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.     

Beitrag zur Charakterisierung und Identifizierung organischer Stoffe niederer molarer Massen in biologischen Kläranlagenabläufen

Contribution to the characterization and identification of low molecular weight organic substances in biologically treated effluent. In a review it is shown that by determination of sum parameters only 25% of low molecular weight substances <1000 Dalton in biologically treated effluent can be divided into different classes of compounds. A method is described to separate the low molecular weight compounds (freeze concentration, electrodialysis, vacuum evaporation, freeze drying) in order to identify single compounds. In a first GC-MS analysis 87 compounds, among them a high proportion of substances with hetero atoms, could be identified. Their concentrations lie between 0,1 and 3 μg/1. Only 3% of the low molecular weight fraction related to the DOC value is covered by the identified substances. Therefore it can be assumed, that a broad variety of low molecular weight compounds in very low concentrations is present in the effluent of biological treatment plants.     

Eignung mikrobieller Biotests für gebundene Schadstoffe Suitability of Microbial Bioassays for Bound Contaminants

Toxicity of soil spiked with water soluble and insoluble contaminants was assessed with bacteria. Three bioassays were used with common soil bacteria. Water soluble fractions from the soils were estimated in microtiterplates via dehydrogenase activity of B. cereus and growth of P. putida. A new method was developed using B. cereus to include effects of soil particles with bound contaminants. As contaminants p-nitrophenol and 2-naphthol were tested in these systems. Results indicated that the concentration of the contaminants in the soil solution is determined by soil properties. Toxicity of water soluble fractions was not only a result of the toxicant's concentration but further modified by soluble components of the soils. It is not possible to assess soil pollution from toxicity of the water soluble fraction in general, because unknown factors of the soluble soil fraction can influence toxicity greatly. The newly developed contact bioassay with B. cereus showed clearly higher toxicity of 2-naphthol estimated in soil suspension than did the comparable test system with the same bacteria using water soluble fractions only. The results stress the importance of including soil particles into bioassay procedures. It was shown that also water insoluble, bound contaminants can act as toxicants. For an estimation of toxicity under conditions more relevant for the environment, the suggested test system can provide useful information to bioavailability of bound contaminants.     

Characterization of Estuarine and Fluvial Dissolved Organic Matter by Thermochemolysis using Tetramethylammonium Hydroxide

Selected samples of dissolved organic matter (DOM) isolated by ultrafiltration (UDOM) have been analyzed by thermochemolysis in the presence of tetramethylammonium hydroxide (TMAH). This technique cleaves ester and ether bonds of bio‐ and geological macromolecules and releases monomer subunits and methylates them in situ as their methyl ethers and methyl esters. Compared with conventional pyrolysis, TMAH thermochemolysis avoids decarboxylation of preexisting carboxylic moieties and produces aromatic acids as their methyl esters. Various phenolic derivatives, which might originate from incorporated lignin‐derived structures, from the highly aliphatic and resistant biopolymer cutan and also from proteinaceous materials, were identified among the products produced from UDOM upon thermochemolysis. The presence of lignin derivatives in UDOM indicates input of organic matter derived from terrestrial sources. Various aromatic acids, perhaps representing the final steps in the oxidation of the side‐chain during microbial oxidation of lignin, were released upon TMAH thermochemolysis, suggesting they are structural constituents of the UDOM. Different ratios of lignin‐derived materials, commonly determined using the CuO oxidation method, such as the Δ value, indicative of the amount of lignin present, the acid/aldehyde ratio (Ad/Al)_G, indicative of the extent of oxidative degradation of the lignin component, and the syringyl/guaiacyl (S/G) and p‐hydroxyphenyl/guaiacyl (P/G) ratios, indicative of the contribution for the different types of lignin, were determined.     

Dissolved Organic Carbon Fractions Formed during Composting of Municipal Solid Waste: Properties and Significance

The properties and transformation of dissolved organic matter (DOM) extracted (10 L water per kilogram compost) from municipal solid waste (MSW) compost at five stages (days 47, 77, 105, 126, and 187) of composting were investigated. The DOM was fractionated into hydrophobic or hydrophilic neutrals, acids, and bases. The unfractionated DOM, the hydrophobic acids and neutrals (HoA and HoN, respectively), and the hydrophilic neutrals (HiN) fractions were studied using solid-state ¹³C-NMR, FTIR, and DRIFT spectroscopy. The HoA fraction was found to be the dominant (percentage of total DOM) hydrophobic fraction, exhibiting a moderate increase during composting. The HoN fraction increased sharply from less than 1% to 18% of the total DOM during 187 days of composting, while the hydrophobic bases (HoB) exhibited the opposite trend. The HiN represented the major fraction of the hydrophiles up to 120 days of composting, decreasing thereafter by 38%. The relative concentration of the hydrophilic acids and bases (HiA and HiB, respectively) exhibited no consistent trend during composting. DRIFT spectra of the unfractionated DOM taken from the composting MSW revealed a decreasing level of polysaccharide structures with time. The ¹³C-NMR and FTIR spectra of the HoA fraction exhibited a polyphenol-humic structure, whereas the HoN spectra exhibited strong aliphatic features. The spectra of the HiN fraction confirmed its polysaccharide nature. During the final stage of composting, the DOM concentration was steady, while a relative decrease of HiN concomitant with an increase of HoA and HoN fractions was observed. These indicate that the DOM contained a low concentration of biodegradable organic matter and a higher content of macromolecules related to humic substances. The biological significance and heavy metal binding of these fractions are being studied based on earlier observations showing enhanced plant growth in the presence of DOM extracted from mature as opposed to immature compost.     

Tracing of Peroxidase Activity in Humic Lake Water

An enzyme assay was developed for studies on peroxidase activities in humic lake water. 3,4-Dimethoxybenzyl alcohol (veratryl alcohol, VeraOH) was used as tracer substrate, and peroxidase (EC 1.11.1.7) activity was measured by high-performance liquid chromatography. The chemical stability of VeraOH and its application as peroxidase substrate was tested under light and dark conditions, different hydrogen peroxide (H₂O₂) concentrations and humic matter contents. VeraOH was stable under low UV radiation at in situ conditions in lake water (<0.010...0.25 kJ m^–2 d^–1), laboratory conditions (<0.05...0.30 kJ m^–2 d^–1), and low (1...100 μM) H₂O₂ concentrations. However, peroxides oxidized VeraOH above 1...10 mM H₂O₂ concentration in sterile Millipore-Q and humic lake water. Dark incubations showed little VeraOH oxidation products. The developed peroxidase assay was tested in the growth medium of Phanerochaete chrysosporium and a bacteria isolate (P.M.D. 20.4.3.1) from mesohumic lake Pääjärvi. Peroxidase activities were also measured in natural microbial communities under standard laboratory and under in situ conditions in humic lake water. Incubation times of about 5 to 12 days were usually needed to record significant (P < 0.05) peroxidase activities, in lake waters. In situ peroxidase activities varied in pelagial surface water (0...0.5 m) on a seasonal scale between 74 nmol L^–1 h^–1 and 273 nmol L^–1 (mean: 176 nmol L^–1 h^–1) and within the water column between 110 nmol L^–1 h^–1 and 800 nmol L^–1 h^–1 (mean: 500 nmol L^–1 h^–1) in polyhumic lake Mekkojärvi.     

Urease-Hemmtest für wirkungsbezogene Untersuchungen der Bindungsformen von Kupfer in Wässern und Bodeneluaten

Urease Inhibition Test for Effect-related Investigations of Copper Speciation in Water and Soil Extracts In 80 different water samples, the inhibition effect of cupric ions was measured by application of a simple urease inhibition test. In relation to the total copper concentration, the inhibition permits an effect-related classification of drinking water, rain water, spring water, surface water, leakage water, wastewater, and soil extracts. Drinking water contains high parts of cupric ions in the free hydrated form. Their copper content results in high urease inhibition and is caused by using copper pipes in house plumbing. In leakage water, wastewater, and soil extracts, cupric ions are mainly complexed by organic compounds in the sample matrix. Hence, the inhibition effects are low. In soil extracts, the urease inhibition increased after an UV-decomposition procedure, but a decreasing effect was observed for a surface and a drinking water sample, respectively. The chemical oxygen demand and UV-absorbance at 255 nm measured before and after the procedure were correlated to the content of organic compounds, i.e., humic acid. In most samples, the organic matrix was not decomposed completely. The oxidizing decomposition apparently forms further complexing agents, which suppress the inhibition effect of cupric ions. In the rain water samples investigated, the content of organic compounds depended on rain intensity. The chemical oxygen demand and UV-absorbance, respectively, were low for rain water collected during strong rain periods and increased with decreasing rain intensity. The rain water trickled over sheet-lead roof showed an increase of urease inhibition compared with the simultaneously collected rain. Consequently, a decomposition of organic matrix could be demonstrated in rain water by use of the urease inhibition test.     

Formation of Humic-like Substances in Mixed and Pure Cultures of Aquatic Microorganisms

A natural decay of plant and animal biomass in aquatic environments results in the release of different simple structured organic substances into water. In our laboratory experiments we investigated whether some of them, e.g., starch or peptone, and also a complex yeast biomass can be transformed into humic-like substances by natural assemblages and some pure cultures of aquatic microorganisms. After a 6 months incubation most of the cultural media enriched with those natural organic substrates turned dark in color and humic-like substances (HS) could be isolated. However, the original substrate organic carbon was mainly mineralized in microbial cultures, and only about 3 % C was converted into HS. Total yields of HS differed in dependence of the individual substrate used (peptone > yeast > starch), the origin of inoculum (river > lake > groundwater), and the incubation temperature (20°C > 10°C). According to their elemental composition, and their spectroscopical and electrophoretic characteristics the microbially formed HS resembled natural aquatic humic matter, but were higher in aliphatic constituents (carbohydrates, peptides) and lower in aromatic structures. HS-like substances were also obtained from pure cultures of a bacterium Bacillus sphaericus and a fungus Cladosporium cladosporioides. The ¹³C and ¹⁵N isotopic contents of the microbially produced HS differed in dependence on the microbial inoculum and the type of organic substrates used, but in general they indicated anabolic processes to be mainly responsible for the humification of the simple organic substrates used in our experiments.     

Biochemical Characterization of Refractory Organic Substances

The characterization of refractory organic substances (ROS) is very complicated because of their heterogeneous structure. Size-exclusion chromatography with continuous detection of dissolved nitrogen (LC-DN), dissolved organic carbon (LC-DOC), and UV-absorbance (LC-UV) is a very useful analytical tool for the characterization of changes of ROS in natural aquatic systems and in technological treatment. The effect of natural, oxidative, and biochemical processes on formation and removal of ROS is described. Additionally the role of hydrolysable carbohydrates in the composition of ROS is presented.     

Einflußfaktoren auf die Freisetzung von DOC und AOX unter ehemaligen Abwasserversickerungsflächen

Factors Influencing the Release of DOC and AOX out of Former Wastewater Infiltration Soils The influence of soil and of infiltration water quality on the release of dissolved organic carbon (DOC) and adsorbable organic halogens (AOX) from two former wastewater infiltration sites was investigated in laboratory column studies. Desorption was the most important factor influencing release processes. It depends on the amount of sorbent and sorbate and the strength of binding. Therefore, for constant irrigation rates a higher soil organic carbon content coincided with higher DOC-contents of the column effluent, while the irrigation water quality was of minor importance. Within one system the dependencies of AOX release and DOC mobilization were found to be similar. Transferring these dependencies on other systems has its limitations because of a different binding quality between sorbent and sorbate.