dichlorophenolindophenol (DCIP) as a redox color indicator, the yeast Saccharomyces cerevisiae, and a temperature-controlling system providing a three-consecutive-stir unit. The absorbance of DCIP decreased due to the metabolism of organic substances in aqueous samples by S. cerevisiae. Under optimum conditions, a calibration curve for glucose glutamic acid concentration between 1.1 and 22mg O(2) L(-1) (r=0.988, six points, n=3) was obtained when the incubation mixture was incubated for 10min at 30 degrees C. The reproducibility of the optical responses in the calibration curve was 1.77% (average of relative standard deviations; RSD(av)). Subsequently, the characterization of this method was studied. The optical responses to pure organic substances and the influence of chloride ions, artificial seawater, and heavy metal ions on the sensor response were investigated before use with real samples. Measurements of real samples using river water were performed and compared with those obtained using the BOD(5) method. Finally, stable responses were obtained for 36 days when the yeast cell suspension was stored at 4 degrees C (response reduction, 89%; RSD(av) value for 9 testing days, 8.4%).     

短程硝化活性污泥微生物群落结构及羟胺代谢对短程硝化的影响

活性污泥法随着技术的成熟,已应用在高氨氮污水/废水处理中,通过不断发展衍生出的很多新型工艺也成为研究热点,短程硝化反应作为代表已逐渐体现出优越性。短程硝化能达到高效净化污水的目的,其反应中的代谢产物羟胺也和微生物类群及反应产物之间有着至关重要的影响。反应器中活性污泥的微生物群落结构和动态密切相关,探究微生物群落结构能帮助生物强化、优化参数,提高脱氮效率。本文主要总结了近年来有关短程硝化/半短程硝化活性污泥微生物群落组成与结构及其与反应器处理效率之间的关系,以及羟胺代谢对短程硝化的影响等方面的研究进展,这些研究加深了对微生物群落结构和污水处理工艺之间的认识,但充分发掘生物信息、提高工艺效能之路仍然充满挑战,还需利用氮平衡方法、Real-time PCR法等多种生物技术手段对短程硝化进行全方位研究,为实践提供坚实的理论基础。     

Stopped-flow system with ozonizer for the estimation of low biochemical oxygen demand in environmental samples

The stopped-flow system with an ozonizer was developed to estimate low biochemical oxygen demand (BOD) in rivers. Rivers contain many biopersistent organic compounds such as humic acid, lignin, and gum arabic. Free radicals generated by self-decomposition of ozone were used as powerful oxidants to split organic compounds. Ozonysis of the samples was carried out by 42.4 g N⁻¹ m⁻³ ozone for 3 min at pH 7.0. Artificial wastewater (AWW) solutions were employed as standard solutions for the calibrations of the BOD sensor. At a BOD of 1 mg l⁻¹, the sensor response after ozonation was 1.6-fold higher than that before ozonation. The response time of the BOD sensor was only 5 min, being independent of the concentrations, and the lower detection limit was 0.5 mg l⁻¹ BOD. The degradations of lignin and tannic acid by ozonation were 54.1 and 42.3%, respectively. In the biosensor responses by ozonation, lignin, gum arabic, and surfactant increased by double or more compared with previous responses. BOD in rivers was estimated using the stopped-flow system. Environmental samples pretreated with ozone gave high responses to the biosensor that were similar to those of the conventional BOD₅ method. Accordingly, a good correlation between the sensor and the conventional BOD₅ was obtained (r = 0.989). The system has to evolve the highly sensitive BOD determination.     

Optimizations of particle size and pulp density for solubilization of rock phosphate by a microbial consortium from activated sludge

Microbial solubilization of rock phosphate is getting more and more attention recently. However, the microorganisms used in previous studies were mostly single or known species, and seldom studies focused on the mixed microorganisms or microbial consortia from natural environments. In this study, a microbial consortium taken from activated sludge was used to solubilize two different mid-low-grade rock phosphates. The results showed that the microbial consortium could effectively solubilize the rock phosphates in National Botanical Research Institute’s phosphate growth medium and released soluble phosphorus in the broth. The biomass increased gradually, whereas the pH decreased sharply during the solubilizing process. The maximum phosphorus solubilization was recorded at particle size of 150?µm. Higher or lower than this optimal particle size, the phosphorus solubilization decreased. The phosphorus solubilization gradually decreased with a larger pulp density from 1 to 5%, and the optimal pulp density was 1%. The solubilization level of microbial consortium varied with different rock phosphates. The results revealed that the soluble phosphorus released from high-silicon ore was higher than which from high-magnesium ore. A strong positive correlation between biomass and phosphorus solubilization in the broth was observed from regression analysis results, and the phosphorus solubilization also had a significant negative correlation with pH in the broth.     

Granular activated carbon single-chamber microbial fuel cells (GAC-SCMFCs): A design suitable for large-scale wastewater treatment processes

As an emerging biotechnology capable of removing contaminants and producing electricity, microbial fuel cells (MFCs) hold a promising future in wastewater treatment. However, several main problems, including the high internal resistance (R_in), low power output, expensive material, and complicated configuration have severely hindered the large-scale application of MFCs. The study targeted these challenges by developing a novel MFC system, granular activated carbon single-chamber MFC, termed as GAC-SCMFC. The batch tests showed that GAC was a good substitute for carbon cloth and GAC-SCMFCs generated high and stable power outputs compared with the traditional two-chamber MFCs (2CMFCs). Critical operational parameters (i.e. wastewater substrate concentrations, GAC amount, electrode distance) affecting the performance of GAC-SCMFCs were examined at different levels. The results showed that the R_in gradually decreased from 60 Ω to 45 Ω and the power output increased from 0.2 W/m³ to 1.2 W/m³ when the substrate concentrations increased from 100 mg/L to 850 mg/L. However, at high concentrations of 1000–1500 mg/L, the power output leveled off. The R_in of MFCs decreased 50% when the electrode distance was reduced from 7.5 cm to 1 cm. The highest power was achieved at the electrode distance of 2 cm. The power generation increased with more GAC being added in MFCs due to the higher amount of biomass attached. Finally, the multi-anode GAC-SCMFCs were developed to effectively collect the electrons generated in the GAC bed. The results showed that the current was split among the multiple anodes, and the cathode was the limiting factor in the power production of GAC-SCMFCs.     

Harnessing the power of enzymes for environmental stewardship

Enzymes are versatile catalysts with a growing number of applications in biotechnology. Their properties render them also attractive for waste/pollutant treatment processes and their use might be advantageous over conventional treatments. This review highlights enzymes that are suitable for waste treatment, with a focus on cell-free applications or processes with extracellular and immobilized enzymes. Biological wastes are treated with hydrolases, primarily to degrade biological polymers in a pre-treatment step. Oxidoreductases and lyases are used to biotransform specific pollutants of various nature. Examples from pulp and paper, textile, food and beverage as well as water and chemical industries illustrate the state of the art of enzymatic pollution treatment. Research directions in enzyme technology and their importance for future development in environmental biotechnology are elaborated. Beside biological and biochemical approaches, i.e. enzyme prospection and the design of enzymes, the review also covers efforts in adjacent research fields such as insolubilization of enzymes, reactor design and the use of additives. The effectiveness of enzymatic processes, especially when combined with established technologies, is evident. However, only a limited number of enzymatic field applications exist. Factors like cost and stability of biocatalysts need to be addressed and the collaboration and exchange between academia and industry should be further strengthened to achieve the goal of sustainability.