Dimethylsulfoxidreduktase-Aktivität Charakterisierung der mikrobiellen Biomasseaktivität im Schlamm einer belüfteten Abwasserteichanlage

Dimethyl Sulfoxide Reduction Method for the Characterization of Biomass Activity in Sludge of an Aerated Lagoon The microbial biomass activity was studied in sludge from a wastewater lagoon (Hatzbachtal) which consisted of 4 aerated ponds and one polishing pond. The lagoon was characterized by low BOD₅ loading and high dilution because of water from the surroundings. Sludge samples were taken from 4 sampling sites of each aerated pond and from one site of the polishing pond. The biomass activity in the sludge samples was analysed with the help of dimethyl sulfoxide reductase (DRA) and dehydrogenase (DHA) activity. In addition, C_org-content was also determined. The effect of different reaction parameters on the DRA was studied in relation to incubation period, temperature and atmosphere as well as sludge weight and pH value. The results presented here show that a linear increase in the DRA occured for the sludge weight ranging from 0.1 to 0.5 g, incubation period from 0.5 to 11 h and incubation temperature from 20 to 50 °C. The pH spectrum from acidic to neutral did not effect the DMS formation in sludge. Although the values for DRA were always higher in anaerobic incubation of sludge than in aerobic incubation, the courses of the DRA in both conditions were parallel. Comparing the biomass activity in sludge from different sampling sites of the aerated ponds, the average C_org-content was found to be reflected by the DRA and DHA. Further, the variation in the DRA could represent also the influence of oxygen concentration. Therefore, a reduction in biomass activities indicated a decline in the availability of oxygen.     

Grundlagen für die Dimensionierung der Belüftung bei Belebtschlammanlagen

Summary The generally accepted theory for the calculation of the oxygen consumption caused by the microbes in an aeration basin assumes that the total of oxygen utilizedq _g is the sum of the endogenous respiration of the sludgeq _e plus a quantity for substrate respirationq _s . This theory—though physiologically probably not quite correct—, is very convenient because it allows a causal calculation of the necessary oxygen input into an activated sludge plant. Extensive data on endogenous respiration of activated sludge are presented (Table 1 and 5) and are correlated with sludge load and sludge age (Fig. 4 and 5). The term substrate respiration is discussed and a calculation method, based on the removal of organic C by the mixed liquor, is proposed. In view of the fact that nitrification is an inevitable reaction in activated sludge treatment (its extent being strictly a function of the operation parameters of a plant), oxygen consumption for the oxidation of ammonia has to be included into the term of substrate respiration. The amount of this oxygen fraction has to be predicted from the degree of nitrification to be expected at the desired operation conditions. The following formula for the calculation of the oxygen input into a completely mixed aeration basin is proposed (explanations of symbols see annex): or This same calculation method may be applied stepwise for aeration basins of the tubular reactor type (longitudinal flow basins) considering the concentration decrease of the substrates along the length profile of the basin (the term for endogenous respiration can be assumed constant for the entire aeration time).

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The Influence of the TTC Concentration on the Determination of Activated Sludge Activity

For a correct determination of the activated sludge activity with the aid of TTC an optimum TTC concentration is required not only in the solution, but also as related to the biomass. Since TTC has toxic effects on bacteria, the selection of the optimum TTC concentration is connected with problems: in the suboptimal range the TTC concentration as referred to the existing bioactivity is too low, in the supraoptimal range the toxic effect does not permit any correct determination of activity. A generally optimum range cannot be stated due to the very different activities and sensitivities of the present bacterial populations. Therefore, the application of the TTC methods requires the simultaneous parallel application of different TTC concentrations and the determination of the activity in the plateau range of the relation between the TTC concentration and the formazan concentration formed.     

Kurzzeitermittlung der Ablaufqualität von Belebtschlammanlagen anhand der Atmungsaktivität

For checking the quality of the effluent from an activated sludge plant for waste-waters from the sulphite pulp production the respiration of an activated sludge sample in air-saturated water is determined initially without the addition of substrate (ground respiration) and after that with an addition of substrate (methanol, substrate respiration) by means of the Clark-electrode. The measuring device is designed by means of zero displacement in such a way that the difference of the oxygen concentration is indicated by a time relay as the difference quotient. Between the ground respiration and the effluent concentration of BOD₅ exists a significant positive regression, which can be used for controlling the plant. The determination of the substrate respiration serves only for checking with respect to perhaps given inhibition of the conversion of matter by the activated sludge. The determination of the activity of the activated sludge sample takes only 20 min; inclusive of the required determination of the dry matter of sludge, the technique provides a well reproducible measured value for the biochemical conversion in the activated sludge plant within 1… 1.5 h.     

Einfluß einiger Umweltfaktoren auf die N2O-Bildung durch Belebtschlamm unter anoxischen Bedingungen Influence of Some Environmental Conditions on N2O-Release by Activated Sludge under Anoxic Conditions

The release of nitrous oxide during denitrification in activated sludge is influenced by various environmental factors. Experiments with different electron acceptors showed that under anoxic conditions increasing concentrations of nitrate as well as nitrite enhance the production of N₂O. The N₂O concentration in the gas phase increases for 2 h, then decreases. Nitrite causes a higher release of N₂O than nitrate. Acetate in high concentration can stimulate N₂O-release. Without addition of acetate nitrate and nitrite reduction are delayed, and hence N₂O-release and consumption are much slower. In activated sludge sulfide can play a role as an inhibitor of the nitrous oxide reductase (like in soil and pure cultures), but is inactivated very rapidly. Repeated addition of sulfide can cause a new release of nitrous oxide. At low pH (less than 6.5) denitrification is slightly inhibited and N₂O-release is 100 times higher than at pH 7.0 to 8.0. Under oxygen-limiting conditions N₂O-release and denitrification starts at 0.5–1.0 mg/L DO, but it is not always accompanied by accumulation of nitrite. Further reduction of the oxygen concentration reduces N₂O-release.     

Partitioning of Fluoroquinolones on Wastewater Sludge

Laboratory‐scale batch experiments were conducted to investigate the adsorption behavior of eight fluoroquinolones (FQs) on aerobic, anoxic, and anaerobic sludge, under different adsorpiton time, pH, and temperature conditions. Results indicated that adsorption of FQs onto all sludge was a physical sorption process. The relationship of the partitioning coefficient (K_d) and the octanol/water partition coefficient (K_ow) for each FQ was established. The adsorbed fraction of FQs on sludge could then be predicted with the K_d. It was calculated that about 50–72% of the FQs were adsorbed on the sludge. Therefore, the adsorption effect must be considered when studying the fate and occurrence of FQs in wastewater treatment systems.     

Treatment of Malting Wastewater in a Granular Sludge Sequencing Batch Reactor (SBR)

Aerobic granular sludge was successfully cultivated in a sequencing batch reactor (SBR) treating wastewater from the malting process with a high content of particulate organic matter. At an organic loading rate of 3.2 kg/(m³ d) COD_total and an influent particle concentration of 0.95 g/L MLSS an average removal of 50% in COD_total and 80% in COD_dissolved could be achieved. A comparison of granular and flocculent sludge grown under the same operating conditions showed no significant difference in removal efficiency although granules exhibited a higher metabolic activity in terms of specific oxygen uptake rate (r_{O2, X}). Two distinct mechanisms of particle removal were observed for granular sludge: during initial granule formation, particles were incorporated into the biofilm matrix. For mature granules, a high level of protozoa growth on the granule surface accounted for the ability to remove particulate COD. Combined evaluation of the development in MLSS content and sludge bed settling rate (i.e., mean derivative of the normalized sludge volume) was found to be an adequate method for monitoring the characteristic settling properties of a granulizing sludge bed. By means of this method, a distinct substrate gradient out of several operating conditions was concluded to have the biggest impact on the formation of aerobic granular sludge.     

Degree of Sulfate‐Reducing Activities on COD Removal in Various Reactor Configurations in Anaerobic Glucose and Acetate‐fed Reactors

Sulfate‐reduction data from various anaerobic reactor configurations, e. g., upflow anaerobic sludge blanket reactor (UASBR), completely stirred tank reactor (CSTR), and batch reactor (BR) with synthetic wastewaters, having glucose and acetate as the substrates and different levels of sulfate, were evaluated to determine the level of sulfate‐reducing activity by sulfate‐reducing bacteria coupled to organic matter removal. Anaerobic reactors were observed for the degree of competition between sulfate‐reducing sulfidogens and methane producing bacteria during the degradation of glucose and acetate. Low sulfate‐reducing activity was obtained with a maximum of 20% of organic matter degradation with glucose‐fed upflow anaerobic sludge bed reactors (UASBRs), while a minimum of 2% was observed with acetate‐fed batch reactors. The highest sulfate removal performance (72–89%) was obtained from glucose fed‐UASB reactors, with the best results observed with increasing COD/SO₄ ratios. UASB reactors produced the highest level of sulfidogenic activity, with the highest sulfate removal and without a performance loss. Hence, this was shown to be the optimum reactor configuration. Dissolved sulfide produced as a result of sulfate reduction reached 325 mg/L and 390 mg/L in CST and UASB reactors, respectively, and these levels were tolerated. The sulfate removal rate was higher at lower COD/SO₄ ratios, but the degree of sulfate removal improved with increasing COD/SO₄ ratios.     

氧化还原条件对红枫湖沉积物磷释放影响的微尺度分析

选取贵州红枫湖为研究对象,在实验室条件下模拟了自然、好氧和厌氧条件下沉积物内源磷的释放过程,联合应用微电极技术和沉积物磷形态分析对沉积物—水界面开展了微尺度观测与研究.结果表明,厌氧条件下红枫湖沉积物总磷含量显著降低,且主要是NaOH提取态磷(NaOH-P)和残渣态磷(rest-P)含量降低所致,厌氧条件下沉积物孔隙水中磷酸盐浓度明显升高,而好氧条件下沉积物孔隙水磷酸盐浓度显著降低,反映厌氧条件显著促进了红枫湖沉积物磷释放.厌氧条件下沉积物内部溶解氧浓度下降、硫还原活动增强可能是导致NaOH-P释放的主要原因.O_2浓度的降低加速了沉积物还原作用并产生大量H2S,进而与二价铁离子形成硫化亚铁沉淀,最终导致NaOH-P(Fe-P)释放到孔隙水中.好氧条件向厌氧条件的转换可通过改变沉积物内部pH值分布和微生物活动促使rest-P释放:厌氧条件下,厌氧微生物不仅可以消耗硫酸根产生H_2S,导致pH值降低,还可消耗有机质,将有机磷转变为无机磷.上述研究结果表明,沉积物—水界面氧化还原环境可影响沉积物氧渗透深度、pH值分布、微生物活动、硫循环以及有机质降解过程,进而控制沉积物磷的形态转化与释放.联合应用微电极技术和沉积物磷形态分析对湖泊沉积物—水界面开展微尺度观测研究是揭示沉积物内源磷释放机制与控制因素的有效途径.     

A Contribution to the Mechanism of Elimination of Dimethylformamide and Dimethylamine by a Heterogenous Culture of Microorganisms

In a batch experiment with activated sludge from a large-scale plant the hydrolytical and biochemical degradation of dimethylformamide (DMF) and its reaction product dimethylamine (DMA) and the conversion connected with this under aerobic and anaerobic conditions is investigated. Parallel to the hydrolysis of DMF the biochemical degradation of DMA occurs. The extensive conversion of these substrates is followed by the nitrification of the ammoniumions formed, and under subsequent anaerobic conditions with methanol as the carbon source also denitrification can be achieved. If the oxygen supply of the activated sludge is insufficient, however, also DMA can be used as a carbon source for denitrification, ammonium-ions being released from DMA in addition. In the batch experiment the elimination rates referred to nitrogen were 7 … 14 mg/g · h for DMF, 0.8 … 1.7 mg/g · h for DMA, 0.3 … 0.6 mg/g · h for NH and 0.4 to 1.3 mg/g · h for nitrate in the presence of methanol. DMF and DMA are obviously not suitable as the only carbon source for denitrification.     

Kurzzeit-Toxizitätstests zur Messung der Toxizität von Chemikalien gegenüber Belebtschlamm

Short-term Toxicity Tests for the Measurement of Toxicity of Chemicals Towards Activated Sludge. To avoid severe damage to the activated sludge of a wastewater treatment plant it is necessary to control the sludge activity by means of biological test systems. Especially the short-term respiration assay and the dehydrogenase assay with resazurin as a redox-dye seem to be appropriate for this monitoring program. The practical performance of both test systems was optimized in some essential points. With the help of several reference substances both monitoring systems were tested for practical use as short-term toxicity tests. The results of the investigations showed that the inhibitory concentrations measured in both test systems were partially very different. In monitoring experiments with laboratory-scale wastewater treatment plants it could be shown that a positive correlation between respiration activity and dehydrogenase activity is not always given. Furthermore, a pH-dependent toxicity could be demonstrated for several substituted phenols.     

Studies on the Behaviour of Water-endangering Substances in Plants for the Biological Cleaning of Municipal Waste Waters. Part 1: Bromoxynil

The behaviour of the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), labelled with ⁸²Br, is investigated in an activated sludge plant on the laboratory scale with municipal wastewater. Only 5% of bromoxynil present in a concentration of 5 mg/l are degraded after preadaption of activated sludge for some weeks. The known hydrolysis products 3,5-dibromo-4-hydroxybenzamide and 3,5-dibromo-4-hydroxybenzoic acid were identified as degradation products. In addition, 1.9±0.9% of the quantity of ⁸²Br appeared in the solution as Br^? by debromination. Moreover, a pronounced adsorption of the active substance on activated sludge occurs. Its activity amounts to about 130% of the initial activity of the solution.     

In situ enzyme activities of filamentous scum bacteria in municipal activated sludge wastewater treatment plants

In situ phosphatase, esterase, lipase, and β‐glucuronidase activities were investigated in filamentous scum bacteria by ELF (enzyme labeled fluorescence)‐technology. Microthrix parvicella exhibited significant activities for all four enzymes with lipase activity being the highest. In situ activities were considerably higher in activated sludge as compared to scum indicating M. parvicella growth to occur mainly in the sludge fraction. “Nocardioform actinomycetes” showed significant activities for phosphatase, esterase, and β‐glucuronidase, lipase activity was only moderate. Activities revealed to be similar for activated sludge and scum. As population densities of enzyme‐active actinomycetes were noticeably higher in scum they presumably find good growth conditions in the scum layer. Enzyme activities in Nostocoida limicola morphotypes were only low with no lipase activity being detected. Due to their phylogenetic diversity activity assignment should be combined with fluorescence in situ hybridization (FISH). Numerous type 0041/0675 and type 1851 filaments showed all but lipase activity. However, some of their attached bacteria revealed to be lipase‐active. Only a few morphotype 0092 filaments revealed phosphatase, esterase, and β‐glucuronidase activity. ELF investigations proved suitable for monitoring in situ filamentous activity. Present imponderabilities of Eikelboom morphotype phylogenetic affiliation are discussed.     

Tidal pumping and biogeochemical processes: Dissolution within the tidal capillary fringe of eogenetic coastal carbonates

Growing evidence suggests microbial respiration of dissolved organic carbon (DOC) may be a principal driver of subsurface dissolution and cave formation in eogenetic carbonate rock. Analyses of samples of vadose zone gasses, and geochemical and hydrological data collected from shallow, uncased wells on San Salvador Island, Bahamas, suggest tidally varying water tables may help fuel microbial respiration and dissolution through oxygenation. Respiration of soil organic carbon transported to water tables generates dysaerobic to anaerobic groundwater, limiting aerobic microbial processes. Positive correlations of carbon dioxide (CO₂), radon-222 (²²²Rn) and water table elevation indicate, however, that tidal pumping of water tables pulls atmospheric air that is rich in oxygen, and low in CO₂ and ²²²Rn, into contact with the tidal capillary fringe during falling tides. Ratios of CO₂ and O₂ in vadose gas relative to the atmosphere indicate this atmospheric oxygen fuels respiration within newly-exposed, wetted bedrock. Deficits of expected CO₂ relative to O₂ concentrations indicate some respired CO₂ is likely removed by carbonate mineral dissolution. Tidal pumping also appears capable of transferring oxygen to the freshwater lens, where it could also contribute to respiration and dissolution; dissolved oxygen concentrations at the water table are at least 5% saturated and decline to anaerobic conditions 1–2 m below. Our results demonstrate how tidal pumping of air to vadose zones can drive mineral dissolution reactions that are focused near water tables and may contribute to the formation of laterally continuous vuggy horizons and potentially caves. © 2020 John Wiley & Sons, Ltd.     

Removal of Hydrogen Sulfide by Physico‐Biological Filter Using Mixed Rice Husk Silica and Dried Activated Sludge

This study investigated the effectiveness of a new packing material, namely mixed rice husk silica with dried activated sludge for removing H₂S. Dried sewage sludge was collected from Putrajaya sewage treatment plant in Malaysia. Rice husk silica was prepared at temperature of 800°C, after acid leaching and mixed with dried sewage sludge to be utilized in a polyvinyl chloride filter. The system was operated under variable conditions of two parameters, different inlet gas concentration and different inlet flow rate. H₂S was passed through the filter with one liter of the packing material. More than 99.96% removal efficiency (RE) with empty bed residence time (EBRT) of 90–45 s and 300 ppm inlet concentration was observed. However, the RE decreased to 96.87% with the EBRT of 30 s. The maximum elimination capacity (EC) of 52.32 g/m³/h was obtained with the RE of 96.87% and H₂S mass loading rate of 54 g/m³/h, while at the RE of 99.96%, maximum EC was 26.99 g/m³/h with the H₂S mass‐loading rate of 27 g/m³/h. A strong significant correlation between increasing of H₂S mass loading rate and pressure drop was also detected (p < 0.01). Maximum pressure drop was 3.0 mm H₂O after 53 days of operating time, the EBRT of 30 s, and 54 g/m³/h of H₂S loading rate. These observations suggest that the mixture of rice husk silica with dried activated sludge is a suitable physico‐biological filter for H₂S removal.     

Optimierung des Belebungsverfahrens durch Minimierung des Volumens von Belebungs- und Nachklärbecken

The separate examination of the processes of the settling and thickening of activated sludge allows us to minimize the volumes of aeration tanks and final clarification tanks. The following initial quantities are given: wastewater quantity, inflow and outflow concentrations of BOD₅, wastewater temperature and reflux ratio of activated sludge and, in addition, three parameters of the thickening behaviour of activated sludge as well as a parameter for the bioactivity of sludge, which are to be determined in an experimental plant. From this, the optimum concentration of the activated sludge in the aeration tank is calculated at which the total of volumes for aeration tank and final clarification tank becomes a minimum. The process is carried out numerically by an example.     

Degradation of Disperse Blue E‐4R in Aqueous Solution by Zero‐Valent Iron/Ozone

Degradation of an anthraquinone dye, disperse blue E‐4R, by zero‐valent iron (ZVI)/ozone (O₃) was carried out in a series of laboratory‐scale experiments. The obtained results indicated that this method was much more effective than single ZVI or single O₃ at removal of color, chemical oxygen demand, total organic carbon, and adsorbable organic halogen. The effect of several related operational parameters, including O₃ dosage, zero valent iron dosage, temperature, pH value, and ZVI particle size were also discussed. Finally, we tried to decontaminate some actual samples with this method, which showed high treatment efficiency to the sample pretreated by conventional activated sludge.     

Toxic Effects of Copper‐Based and Synthetic Organic Pesticides on Activated Sludge

Toxic effects of five commonly used pesticides on the biomass of a municipal activated sludge system were determined on the basis of the reduction in the oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR). Toxicity levels of the selected pesticides were determined by employing a modified OECD 209 (Organisation for Economic Cooperation and Development) method which was performed as batch experiments using a respirometer. Copper sulphate (CuSO₄ · 5 H₂O), copper oxychloride (Cu₂Cl(OH)₃), copper calcium oxychloride (CaCu₃Cl₂(OH)₆) as copper‐based pesticides and chlorsulphuron (C₁₂H₁₂ClN₅O₄S), 2,4‐dichlorophenoxyacetic acid (2,4‐D) (C₈H₆Cl₂O₃) as synthetic organic pesticides were selected for the experiments. The EC₅₀ values were determined to be 78, 249 and 281 mg/L for CuSO₄ · 5 H₂O, Cu₂Cl(OH)₃ and CaCu₃Cl₂(OH)₆, respectively. Corresponding values for C₁₂H₁₂ClN₅O₄S and 2,4‐D were 860 and 3664 mg/L, respectively. Results indicated that toxicity effects of copper‐based pesticides were higher than that of synthetic organic pesticides. CuSO₄ · 5 H₂O was found to exert the highest toxicity among the copper‐based pesticides, whereas, C₁₂H₁₂ClN₅O₄S was determined to be the most toxic among the organic pesticides on activated sludge biomass.     

Use of Chemotaxonomy To Study the Influence of Benzalkonium Chloride on Bacterial Populations in Biodegradation Testing

Contradictory results are reported for the behaviour of quaternary ammonium compounds (QACs) in sewage treatment plants (STPs). QACs may sorb onto activated sludge. Only little information is available with respect to effects of QACs against bacteria in STPs. Only 5 to 15 % of bacteria present in sewage sludge can be detected by means of culture dependent microbiological methods. The shift of the bacterial populations due to effects of test compounds have not been studied up to now with culture independent methods. The microbial populations shift was studied in situ using culture independent chemotaxonomy profiling ubiquinones and polyamines. Additionally, toxic effects of QACs against bacteria present in the test vessels of the Zahn‐Wellens test (OECD 302 B) were assessed with a toxicity control in the test. The ubiquinone profiles representing changes in Gram‐negative populations mainly showed that the activated sludge was affected only in test vessels containing benzalkonium chloride. According to chemotaxonomy Acinetobacter or/and some members of Pseudomonas spp. have been selected by benzalkonium chloride after some adaptation period (8 to 12 days).     

Auswahl der Inkubationszeit bei Bestimmung der Belebtschlammaktivität mit Hilfe des TTC-Tests

The general assumption that the quantity of the formazan formed is linearly proportional to the time of incubation cannot be confirmed in practice. A formation of formazan which is too low initially is due to the initially passive transport of TTC to the sites of action. A formazan formation which is initially very high occurs through non-enzymatic reduction of TTC by reducing substances in the environment outside the bacterial cells. During longer periods of incubation, especially during toxicological tests, the substrate may get exhausted, so that less formazan will be formed. For the correct determination of the dehydrogenase activity by means of TTC it is necessary to determine the period of incubation within which the increase of formazan is linearly proportional to time. The dehydrogenase activity can be exactly determined only from this period of time of incubation.